bluuebunny/crossref_metadata_2025 huggingface dataset:
Glycosylation-independent targeting enhances enzyme delivery to lysosomes and decreases storage in mucopolysaccharidosis type VII mice
Authors: LeBowitz Jonathan H. , Grubb Jeffrey H. , Maga John A. , Schmiel Deborah H. , Vogler Carole , Sly William S.
Published: March 2004

Enzyme-replacement therapy is an established means of treating lysosomal storage diseases. Infused therapeutic enzymes are targeted to lysosomes of affected cells by interactions with cell-surface receptors that recognize carbohydrate moieties, such as mannose and mannose 6-phosphate, on the enzymes. We have tested an alternative, peptide-based targeting system for delivery of enzymes to lysosomes in a murine mucopolysaccharidosis type VII (MPS VII) model. This strategy depends on the interaction of a fragment of insulin-like growth factor II (IGF-II), with the IGF-II binding site on the bifunctional, IGF-II cation-independent mannose 6-phosphate receptor. A chimeric protein containing a portion of mature human IGF-II fused to the C terminus of human β-glucuronidase was taken up by MPS VII fibroblasts in a mannose 6-phosphate-independent manner, and its uptake was inhibited by the addition of IGF-II. Furthermore, the tagged enzyme was delivered effectively to clinically significant tissues in MPS VII mice and was effective in reversing the storage pathology. The tagged enzyme was able to reduce storage in glomerular podocytes and osteoblasts at a dose at which untagged enzyme was much less effective. This peptide-based, glycosylation-independent lysosomal targeting system may enhance enzyme-replacement therapy for certain human lysosomal storage diseases.
Serum leptin level is a regulator of bone mass
Authors: Elefteriou F. , Takeda S. , Ebihara K. , Magre J. , Patano N. , Ae Kim C. , Ogawa Y. , Liu X. , Ware S. M. , Craigen W. J. , Robert J. J. , Vinson C. , Nakao K. , Capeau J. , Karsenty G.
Published: March 2004

Leptin is a powerful inhibitor of bone formationin vivo. This antiosteogenic function involves leptin binding to its receptors on ventromedial hypothalamic neurons, the autonomous nervous system and β-adrenergic receptors on osteoblasts. However, the mechanisms whereby leptin controls the function of ventromedial hypothalamic antiosteogenic neurons remain unclear. In this study, we compared the ability of leptin to regulate body weight and bone mass and show that leptin antiosteogenic and anorexigenic functions are affected by similar amounts of leptin. Using a knock-in ofLacZin theleptinlocus, we failed to detect any leptin synthesis in the central nervous system. However, increasing serum leptin level, even dramatically, reduced bone mass. Conversely, reducing serum-free leptin level by overexpressing a soluble receptor for leptin increased bone mass. Congruent with these results, the high bone mass of lipodystrophic mice could be corrected by restoring serum leptin level, suggesting that leptin is an adipocyte product both necessary and sufficient to control bone mass. Consistent with the high bone mass phenotype of lipodystrophic mice, we observed an advanced bone age, an indirect reflection of premature bone formation, in lipodystrophic patients. Taken together, these results indicate that adipocyte-derived circulating leptin is a determinant of bone formation and suggests that leptin antiosteogenic function is conserved in vertebrates.
NGEP, a gene encoding a membrane protein detected only in prostate cancer and normal prostate
Authors: Bera Tapan K. , Das Sudipto , Maeda Hiroshi , Beers Richard , Wolfgang Curt D. , Kumar Vasantha , Hahn Yoonsoo , Lee Byungkook , Pastan Ira
Published: March 2004

We identified a gene (NGEP) that is expressed only in prostate cancer and normal prostate. The two NGEP transcripts are 0.9 kb and 3.5 kb in size and are generated by a differential splicing event. The short variant (NGEP-S) is derived from four exons and encodes a 20-kDa intracellular protein. The long form (NGEP-L) is derived from 18 exons and encodes a 95-kDa protein that is predicted to contain seven-membrane-spanning regions.
In situ
hybridization shows that NGEP mRNA is localized in epithelial cells of normal prostate and prostate cancers. Immunocytochemical analysis of cells transfected with NGEP cDNAs containing a Myc epitope tag at the carboxyl terminus shows that the protein encoded by the short transcript is localized in the cytoplasm, whereas the protein encoded by the long transcript is present on the plasma membrane. Because of its selective expression in prostate cancer and its presence on the cell surface, NGEP-L is a promising target for the antibody-based therapies of prostate cancer.
Multivariate proteomic analysis of murine embryonic stem cell self-renewal versus differentiation signaling
Authors: Prudhomme Wendy , Daley George Q. , Zandstra Peter , Lauffenburger Douglas A.
Published: March 2004

A number of extracellular stimuli, including soluble cytokines and insoluble matrix factors, are known to influence murine embryonic stem cell self-renewal and differentiation behavioral responses via intracellular signaling pathways, but their net effects in combination are difficult to understand. To gain insight concerning key intracellular signals governing these behavioral responses, we employ a multivariate systems analysis of proteomic data generated from combinatorial stimulation of mouse embryonic stem cells by fibronectin, laminin, leukemia-inhibitory factor, and fibroblast growth factor 4. Phosphorylation states of 31 intracellular signaling network components were obtained across 16 different stimulus conditions at three time points by quantitative Western blotting, and partial-least-squares modeling was used to determine which components were most strongly correlated with cell proliferation and differentiation rate constants obtained from flow cytometry measurements of Oct-4 expression levels. This data-driven, multivariate (16 conditions × 31 components × 3 time points = ≈1,500 values) proteomic approach identified a set of signaling network components most critically associated (positively or negatively) with differentiation (Stat3, Raf1, MEK, and ERK), proliferation of undifferentiated cells (MEK and ERK), and proliferation of differentiated cells (PKBα, Stat3, Src, and PKCε). These predictions were found to be consistent with previous
in vivo
literature, along with direct
in vitro
test here by a peptide inhibitor of PKCε. Our results demonstrate how a computational systems biology approach can elucidate key sets of intracellular signaling protein activities that combine to govern cell phenotypic responses to extracellular cues.
Regulation of expression of Bcl-2 protein family member Bim by T cell receptor triggering
Authors: Sandalova Elena , Wei Cheng-Hong , Masucci Maria G. , Levitsky Victor
Published: March 2004

Bim, a proapoptotic BH3-only member of the Bcl-2 protein family, is required for central and peripheral deletion of T lymphocytes. Mechanisms regulating Bim activity in T cells remain poorly understood. We show that expression of Bim is up-regulated in human T cells after polyclonal or specific T cell receptor triggering. Induction of Bim was affected by the agonistic potency of MHC:peptide ligands. Peptides that failed to induce Bim expression, failed to induce apoptosis in specific T cells, whereas partially agonistic ligands, which trigger death receptor-independent activation-induced cell death (AICD), induced Bim, but were inefficient in up-regulating Bcl-XL. Activation of protein kinase C and calcineurin appeared to be necessary and sufficient for Bim up-regulation after T cell receptor ligation. Immunosuppressive drugs known to prevent T cell deletionin vivo, such as cyclosporin A or FK506, blocked Bim up-regulation and rescued T cells from death receptor-independent AICD, whereas rapamycin, which allows the development of stable immunological tolerance, did not exhibit these activities. These results define a new mode of Bim regulation, strongly implicate Bim as a mediator of AICD, and suggest that Bim up-regulation can be targeted to influence the outcome of specific immune responses.
Determinants of liposome fusion mediated by synaptic SNARE proteins
Authors: Schuette Christina G. , Hatsuzawa Kiyotaka , Margittai Martin , Stein Alexander , Riedel Dietmar , Küster Petra , König Marcelle , Seidel Claus , Jahn Reinhard
Published: March 2004

Synaptic exocytosis requires the soluble
N
-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins syntaxin 1, SNAP-25, and synaptobrevin (VAMP). Assembly of the SNAREs into a stable core complex is supposed to catalyze membrane fusion, and proteoliposomes reconstituted with synaptic SNARE proteins spontaneously fuse with each other. We now show that liposome fusion mediated by synaptic SNAREs is inhibited by botulinum neurotoxin E (BoNT/E) but can be rescued by supplementing the C-terminal portion of SNAP-25. Furthermore, fusion is prevented by a SNAP-25-specific antibody known to block exocytosis in chromaffin cells, and it is competed for by soluble fragments of the R-SNAREs synaptobrevin 2, endobrevin/VAMP-8, and tomosyn. No accumulation of clustered vesicles is observed during the reaction. Rapid artificial clustering of SNARE-containing proteoliposomes enhances the fusion rate at low but not at saturating liposome concentrations. We conclude that the rate of liposome fusion is dominated by the intrinsic properties of the SNAREs rather than by the preceding docking step.
Mice deficient for soluble adenylyl cyclase are infertile because of a severe sperm-motility defect
Authors: Esposito Gloria , Jaiswal Byjay S. , Xie Fang , Krajnc-Franken Magda A. M. , Robben Tamara J. A. A. , Strik Ankie M. , Kuil Cor , Philipsen Ria L. A. , van Duin Marcel , Conti Marco , Gossen Jan A.
Published: March 2004

To acquire the ability to fertilize, spermatozoa undergo complex, but at present poorly understood, activation processes. The intracellular rise of cAMP produced by the bicarbonate-dependent soluble adenylyl cyclase (sAC) has been suggested to play a central role in initiating the cascade of the events that culminates in spermatozoa maturation. Here, we show that targeted disruption of the sAC gene does not affect spermatogenesis but dramatically impairs sperm motility, leading to male sterility. sAC mutant spermatozoa are characterized by a total loss of forward motility and are unable to fertilize oocytes
in vitro
. Interestingly, motility in sAC mutant spermatozoa can be restored on cAMP loading, indicating that the motility defect observed is not caused by a structural defect. We, therefore, conclude that sAC plays an essential and nonredundant role in the activation of the signaling cascade controlling motility and, therefore, in fertility. The crucial role of sAC in fertility and the absence of any other obvious pathological abnormalities in sAC-deficient mice may provide a rationale for developing inhibitors that can be applied as a human male contraceptive.
Combination of rapamycin and protein tyrosine kinase (PTK) inhibitors for the treatment of leukemias caused by oncogenic PTKs
Authors: Mohi M. Golam , Boulton Christina , Gu Ting-Lei , Sternberg David W. , Neuberg Donna , Griffin James D. , Gilliland D. Gary , Neel Benjamin G.
Published: March 2004

Abnormal protein tyrosine kinases (PTKs) cause many human leukemias. For example, BCR/ABL causes chronic myelogenous leukemia (CML), whereas FLT3 mutations contribute to the pathogenesis of acute myelogenous leukemia. The ABL inhibitor Imatinib (Gleevec, STI571) has remarkable efficacy for treating chronic phase CML, and FLT3 inhibitors (e.g., PKC412) show similar promise in preclinical studies. However, resistance to PTK inhibitors is a major emerging problem that may limit long-term therapeutic efficacy. Development of rational combination therapies will probably be required to effect cures of these and other neoplastic disorders. Here, we report that the mTOR inhibitor rapamycin synergizes with Imatinib against BCR/ABL-transformed myeloid and lymphoid cells and increases survival in a murine CML model. Rapamycin/Imatinib combinations also inhibit Imatinib-resistant mutants of BCR/ABL, and rapamycin plus PKC412 synergistically inhibits cells expressing PKC412-sensitive or -resistant leukemogenic FLT3 mutants. Biochemical analyses raise the possibility that inhibition of 4E-BP1 phosphorylation may be particularly important for the synergistic effects of PTK inhibitor/rapamycin combinations. Addition of a mitogen-activated protein kinase kinase inhibitor to rapamycin or rapamycin plus PTK inhibitor further increases efficacy. Our results suggest that simultaneous targeting of more than one signaling pathway required by leukemogenic PTKs may improve the treatment of primary and relapsed CML and/or acute myelogenous leukemia caused by FLT3 mutations. Similar strategies may be useful for treating solid tumors associated with mutant and/or overexpressed PTKs.
Disruption of forkhead transcription factor (FOXO) family members in mice reveals their functional diversification
Authors: Hosaka Taisuke , Biggs William H. , Tieu David , Boyer Antonia D. , Varki Nissi M. , Cavenee Webster K. , Arden Karen C.
Published: March 2004

Genetic analysis in
Caenorhabditis elegans
has uncovered essential roles for DAF-16 in longevity, metabolism, and reproduction. The mammalian orthologs of DAF-16, the closely-related FOXO subclass of forkhead transcription factors (FKHR/FOXO1, FKHRL1/FOXO3a, and AFX/FOXO4), also have important roles in cell cycle arrest, apoptosis and stress responses
in vitro
, but their
in vivo
physiological roles are largely unknown. To elucidate their role in normal development and physiology, we disrupted each of the
Foxo
genes in mice.
Foxo1
-null embryos died on embryonic day 10.5 as a consequence of incomplete vascular development.
Foxo1
-null embryonic and yolk sac vessels were not well developed at embryonic day 9.5, and
Foxo1
expression was found in a variety of embryonic vessels, suggesting a crucial role of this transcription factor in vascular formation. On the other hand, both
Foxo3a
- and
Foxo4
-null mice were viable and grossly indistinguishable from their littermate controls, indicating dispensability of these two members of the Foxo transcription factor family for normal vascular development.
Foxo3a
-null females showed age-dependent infertility and had abnormal ovarian follicular development. In contrast, histological analyses of
Foxo4
-null mice did not identify any consistent abnormalities. These results demonstrate that the physiological roles of
Foxo
genes are functionally diverse in mammals.
Molecular chaperones function as steroid receptor nuclear mobility factors
Authors: Elbi Cem , Walker Dawn A. , Romero Guillermo , Sullivan William P. , Toft David O. , Hager Gordon L. , DeFranco Donald B.
Published: March 2004

Live cell imaging has revealed the rapid mobility of steroid hormone receptors within nuclei and their dynamic exchange at transcriptionally active target sites. Although a number of other proteins have been shown to be highly mobile within nuclei, the identity of soluble factors responsible for orchestrating nuclear trafficking remains unknown. We have developed a previously undescribed
in situ
subnuclear trafficking assay that generates transcriptionally active nuclei, which are depleted of soluble factors required for the nuclear mobility of glucocorticoid (GR) and progesterone receptors (PR). Using this system and a fluorescence recovery after photobleaching technique, we demonstrate that nuclear mobility of GR recovered on incubation with reticulocyte lysate was inhibited by geldanamycin, a drug that blocks the chaperone activity of heat-shock protein 90. Direct proof of molecular chaperone involvement in steroid receptor subnuclear trafficking was provided by the ATP-dependent recovery of nuclear mobility of GR and PR on incubation with various combinations of purified chaperone and/or cochaperone proteins. Additionally, for both receptors, the inclusion of hormone during the recovery period leads to a retardation of nuclear mobility. Thus, our results provide a description of soluble nuclear mobility factors and furthermore demonstrate a previously unrecognized role for molecular chaperones in the regulation of steroid receptor function within the nucleus.
Glutamate-183 in the conserved TGES motif of domain A of sarcoplasmic reticulum Ca
2+
-ATPase assists in catalysis of
E
2
/
E
2
P partial reactions
Authors: Clausen Johannes D. , Vilsen Bente , McIntosh David B. , Einholm Anja P. , Andersen Jens Peter
Published: March 2004

The recently determined crystal structures of the sarcoplasmic reticulum Ca
2+
-ATPase show that in the
E
1
Ca
2
form, domain A is almost isolated from the other cytoplasmic domains, P and N, whereas in
E
2
, domain A has approached domains P and N, with E183 of the highly conserved P-type ATPase signature sequence TGES in domain A now being close to the phosphorylated aspartate in domain P, thus raising the question whether E183 acquires a catalytic role in
E
2
and
E
2
P conformations. This study compares the partial reactions of mutant E183A and wild-type Ca
2+
-ATPase, using transient and steady-state kinetic measurements. It is demonstrated that dephosphorylation of the
E
2
P phosphoenzyme intermediate, as well as reverse phosphorylation of
E
2
with P
i
, is severely inhibited in the mutant. Furthermore, the apparent affinity of
E
2
for the phosphoryl transition state analog vanadate is reduced by three orders of magnitude, consistent with a destabilization of the transition state complex, and the mutant displays reduced apparent affinity for P
i
in the
E
2
form. The
E
1
Ca
2
conformation, on the other hand, shows normal phosphorylation with ATP and normal Ca
2+
binding properties, and the rates of the conformational transitions
E
1
PCa
2
→
E
2
P and
E
2
→
E
1
Ca
2
are only 2- to 3-fold reduced, relative to wild type. These results, which likely can be generalized to other P-type ATPases, indicate that E183 is critical for the phosphatase function of
E
2
and
E
2
P, possibly interacting with the phosphoryl group or attacking water in the transition state complex, but is of little functional importance in
E
1
and
E
1
P.
Two structural features of λ integrase that are critical for DNA cleavage by multimers but not by monomers
Authors: Lee Sang Yeol , Aihara Hideki , Ellenberger Tom , Landy Arthur
Published: March 2004

Despite many years of genetic and biochemical studies on the λ integrase (Int) recombination system, it is still not known whether the Int protein is competent for DNA cleavage as a monomer. We have addressed this question, as part of a larger study of Int functions critical for the formation of higher-order complexes, by isolating “multimer-specific” mutants. We identify a pair of oppositely charged residues, E153 and R169, that comprise an intermolecular salt bridge within a functional Int multimer. Mutation of either of these residues significantly reduces both the cleavage of full-
att
sites and the resolution of Holliday junctions without compromising the cleavage of half-
att
site substrates. Allele-specific suppressor mutations were generated at these residues. Their interaction with wild-type Int on preformed Holliday junctions indicates that the mutated residues comprise an intermolecular salt bridge. We have also shown that the most C-terminal seven residues of Int, which comprise another previously identified subunit interface, inhibit DNA cleavage by monomeric but not multimeric Int. Taken together, our results lead us to conclude that Int can cleave DNA as a monomer. We also identify and discuss unique structural features of Int that act negatively to reduce its activity as a monomer and other features that act positively to enhance its activity as a multimer.
Reengineered salivary glands are stable endogenous bioreactors for systemic gene therapeutics
Authors: Voutetakis Antonis , Kok Marc R. , Zheng Changyu , Bossis Ioannis , Wang Jianghua , Cotrim Ana P. , Marracino Natanya , Goldsmith Corinne M. , Chiorini John A. , Loh Y. Peng , Nieman Lynnette K. , Baum Bruce J.
Published: March 2004

The use of critical-for-life organs (e.g., liver or lung) for systemic gene therapeutics can lead to serious safety concerns. To circumvent such issues, we have considered salivary glands (SGs) as an alternative gene therapeutics target tissue. Given the high secretory abilities of SGs, we hypothesized that administration of low doses of recombinant adeno-associated virus (AAV) vectors would allow for therapeutic levels of transgene-encoded secretory proteins in the bloodstream. We administered 10
9
particles of an AAV vector encoding human erythropoietin (hEPO) directly to individual mouse submandibular SGs. Serum hEPO reached maximum levels 8-12 weeks after gene delivery and remained relatively stable for 54 weeks (longest time studied). Hematocrit levels were similarly increased. Moreover, these effects proved to be vector dose-dependent, and even a dosage as low as 10
8
particles per animal led to significant increases in hEPO and hematocrit levels. Vector DNA was detected only within the targeted SGs, and levels of AAV copies within SGs were highly correlated with serum hEPO levels (
r
= 0.98). These results show that SGs appear to be promising targets with potential clinical applicability for systemic gene therapeutics.
RETRACTED: Structure of the multidrug resistance efflux transporter EmrE from
Escherichia coli
Authors: Ma Che , Chang Geoffrey
Published: March 2004

Multidrug resistance efflux transporters threaten to reverse the progress treating infectious disease by extruding a wide range of drug and other cytotoxic compounds. One such drug transporter, EmrE, from the small multidrug resistance family, utilizes proton gradients as an energy source to drive substrate translocation. In an effort to understand the molecular structural basis of this transport mechanism, we have determined the structure of EmrE from
Escherichia coli
to 3.8 Å. EmrE is a tetramer comprised of two conformational heterodimers related by a pseudo two-fold symmetry axis perpendicular to the cell membrane. Based on the structure and biochemical evidence, we propose a mechanism by which EmrE accomplishes multidrug efflux by coupling conformational changes between two heterodimers with proton gradient. Because of its simplicity and compact size, the structure of EmrE can serve as an ideal model for understanding the general structural basis of proton:drug antiport for other drug efflux systems.
Categorization in the monkey hippocampus: A possible mechanism for encoding information into memory
Authors: Hampson Robert E. , Pons Tim P. , Stanford Terrence R. , Deadwyler Sam A.
Published: March 2004

The mammalian hippocampus processes sensory information into memory. The neurobiological basis of this representation, as well as the type of information that is encoded, is central to understanding how memories are formed. Normally, there is an infinite amount of information that could be encoded for any given stimulus. Thus, the question arises as to how the hippocampus selects and encodes features of a given stimulus. Here, we show that neurons in the hippocampus of the monkey appear to categorize types of visual stimuli presented in a delayed-match-to-sample memory task. By extracting unique combinations of features, these category cells are able to encode aspects of behaviorally important images instead of encoding all visual details. The subject is then able to rapidly select an appropriate response to that stimulus when distracting stimuli are presented simultaneously, thereby facilitating performance. Moreover, across animals, this specific type of encoding differed considerably. Just as in humans, different monkeys attended to and selected different aspects of the same stimulus image, most likely reflecting different histories, strategies, and expectations residing within individual hippocampal networks.
Engineering biological structures of prescribed shape using self-assembling multicellular systems
Authors: Jakab Karoly , Neagu Adrian , Mironov Vladimir , Markwald Roger R. , Forgacs Gabor
Published: March 2004

Self-assembly is a fundamental process that drives structural organization in both inanimate and living systems. It is in the course of self-assembly of cells and tissues in early development that the organism and its parts eventually acquire their final shape. Even though developmental patterning through self-assembly is under strict genetic control it is clear that ultimately it is physical mechanisms that bring about the complex structures. Here we show, both experimentally and by using computer simulations, how tissue liquidity can be used to build tissue constructs of prescribed geometry
in vitro
. Spherical aggregates containing many thousands of cells, which form because of tissue liquidity, were implanted contiguously into biocompatible hydrogels in circular geometry. Depending on the properties of the gel, upon incubation, the aggregates either fused into a toroidal 3D structure or their constituent cells dispersed into the surrounding matrix. The model simulations, which reproduced the experimentally observed shapes, indicate that the control parameter of structure evolution is the aggregate–gel interfacial tension. The model-based analysis also revealed that the observed toroidal structure represents a metastable state of the cellular system, whose lifetime depends on the magnitude of cell–cell and cell–matrix interactions. Thus, these constructs can be made long-lived. We suggest that spherical aggregates composed of organ-specific cells may be used as “bio-ink” in the evolving technology of organ printing.
CIS display:
In vitro
selection of peptides from libraries of protein–DNA complexes
Authors: Odegrip Richard , Coomber David , Eldridge Bill , Hederer Rosemarie , Kuhlman Philip A. , Ullman Christopher , FitzGerald Kevin , McGregor Duncan
Published: March 2004

We describe the development of an
in vitro
library selection system (CIS display) that exploits the ability of a DNA replication initiator protein (RepA) to bind exclusively to the template DNA from which it has been expressed, a property called cis-activity. A diverse peptide library is created by ligation of DNA fragments of random sequence to a DNA fragment that encodes RepA. After
in vitro
transcription and translation, a pool of protein–DNA complexes is formed where each protein is stably associated with the DNA that encodes it. These complexes are amenable to the affinity selection of ligands to targets of interest. Here we show that RepA is a highly faithful cis-acting DNA-binding protein and demonstrate that libraries encoding >10
12
random 18-mer peptides can be constructed and used to isolate peptides that bind specifically to disparate targets. The use of DNA to encode the displayed peptides offers advantages over
in vitro
peptide display systems that use mRNA.
Cysteinyl leukotriene 1 receptor controls the severity of chronic pulmonary inflammation and fibrosis
Authors: Beller Thomas C. , Friend Daniel S. , Maekawa Akiko , Lam Bing K. , Austen K. Frank , Kanaoka Yoshihide
Published: March 2004

The cysteinyl leukotrienes (cys-LTs), leukotriene (LT) C4, LTD4, and LTE4, are smooth muscle constrictors that signal via the CysLT1receptor. Here we report that the cys-LTs play an important role in chronic pulmonary inflammation with fibrosis induced by bleomycin in mice. Targeted disruption of LTC4synthase, the pivotal enzyme for cys-LT biosynthesis, protected significantly against alveolar septal thickening by macrophages and fibroblasts and collagen deposition. In contrast, targeted disruption of the CysLT1receptor significantly increased both the concentration of cys-LTs in the bronchoalveolar lavage fluid and the magnitude of septal thickening as defined by morphology, digital image analysis, and deposition of reticular fibers. These findings change our understanding of the pathobiology mediated by the cys-LTs by revealing their role in chronic inflammation with fibrosis, likely via the CysLT2receptor, and by uncovering a dual role for the CysLT1receptor, namely proinflammatory acute constriction of smooth muscle and antiinflammatory counteraction of chronic injury.
Pseudomonas syringaepv.tomatocells encounter inhibitory levels of water stress during the hypersensitive response ofArabidopsis thaliana
Authors: Wright Catherine A. , Beattie Gwyn A.
Published: March 2004

During plant defense against bacterial pathogens, the hypersensitive response (HR) functions to restrict pathogen growth and spread. The mechanisms driving this growth restriction are poorly understood. We used a water stress-responsive transcriptional fusion to quantify the water potential sensed by individualPseudomonas syringaepv.tomatoDC3000 cells during infection ofArabidopsis thalianaleaves. A nonpathogenic DC3000hrcCmutant defective in type III secretion, as well as the saprophytePseudomonas fluorescensA506, sensed water potentials of -0.3 to -0.4 MPa at 48 h postinfiltration (hpi). During pathogenesis, DC3000 sensed lower water potentials (-0.4 to -0.9 MPa), demonstrating that it can modify the intercellular environment, and these water potentials were associated with optimal DC3000 growth in culture. During the HR, DC3000 cells sensed water potentials (-1.6 to -2.2 MPa) that were low enough to prevent cell division in the majority of cells in culture. This water potential decrease occurred within only 4 hpi and was influenced by avirulence gene expression, withavrRpm1expression associated with lower water potentials thanavrRpt2oravrBexpression at 48 hpi. The population sizes of the DC3000 variants tested were significantly correlated with the apoplastic water potential at 48 hpi, with a decrease of -0.9 MPa associated with a 10-fold decrease in cells per gram of leaf. These results suggest that the apoplastic water potential is a determinant of endophytic bacterial population size, and water stress, resulting from high osmolarity or tissue desiccation, is at least one factor restricting bacterial growth during the HR.
DrosophilaStem-Loop Binding Protein Intracellular Localization Is Mediated by Phosphorylation and Is Required for Cell Cycle-regulated Histone mRNA Expression
Authors: Lanzotti David J. , Kupsco Jeremy M. , Yang Xiao-Cui , Dominski Zbigniew , Marzluff William F. , Duronio Robert J.
Published: March 2004

Stem-loop binding protein (SLBP) is an essential component of the histone pre-mRNA processing machinery. SLBP protein expression was examined during Drosophila development by using transgenes expressing hemagglutinin (HA) epitope-tagged proteins expressed from the endogenous Slbp promoter. Full-length HA-dSLBP complemented a Slbp null mutation, demonstrating that it was fully functional. dSLBP protein accumulates throughout the cell cycle, in contrast to the observed restriction of mammalian SLBP to S phase. dSLBP is located in both nucleus and cytoplasm in replicating cells, but it becomes predominantly nuclear during G2. dSLBP is present in mitotic cells and is down-regulated in G1 when cells exit the cell cycle. We determined whether mutation at previously identified phosphorylation sites, T120 and T230, affected the ability of the protein to restore viability and histone mRNA processing to dSLBP null mutants. The T120A SLBP restored viability and histone pre-mRNA processing. However, the T230A mutant, located in a conserved TPNK sequence in the RNA binding domain, did not restore viability and histone mRNA processing in vivo, although it had full activity in histone mRNA processing in vitro. The T230A protein is concentrated in the cytoplasm, suggesting that it is defective in nuclear targeting, and accounting for its failure to function in histone pre-mRNA processing in vivo.
Joint Ventures, Universities, and Success in the Advanced Technology Program
Authors: Darby Michael R. , Zucker Lynne G. , Wang Andrew
Published: March 2004

America's most innovative firms (with 40%+ of U.S. patents assigned to U.S. entities during 1988–96) participate, often repeatedly, in the Commerce Department's Advanced Technology Program (ATP). Participation significantly increases firms’innovation (patenting) while receiving ATP support versus before and after. Firms generally increase patenting by 5–30 patents per year when participating—up 4 to 25% from before participation. University participation in a project increases firm patenting. Funding amount matters for single participants (often startups with small R&D budgets) with positive impact limited to firms with large grants. For joint ventures, participation is more important than funding level. (JEL O3, H5, L24)
Are Faculty Critical? Their Role in University–Industry Licensing
Authors: Thursby Jerry G. , Thursby Marie C.
Published: March 2004

Understanding the nature of the involvement of faculty in university licensing is important for understanding how technology is transferred through licensing as well as more controversial issues, such as the need for university licensing. Using data from a survey of firms that actively license in from universities, the authors explore the importance of faculty in the licensing and development of inventions, as well as how and why they are used and how the use of faculty relates to characteristics of firms. In particular, the authors find that the use of faculty through sponsored research in lieu of a license is closely related to the amount of basic research conducted by firms, whereas the use of faculty within the terms of a license is related to the prevalence of personal contacts between industry research and development researchers and university faculty. (JEL J44, 031)
Economic Competition and Evolution: Are There Lessons from Ecology?
Authors: Tisdell Clem
Published: March 2004

After generally discussing models in ecology and economics that combine competition, optimization, and evolution, this article concentrates on models of intraspecific competition. It demonstrates the importance of diversity/inequalities within populations of species and other environments for the sustainability of their populations, given the occurrence of environmental change. This is demonstrated both for scramble (open‐access) and contest competition. Implications are drawn for human populations and industrial organization. The possibility is raised that within‐industry competition may not always exist between firms in all stages of the development of a new industry. Policy implications are considered. For example, it is argued that policies designed to encourage intense business competition and maximum economic efficiency have the drawback of eventually making industries highly vulnerable to exogenous economic changes. (JEL L100, J100, Q150)
Financial Innovation and Divisia Money in Taiwan: Comparative Evidence from Neural Network and Vector Error‐Correction Forecasting Models
Authors: Binner Jane M. , Gazely Alicia M. , Chen Shu‐Heng , Chie Bin‐Tzong
Published: March 2004

In this article a Divisia monetary index is constructed for the Taiwan economy, and its inflation forecasting potential is compared with that of its traditional simple sum counterpart. The Divisia index is adjusted in two ways to allow for the financial liberalization that Taiwan has experienced since the 1970s. The powerful artificial intelligence technique of neural networks is used and is found to beat the conventional econometric techniques in a simple inflation forecasting experiment. The preferred inflation forecasting model is achieved using networks that employ a Divisia M2 measure of money that has been adjusted to incorporate a learning mechanism to allow individuals to gradually alter their perceptions of the increased productivity of money. The explanatory power of the two innovation‐adjusted Divisia aggregates dominates that of the simple sum counterpart in the majority of cases. (JEL C4, E4, E5)
Global Linkages of Subnational Regions: Coastal Exports and International Networks
Authors: Bardhan Ashok Deo , Guhathakurta Subhrajit
Published: March 2004

This article explores the determinants of exports of the two coastal regions of the continental United States and the differences between them. The authors evaluate the contribution of international sociocultural and business networks to exports of the two major coastal regions of the United States, the West Coast and the East Coast, and examine their geographic trading affinities. The article finds that although transnational business ties play a significant positive role in boosting overall exports of both coasts, social networks of foreign‐born immigrants play a similar role only for the West Coast. The study also suggests that the significance of distance as a proxy for transportation costs is somewhat diminished once these networks are taken into account. Both regions seem to share a propensity for exports to the Asia‐Pacific region. (JEL R0, F10, F20)
Depositor Liquidity and Loss Sharing in Bank Failure Resolutions
Authors: Kaufman George G.
Published: March 2004

Bank failures are widely feared because depositors may suffer losses in the value of their deposits and restrictions in access to their deposits. In the United States, this is not true for insured deposits, which are made fully available to depositors almost immediately. But both problems may occur for uninsured deposits. One way to mitigate liquidity loss to uninsured depositors is to make the estimated recovery value of their deposits quickly available to them by the Federal Deposit Insurance Corporation (FDIC). Such a policy would greatly enhance the FDIC's ability to resolve large bank insolvencies without having to protect uninsured depositors through too‐big‐to‐fail policies. (JEL G21, G28, G10)
Dynamic Relationships among GCC Stock Markets and Nymex Oil Futures
Authors: Hammoudeh Shawkat , Aleisa Eisa
Published: March 2004

Daily relationships among stock markets of the Gulf Cooperation Council (GCC) members, excluding Qatar, form two equilibrium relationships with varying predictive power. The Saudi market leads, followed by Bahrain and United Arab Emirates. Kuwait, which is dominated by momentum traders, and Oman have the weakest links with the other GCC markets. Only the Saudi index can predict—and be predicted by—New York Mercantile Exchange oil futures prices. Therefore these markets are candidates for diversified regional portfolios at the country level. The trading day effect is weak for all GCC markets and oil futures prices but remains consistent with findings for the U.S. stock market. (JEL C22, F3, Q49)
The Education/Growth Relationship: Evidence from Real State Panel Data
Authors: Bensi Michelle T. , Black David C. , Dowd Michael R.
Published: March 2004

This article employs 1963–97 panel data for the 48 contiguous U.S. states (and District of Columbia) to examine the relationship between real personal income and real education expenditures as well as that between real personal income and six measures of real research and development expenditures. Bivariate regressions are employed to determine whether the information content between real education expenditures and real income runs from real income to real education spending or vice versa. The authors find that when data are relative to the U.S. average, the direction of information content runs from real state‐level education expenditures to real state‐level income. (JEL I2, H72)
The Demand for Cigarettes in the Republic of Korea and Implications for Government Policy to Lower Cigarette Consumption
Authors: Kim Songjune , Seldon Barry J.
Published: March 2004

The government of the Republic of Korea has become concerned with reducing cigarette consumption, which historically has been very prevalent. This study contributes to the debate over cigarette policies in South Korea, with implications for other Asian countries with similar economic and cultural backgrounds. The authors estimate cigarette demand in South Korea over the period 1960–97, considering various government policies employed to reduce cigarette consumption. Results of the estimation suggest that taxation appears to be a viable means of reducing cigarette consumption, but methods to inform the public about the health hazards of smoking should be reconsidered and strengthened. (JEL H51, I18, L66)
CFOs and the audit committee
Authors: Morrow John F.
Published: March 2004

The relationship between the CFO and the audit committee has changed from oftentimes one of minimal contact to almost daily interaction as a result of the Sarbanes‐Oxley legislation. This is probably for the best, as the remoteness of audit committees in the past likely contributed to at least some of the corporate dysfunction we have seen in recent years.
The case for strategic real estate outsourcing
Authors: Martin Lauralee
Published: March 2004

Maintaining a constant focus on business priorities is a key factor for management success today. With increasing competition in every business space, any distraction from core business propositions can initiate a slide in market share and profitability. In addition, at a time when challenging economic conditions limit growth opportunities for many companies, controlling costs assumes heightened importance for delivering profitable results.
Digital library and information systems: where are we heading?
Authors: Yeates Robin
Published: March 2004

States that the market for computer systems that manage digital content is currently in turmoil but that the overall direction of both library policy and supplier strategy seems clear – the digital library will play a key role in organizing content and supporting and facilitating access to it. Suggests that digital libraries will become diffuse asset management systems capable of high levels of integration with other learning, management and recreation systems, and that digital libraries will tend increasingly to adopt an ever wider range of platforms and content technologies created for wider commercial and economic purposes.
Web services: what they are and their importance for libraries
Authors: Ineˆs Cordeiro Maria , de Carvalho Joaquim
Published: March 2004

This paper introduces the main ideas and concepts underlying Web services, and their potential for a new generation of Internet‐based distributed applications. A summary of the key standards that are the foundation of Web services technology and its underlying basic model is provided, exploring the potential for interoperability among independently managed systems. This is of special importance for libraries, along with the advantage of reducing the development and deployment costs of distributed applications, and of leveraging the value of IT and data legacy assets. The paper explains developments in Web services technology in the library field and underlines its significance for changes in the library information systems paradigm.
Expanding the role of EAD: providing adequate metadata for digital as well as original archive documents
Authors: Sexton Anna , Turner Chris
Published: March 2004

This article discusses the use of Encoded Archival Description (EAD) as a metadata framework within applications that are primarily concerned with the provision of access to digital forms of archive documents. These digital forms are transcripts encoded using the Text‐Encoding Initiative (TEI) and images. The article argues that EAD, as it currently stands, is focused on the provision of metadata for original archive documents rather than for digital forms of originals, and it explores where metadata about originals and their digital forms converge and diverge. It suggests how the EAD framework can be expanded to allow for the capture of adequate metadata about both types of document and asserts that such expansion enables EAD to act as a more complete and comprehensive metadata framework in online environments. This approach to digitisation relies on the flexibility of XML technology. The article is based on the research undertaken within the LEADERS project (http://www.ucl.ac.uk/leaders‐project/).
From notepad to content management: the evolution of a web publication
Authors: Evans Peter
Published: March 2004

This article describes the evolution of the medium sized (500 page) Biblio Tech Review web site, from conventional management methods to advanced content management. The article covers design considerations, authoring tools, database publishing and content management techniques. Problems and their solution are described. The implications for digital preservation are outlined. Diagrams illustrate site architecture and database structure.
Multi‐disciplinary content management in “Darwin Country” – a case study
Authors: Boyd Peter D.A.
Published: March 2004

“Darwin Country” is a website project giving access to images and information about the archives, paintings, decorative arts, archaeology and scientific collections of Shrewsbury Museums Service and its partners. By April 2002 the website had about 9000 pages and 6000 images. It is grant‐aided by the Resource IT Challenge Fund and the West Midlands Regional Museum Council. This database‐driven website utilises open‐source software to provide multi‐disciplinary content in a “knowledge‐net” environment created and maintained through a Content Management System. However, the author argues that “Content Management” is actually a process that involves more than a piece of software and should start with a vision and formulation of a strategy.
Electronic content development in a community setting
Authors: Raven Claire
Published: March 2004

Content Management has long been a controversial issue in the academic, public and private sectors. But how does the debate apply to community organisations and information about them? This article considers some answers to this based on the experience of Manchester Community Information Network (MCIN), who have amassed a great deal of experience in facilitating the creation of electronic content in a community setting. MCIN is a charity working in the North West of England which has been involved in community development work for about 8 years, using ICT to develop electronic community information and participation. The article will explore issues relating to the first stages of the content life cycle; the creation, editing, storage and publishing of electronic community information.
Creating an in‐house content management system
Authors: Roberts Alexander
Published: March 2004

This paper describes the choice to develop an in‐house content management system for the Coalfield Web Materials and the Mine of Information projects currently running at the University of Wales Swansea.
Linking: where are we now and where are we going?
Authors: Samuels Harry E.
Published: March 2004

Linking in the scholarly environment has built on the basic linking capabilities of the World Wide Web and evolved to make the research process faster and more efficient. There are many components in the current linking environment and they often overlap and frequently inter‐operate. Despite the advances, linking is still in its infancy and future evolution will be rapid, especially as the underlying technologies evolve. This paper describes the current scholarly linking environment, its evolution, its components, and what we have to look forward to in the future.
Selecting a content management system
Authors: White Martin
Published: March 2004

Selecting content management system software cannot be accomplished just by looking at brochures and comparing features, as vendors offer a complex mix of additional functionality around a core set of application modules. To reduce the risks inherent in the deployment of software that will have a significant impact on the way an organization works, it is important to prepare a content management strategy as a precursor to a formal definition of requirements. The overall process can take a year from the decision to explore the benefits of a CMS, to having an organization‐wide deployment.
Teaching and learning with images
Authors: Mowat Elaine
Published: March 2004

Digitisation initiatives have produced a wealth of high quality digital images for educational use. Illustrated by examples from the SCRAN digital library, this article describes some of the ways in which images can be used to support innovative and effective learning and teaching in further and higher education.