bluuebunny/crossref_metadata_2025_split huggingface dataset:
Atypical nasal challenges in patients with idiopathic rhinitis: more evidence for the existence of allergy in the absence of atopy?
Authors: S. Carney A. , Powe D. G. , Huskisson R. S. , Jones N. S. 
Published: March 2003

SummaryBackground The pathophysiology of idiopathic rhinitis is unknown although evidence is accumulating to suggest that many patients may have a localized form of allergic rhinitis in the absence of other atopic symptoms and markers. This study compares detailed nasal challenge results obtained from patients with idiopathic rhinitis to those of atopic and normal controls.Methods Patients with idiopathic rhinitis (n = 23), perennial allergic rhinitis (n = 8) and normal controls (n = 8) underwent a normal saline challenge to exclude hyper‐reactivity and then bilateral nasal allergen challenges. Nasal patency was assessed by anterior active rhinomanometry.Results All of the patients with atopic rhinitis demonstrated positive bilateral allergen challenges. All normal control subjects had bilateral negative challenges. Two patients in the idiopathic group tested positively to saline and were excluded from further study with 62% of the remainder testing positive to allergens. Of the idiopathic patients testing positive, 85% were sensitive to house dust mite.Conclusion A significant proportion of patients with idiopathic rhinitis have positive nasal challenges, the vast majority to house dust mite allergen. These findings add to the weight of evidence that suggests ‘localized allergy’ may exist in the absence of systemic atopic markers.
The involvement of matrix metalloproteinases in basement membrane injury in a murine model of acute allergic airway inflammation
Authors: Kumagai K. , Ohno I. , Imai K. , Nawata J. , Hayashi K. , Okada S. , Senoo H. , Hattori T. , Shirato K. 
Published: March 2003

SummaryBackground Airway remodelling in asthma such as subepithelial fibrosis is thought to be the repair process that follows the continuing injury as of chronic airway inflammation. However, how acute allergic inflammation causes tissue injury in the epithelial basement membrane in asthmatic airways remains unclear. Matrix metalloproteinases (MMPs) capable of degrading almost all of the extracellular matrix components have been demonstrated to be involved in cell migration through the basement membrane in vivo and in vitro.Objective We investigated the alterations of matrix construction and the role of MMPs in matrix degradation in the subepithelium during acute allergic airway inflammation.Methods Airway inflammation, the ultrastructure of the subepithelium and injury of types III and IV collagen in tracheal tissues from ovalbumin (OVA)‐sensitized mice after OVA inhalation with or without the administration of tissue inhibitor of metalloproteinase‐2 (TIMP‐2) and dexamethasone were evaluated by cell counting in bronchoalveolar lavage (BAL) fluids, electron microscopy and immunohistochemistry, respectively.Results The disruption of the lamina densa and matrix construction and the decrease of the immunoreactivity for type IV collagen in subepithelium were observed in association with the accumulation of inflammatory cells in airways 3 days after OVA inhalation. This disorganization of the matrix components in the subepithelium, as well the cellular accumulation, was abolished by the administration of TIMP‐2 and dexamethasone. The immunoreactivity for type IV collagen in the subepithelium in OVA‐inhaled mice returned to the level of that in saline‐inhaled mice 10 days after inhalation in association with a decrease of the cell numbers in the BAL fluid. The immunoreactivity for type III collagen was changed neither 3 nor 10 days after OVA inhalation.Conclusion These results suggest that epithelial basement membrane gets injured by, at least in part, MMPs as a consequence of cell transmigration through the membrane during acute allergic airway inflammation.
Gelatin‐specific cellular immune responses persist for more than 3 years after priming with gelatin containing DTaP vaccine
Authors: Kumagai T. , Kamada M. , Igarashi C. , Yuri K. , Furukawa H. , Nagata N. , Saito A. , Okui T. , Yano S. 
Published: March 2003

SummaryBackground Gelatin‐specific cell‐mediated immunity develops in subjects inoculated with gelatin containing DTaP vaccine. However, it is not yet known whether such established sensitization to gelatin disappears or persists with time.Objective The aim of this study was to follow the patients with gelatin sensitization elicited by DTaP vaccination for their lymphocyte responsiveness and IgE, IgG antibody specific to gelatin over several years and to compare the activities with those at the time of enrolment into the study.Methods We studied 28 subjects who developed positive lymphocyte proliferation test (LPT) after receiving gelatin containing DTaP vaccine and eight subjects who had a negative LPT after inoculation of non‐gelatin DTaP. Determination of IgE, IgG antibodies and specific lymphoproliferative response directed against gelatin were performed at enrolment and on follow up.Results None of the subjects had antibody to gelatin at enrolment and none developed gelatin IgE or IgG during follow‐up. There was no significant difference in the SIs of the subjects receiving gelatin DTaP (P = 0.150, 95% CI, −0.198–0.032), whereas lymphocyte activity to gelatin increased between enrolment and follow‐up in the subjects with non‐gelatin DTaP (P = 0.011, 95% CI, 0.063–0.338).Conclusion Gelatin‐specific lymphocyte activity persists at comparable levels for more than 3 years in subjects who acquire a positive LPT response to gelatin after receiving primary DTaP vaccine containing gelatin. Furthermore, five out of eight subjects initially with negative LPT to gelatin have been shown to acquire specific LPT with time.
Giving tape recordings or written summaries of consultations to people with cancer: a systematic review
Authors: Scott J. Tim , Entwistle Vikki A. , Sowden Amanda J. , Watt Ian 
Published: March 2003

Objective To examine the effects of providing recordings or summaries of consultations to people with cancer and their families. Design Systematic review. Data sources MEDLINE, CINAHL, Cancerlit, EMBASE and other electronic bibliographic databases. Bibliographies of relevant papers. Selection criteria Randomized and non‐randomized controlled trials of the provision of taped recordings or written summaries of consultations to people with cancer and/or their families. Main results Eight randomized controlled trials were found, all involving adult participants. No non‐randomized controlled trials were found. The quality of the studies was generally poor. Between 83% and 96% of people who received recordings or summaries found them useful to remind them of what was said and/or to inform family members and friends about their illness and treatment. Of seven studies that assessed recall of information given during the consultation, four reported better recall among the groups that received recordings or summaries than among control groups. Receiving a recording or summary had no significant effect on anxiety or depression between the groups. None of the included studies assessed survival or health outcomes other than psychological outcomes. Conclusions Wider use of consultation tapes and summary letters could benefit many adults with cancer, without causing additional anxiety or depression, but consideration should be given to individuals’ circumstances and preferences.
Uncoupling of protein‐3 induces an uncontrolled uncoupling of mitochondria after expression in muscle derived L6 cells
Authors: Guerini Danilo , Prati Elisabetta , Desai Urvi , Nick Hans Peter , Flammer Rolf , Grüninger Stephan , Cumin Frederic , Kaleko Machael , Connelly Sheila , Chiesi Michele 
Published: March 2003

The uncoupling proteins (UCPs) are thought to uncouple oxidative phosphorylation in the mitochondria and thus generate heat. One of the UCP isoforms, UCP3, is abundantly expressed in skeletal muscle, the major thermogenic tissue in humans. UCP3 has been overexpressed at high levels in yeast systems, where it leads to the uncoupling of cell respiration, suggesting that UCP3 may indeed be capable of dissipating the mitochondrial proton gradient. This effect, however, was recently shown to be a consequence of the high level of expression and incorrect folding of the protein and not to its intrinsic uncoupling activity. In the present study, we investigated the properties of UCP3 overexpressed in a relevant mammalian host system such as the rat myoblast L6␣cell line. UCP3 was expressed in relatively low levels (< 1 µg·mg−1 membrane protein) with the help of an adenovirus vector. Immunofluorescence microscopy of transduced L6 cells showed that UCP3 was expressed in more than 90% of the cells and that its staining pattern was characteristic for mitochondrial localization. The oxygen consumption of L6 cells under nonphosphorylating conditions increased concomitantly with the levels of UCP3 expression. However, uncoupling was associated with an inhibition of the maximal respiratory capacity of mitochondria and was not affected by purine nucleotides and free fatty acids. Moreover, recombinant UCP3 was resistant to Triton X‐100 extraction under conditions that fully solubilize membrane bound proteins. Thus, UCP3 can be uniformly overexpressed in the mitochondria of a relevant muscle‐derived cell line resulting in the expected increase of mitochondrial uncoupling. However, our data suggest that the protein is present in an incompetent conformation.
Effect of adenosine 5′‐[β,γ‐imido]triphosphate on myosin head domain movements
Authors: Hartvig Nóra , Lõrinczy Dénes , Farkas Nelli , Belagyi Joseph 
Published: March 2003

Conventional and saturation transfer electron paramagnetic resonance spectroscopy (EPR and ST EPR) was used to study the orientation of probe molecules in muscle fibers in different intermediate states of the ATP hydrolysis cycle. A separate procedure was used to obtain ST EPR spectra with precise phase settings even in the case of samples with low spectral intensity.Fibers prepared from rabbit psoas muscle were labeled with isothiocyanate spin labels at the reactive thiol sites of the catalytic domain of myosin. In comparison with rigor, a significant difference was detected in the orientation‐dependence of spin labels in the ADP and adenosine 5′‐[β,γ‐imido]triphosphate (AdoPP[CH2]P) states, indicating changes in the internal dynamics and domain orientation of myosin. In the AdoPP[CH2]P state, approximately half of the myosin heads reflected the motional state of ADP–myosin, and the other half showed a different dynamic state with greater mobility.
Solution structure of the Alzheimer amyloid β‐peptide (1–42) in an apolar microenvironment
Authors: Crescenzi Orlando , Tomaselli Simona , Guerrini Remo , Salvadori Severo , D'Ursi Anna M. , Temussi Piero Andrea , Picone Delia 
Published: March 2003

The major components of neuritic plaques found in Alzheimer disease (AD) are peptides known as amyloid β‐peptides (Aβ), which derive from the proteolitic cleavage of the amyloid precursor proteins. In vitro Aβ may undergo a conformational transition from a soluble form to aggregated, fibrillary β‐sheet structures, which seem to be neurotoxic. Alternatively, it has been suggested that an α‐helical form can be involved in a process of membrane poration, which would then trigger cellular death.Conformational studies on these peptides in aqueous solution are complicated by their tendency to aggregate, and only recently NMR structures of Aβ‐(1–40) and Aβ‐(1–42) have been determined in aqueous trifluoroethanol or in SDS micelles. All these studies hint to the presence of two helical regions, connected through a flexible kink, but it proved difficult to determine the length and position of the helical stretches with accuracy and, most of all, to ascertain whether the kink region has a preferred conformation. In the search for a medium which could allow a more accurate structure determination, we performed an exhaustive solvent scan that showed a high propensity of Aβ‐(1–42) to adopt helical conformations in aqueous solutions of fluorinated alcohols. The 3D NMR structure of Aβ‐(1–42) shows two helical regions encompassing residues 8–25 and 28–38, connected by a regular type I β‐turn. The surprising similarity of this structure, as well as the sequence of the C‐terminal moiety, with those of the fusion domain of influenza hemagglutinin suggests a direct mechanism of neurotoxicity.
Optimizing the delivery systems of chimeric RNA·DNA oligonucleotides
Authors: Liang Li , Liu De‐Pei , Liang Chih‐Chuan 
Published: March 2003

Special oligonucleotides for targeted gene correction have attracted increasing attention recently, one of which is the chimeric RNA·DNA oligonucleotide (RDO) system. RDOs for targeted gene correction were first designed in 1996, and are typically 68 nucleotides in length including continuous RNA and DNA sequences (RNA is 2′‐O‐methyl‐modified). They have a 25 bp double stranded region homologous to the targeted gene, two hairpin ends of T loop and a 5 bp GC clamp, that give the molecule much greater stability [Fig. 1]. One mismatch site in the middle of the double‐stranded region is designed for targeted gene therapy. RDOs have been used recently for targeted gene correction of point mutations both in vitro and in vivo, but many problems must be solved before clinical application. One of the solutions is to optimize the delivery vectors for RDOs. To date, few RDO delivery systems have been used. Therefore, new vectors should be tried for RDO transfer, such as the use of nanoparticles. Additionally, different kinds of modifications should be applied to RDO carrier systems to increase the total correction efficiency in vivo. Only with the development of delivery systems can RDOs be used for gene therapy, and successfully applied to functional genomics.Diagrammatic structure of RDO. RDOs are typically 68 nucleotides in length, including continuous 2′‐O‐methyl‐modified RNA and DNA sequences, a 25 bp double‐stranded region homologous to the targeted gene, two hairpin ends of T loop and a 5 bp GC clamp, which give the molecule greater stability. One mismatch site in the middle of the double‐stranded region is designed for targeted gene therapy.image
Immunochemical and functional characterization of an agonist‐like monoclonal antibody against the M2 acetylcholine receptor
Authors: Elies Rozenn , Fu L. X. Michael , Eftekhari Pierre , Wallukat Gerd , Schulze Wolfgang , Granier Claude , Hjalmarson Åke , Hoebeke Johan 
Published: March 2003

Monoclonal antibodies were raised against a peptide corresponding to the second extracellular loop of the M2 acetylcholine receptor. One of the monoclonal antibodies, B8E5, was selected for further characterization on the basis of its high yield, its isotype (IgG2a), its dissociation kinetics and its agonist‐like activity.The epitope recognized by B8E5 corresponded to the N‐terminal part of the second extracellular loop of the receptor (V‐R‐T‐V‐E‐) as determined by competition immunoassays and epitope scanning. The KA of B8E5 for the target peptide was assessed by surface plasmon resonance (SPR) to be 6.5×107 M−1 by equilibrium and 3.7×107 M−1 by kinetic analysis. B8E5 recognized the M2 acetylcholine receptor on rat cardiac tissue. It only recognized the non‐reduced receptor in immunoblots. The antibody had no effect on antagonist binding but decreased the affinity for the agonist carbachol.B8E5 decreased the beating frequency of neonatal rat cardiomyocytes. The effect was specific since it was blocked by the target peptide and the antagonist atropine. The EC50 of the antibody corresponded to the KA measured by surface plasmon resonance. The physiological effect of the antibody did not lead to desensitization. The Fab fragments had no physiological effect; subsequent addition of anti‐mouse IgG however restored the physiological effect.These results confirm that the N‐terminus of the second extracellular loop is a functional target for antibodies against the M2 acetylcholine receptor. They suggest that the functional epitope is only accessible in the non‐reduced receptor. The antibodies act through a functional dimerization of the receptor.
Sorting of invertase signal peptide mutants in yeast dependent and independent on the signal‐recognition particle
Authors: Rothe Christine , Lehle Ludwig 
Published: March 2003

There is growing evidence that yeast contains two efficient pathways of protein translocation across the endoplasmic reticulum membrane, one dependent on the signal‐recognition particle (SRP) and the other independent. Their specificity, however, is largely obscure. For higher eukaryotes it has been shown that a high average hydrophobicity of the core region with a minimal length around six or seven amino acids, as well as a stabilized α‐helix, are decisive structural features for translocation. Using yeast invertase as a secretory model protein, we have found that mutated signal sequences with Pro or Gly in the core, or having only four hydrophobic amino acids, are not functional in translocation across microsomal membranes of dog pancreas because they do not interact with the SRP. Expression of these mutant variants in Saccharomyces cerevisiae revealed that they are sorted independently of the SRP since translocation was not impaired in an SRP‐deficient yeast strain. In contrast to this, wild‐type invertase is translocated SRP‐dependently in wild‐type cells and shows a decreased translocation in SRP‐deficient cells.By overexpression of Srp54p, but not of Hsc70p, the translocation defect of wild‐type invertase in an SRP54 disruptant is restored. The data indicate that targeting of proteins to the endoplasmic reticulum in Saccharomyces cerevisiae seems to be more flexible than in higher eukaryotes as far as the structural requirements of signal sequences are concerned, and that the route taken is specified by the sequence.
Fibrin selectivity of the isolated protease domains of tissue‐type and vampire bat salivary gland plasminogen activators
Authors: Toschi Luisella , Bringmann Peter , Petri Thomas , Donner Peter , Schleuning Wolf‐Dieter 
Published: March 2003

The activity of vampire bat (Desmodus rotundus) salivary plasminogen activator (D. rotundus PAα1) and to a much lesser extent of tissue‐type plasminogen activator (t‐PA) is stimulated by the presence of fibrin. This cofactor requirement has in the past intuitively been attributed to fibrin binding. We have previously shown that elements of the non‐protease domain of D. rotundus PAα1 could contribute to fibrin stimulation irrespective of fibrin binding.We now demonstrate that the protease domain of D. rotundus PAα1 by itself exhibits fibrin selectivity, i.e. it is 32‐fold stimulated by fibrin but only 1.5‐fold by fibrinogen. To a lesser extent this fibrin selectivity is also shared by the protease domain of t‐PA. Our findings indicate that protein‐protein interactions apart from fibrin binding affect the stimulatory mechanism of fibrin on D. rotundus PAα1 and t‐PA.
Isolation and characterization of an evolutionary precursor of human monoamine oxidases A and B
Authors: Sablin Sergey O. , Yankovskaya Victoria , Bernard Suzanne , Cronin Ciarán N. , Singer Thomas P. 
Published: March 2003

An interesting flavoprotein‐type monoamine oxidase (MAO) was recently isolated from Aspergillus niger and cloned [Schilling, B. & Lerch, K. (1995 a) Biochim. Biophys. Acta 1243, 529−537; Schilling, B. & Lerch, K. (1995 b) Mol. Gen. Genet. 247, 430−438]. The properties of this MAO, as well as a substantial part of its amino acid sequence, resemble those of both MAO A and B from higher animals, raising the possibility that it may be an evolutionary precursor of these mitochondrial enzymes. It differs from MAO A and B in several respects, however, including the fact that it is soluble and of peroxisomal location and that the FAD is non‐covalently attached.We have overexpressed the fungal enzyme (MAO‐N) in Escherichia coli and isolated it in pure form. Since several of the observations of previous workers on MAO‐N could not be reproduced, we have reexamined its substrate specificity, interaction with reversible and irreversible inhibitors and other catalytic and molecular properties. MAO‐N has a considerably higher turnover number on many aliphatic and aromatic amines than either form of the mammalian enzyme. Some aspects of the substrate specificity resemble those of MAO B, while others are similar to MAO A, including biphasic kinetics in double reciprocal plots. Contrary to a previous report [Schilling, B. & Lerch, K. (1995 a) Biochim. Biophys. Acta 1243, 529−537], however, the fungal enzyme does not oxidize serotonin, norepinephrine, dopamine or other biogenic amines. MAO‐N is irreversibly inhibited by stoichiometric amounts of both (−)deprenyl and clorgyline in a mechanism‐based reaction, forming flavocyanine adducts with N5 of the FAD, like the mammalian enzymes, but inactivation is much faster with clorgyline than deprenyl, suggesting a closer resemblance to MAO A than B. The dissociation constants for a large number of reversible competitive inhibitors have been determined for MAO‐N and comparison with similar values for MAO A and B again pointed to a greater similarity to the former than the latter.
Photophysical properties of porphyrin planar aggregates in liposomes
Authors: Ricchelli Fernanda , Gobbo Silvano , Moreno Giuliana , Salet Christian , Brancaleon Lorenzo , Mazzini Alberto 
Published: March 2003

This paper describes studies of some photophysical properties of non‐covalent planar aggregates of hematoporphyrin and protoporphyrin. This porphyrin species has been recently discovered and can be generated in lipid bilayers such as liposomes and inner mitochondrial membranes. The relative weight of this species in different media, as compared to porphyrin monomers and stacked aggregates, has been deduced by fluorescence decay studies. In contrast with what is observed for stacked aggregates, promotion of planar suprastructures can occur both in aqueous and lipid environments. The spectroscopic properties are very similar to those of the corresponding monomers, in particular as regards the shape of the absorption and emission spectra. The fluorescence decay times are generally higher than those of the monomers, and depend on the medium in which the planar aggregates are formed.The photooxidation properties of porphyrin planar aggregates, as revealed by oxygen consumption and histidine photodegradation upon irradiation at 365 nm, were compared to those of the monomers. The extent of the photooxidation process is nearly 20−30 % higher in planar aggregates than in the monomers. In contrast, it is well known that cofacial aggregates are photochemically inert and only monomeric species of porphyrin are efficient photosensitizers. The biological relevance of these findings is discussed.