bluuebunny/crossref_metadata_2025_split huggingface dataset:
Investigation of optimal transduction conditions for baculovirus‐mediated gene delivery into mammalian cells
Authors: Hsu Ching‐Suei , Ho Yi‐Chen , Wang Kuei‐Chun , Hu Yu‐Chen 
Published: September 2004

AbstractAlthough baculovirus‐mediated gene delivery into mammalian cells has been documented in a wealth of the literature, systematic investigation of the optimal transduction conditions remains unavailable. In this work, a transduction protocol using unconcentrated baculovirus is proposed for simple and efficient gene delivery into HeLa cells. We found that ∼75–85% of the cells could be readily transduced and express the reporter protein when virus transduction occurred for 4 h at 25°C using Dulbecco's phosphate‐buffered saline (D‐PBS) as the surrounding solution. This method contrasts with previous protocols in which transduction occurs for 1 h at 37°C using growth medium (e.g., DMEM) as the surrounding solution. Investigation of the physical parameters led to the findings that: 1) baculovirus uptake by HeLa cells continued for at least 4 h in the event of high virus dosage, which led to higher gene expression; 2) the half‐life of baculovirus dramatically decreased at 37°C; 3) EGTA pretreatment did not apparently facilitate the gene delivery when the cells grew to multilayers; and 4) lower transduction efficiency and gene expression were obtained when DMEM was used (in comparison with D‐PBS and TNM‐FH), suggesting that DMEM contains certain inhibitory factors for baculovirus transduction. Our data uncovered several aspects that were not investigated before and the optimized transduction conditions allowed for gene delivery as efficient as that by the protocols commonly employed by others, but eliminated the need for virus ultracentrifugation. The protocol not only represented a simpler approach, but also considerably reduced possible virus inactivation during ultracentrifugation, thus making it easier to convert the baculovirus/mammalian cell system to a tool for eukaryotic protein production on a larger scale. © 2004 Wiley Periodicals, Inc.
Detection of biomolecular interaction between biotin and streptavidin on a self‐assembled monolayer using magnetic nanoparticles
Authors: Arakaki Atsushi , Hideshima Sho , Nakagawa Takahito , Niwa Daisuke , Tanaka Tsuyoshi , Matsunaga Tadashi , Osaka Tetsuya 
Published: September 2004

AbstractFor developing a magnetic bioassay system, an investigation to determine the presence of a specific biomolecular interaction between biotin and streptavidin was done using magnetic nanoparticles and a silicon substrate with a self‐assembled monolayer. Streptavidin was immobilized on the magnetic particles, and biotin was attached to the monolayer‐modified substrate. The reaction of streptavidin‐modified magnetic particles on the biotin‐modified substrate was clearly observed under an optical microscope. The magnetic signals from the particles were detected using a magnetic force microscope. The results of this study demonstrate that the combination of a monolayer‐modified substrate with biomolecule‐modified magnetic particles is useful for detecting biomolecular interactions in medical and diagnostic analyses. © 2004 Wiley Periodicals, Inc.
Densonucleosis virus purification by ion exchange membranes
Authors: Specht Rachel , Han Binbing , Wickramasinghe S. Ranil , Carlson Jonathan O. , Czermak Peter , Wolf Anne , Reif Oscar‐Werner 
Published: September 2004

AbstractPreparative chromatography is widely used in the downstream purification of biopharmaceutical products. Replacement of resins by membranes as chromatographic supports, overcomes many of the limitations associated with resin‐based chromatography such as high‐pressure drops, slow processing rates due to pore diffusion and channeling of the feed through the bed. In particular, adsorptive membranes may be ideally suited for virus capture. Virus capture is critical in a number of applications. In gene therapy and vaccine production, large‐scale purification of virus vectors is often essential. In the manufacture of biopharmaceuticals, validation of virus clearance is critical.Here results for purification of Aedes aegypti densonucleosis virus (AeDNV) using anion and cation exchange membranes are presented. AeDNV is a non‐enveloped, single‐stranded mosquito‐specific parvovirus. Virus particles are around 20 nm in size. AeDNV could find potential applications in integrated vector‐borne disease control programs. In addition, capture of parvovirus for validation of virus clearance in the manufacture of biopharmaceuticals is of commercial importance.By adjusting the pH of the feed stream, AeDNV particles may be adsorbed by both anion and cation exchange membranes. However, strongly basic anion exchange membranes were the most effective in adsorbing AeDNV particles. Adsorption and subsequent elution of AeDNV by anion exchange membranes leads to significant virus concentration. Dynamic and static capacities for anion exchange membranes were similar. Further, a sharp elution curve was obtained suggesting that pore diffusional resistances are insignificant. The adsorption of AeDNV particles by anion exchange membranes may be described by a linear isotherm. © 2004 Wiley Periodicals, Inc.
Simple and extractionless high‐performance liquid chromatographic determination of rosiglitazone in human plasma and application to pharmacokinetics in humans
Authors: Kim Kyoung‐Ah , Park Ji‐Young 
Published: September 2004

AbstractA simple and extractionless HPLC method using ﬂuorescence detection was developed for the determination of rosiglitazone in human plasma. After deproteinization using perchloric acid the plasma samples were directly injected onto the HPLC system. The mobile phase was composed of acetonitrile (52%) and 20 mm ammonium acetate (48%, pH 7.5), and analysis was run at a ﬂow rate of 0.2 mL/min with the detector operating at 247 nm for excitation wavelength and at 367 nm for emission wavelength, respectively. The method has a mean recovery of 97%, while the intra‐day and inter‐day precisions were all less than 7%. This method is simple, speciﬁc, sensitive and requires only a small plasma volume with short analytical time, and is suitable for the determination of plasma rosiglitazone in routine measurements for pharmacokinetic studies. Copyright © 2004 John Wiley & Sons, Ltd.
QoS guarantee in telecommunication networks: technologies and solutions
Authors: Fortunato Enzo , Marchese Mario , Mongelli Maurizio , Raviola Annamaria 
Published: September 2004

AbstractThe aim of this paper is to show the effect of the presence of specific management functions within a network that offers quality of service (QoS). The objective is not privileging a particular technology but to highlight the importance to know which control functions a solution may use, which performance limits the functions have and what can be a realistic user expectation.The paper focuses on the meaning of QoS and on the applications requiring quality, then describes QoS solutions including transport technologies, QoS‐oriented technologies, parameters and management functions. In more detail, the effect on QoS provision of the following issues is investigated and discussed concerning the possibility (or not) to aggregate and differentiate traffic, the implementation of call admission control and of traffic filtering to limit flows to their committed rates. Again, the conclusions should not be considered a merit mark about technology, but only an investigation about: what users and customers should expect by the technologies using specific control functions evidencing that the real limitations are not imposed by a specific technology, whose features may be changed and extended, but by the application of control functions that can guarantee requirements' matching. Copyright © 2004 John Wiley & Sons, Ltd.
Inhibitory effects of novel tetrahydropyridine derivatives on nitric oxide and reactive oxygen species production in glioma cells
Authors: Yoon Kyoung Jin P. , Redda Kinfe K. , Mazzio Elizabeth , Soliman Karam F.A. 
Published: September 2004

AbstractThe immune response in the central nervous system involves the degeneration of neurological tissue. The therapeutic use of nonsteroidal anti‐inflammatory agents (NSAIDs) can be effective in reducing inflammation and associated neurodegeneration. The current study was designed to examine the effects of previously synthesized N‐(substituted benzoylamino)‐4‐ethyl‐1,2,3,6‐tetrahydropyridines (THPs) on free radical and nitric oxide (NO) generation in C6 glioma cells activated with bacterial endotoxin [lipopolysaccharide (LPS): Escherichia coli 0111:B4] and interferon‐γ. Stimulated C6 cells exhibited elevated iNOS protein expression, an increase of reactive oxygen species and NO2−. All parameters were attenuated significantly by indomethacin and various THPs (4‐Ome, 3‐CF3, 4‐Me, 4‐Et, 4‐t‐butyl, and 4‐n‐butyl). The results indicate a possible role for THP derivatives as anti‐inflammatory agents. Drug Dev. Res. 61:189–196, 2004. © 2004 Wiley‐Liss, Inc.
Augmentation of all‐trans‐retinoic acid concentration in plasma by preventing inflammation responses induced by atRA‐loaded microspheres with concurrent treatment of dexamethasone
Authors: Park Kyeongsoon , Choi Yongdoo , Kim Sang Yoon , Byun Youngro 
Published: September 2004

AbstractAll‐trans retinoic acid (atRA)‐loaded microspheres severely induce inflammatory responses after microsphere implantation. Fibroblasts and a thick band of fibrous capsule resulting from the inflammatory responses could hamper drug permeation to the bloodstream because fibroblasts actively metabolize atRA into polar metabolites and the thick fibrous capsule acts as a diffusion barrier. In the present study, we investigated whether the fibroblast proliferation and collagen deposition induced by atRA released from microspheres might affect the atRA concentration in plasma and atRA metabolism with or without treatment of dexamethasone as an anti‐inflammatory drug. After subcutaneous injection of atRA‐loaded microspheres in rats, it was observed that atRA‐loaded microspheres induced severe inflammatory responses and stimulated fibroblast proliferation and collagen deposition in fibrous capsules. On the other hand, the orally treated dexamethasone effectively prevented inflammatory responses in a dose‐dependent manner and suppressed about 49% of the number of fibroblasts and collagen deposition in fibrous capsules at 14 days. In addition, after the treatment of dexamethasone, the atRA concentration in plasma was increased, and its metabolism was decreased approximately by 40% at 7 days, compared to the group treated alone with atRA‐loaded microspheres. In conclusion, the concurrent treatment of dexmethasone with atRA‐loaded microspheres could prevent inflammatory responses and metabolism of atRA, thereby maintaining the atRA concentration in plasma for longer periods in the therapeutic range. Drug Dev. Res. 61:197–206, 2004. © 2004 Wiley‐Liss, Inc.
Effects of a new 1,3,4‐thiadiazolium mesoionic compound, MI‐D, on the acute inflammatory response
Authors: Cardoso Júlio C. , Cadena Sílvia M. S. C. , Zampronio Aleksander , Arruda Ana Maria S. , Carnieri Eva G. S. , Echevarria Aurea , Constantin Jorgete , Bracht Adelar , Oliveira Maria Benigna M. 
Published: September 2004

AbstractA new mesoionic compound, 4‐phenyl‐5‐(4‐nitro‐cinnamoyl)‐1,3,4‐thiadiazolium‐2‐phenylamine (MI‐D), is described along with some of its biological properties. Its effects on hepatic metabolism, on O and nitric oxide (NO) production, and in in vivo models for potential antinociceptive, antipyretic, and antiinflammatory activities were determined. In perfused rat liver, MI‐D (25 µM) stimulated glycogenolysis (95%), and inhibited oxygen uptake (37%) with affecting glycolysis. In phorbol 12‐myristate 13‐acetate‐stimulated macrophages, O generation was reduced (95%) by MI‐D (15 µM), whereas the production of NO was unaffected. MI‐D (2 mg/kg) inhibited (55%) the number of abdominal writhings induced by acetic acid. At 1 mg/kg, MI‐D inhibited the febrile response (5 h) induced by lipopolysaccharide (LPS) and was also effective against a preexisting febrile response. Treatment with MI‐D (1 mg/kg) reduced by 67% prostaglandin (PGE2) levels in the cerebrospinal fluid of LPS‐exposed mice, and at a higher dose (8 mg/kg) MI‐D inhibited paw edema formation (2 h) induced by carrageenan. MI‐D has a spectrum of activities similar to other nonsteroidal antiinflammatory drugs, qualifying it as a potential anti‐inflammatory drug. Drug Dev. Res. 61:207–217, 2004. © 2004 Wiley‐Liss, Inc.
Characterization of the anti‐platelet actions of FK419, a novel non‐peptide antagonist of platelet GPIIb/IIIa
Authors: Mihara Kayoko , Aoki Toshiaki , Moriguchi Akira , Yamamoto Hiroko , Maeda Masashi , Tojo Nobuteru , Yamanaka Toshio , Ohkubo Mitsuru , Matsuoka Nobuya , Seki Jiro , Mutoh Seitaro 
Published: September 2004

AbstractThe anti‐platelet properties of FK419 ((S)‐2‐acetylamino‐3‐[(R)‐[1‐[3‐(piperidin‐4‐yl)propionyl]piperidin‐3‐ylcarbonyl] amino]propionic acid), a novel non‐peptide GPIIb/IIIa antagonist, were compared in a variety of experimental settings, both in vitro and in vivo, with other GPIIb/IIIa antagonists including xemilofiban, lamifiban, tirofiban, and FK633. Receptor binding studies suggested that FK419 had potent GPIIb/IIIa antagonistic activity that is comparable with those of reference antagonists. FK419 effectively inhibited human platelet aggregation, regardless of agonist stimuli (IC50 = 35–170 nM). FK419 demonstrated in vitro species‐dependent anti‐platelet activity, with higher potency in human than in dog, guinea pig, or rat tissue, and dose‐dependently inhibited ex vivo platelet aggregation in dogs and guinea pigs. In contrast to other antagonists, FK419 minimally affected template bleeding time at doses that completely inhibited platelet aggregation in canines. These results demonstrate that FK419 is a novel, potent, and selective GPIIb/IIIa antagonist that safely inhibits platelet aggregation in vivo, suggesting that it may be a promising anti‐platelet agent for thrombotic diseases. Drug Dev. Res. 61:233–241, 2004. © 2004 Wiley‐Liss, Inc.
Bacillus Calmette Guérin triggers the IL‐12/IFN‐γ axis by an IRAK‐4‐ and NEMO‐dependent, non‐cognate interaction between monocytes, NK, and T lymphocytes
Authors: Feinberg Jacqueline , Fieschi Claire , Doffinger Rainer , Feinberg Max , Leclerc Tony , Boisson‐Dupuis Stéphanie , Picard Capucine , Bustamante Jacinta , Chapgier Ariane , Filipe‐Santos Orchidée , Ku Cheng‐Lung , de Beaucoudrey Ludovic , Reichenbach Janine , Antoni Guillemette , Baldé Ramatoulaye , Alcaïs Alexandre , Casanova Jean‐Laurent 
Published: September 2004

AbstractThe IL‐12/IFN‐γ axis is crucial for protective immunity to Mycobacterium in humans and mice. Our goal was to analyze the relative contribution of various human blood cell subsets and molecules to the production of, or response to IL‐12 and IFN‐γ. We designed an assay for the stimulation of whole blood by live M. bovis Bacillus Calmette‐Guérin (BCG) alone, or BCG plus IL‐12 or IFN‐γ, measuring IFN‐γ and IL‐12 levels. We studied patients with a variety of specific inherited immunodeficiencies resulting in a lack of leukocytes, or T, B, and/or NK lymphocytes, or polymorphonuclear cells, or a lack of expression of key molecules such as HLA class II, CD40L, NF‐κB essential modulator (NEMO), and IL‐1 receptor‐associated kinase‐4 (IRAK‐4). Patients with deficiencies in IL‐12p40, IL‐12 receptor β1 chain (IL‐12Rβ1), IFN‐γR1, IFN‐γR2, and STAT‐1 were used as internal controls. We showed that monocytes were probably the main producers of IL‐12, and that NK and T cells produced similar amounts of IFN‐γ. NEMO and IRAK‐4 were found to be important for IL‐12 production and subsequent IFN‐γ production, while a lack of CD40L or HLA class II had no major impact on the IL‐12/IFN‐γ axis. The stimulation of whole blood by live BCG thus triggers the IL‐12/IFN‐γ axis by an IRAK‐4‐ and NEMO‐dependent, non‐cognate interaction between monocytes, NK, and T lymphocytes.
Commentary: New insights into the complexity of phosphatidylinositol lipid signaling in B lymphocytes
Authors: Tybulewicz Victor L. J. 
Published: September 2004

AbstractStimulation of B cells through the B cell antigen receptor (BCR) leads to the activation of numerous intracellular signaling pathways including phosphoinositide (PI) 3‐kinases (PI3K). These generate the 3‐PI lipids which in turn act as critical second messengers. Gene‐targeting experiments had shown that B cells deficient in p85α, an adaptor protein required for PI3K function, were defective in their ability to proliferate in response to BCR stimulation. In this issue of the European Journal of Immunology, Hess et al. report analysis of intracellular signaling pathways in p85α‐deficient B cells and show that in the absence of p85α there is a partial impairment in BCR‐induced calcium flux, and a reduction in activation of the transcription factor NF‐κB. Unexpectedly, they show that while the BCR‐induced phosphorylation of the PI3K‐dependent kinase Akt is reduced in p85α‐deficient cells, the phosphorylation of two downstream targets of Akt – FOXO1 and ribosomal protein S6 – is largely unaffected. Furthermore, they show that treatment of wild‐type B cells with PI3K inhibitors had a more profound effect than disruption of the p85α gene. Taken together, these results indicate that in the absence of p85α, there is still significant residual PI3K activity. These results highlight the need for careful measurement of PI3K activity in gene‐targeted mice and cells, by directly measuring levels of the 3‐PI lipids.See accompanying article: http://dx.doi.org/10.1002/eji.200425326
Fundamental differences in spontaneous synaptic inhibition between deep and superficial layers of the rat entorhinal cortex
Authors: Woodhall Gavin L. , Bailey Sarah J. , Thompson Sarah E. , Evans D. Ieuan P. , Jones Roland S.G. 
Published: September 2004

AbstractWe have previously shown that there are clear differences between spontaneous excitatory synaptic currents recorded in layers V and II of the rat entorhinal cortex (EC) in vitro, and have suggested that these might contribute to a more pronounced susceptibility of the deeper layer to epileptogenesis. In the present study, we have made a detailed comparison of spontaneous synaptic inhibition between the two layers by recording spontaneous inhibitory synaptic currents (sIPSCs) using whole‐cell patch‐clamp techniques in EC slices. Pharmacological studies indicated that sIPSCs were mediated exclusively by γ‐aminobutyric acid (GABA)A receptors. There was little difference in average amplitudes, rise or decay times of sIPSCs in layer II compared with layer V. However, in the former, events occurred at 4–5 times the frequency seen in the latter, and frequencies of ≤40 Hz were not uncommon. When activity‐independent, miniature IPSCs were isolated in tetrodotoxin (TTX), the frequency in layer V was more than halved, but in layer II only a small reduction was seen, and the frequency remained very high. In terms of kinetics, while averaged sIPSCs in each layer were very similar, detailed comparison of individual sIPSCs within layers revealed distinct differences, possibly reflecting inputs from different subtypes of interneurons or inputs at different somatodendritic locations. In layer V, sIPSCs could be divided into three groups, one with slow rise and decay kinetics and a second with fast rise kinetics, further distinguished into two groups by either fast or slow decay kinetics. The distinction between events in layer II was simpler, one group having both fast rise and decay times and the second with both parameters much slower. Finally, IPSCs could occur in high‐frequency bursts in both layers, although these were much more prevalent in layer II. The results are discussed in terms of the overall level of background inhibition in the two layers, as well as how this might relate to their susceptibilities to epileptogenesis. © 2004 Wiley‐Liss, Inc.
Improving mother–child interaction in low‐income Turkish–Dutch families: A study of mechanisms mediating improvements resulting from participating in a home‐based preschool intervention program
Authors: Tuijl Cathy van , Leseman Paul P.M. 
Published: September 2004

AbstractThis study examined whether the effects on cognitive and language outcomes of a recently developed home‐based educational intervention program, Opstap Opnieuw, for 4–6‐years‐old disadvantaged children could be explained by improved mother–child interaction. The present sample (n=30) was drawn from a larger sample of Turkish–Dutch families (n=181) for which in a previous study significant effects of Opstap Opnieuw were found on children's (first) language and cognitive pre‐math skill, 5 months after the program ended. The present study focused on two facets of interaction quality as possible mediators of these program effects: the mean cognitive distancing level of mothers' communication and instruction behaviour as an indicator of the cognitive and verbal stimulation provided, and the degree of cooperation as an indicator of mothers' social‐emotional support to their children. Both measures were based on systematic observation of mother–child interaction during sorting tasks. Participation in the program appeared to improve mothers' social‐emotional support behaviour substantially, but not their cognitive distancing behaviour. For Turkish (first language) vocabulary, about half of the program effect appeared to be mediated by the improved social‐emotional support. For cognitive pre‐mathematical skills, two‐thirds of the program effect appeared to be mediated by improved social‐emotional support. Mothers' cognitive distancing was moderately‐strongly related to children's vocabulary development, but did not mediate program effects. Some implications of the results are discussed. Copyright © 2004 John Wiley & Sons, Ltd.
Large‐scale molecular dynamics simulations of HLA‐A*0201 complexed with a tumor‐specific antigenic peptide: Can the α3 and β2m domains be neglected?
Authors: Wan Shunzhou , Coveney Peter , Flower Darren R. 
Published: September 2004

AbstractLarge‐scale massively parallel molecular dynamics (MD) simulations of the human class I major histocompatibility complex (MHC) protein HLA‐A*0201 bound to a decameric tumor‐specific antigenic peptide GVYDGREHTV were performed using a scalable MD code on high‐performance computing platforms. Such computational capabilities put us in reach of simulations of various scales and complexities. The supercomputing resources available for this study allow us to compare directly differences in the behavior of very large molecular models; in this case, the entire extracellular portion of the peptide–MHC complex vs. the isolated peptide binding domain. Comparison of the results from the partial and the whole system simulations indicates that the peptide is less tightly bound in the partial system than in the whole system. From a detailed study of conformations, solvent‐accessible surface area, the nature of the water network structure, and the binding energies, we conclude that, when considering the conformation of the α1–α2 domain, the α3 and β2m domains cannot be neglected. © 2004 Wiley Periodicals, Inc. J Comput Chem 25: 1803–1813, 2004