bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fiber diffraction study of spicules from marine sponges
Authors: Croce Gianluca , Frache Alberto , Milanesio Marco , Viterbo Davide , Bavestrello Giorgio , Benatti Umberto , Giovine Marco , Amenitsch Heinz 
Published: October 2003

AbstractA synchrotron radiation fiber diffraction structural study of the axial filament of siliceous spicules from two species of marine sponges (the Demosponge Geodia cydonium and the Hexactinellid Scolymastra joubini) was carried out. The sharpness of the spots in the diffraction patterns indicated that the protein units in the filament of both samples were highly organized. A possible explanation is that the arrangement of the protein units is similar to that of the pores in highly ordered siliceous mesoporous materials. Nevertheless, the diffraction patterns are quite different for the two types of spicules. The pattern of G. cydonium is consistent with a regular 2D hexagonal lattice of protein units in the direction perpendicular to the spicule axis, with a repeating distance of 5.8 nm; the units are linked to form fibers along the axis. The pattern of S. joubini indicates the presence of two different 2D lattices in which the repeating protein units are inclined by +50° and –50° with respect to the elongation axis; the distance between the units increases to 8.4 nm. This 2D model is consistent with hexagonal packing of spirally oriented cylindrical protein units elongated along the filament axis. Microsc. Res. Tech. 62:378–381, 2003. © 2003 Wiley‐Liss, Inc.
Combination with CD/5‐FC gene therapy enhances killing of human bladder‐cancer cells by radiation
Authors: Zhang Zhujun , Shirakawa Toshiro , Hinata Nobuyuki , Matsumoto Akira , Fujisawa Masato , Okada Hiroshi , Kamidono Sadao , Matsuo Masafumi , Gotoh Akinobu 
Published: October 2003

AbstractBackgroundResistance to radiation and chemotherapy is a significant obstacle to the treatment of advanced bladder cancer. Gene therapy combined with radiation represents a new approach to cancer treatment. In the present study, we investigated whether adenovirally directed, cytosine deaminase (CD)/5‐fluorocytosine (5‐FC) gene therapy could induce cell toxicity and radiosensitization through the intracellular production of 5‐fluorouracil (5‐FU) in bladder‐cancer cells.MethodsThree human bladder‐cancer cell lines, KK47 (wild‐type p53+), T24 (p53 mutated) and 5637 (p53 mutated), were investigated. A recombinant adenovirus vector containing the CD gene (Ad‐RSV‐CD) was used. Cells were infected with Ad‐RSV‐CD and treated with 5‐FC. Forty‐eight hours after infection, the cells were irradiated and cytotoxicity assays performed to determine the extent of increase in in vitro cytotoxicity. A KK47 subcutaneous tumor‐xenografts model was used in an animal study to examine the tumor growth inhibitory effect of this combination therapy. Ad‐RSV‐CD was directly injected into the tumor and daily 5‐FC was intraperitoneally injected. Forty‐eight hours after injection of Ad‐RSV‐CD, the tumor was irradiated. The tumor volume was measured every day.ResultsIn all three cell lines, the combination treatment enhanced the cell killing of human bladder‐cancer cells in vitro. It also enhanced the tumor‐growth inhibition in the KK47 tumor model.ConclusionsIn the present study, we demonstrated that CD/5‐FC gene therapy combined with radiation therapy enhances cell killing of human bladder‐cancer cells in in vitro and in vivo animal models. Copyright © 2003 John Wiley & Sons, Ltd.
Coxsackie‐and‐adenovirus receptor mRNA expression in human heart failure
Authors: Sasse A. , Wallich M. , Ding Z. , Goedecke A. , Schrader J. 
Published: October 2003

AbstractBackgroundAdenoviral vectors are widely used as gene‐transfer vehicles in experimental and clinical studies. Since virus incorporation and transfection efficacy depend to a large extent on the concentration of the coxsackie‐and‐adenovirus (CAR) receptor on target cells the aim of this study was to quantify the CAR‐receptor concentration in various human cardiomyopathies.MethodsAfter RNA isolation from myocardial biopsies obtained during surgical procedures, cDNA was generated by reverse transcription. The relative RNA content was analyzed by quantitative PCR using glyceraldehydes‐3‐phosphate dehydrogenase (GAPDH) as a standard reference. The cardiomyopathies (CM) analyzed were categorized according to their etiology in dilated CM (DCM, n = 28), ischemic CM (ICM, n = 52), CM in mitral valve disease (MVCM, n = 32) and aortic valve disease (AVCM, n = 32). Data were related to non‐cardiomyopathic tissue from donor hearts (non‐CM, n = 64).ResultsCompared with non‐CM hearts DCM showed a 34‐fold (±5.4) increase in CAR mRNA concentration, in ICM CAR mRNA was elevated by a factor of 12 (±4.3), in MVCM by 27 (±7) and AVCM by factor 47 (±9.3) (ANOVA p < 0.001). Compared with the expression in rat hearts CAR levels were found to be similar to those in human ICM.ConclusionsThese results show that cardiomyopathies associated with heart failure transcribe substantially higher levels – on average by a factor of 30 – of CAR‐mRNA than non‐failing control hearts. Myocardial gene transfer using adenoviral vectors should therefore be facilitated in human cardiomyopathies and may present a promising approach for therapeutic interventions. Copyright © 2003 John Wiley & Sons, Ltd.
Chemotherapy increases transgene expression in leukemic cells
Authors: Vereecque Rodolphe , Saudemont Aurore , Depil Stéphane , Quesnel Bruno 
Published: October 2003

AbstractBackgroundPatients with acute myeloid leukemia (AML) often obtain complete remission with chemotherapy but the majority of patients relapse. Combining chemotherapy and gene therapy appears to be a promising approach; however, the effects of chemotherapy on transgene expression in leukemic cells have not yet been investigated.MethodsDA1‐3b leukemic cells were transfected with pCDNA3 plasmids carrying GM‐CSF or LacZ cDNA. The leukemic K562 cell line and primary cultured AML cells were transduced with an Ad5.CMV‐LacZ adenoviral vector. Cells were then incubated with various concentrations of daunorubicin (DNR) and cytosine arabinoside (Ara‐C), and expression of the transgene was measured. Murine DA1‐3b‐pCDNA3/LacZ leukemic cells were also injected into syngeneic C3H/Hej mice.ResultsIn the cells carrying pCDNA3, DNR and Ara‐C dramatically increased expression of the LacZ and GM‐CSF transgenes. Over‐expression depended on drug dose and was due to increased transcription. Enhancement was also observed in K562 cells and in some primary cultured AML samples transduced with the Ad5.CMV‐LacZ adenovirus. Addition of N‐acetyl‐L‐cysteine inhibited the over‐expression, suggesting that reactive oxygen species were involved in activating the CMV promoter. In the A549 lung carcinoma cell line transduced with Ad5.CMV‐LacZ, Ara‐C had only a minor effect, and DNR had a detrimental effect, suggesting that expression depends on cell type. In vivo experiments in which mice received DA1‐3b‐pCDNA3/LacZ leukemic cells, and were then treated with Ara‐C, also showed increased transgene expression in these leukemic cells.ConclusionsIn leukemic cells, chemotherapeutic agents can induce over‐expression of transgenes. This suggests a promising combined strategy for the treatment of acute leukemia. Copyright © 2003 John Wiley & Sons, Ltd.
Rational development of a HIV‐1 gene therapy vector
Authors: Anson D. S. , Fuller M. 
Published: October 2003

AbstractBackgroundHIV‐1 provides an attractive option as the basis for gene transfer vectors due to its ability to stably transduce non‐cycling cell populations. In order to fully utilise the promise of HIV‐1 as a vector it is important that the effects of viral cis sequence elements on vector function are carefully delineated.MethodsIn this study we have systematically evaluated the effect of various cis elements from the HIV‐1 YU‐2 genome that have been implicated as either affecting vector performance, or HIV‐1 replication, on the efficiency of vector production (titre and infectivity). As a measure of the relative safety of vectors their propensity to inadvertently transfer the gagpol gene to transduced cells was assessed.ResultsSequences that were found to increase vector titre were from the 5′ end of the gag gene, from the 5′ and 3′ ends of the env gene, from immediately upstream of the polypurine tract, and the central polypurine tract. The substitution of the HIV‐1 RRE with heterologous RNA transport elements, or the deletion of the RRE, resulted in greatly reduced vector titres. RNA analysis suggested that the role of the Rev/RRE system extends beyond simply acting as an RNA nuclear export signal. The relative safety of different vector designs was compared and an optimal construct selected.ConclusionsBased on our results we have constructed a vector that is both more efficient, and has better safety characteristics, than the widely used pHR′ HIV‐1 vector construct. Copyright © 2003 John Wiley & Sons, Ltd.
DNA transfection of mononuclear cells in muscle tissue
Authors: Grønevik Eirik , Tollefsen Stig , Sikkeland Liv Ingunn Bjoner , Haug Terje , Tjelle Torunn Elisabeth , Mathiesen Iacob 
Published: October 2003

AbstractBackgroundGenes encoding non‐self proteins may be injected into skeletal muscles in vivo to obtain induction of cellular and humoral immune responses against the encoded antigens (DNA vaccination). Bone marrow derived professional antigen‐presenting cells (APCs) play a key role in the induction of immunity by DNA vaccination. In the present work we have investigated whether the APCs are transfected by DNA injection into muscle.MethodsDNA encoding green fluorescent protein (GFP) was injected into rat and mouse limb muscle and followed by electroporation. Whole mount muscle tissue with GFP‐positive mononuclear cells (MNCs) were treated with immunocytochemical markers specific for leukocytes, and studied with fluorescent microscopy. To detect transfected cells migrating to peripheral lymphoid tissue RT‐PCR was applied on RNA isolated from the draining popliteal lymph node and spleen. Lymphoid tissue was also analyzed with real‐time PCR for distribution of the injected plasmid.ResultsMNCs were transfected after intramuscular DNA injection, and, following DNA injection with electroporation, the number of GFP‐positive MNCs increased 6‐fold in rats and 14‐fold in mice. None of the GFP‐positive MNCs were stained with leukocyte‐specific antibodies. Even though GFP encoding DNA was detected in the popliteal lymph node, no RNA encoding GFP was found in the lymph node or spleen. However, MHC II‐positive cells in the muscle tissue appeared preferentially around the transfected MNCs.ConclusionsMany MNCs in the muscle are transfected after intramuscular DNA injection. Electroporation significantly increases the number of transfected MNCs. None of the observed transfected MNCs however were leukocytes. MHC II‐positive cells accumulated around transfected MNCs; this suggests that transfer of antigen from transfected MNCs to APCs may contribute to the immune response. Copyright © 2003 John Wiley & Sons, Ltd.
Functional delivery of large genomic DNA to human cells with a peptide‐lipid vector
Authors: White Robert E. , Wade‐Martins Richard , Hart Stephen L. , Frampton Jon , Huey Bryan , Desai‐Mehta Ami , Cerosaletti Karen M. , Concannon Patrick , James Michael R. 
Published: October 2003

AbstractBackgroundNonviral gene transfer vectors have the potential to deliver much larger DNA constructs than current viral vectors but suffer from a low transfection efficiency. The LID vector, composed of Lipofectin (L), an integrin‐targeting peptide (I) and DNA (D), is a highly efficient synthetic vector, both in vitro and in vivo, which may allow the transfer of genomic loci for gene therapy.MethodsTransfection efficiencies were quantitated using the green fluorescent protein (GFP) reporter. Expression of a large genomic locus (NBS1 [Nijmegen breakage syndrome], encoding nibrin) was assessed by immunofluorescence.ResultsWe report a systematic study of the parameters influencing delivery of BAC‐based plasmids ranging in size from 12 to 242 kb using the LID vector. We showed 60% of cells were transfected with the smaller plasmids while plasmids up to 242 kb were consistently delivered to over 10% of cells. The number of transfected cells was related to number of plasmids in the transfection complex independent of plasmid size. Atomic force microscopy showed that LID particle size increased with plasmid size consistent with one plasmid molecule per particle. When LID vectors were used to deliver the NBS1 gene as a 143 kb construct to primary NBS cells, at least 57% of cells expressing GFP also expressed functional nibrin.ConclusionsWe show that LID vectors represent a promising tool for the transfer of complete genomic loci. Copyright © 2003 John Wiley & Sons, Ltd.
Enhancing adenovirus‐mediated gene transfer in vitro and in vivo by addition of protamine and hydrocortisone
Authors: Lin Tongyu , Gu Jian , Zhang Lidong , Davis John J. , Huang Xuefeng , Cabbini Giovanni , Ji Lin , Fang Bingliang 
Published: October 2003

AbstractBackgroundInclusion of positively charged polymers such as protamine in adenovector formulations has been reported to improve the efficiency of adenovirus‐mediated gene transfer in vitro and in vivo. On the other hand, corticosteroids are known to inhibit inflammation and thus might be useful in minimizing vector‐related toxicity. In this study, we evaluated the combined effect of protamine sulfate and hydrocortisone on the efficiency of adenovirus‐mediated gene transfer in vitro and in vivo.MethodsProtamine and hydrocortisone at different concentrations were added to adenovector formulations. In vitro transgene expression with or without inclusion of protamine and hydrocortisone was evaluated in the breast cancer cell lines MDA‐MB‐231 and MCF7 and the lung cancer cell lines A549 and H460. In vivo transgene expression in the mouse lung was determined after aerosolized vector delivery.ResultsThe combination of 2 µg/ml protamine and 125 ng/ml hydrocortisone significantly increased transgene expression in vitro in all the cell lines tested. Protamine is only effective when it is added to cells before or together with adenovectors, whereas hydrocortisone is effective when it is added to cells before, together with, or after adenovectors. Inclusion of protamine and hydrocortisone also augmented apoptosis induction caused by adenovectors expressing proapoptotic genes in cancer cells. Moreover, protamine and hydrocortisone dramatically enhanced transgene expression in the mouse lung after aerosolized vector delivery.ConclusionsInclusion of protamine and hydrocortisone in adenovector formulations can improve adenovector‐mediated gene expression and may be useful for clinical applications of current adenovirus‐mediated gene therapy. Copyright © 2003 John Wiley & Sons, Ltd.
Modulation of renal glomerular disease using remote delivery of adenoviral‐encoded solubletype II TGF‐β receptor fusion molecule
Authors: Haviv Yosef S. , Takayama Koichi , Nagi Peter A. , Tousson Albert , Cook William , Wang Minghui , Lam John T. , Naito Seiji , Lei Xiaosheng , Carey Delicia E. , Curiel David T. 
Published: October 2003

AbstractBackgroundSystemic adenoviral (Ad) gene therapy for renal disorders is largely hampered by the unique architecture of the kidney. Consequently, currently available Ad vectors are of only limited therapeutic utility in the context of glomerular and fibroproliferative renal diseases.MethodsThe Ad vectors studied in the context of blocking renal fibrosis were AdTβ‐ExR and AdCATβ‐TR. AdTβ‐ExR encodes a chimeric soluble molecule comprising the entire ectodomain of the human type II TGF‐β receptor, genetically fused to the Fc fragment of the human IgG1 (sTβRII), while AdCATβ‐TR encodes only the dominant‐negative truncated ectodomain of the human type II TGF‐β receptor. The biologic activity of the type II TGF‐β receptor was evaluated in vitro by its ability to inhibit cellular proliferation and in vivo in a unilateral ureter obstruction fibrosis model. Renal targeting with sTβRII was evaluated immunohistochemically after intramuscular (IM) delivery of AdTβ‐ExR. The renal antifibrotic effect of the Ad vectors was evaluated in a lupus murine model with both light and electron microscopy and urinalysis.ResultssTβRII was detected in the glomeruli after remote IM injection of AdTβ‐ExR, but not the control AdCATβ‐TR, indicating renal deposition of the heterologous soluble fusion protein after its expression in the muscle and secretion into the circulation. AdTβ‐ExR, but not AdCATβ‐TR, could transiently inhibit mesangial expansion, glomerular hypercellularity, proteinuria and cortical interstitial fibrosis in a murine lupus model. However, the autoimmune renal disease eventually surpassed the antifibrotic effect.ConclusionsThese results indicate the superiority of a soluble type II TGF‐β receptor over a dominant‐negative, non‐soluble type II TGF‐β receptor in the context of blocking renal fibrosis in murine models. Copyright © 2003 John Wiley & Sons, Ltd.
Myocardial injection of CA promoter‐based plasmid mediates efficient transgene expression in rat heart
Authors: Huang Jianhua , Ito Yoshinori , Kobune Masayoshi , Sasaki Katsunori , Nakamura Kiminori , Dehari Hironari , Takahashi Kazuhiro , Ikeda Katsuya , Uchida Hiroaki , Kato Kazunori , Hamada Hirofumi 
Published: October 2003

AbstractBackgroundAlthough naked plasmid injection is the safest and most convenient method for gene delivery, a major limitation of this approach is currently poor transgene expression. The CA promoter (chicken β‐actin promoter with cytomegalovirus, CMV, enhancer) is one of the strongest transcriptional control modules found; however, it is uncertain whether a CA promoter‐based vector is efficient enough for naked gene therapy in a cardiovascular context.MethodsThe β‐galactosidase (LacZ) expression provided by CA promoter plasmid (pCAZ2) injection into the skeletal muscle or the heart of Lewis rats was compared with CMV promoter plasmid or adenoviral vector (AxCAZ3). The effect of Simian virus 40 of the replication origin (SV40ori) deletion from pCAZ2 on transgene expression was also evaluated.ResultspCAZ2 showed the highest LacZ expression in both skeletal muscle and heart in comparison with the CMV promoter‐based vector 5 days after naked plasmid injection. LacZ expression in the heart obtained using 20 µg of pCAZ2 was almost equivalent to that shown with AxCAZ3 at 6.0 × 109 optical particle units. The time course of transgene expression driven by CMV and CA promoters in the heart were similar, with the CA promoter providing significantly higher gene expression than the CMV promoter across all time points examined. SV40ori deletion from pCAZ2 did not affect transgene expression in either skeletal muscle or heart.ConclusionsTransgene expression mediated by naked CA promoter‐based plasmid injection was shown to be quite efficient in the heart. We propose that the CA promoter vector is suitable for myocardial gene therapy. Copyright © 2003 John Wiley & Sons, Ltd.
Enhanced antitumor activity of a combination of MBD2‐antisense electrotransfer gene therapy and bleomycin electrochemotherapy
Authors: Ivanov Marie‐Agnès , Lamrihi Badia , Szyf Moshe , Scherman Daniel , Bigey Pascal 
Published: October 2003

AbstractBackgroundMBD2 is a methylated DNA‐binding protein that has been previously suggested to have transcriptional silencing as well as DNA demethylase activities. We have previously shown that electrotransfer of an MBD2‐antisense encoding plasmid inhibits tumor growth in vivo. In this study we tested whether a combination of MBD2‐antisense gene therapy and bleomycin chemotherapy has an augmented antitumor effect in comparison with either monotherapy.MethodsMice bearing human non‐small‐cell lung carcinoma line H1299 xenoplants were treated with electrotransfer of either bleomycin or MBD2‐antisense expression plasmid or a combination of both therapies and tumor growth following treatment was monitored.ResultsA combination of electrotransfer of MBD2‐antisense and bleomycin electrochemotherapy has an additive inhibitory effect on the rate of tumor growth and a synergistic effect on the number of tumor‐free animals when compared with either monotherapy.ConclusionsOur results suggest that a combination of MBD2‐antisense electrotransfer gene therapy and chemotherapy with bleomycin is a candidate new approach to anticancer therapy. Copyright © 2003 John Wiley & Sons, Ltd.
Epidemiology of primary dystonias in Japan: Comparison with Western countries
Authors: Matsumoto Shinichi , Nishimura Masataka , Shibasaki Hiroshi , Kaji Ryuji 
Published: October 2003

AbstractWe performed epidemiological studies of primary dystonia in the city of Kyoto. The prevalence was at least 10.1 per 100,000 persons, which was similar to that in Western countries. Facial dystonia was more common than other types, which contrasts with that reported in Europe. Age of onset for both genders was in agreement with that in other countries. © 2003 Movement Disorder Society
Emotion recognition in patients with idiopathic Parkinson's disease
Authors: Yip James T.H. , Lee Tatia M.C. , Ho Shu‐Leong , Tsang Kin‐Lun , Li Leonard S.W. 
Published: October 2003

AbstractEmotion recognition (ER) was examined in 64 patients with idiopathic Parkinson's disease (PD; 56 bilateral and 8 right‐sided) and 64 matched healthy volunteers. Participants were administered an ER battery, consisting of the following subscores: overall ER (OER), overall facial ER, facial emotion identification (FEI) and discrimination, overall prosodic ER, and prosodic emotion identification (PEI) and discrimination. Measures of visuospatial functions, auditory attention, and depression were also administered. After controlling for visuospatial functions, auditory attention and depression, results indicated that patients with bilateral PD had poorer performance on all ER subscores, regardless of the modality and type of experimental task involved, relative to healthy volunteers. However, patients with right‐sided PD had difficulty on FEI and PEI only. Whereas none of the clinical variables examined in this study predicted any of the ER subscores, visual organization and auditory attention positively predicted OER in patients with PD. In addition, visual organization also positively predicted FEI in these patients. Implications are discussed in terms of the neural substrates underlying ER. © 2003 Movement Disorser Society
Is FDG‐PET a useful tool in clinical practice for diagnosing corticobasal ganglionic degeneration?
Authors: Coulier Ilse M.F. , de Vries Joeke J. , Leenders Klaus L. 
Published: October 2003

AbstractSeven consecutive patients were suspected to suffer from corticobasal ganglionic degeneration (CBGD) and were studied with [18]F‐fluorodeoxyglucose (FDG) PET imaging of the brain. At the time of their FDG‐PET scan, 4 of 7 patients fulfilled the clinical criteria of CBGD as proposed by Lang and associates [In Calne DB, 1994; Neurodegenerative disease. Philadelphia: Saunders]. For 2 of these 4 patients, however, an alternative clinical diagnosis was also considered. Three of the seven patients underwent an FDG‐PET scan when their clinical features were not yet developed sufficiently to confirm a clinical diagnosis of CBGD. Simple visual analysis of the FDG‐PET scans was carried out. All 7 patients showed an asymmetrical pattern of glucose metabolism that was demonstrated in previous studies to be characteristic in patients who had a clinical diagnosis of CBGD. The PET results helped to confirm the clinical suspicion of CBGD in 2 patients and to rule out other diagnoses in 2 other patients. For 3 patients with no sufficient symptoms to diagnose CBGD, the pattern of glucose metabolism was characteristically asymmetrical. A probable diagnosis of CBGD was made in these patients. Our results suggest that routine visual inspection of a cerebral FDG‐PET scan is a useful tool to confirm suspicion of the clinical diagnosis of probable CBGD, to differentiate from other hypokinetic‐rigid syndromes, and to support a diagnosis CBGD in patients who do not (yet) sufficiently fulfill the clinical criteria. © 2003 Movement Disorder Society