bluuebunny/crossref_metadata_2025_split huggingface dataset:
Conformational fluctuations and electronic properties in myoglobin
Authors: Aschi Massimiliano , Zazza Costantino , Spezia Riccardo , Bossa Cecilia , Di Nola Alfredo , Paci Maurizio , Amadei Andrea 
Published: March 2004

AbstractIn this article we use the recently developed perturbed matrix method (PMM) to investigate the effect of conformational fluctuations on the electronic properties of heme in Myoglobin. This widely studied biomolecule has been chosen as a benchmark for evaluating the accuracy of PMM in a large and complex system. Using a long, 80‐ns, molecular dynamics simulation and unperturbed Configuration Interaction (CISD) calculations in PMM, we reproduced the main spectroscopic features of deoxy‐Myoglobin. Moreover, in line with our previous results on a photosensitive protein, this study reveals a clear dynamical coupling between electronic properties and conformational fluctuations, suggesting that this correlation could be a general feature of proteins. © 2004 Wiley Periodicals, Inc. J Comput Chem 7: 974–984, 2004
Applied cognitive psychology: Implications of cognitive psychology for clinical psychology and psychotherapy
Authors: Eysenck Michael W. 
Published: March 2004

AbstractCognitive psychology has made numerous contributions to clinical psychology, and these contributions are considered especially with reference to the anxiety disorders. It is argued that there are four major contributions that can be identified. First, the cognitive approach has led to the development of complex models showing the main cognitive processes and structures of relevance to an understanding of anxiety disorders. Second, controlled laboratory studies permit a more detailed investigation of cognitive biases in anxious patients than generally is feasible in more naturalistic settings. Third, the cognitive approach provides relevant evidence with respect to the issue of whether cognitive biases play a role in the development and maintenance of anxiety disorders. Fourth, the enhanced understanding of the anxiety disorders that has arisen from the cognitive approach has had beneficial effects on therapeutic practice in a number of significant ways. In sum, it is claimed that clinical psychology has benefited considerably from cognitive theory and research. © 2004 Wiley Periodicals, Inc. J Clin Psychol.
Cognitive factors in health psychology and behavioral medicine
Authors: Montgomery Guy H. 
Published: March 2004

AbstractResearch in health psychology and behavioral medicine has increased exponentially over the past three decades. Prevention and control efforts across a variety of diseases have recognized that individual difference factors, belief systems, attitudes, behaviors, and environments can affect an individual's health. This work continues, as there has been no abatement in the rate at which aversive, chronic, and life‐threatening diseases continue to affect millions of Americans each year. The purpose of this review is to discuss the influence of cognitive factors on health psychology and behavioral medicine using the cancer literature as an example. The main discussion focuses on the impact of cognitive constructs on behavioral medicine and health psychology in cancer, which appears to be widespread. The article concludes with some implications for future research in the area. © 2004 Wiley Periodicals, Inc. J Clin Psychol.
An essential role for the His‐8 residue of the SDF‐1α–chimeric, tropism‐redirected Env protein of the Moloney murine leukemia virus in regulating postbinding fusion events
Authors: Katane Masumi , Fujita Rika , Takao Eiko , Kubo Yoshinao , Aoki Yasunobu , Amanuma Hiroshi 
Published: March 2004

AbstractBackgroundTo use retroviral vectors for the cell‐specific delivery of genes, it is necessary to redirect their receptor tropism to cell‐specific receptors. Previously, we reported that a Moloney murine leukemia virus (MLV) retroviral vector containing a human stromal‐derived factor‐1α (SDF‐1α)–chimeric envelope protein (Env) (S3) acquired the ability to transduce human cells via CXCR4, the cognate receptor for SDF‐1α, while retaining the ability to transduce mouse cells via mCAT1.MethodsWe constructed expression plasmids for derivatives of the S3 Env protein; S3‐D84K containing an Asp‐84‐to‐Lys (D84K) substitution, S3‐H8R‐D84K containing D84K and an additional His‐8‐to‐Arg substitution, and S3‐D84K‐RY containing D84K and additional Gln‐227‐to‐Arg plus Asp‐243‐to‐Tyr substitutions which have been suggested to suppress the loss of function of His‐8. Cellular expression, virion incorporation, and entry functions of these derivatives were investigated.ResultsAll three derivatives were incorporated into virions. The S3‐D84K vector lost its ecotropism, but could transduce CXCR4‐expressing human and mouse cells at titers of 103 to 104 colony‐forming units (cfu)/ml. The S3‐H8R‐D84K vector did not show transduction, although its Env protein could bind to CXCR4. The transduction titer of the S3‐D84K‐RY vector via CXCR4 was slightly lower than that of the S3‐D84K vector. These results indicate that the His‐8 residue of the S3‐D84K Env protein is indispensable and may be fully functional in postbinding membrane fusion.ConclusionsInsertion of a ligand at Pro‐79 of the Moloney MLV Env protein has proved to be a valuable strategy for constructing direct targeting retroviral vectors, since it permits the formation of a redirected Env protein without ecotropism, and it does not disrupt the function of the essential His‐8 residue. Copyright © 2004 John Wiley & Sons, Ltd.
LIM kinase‐2 targeting as a possible anti‐metastasis therapy
Authors: Suyama Eigo , Wadhwa Renu , Kawasaki Hiroaki , Yaguchi Tomoko , Kaul Sunil C. , Nakajima Motowo , Taira Kazunari 
Published: March 2004

AbstractBackgroundMetastatic properties of tumors involve movement of cancerous cells from one place to another and tissue invasion. Metastatic cells have altered cell adhesion and movement that can be examined by in vitro chemotaxis assays. The Rho/ROCK/LIM kinase pathway is one of the major signaling pathways involved in tumor metastasis. It is involved in the regulation of the actin cytoskeleton. Using the randomized ribozyme library, we initially found that metastatic human fibrosarcoma cells harboring ribozyme specific for ROCK lose their metastatic properties. In this study, we have determined the effect of ribozymes specific for LIM kinase‐2 on metastatic and proliferative phenotypes of human fibrosarcoma cells.MethodsWe attempted to target LIM kinase‐2 (LIMK‐2) expression by hammerhead ribozymes (Rz) in human metastatic fibrosarcoma cells. An effective ribozyme was selected based on the expression analysis. Cells were stably transfected with Rz specifically effective for LIMK‐2 and were examined for metastatic and proliferative properties.ResultsAnalyses of cellular phenotypes such as cell proliferation, cell migration and colony‐forming efficiency revealed that the suppression of LIMK‐2 expression in human fibrosarcoma cells limits their migration and dense colony‐forming efficiency without affecting cell proliferation rate or viability.ConclusionsSpecific targeting of metastatic and malignant properties of tumor cells by LIMK‐2 ribozyme may serve as an effective therapy for invasive tumors with minimum effect on the surrounding normal cells. Copyright © 2004 John Wiley & Sons, Ltd.
Lactosylated polyethylenimine for gene transfer into airway epithelial cells: role of the sugar moiety in cell delivery and intracellular trafficking of the complexes
Authors: Grosse Stéphanie , Aron Yolande , Honoré Isabelle , Thévenot Guiti , Danel Claire , Roche Annie‐Claude , Monsigny Michel , Fajac Isabelle 
Published: March 2004

AbstractBackgroundAs we have previously shown that lactosylated polyethylenimine (PEI) is the most efficient glycosylated PEI for gene transfer into human airway epithelial cells in primary culture, we have studied here the role of the lactose residue in the enhancement of gene transfer efficiency observed with lactosylated PEI as compared with unsubstituted PEI in immortalized (ΣCFTE29o‐ cells) and primary human airway epithelial cells.Methods and resultsAfter three transfections of 1 h performed daily, 60% of ΣCFTE29o‐ cells were transfected with lactosylated PEI, whereas 25% of cells were transfected with unsubstituted PEI (p < 0.05). Cell viability was 1.8‐fold greater with lactosylated PEI as compared with unsubstituted PEI (p < 0.05). As assessed by flow cytometry, the cellular uptake of lactosylated complexes was greater than that of complexes made with unsubstituted PEI (p < 0.05) and involved mostly a receptor‐mediated endocytosis. The study of the intracellular trafficking in airway epithelial cells of complexes showed an endosomal and lysosomal accumulation of lactosylated complexes. In the presence of a proton pump inhibitor, the level of lactosylated and unsubstituted PEI‐mediated gene expression was reduced more than 20‐fold, whereas the cell viability increased to almost 100%. For both complexes, a nuclear localization was observed for less than 5% of intracellular complexes.ConclusionsOur results show that the greater gene transfer efficiency observed for lactosylated complexes may be attributed to a higher amount of lactosylated complexes incorporated by airway epithelial cells and a lower cytotoxicity that might be related to reduced endosomolytic properties. However, the lactose residues substituting the PEI did not promote the entry of the plasmid into the nucleus. Copyright © 2004 John Wiley & Sons, Ltd.
Biodistribution of the RD114/mammalian type D retrovirus receptor, RDR
Authors: Green Bronwyn J. , Lee C. Soon , Rasko John E. J. 
Published: March 2004

AbstractBackgroundThe limited expression of viral receptors on target cells is a recognized barrier to therapeutic gene transfer. Previous analysis of receptor expression has been performed using indirect methods due to a lack of receptor‐specific antibodies.MethodsIn this report we have used anti‐RDR antiserum to provide direct histochemical and flow cytometric analysis of the expression of RDR, which is the cognate receptor for RD114‐pseudotyped vectors as well as being a neutral amino acid transporter.ResultsRDR was present on a range of normal tissues with relevance to gene therapy including: colon, testis, ovary, bone marrow and skeletal muscle. It was also highly expressed on immature cells present in the squamous epithelia of skin, cervix, nasal mucosa, bronchus and tonsil. Of relevance to possible germline gene transfer, we demonstrated a lack of RDR expression on male or female germ cells. RDR expression on mature hemopoietic cell subsets showed up to 5‐fold variability between individuals within each lineage—with some individuals expressing low levels of RDR across all blood lineages. Both myeloid and monocytic lineages contained the highest fraction of cells expressing RDR, whereas lymphoid lineages showed the lowest. Coexpression of CD34 and RDR ranged from 2.04 to 0.44% in G‐CSF‐mobilized peripheral blood samples.ConclusionsAs a means to optimize gene transfer protocols, biodistribution studies such as these are fundamental to enable targeting of the virus receptor most abundantly expressed on relevant populations. The inter‐individual variation of receptor expression seen here also raises the possible requirement for tailor‐made gene therapy protocols. Copyright © 2004 John Wiley & Sons, Ltd.
Transgene expression in hypertrophied and aged skeletal muscle in vivo by lentivirus delivery
Authors: Ouyang Jun , Alway Stephen E. 
Published: March 2004

AbstractBackgroundHuman immunodeficiency virus type 1 (HIV‐1)‐based lentiviral vectors can infect dividing and non‐dividing cells. The purpose of this study was to investigate the efficacy of lentiviral transfer of the green fluorescent protein (eGFP) gene into post‐mitotic quail skeletal muscle and into fibers undergoing hypertrophy and hyperplasia via addition of satellite cell nuclei.MethodsA vesicular stomatitis virus G‐protein (pMFG)‐pseudotyped lentiviral vector encoding the eGFP gene coding sequence, flanked by two long terminal repeats (LTRs) (pHR‐CMV‐eGFP‐Wsin‐18) and an optional plasmid (pRSVrev), was generated in 293T cells by means of the three‐plasmid system. The resulting lentiviral vector, LtV/eGFP, was injected in vivo into the patagialis muscles of young (6 weeks of age) and senescent (100 weeks of age) Japanese quail. Polybrene (10 µg/muscle) was injected in the right patagialis muscles of some birds. A weight was attached to the left wing of birds immediately following LtV/eGFP injection for 7, 14 or 21 days to induce hypertrophy in the patagialis muscle via satellite cell activation. A time‐released bromodeoxyuridine (BrdU) pellet was implanted in some birds to identify satellite cells that had proliferated during stretch‐overload. Control and experimental muscles were examined 7, 14 or 21 days following injection and wing loading.ResultsSeven days post‐injection proved to be insufficient to visualize and quantify the eGFP signal. EGFP‐positive fibers were evident 14 and 21 days after injection of LtV/eGFP. As expected, wing weighting resulted in hypertrophy of the patagialis of senescent and young birds and an increase in collagen and other connective tissue in muscles of old birds. Nevertheless, this did not interfere with expression levels or intensity of the eGFP vector because the area of eGFP‐positive fibers normalized to total muscle area was similar in control and overloaded muscles, and in muscles that were injected with the lentivirus vector but not loaded. Polybrene improved eGFP expression by 41% as compared with muscles that received only the lentivirus.ConclusionsThese findings provide the first evidence that this lentivirus vector is an efficient tool for gene transfer and expression in mature (non‐growing) fibers of adult birds, sarcopenic fibers of senescent birds and hypertrophying skeletal muscle fibers of both young adult and old birds. This system could be useful for tissue engineering, or treating aging‐associated muscle‐wasting diseases such as sarcopenia, especially when exercise/overload are added to gene therapy approaches to combat muscle loss with aging. Copyright © 2004 John Wiley & Sons, Ltd.
Expression of HIV‐1 integrase in CEM cells inhibits HIV‐1 replication
Authors: van Griensven Johan , Zhan Xiaojing , Van Maele Bénédicte , Pluymers Wim , Michiels Martine , De Clercq Erik , Cherepanov Peter , Debyser Zeger 
Published: March 2004

AbstractBackgroundHIV‐1 integrase (IN) is an interesting target for the gene therapy of AIDS. Although the in vivo functions are not well characterized, it is thought that IN has pleiotropic effects and plays a central role in the interplay between the virus and the host cell. Expression of IN in mammalian cells has proven difficult. We have previously established a 293T‐derived cell line that stably expresses high levels of HIV‐1 IN from a synthetic gene. We now have constructed CEM‐derived cell lines stably expressing the enzyme or its different domains and studied the impact of IN expression on HIV‐1 replication.MethodsThe CEM cell lines were selected following transduction with a retroviral vector encoding the full‐length IN, the N‐terminal domain, the catalytic core or the C‐terminal domain. Stable IN expression in CEM cell lines was verified by Western blotting. The impact of IN expression on HIV‐1 replication and HIV‐1 vector transduction was studied.ResultsA marked inhibitory effect on HIV‐1 replication was observed in CEM cells expressing IN. Expression of IN interfered with both particle production and integration. Expression of the N‐terminal domain alone was sufficient for the inhibiting of HIV‐1 replication.ConclusionsExpression of IN in CEM cells inhibits HIV‐1 replication by a cumulative inhibitory effect on integration and particle production, in accord with the known pleiotropic interactions of IN. The inhibition of HIV‐1 replication in CEM cells expressing the N‐terminal domain of IN may lead to a novel approach for the gene therapy of AIDS. Copyright © 2004 John Wiley & Sons, Ltd.
Adenovirus vectors targeting αV integrin or heparan sulfate receptors display different distribution of transgene activity after intramuscular injection
Authors: Cirielli Corrado , Serino Francesco , Straino Stefania , Toietta Gabriele , Abeni Damiano , Ventoruzzo Giorgio , Orlando Giuseppe , Mazzanti Paola , Melillo Guido , Whickham Thomas J. , Kovesdi Imre , Biglioli Paolo , Gaetano Carlo , Capogrossi Maurizio C. 
Published: March 2004

AbstractBackgroundModification of the fiber proteins in replication‐deficient adenoviral (Ad) vectors through incorporation of specific receptor‐binding motifs may represent a strategy to enhance their tissue targeting capabilities.MethodsIn this study, we compared an unmodified Ad (GV10) with two mutated vectors obtained by insertion of specific target sequences that redirect binding, either toward αV integrin (RGD) or heparan sulfate (UTV) cellular receptors, for reporter gene expression spatial distribution in the rabbit skeletal muscle. In a first series of experiments, injection volume was kept constant and activity of a lacZ transgene was evaluated 48 h after injection of the Ad vectors at different doses. In separate experiments, the effects of different volumes of injection at a constant dose of Ad vector were monitored.ResultsAll vectors evaluated showed a significant increase in the number of lacZ‐positive muscle segments, with increasing vector dose. However, in muscles treated with the UTV vector, fewer muscle fibers were β‐gal‐positive than in GV10 or RGD vector treated animals. In fact, total β‐gal activity increased in a dose‐dependent fashion in the GV10‐ and RGD‐treated muscles, but not in the UTV‐treated ones. Remarkably, in samples from UTV‐treated animals, a volume‐dependent enhancement of transgene expression was observed during experiments performed at the same dose and different injection volumes.ConclusionsThe results of the present study demonstrate that altering Ad affinity for cellular receptors modulates the level and distribution of transgene activity, conferring characteristics that may allow for treatment customization. Copyright © 2004 John Wiley & Sons, Ltd.
Suppression of tumor growth, invasion and angiogenesis of human gastric cancer by adenovirus‐mediated expression of NK4
Authors: Heideman Daniëlle A. M. , van Beusechem Victor W. , Bloemena Elisabeth , Snijders Peter J. F. , Craanen Mikael E. , Offerhaus G. Johan A. , Derksen Patrick W. B. , de Bruin Michiel , Witlox M. Adhiambo , Molenaar Bonnie , Meijer Chris J. L. M. , Gerritsen Winald R. 
Published: March 2004

AbstractBackgroundTo improve the prognosis of patients with gastric cancer it is important to develop novel treatment modalities targeting the malignant behavior of tumor cells. Concerning this, NK4, which acts as HGF‐antagonist and angiogenesis inhibitor, might be a potential therapeutic agent for gastric cancer. The HGF‐c‐MET pathway plays a pivotal role in gastric tumor growth, invasion, metastasis and angiogenesis. Therefore, the current study investigates whether adenoviral vector‐mediated NK4 gene therapy has therapeutic potential for gastric cancer.MethodsExpression of HGF and c‐MET in normal and (pre‐)malignant gastric tissue was studied by immunohistochemistry. The effects of adenoviral vector‐mediated expression of NK4 on the biological behavior of gastric cancer cells were studied in vitro and in vivo.ResultsThe majority of gastric cancers, i.e. 76%, express c‐MET and in all carcinomas HGF is expressed in either tumor or stromal cells. Normal gastric epithelial cells do not express either of these proteins. Transduction of gastric cancer cells with the replication‐deficient adenoviral vector AdCMV.NK4 resulted in efficient production and secretion of NK4. Consequently, proliferation, migration and invasion of gastric cancer cells were significantly inhibited. In addition, significantly reduced proliferation of vascular endothelial cells and efficient inhibition of angiogenesis were achieved. Finally, treatment of established human gastric tumor xenografts with AdCMV.NK4 resulted in significant tumor growth delay and significant reduction of intratumoral microvessel density.ConclusionsThe present study shows that adenoviral vector‐mediated expression of NK4 is a promising strategy to treat human gastric cancer by simultaneous interfering with primary tumor growth, metastasis and angiogenesis. Copyright © 2004 John Wiley & Sons, Ltd.
Polymer‐coated polyethylenimine/DNA complexes designed for triggered activation by intracellular reduction
Authors: Carlisle Robert C. , Etrych Tomas , Briggs Simon S. , Preece Jon A. , Ulbrich Karel , Seymour Leonard W. 
Published: March 2004

AbstractBackgroundSite‐specific gene delivery requires vectors that combine stability in the delivery phase with substantial biological activity within target cells. The use of biological trigger mechanisms provides one promising means to achieve this, and here we report a transfection trigger mechanism based on intracellular reduction.MethodsPlasmid DNA was condensed with thiolated polyethylenimine (PEI‐SH) and the resulting nanoparticles surface‐coated using thiol‐reactive poly[N‐(2‐hydroxypropyl)methacrylamide] (PHPMA) with 2‐pyridyldisulfanyl or maleimide groups, forming reducible disulphide‐linked or stable thioether‐linked coatings, respectively.ResultsBoth sets of polymer‐coated complexes had similar size and were stable to a 250‐fold excess of the polyanion poly(aspartic acid) (PAA). Reduction with dithiothreitol (DTT) allowed complete release of DNA from disulphide‐linked coated complexes, whereas complexes with thioether‐linked coating remained stable. Disulphide‐linked complexes showed 40–100‐fold higher transfection activity than thioether‐linked ones, and activity was selectively further enhanced by boosting intracellular glutathione using glutathione monoethyl ester or decreased using buthionine sulfoximine. The chloroquine‐ and serum‐independent transfection activity of disulphide‐linked coated complexes suggests this system may provide a viable trigger mechanism to enable site‐specific transfection in complex biological settings.ConclusionsLinkage of hydrophilic polymer coating to PEI/DNA complexes via reducible disulphide bonds offers a means of fulfilling the contradictory requirements for extracellular stability and intracellular activity. Copyright © 2004 John Wiley & Sons, Ltd.