bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of Hox genes in the Bichir, Polypterus palmas
Authors: Ledje Christina , Kim Chang‐Bae , Ruddle Francis H. 
Published: October 2002

AbstractIt has been suggested that the increase in the number of Hox genes may have been one of the key events in vertebrate evolution. Invertebrates have one Hox cluster, while mammals have four. Interestingly, the number of Hox gene clusters is greater in the teleost fishes, zebrafish and medaka, than in mouse and human. The greater number of Hox clusters in the teleosts suggests that Hox gene duplication events have occurred during the radiation of ray‐finned fishes. The question is when the Hox gene duplication event(s) that lead to seven Hox clusters in the teleosts actually occurred.We have addressed this question by studying the Hox genes in the bichir, Polypterus palmas. A preliminary PCR‐estimation of the number of Hox genes suggests that Polypterus has five different Hox9 cognate group genes, which may be an indication of more than four Hox clusters in the bichir. J. Exp. Zool. (Mol. Dev. Evol.) 294:107–111, 2002. © 2002 Wiley‐Liss, Inc.
Non‐viral vector‐mediated uptake, distribution, and stability of chimeraplasts in human airway epithelial cells
Authors: de Semir David , Petriz Jordi , Avinyó Anna , Larriba Sara , Nunes Virginia , Casals Teresa , Estivill Xavier , Aran Josep M. 
Published: October 2002

AbstractBackgroundChimeraplasty is a novel methodology that uses chimeric RNA/DNA oligonucleotides (chimeraplasts) to stimulate genomic DNA repair. Efficient uptake and nuclear localization of intact chimeraplasts are key parameters to achieve optimal correction of mutation defects into specific cell types.MethodsA 5′‐end FITC‐labeled 68‐mer RNA/DNA oligonucleotide was complexed with the polycation polyethylenimine (PEI) and the cationic lipids Cytofectin and GenePorter. Flow cytometry was employed to evaluate chimeraplast uptake under different conditions. Intracellular chimeraplast distribution and co‐localization with endocytosis markers were assessed by confocal microscopy. Relative quantification of chimeraplast metabolism was performed by denaturing PAGE and GeneScan™ analysis.ResultsIn airway epithelial cells, optimized chimeraplast uptake reached near 100% efficiency with the carriers tested. However, chimeraplast nuclear localization could only be achieved using PEI or Cytofectin. Chimeraplast/GenePorter lipoplexes were retained in the cytoplasm. PEI polyplexes and Cytofectin lipoplexes displayed different uptake rates and internalization mechanisms. Chimeraplast/PEI polyplexes were internalized at least partially by fluid‐phase endocytosis. In contrast, phagocytosis may have contributed to the internalization process of large‐sized chimeraplast/Cytofectin lipoplexes. Moreover, significant chimeraplast degradation was detected 24 h after transfection with both PEI polyplexes and Cytofectin lipoplexes, although the latter seemed to confer a higher degree of protection against nuclease degradation.ConclusionBoth Cytofectin and PEI are efficient for chimeraplast nuclear uptake into airway epithelial cells. However, despite the distinct structures and trafficking pathways of the corresponding complexes, none of them could prevent nuclease‐mediated metabolism of the chimeric oligonucleotides. These findings should be taken into account for future investigations of chimeraplast‐mediated gene repair in airway epithelial cells. Copyright © 2002 John Wiley & Sons, Ltd.
High efficiency transfection of porcine vascular cells in vitro with a synthetic vector system
Authors: Parkes Richard , Meng Qing‐Hai , Elena Siapati K. , McEwan Jean R. , Hart Stephen L. 
Published: October 2002

AbstractBackgroundGene therapy strategies for the treatment of vascular disease such as the prevention of post‐angioplasty restenosis require efficient, non‐toxic transfection of vascular cells. In vitro studies in these cells contribute to vector development for in vivo use and for the evaluation of genes with therapeutic potential. The aim of this project was to evaluate a novel synthetic vector consisting of a liposome (L), an integrin targeting peptide (I), and plasmid DNA (D), which combine to form the LID vector complex.MethodsCultures of porcine smooth muscle cells and endothelial cells were established and then transfected with the LID vector, using the reporter genes luciferase and green fluorescent protein and the metalloprotease inhibitor TIMP‐1.ResultsThe LID vector system transfected primary porcine vascular smooth muscle cells and porcine aortic endothelial cells with efficiency levels of 40% and 35%, respectively. By increasing the relative DNA concentration four‐fold, incubation periods as short as 30 min achieved the same levels of luciferase transgene expression as 4 h incubations at lower DNA concentrations. The transfection did not affect cell viability as measured by their proliferative potential. Serum levels of up to 20% in the transfection medium had no adverse affect on the efficiency of transfer and gene expression in either cell type. Transfections with the cDNA for TIMP‐1 produced protein levels that peaked at 130 ng/ml per 24 h and persisted for 14 days at 10 ng/ml per 24 h.ConclusionThis novel vector system has potential for studies involving gene transfer to cardiovascular cells in vitro and in vivo. Copyright © 2002 John Wiley & Sons, Ltd.
A short synthesis of [14C]‐labelled levamisole and its major metabolite
Authors: Janssen Cor G.M. , Thijssen Jos B.A. , Verluyten Willy L.M. 
Published: October 2002

AbstractLevamisole (I) is the levo isomer of tetramisole. It is a broad spectrum anthelmintic which is used extensively as a veterinary drug for food producing animals. Metabolism and environmental studies necessitated the synthesis of 14C‐labelled levamisole (6) and of its major metabolite (12).14C‐Tetramisole was obtained in two steps from 14C‐thiourea. Resolution via salt formation and crystallization afforded 14C‐levamisole and 14C‐dexamisole. Racemisation followed again by resolution made it possible to improve the over‐all radiochemical yield of 14C‐levamisole to 51.0%. The compound had a specific activity of 73.6 MBq/mmol, a HPLC purity of 99.8% and an enantiomeric excess of 99.4%.The 14C‐labelled metabolite of levamisole (II) was obtained from 14C‐potassium cyanate in 4 consecutive steps. Resolution via chiral HPLC afforded the desired compound in a 16.2% overall radiochemical yield. It had a specific activity of 895 MBq/mmol, a HPLC purity of 98.7% and an enantiomeric excess of 100%. Copyright © 2002 John Wiley & Sons, Ltd.
Synthesis of PNU‐159548 labelled with 14C and 2H
Authors: Felicini C. , Fontana E. 
Published: October 2002

AbstractThe cytotoxic antitumor compound PNU‐159548 (1) has been labelled with 14C and 2H. A three‐step sequence starting from [14‐14C]idarubicin (2a) led to radiochemically pure (>98%) [14‐14C]PNU‐159548 with a specific activity of 1.13 GBq/mmol. The synthesis of [2H4]PNU‐159548 was carried out in a similar manner starting from [1,1,2,2‐2H4]2‐bromoethanol (3b). Copyright © 2002 John Wiley & Sons, Ltd.
Synthesis of [14C]‐ and [35S]‐labelled PI‐88 for pharmacokinetic and tissue distribution studies
Authors: Ferro Vito , Li Caiping , Wang Bin , Fewings Kym , King Andrew R. , Hammond Edward , Creese Brian R. 
Published: October 2002

AbstractPI‐88, uniformly labelled with [14C] was prepared by incorporating D‐[14C]glucose into the fermentation of Pichia (Hansenula) holstii NRRL Y‐2448 under controlled conditions to produce [14C]‐labelled extracellular phosphomannan. Subsequent acid catalyzed hydrolysis gave the [14C]‐labelled oligosaccharide phosphate fraction which was sulfonated with excess sulfur trioxide pyridine complex to give [14C]PI‐88. [35S]‐labelled PI‐88 was similarly prepared by the sulfonation of unlabelled oligosaccharide phosphate fraction with [35S] sulfur trioxide pyridine complex. Copyright © 2002 John Wiley & Sons, Ltd.
Vaginal dysplastic lesions in women with hysterectomy and receiving radiotherapy are linked to high‐risk human papillomavirus
Authors: Barzon Luisa , Pizzighella Sergio , Corti Luigi , Mengoli Carlo , Palù Giorgio 
Published: October 2002

AbstractPatients undergoing radiotherapy for advanced cervical and endometrial cancer bear a considerable risk of developing vaginal preneoplastic lesions. Radiotherapy itself has been considered to have a role in the pathogenesis of vaginal dysplasia, although human papillomavirus (HPV) involvement has also been suggested. A series of 88 patients who underwent hysterectomy and were irradiated for gynecological cancer, including 43 with postradiation vaginal dysplasia at colposcopy and 45 without vaginal lesions, were included in this study. Detection and genotyping of HPV DNA in vaginal scraping were carried out by a PCR‐based method and compared with colposcopic and cytological findings and with other clinical and laboratory data. Forty‐two (97.7%) colposcopy‐positive subjects and 6 (13.3%) colposcopically‐negative patients were PCR‐positive for high‐risk HPV DNA (P < 0.000001). Twenty‐two out of the 43 patients with colposcopic lesions showed an abnormal Papanicolau (PAP) test. Cytologic examination was negative in all colposcopically negative women. Type 16 HPV DNA was more frequent in patients with high‐grade squamous intraepithelial lesions and in patients treated with external radiotherapy, whereas other types of high‐risk HPV were more common in patients with low‐grade lesions and in those treated with brachytherapy. When considering colposcopy as the standard for diagnosing vaginal dysplasia, HPV DNA testing was more sensitive than the PAP test. However, the specificity of the PAP test was higher with no false‐positive case. In conclusion, vaginal preneoplastic changes in women post‐hysterectomy and receiving radiotherapy for cervical, endometrial, and vaginal cancer represent an HPV‐related nosologic entity. Whereas colposcopic examination can detect these preneoplastic lesions, HPV genotyping is a sensitive, inexpensive, and noninvasive method that may complement colposcopy and the PAP test. J. Med. Virol. 67:401–405, 2002. © 2002 Wiley‐Liss, Inc.
Combined detection and genotyping of Chikungunya virus by a specific reverse transcription‐polymerase chain reaction
Authors: Hasebe F. , Parquet M.C. , Pandey B.D. , Mathenge E.G.M. , Morita K. , Balasubramaniam V. , Saat Z. , Yusop A. , Sinniah M. , Natkunam S. , Igarashi A. 
Published: October 2002

AbstractA reverse transcription‐polymerase chain reaction (RT‐PCR) was developed for the detection of Chikungunya virus infection. Based on the nonstructural protein 1 (nsP1) and glycoprotein E1 (E1) genes of Chikungunya, two primer sets were designed. Total RNA were extracted from the cell culture fluid of Aedes albopictus C6/36 cells inoculated with the S27 prototype virus, isolated in Tanzania in 1953, and the Malaysian strains (MALh0198, MALh0298, and MALh0398), isolated in Malaysia in 1998. For both sets of RNA samples, the expected 354‐ and 294‐base pair (bp) cDNA fragments were amplified effectively from the nsP1 and E1 genes, respectively. Phylogenetic analysis was conducted for the Malaysian strain and other virus strains isolated from different regions in the world endemic for Chikungunya, using partial E1 gene sequence data. The Malaysian strains isolated during the epidemics of 1998 fell into a cluster with other members of the Asian genotype. J. Med Virol. 67:370–374, 2002. © 2002 Wiley‐Liss, Inc.
Norwalk‐like virus 95/96‐US strain is a major cause of gastroenteritis outbreaks in Australia
Authors: White Peter A. , Hansman Grant S. , Li Angelina , Dable Joanne , Isaacs Monica , Ferson Mark , McIver Christopher J. , Rawlinson William D. 
Published: October 2002

AbstractNorwalk‐like viruses (NLVs) were detected using a nested reverse transcriptase‐polymerase chain reaction (RT‐PCR) with primers directed to the RNA polymerase region. Samples were examined from 11 separate outbreaks of gastroenteritis and five sporadic cases of childhood gastroenteritis between 1997 and 2000. Phylogenetic analysis of the 298 bp sequences showed that all strains belong to NLV genogroup II and the majority of the sequenced isolates (30/36) were members of the 95/96‐US subset of strains associated with outbreaks recorded worldwide between 1995 and 1996. This was confirmed by analysis of the full length capsid region of a representative Australian isolate. This study demonstrates the usefulness of targeting primers for NLVs to the predominant circulating genotype(s) and confirms the spread of this subtype globally, including the Southern Hemisphere. J. Med. Virol. 68:113–118, 2002. © 2002 Wiley‐Liss, Inc.
Electromagnetic field representations and computations in complex structures II: alternative Green's functions
Authors: Felsen Leopold B. , Mongiardo Mauro , Russer Peter 
Published: October 2002

AbstractIn this second paper of the three‐part sequence, we deal with alternative Green's function (GF) representations for the subdomain (SD) problem in the complexity architecture of Part I [1]. The relevant GFs for systematic analytic modelling are those associated with at least partially vector and co‐ordinate separable boundary conditions. Such ‘canonical’ GFs, when ‘matched’ to a real problem, can form background kernels which simplify the numerical complexity of real‐problem exact integral equations. The analytic machinery involves Sturm–Liouville (SL) theory for the reduced one‐dimensional (1D) spectral GF problems resulting from separation of variables in various co‐ordinates, set in its most general form in the complex spectral wavenumber domain. Spectral synthesis in the complex spectral wavenumber planes for 2D and 3D co‐ordinate‐separable full GFs lays the foundation for direct construction (via contour deformations, branch point, and pole residue evaluations) of alternative field representations, and their correspondingly different wave‐physical phenomenologies. Illustrative examples show the connection between the canonical GFs and their network representations. Copyright © 2002 John Wiley & Sons, Ltd.
TLM representation of the hyperbolic heat conduction equation
Authors: Pulko S. H. , Wilkinson A. J. , Saidane A. 
Published: October 2002

AbstractMany heat transfer situations are adequately described by the parabolic thermal diffusion equation. However, in situations in which very rapid heating occurs or in slower heating regimes for particular materials, the hyperbolic heat conduction equation is a better representation. Here, a parameterized nodal structure for transmission line modelling (TLM) representation of hyperbolic heat conduction processes is devised. A TLM model based on the nodal structure is implemented and temperature field predicted by the model are compared with analytical results for the same physical situation. Copyright © 2002 John Wiley & Sons, Ltd.
Specific up‐regulation of GADD153/CHOP in 1‐methyl‐4‐phenyl‐pyridinium‐treated SH‐SY5Y cells
Authors: Conn Kelly J. , Gao Wen‐Wu , Ullman M. David , McKeon‐O'Malley Catherine , Eisenhauer Patricia B. , Fine Richard E. , Wells John M. 
Published: October 2002

AbstractGrowth arrest DNA damage‐inducible 153 (GADD153) expression was increased in 1‐methyl‐4‐phenyl‐pyridinium (MPP+)‐treated human SH‐SY5Y neuroblastoma cells as determined by gene microarray analysis. GADD153 expression increased after 24 hr of MPP+ (1 mM) exposure and preceded activation of caspase 3. Comparison of GADD153 expression among cultures treated with other toxins whose primary mode of action is either via mitochondrial impairment (rotenone) or via oxidative stress (6‐hydroxydopamine or hydrogen peroxide) showed that GADD153 was uniquely up‐regulated by MPP+. Together these data suggest that a cellular mechanism distinct from mitochondrial impairment or oxidative stress contributes significantly to the up‐regulation of GADD153 by MPP+ and that GADD153 may function as an inducer of apoptosis following MPP+ exposure. Published 2002 Wiley‐Liss, Inc.
Elevation of brain glutathione by γ‐glutamylcysteine ethyl ester protects against peroxynitrite‐induced oxidative stress
Authors: Drake Jennifer , Kanski Jaroslaw , Varadarajan Sridhar , Tsoras Maria , Butterfield D. Allan 
Published: October 2002

AbstractElevation of glutathione (GSH) has been recognized as an important method for modulating levels of reactive oxygen species (ROS) in the brain. We investigated the antioxidant properties of γ‐glu‐cys‐ethyl ester (GCEE) in vitro and its ability to increase GSH levels upon in vivo i.p. injection. GCEE displays antioxidant activity similar to GSH as assessed by various in vitro indices such as hydroxyl radical scavenging, dichlorofluorescein fluorescence (DCF), protein specific spin labeling, glutamine synthetase (GS) activity, and protein carbonyls. Intraperitoneal injection of GCEE to gerbils resulted in a 41% increase in brain total GSH levels in vivo as determined by the DTNB‐GSH reductase recycling method. Gerbils injected with buthionine sulfoximine (BSO), an inhibitor of γ‐glutamylcysteine synthetase, had 40% less total brain glutathione. Gerbils injected with BSO followed by a GCEE injection had GSH levels similar to vehicle‐injected controls, suggesting that GCEE upregulates GSH biosynthesis by providing γ‐glutamylcysteine and not cysteine. Cortical synaptosomes from GCEE‐injected animals were less susceptible to peroxynitrite‐induced oxidative damage as assessed by DCF fluorescence, protein‐specific spin labeling, and GS activity. These experiments suggest that GCEE is effective in increasing brain GSH levels and may potentially play an important therapeutic role in attenuating oxidative stress in neurodegenerative diseases associated with oxidative stress such as Alzheimer disease. © 2002 Wiley‐Liss, Inc.
Regulation of the ferritin H subunit by vitamin B12 (cobalamin) in rat spinal cord
Authors: Cairo Gaetano , Ronchi Raffaella , Buccellato Francesca R. , Veber Daniela , Santambrogio Paolo , Scalabrino Giuseppe 
Published: October 2002

AbstractCobalamin‐deficient (Cbl‐D) central neuropathy is a pure myelinolytic disease, in which gliosis is also observed. Iron is abundant in the mammalian central nervous system, where it is required for various essential functions including myelinogenesis. It is predominantly located in the white matter and oligodendrocytes, which also actively synthesize the major iron proteins (e.g., ferritin, transferrin). We investigated the expression of the main proteins of iron metabolism in the spinal cord (SC) of totally gastrectomized Cbl‐D rats 2 months after surgery (i.e., when the Cbl‐D status has become severe). There were no significant changes in iron content, the activity of iron regulatory proteins, or the expression of transferrin or its receptor in the SC. We observed a significant decrease in the levels of both H and L ferritin subunits, with a more marked reduction in the latter. Post‐operative cobalamin replacement therapy normalized only the H‐ferritin subunits, and only in the SC. Our results therefore suggest that permanent cobalamin deficiency affects iron metabolism in the rat SC preferentially from a functional point of view, because H‐ferritin is known to be involved in the uptake and release of iron. © 2002 Wiley‐Liss, Inc.
p35/Cdk5 pathway mediates soluble amyloid‐β peptide‐induced tau phosphorylation in vitro
Authors: Town Terrence , Zolton Joseph , Shaffner Reed , Schnell Billy , Crescentini Robert , Wu Yajuan , Zeng Jin , DelleDonne Anthony , Obregon Demian , Tan Jun , Mullan Mike 
Published: October 2002

AbstractAlzheimer's disease (AD) is pathologically characterized by deposition of amyloid‐β peptides (Aβ) as senile plaques and by the occurrence of neurofibrillary tangles (NFTs) composed primarily of hyperphosphorylated tau protein. Activation of cyclin‐dependent kinase 5 (Cdk5) via its potent activator p25 has recently been shown to promote phosphorylation of tau at AD‐specific phosphoepitopes, and increased cleavage of p35 to p25 has been demonstrated in AD patients, suggesting that Cdk5 may represent a pathogenic tau protein kinase. We were interested in the potential effect of soluble forms of Aβ on Cdk5‐mediated AD‐like tau phosphorylation, insofar as previous studies of human biopsies and aged canine and primate brains have shown that dystrophic neurites appear before the formation of neuritic plaques. We transfected N2a cells with a p35 vector (N2a/p35 cells) and, after differentiation, challenged these cells with Aβ1–42 peptide in soluble form (sAβ1–42). Results show that sAβ1–42 at relatively low levels (1–5 μM) dose‐dependently increases tau phosphorylation at AD‐specific phosphoepitopes in differentiated N2a/p35 cells compared with controls, an effect that is blocked by antisense oligonucleotides against p35. sAβ1–42‐induced tau phosphorylation is concomitant with an increase in both p25 to p35 ratio and Cdk5 activity (but not protein levels). Additionally, blockade of L‐type calcium channels or inhibition of calpain completely abolishes this effect. Taken together, these data indicate that sAβ is a potent activator of the p25/Cdk5 pathway, resulting in promotion of AD‐like tau phosphorylation in vitro. © 2002 Wiley‐Liss, Inc.
Adenoviral expression of protein‐L‐isoaspartyl methyltransferase (PIMT) partially attenuates the biochemical changes in PIMT‐deficient mice
Authors: Ogawara Midori , Takahashi Mayumi , Shimizu Takahiko , Nakajima Mitsunari , Setoguchi Yasuhiro , Shirasawa Takuji 
Published: October 2002

AbstractProtein‐L‐isoaspartyl methyltransferase (PIMT) is a putative protein repair enzyme, which methylates the α‐carboxyl group of atypical L‐isoaspartyl residues in aged proteins and converts them to normal L‐aspartyl residues. Two splicing variants, PIMT‐I and PIMT‐II, have been reported, although their biological functions and specific subcellular substrates are still to be defined. We and another group have previously showed that PIMT‐deficient mice succumbed to fatal epileptic seizures associated with an abnormal accumulation of isoaspartate (IsoAsp) in the brain. In the present study, we prepared two recombinant adenovirus vectors that contained PIMT‐I or PIMT‐II, respectively, in order to investigate the differential biological roles of PIMT‐I and PIMT‐II. These recombinant viruses differentially conferred PIMT‐I or PIMT‐II expressions in cultured neurons. Biochemical analyses showed that either of PIMT‐I or PIMT‐II effectively repaired the damaged proteins in PIMT‐deficient neurons, but the concomitant expression failed to show an additive effect in the repair of IsoAsp. These results suggested that PIMT‐I and PIMT‐II might share a common biological function and/or subcellular substrates. In addition, we administered an adeno‐PIMT‐I vector into the brain of PIMT‐deficient mice at embryonic day 14.5 by an exo‐utero method to assess the biological effects in vivo. The result showed that recombinant adeno‐PIMT improved the symptoms of PIMT‐deficient mice in vivo, but only partially repaired IsoAsp in damaged proteins. The gene therapy presented in this report provided a better prognosis for the survival of PIMT‐deficient mice than the previously reported anti‐epileptic drug therapy. The results suggested a new reagent for gene therapy applicable to ageing‐associated neurodegenerative disorders. © 2002 Wiley‐Liss, Inc.