bluuebunny/crossref_metadata_2025 huggingface dataset:
On‐column digestion of proteins in aqueous‐organic solvents
Authors: Slysz Gordon W. , Schriemer David C.
Published: April 2003

Proteolytic digestion is an important step in protein identification by peptide mass mapping and tandem mass spectrometry (MS/MS)‐based peptide sequencing. Traditional methods of protein digestion require extended incubation times and have difficulty with proteolytically resistant proteins. Here, we describe a method in which a protein solution was combined with a mixed aqueous‐organic solution (methanol, isopropanol, or acetonitrile) and passed through a microcolumn containing immobilized trypsin. Myoglobin sequence coverage was high (>85%) in all three solvents, and differences in spectra were seen among the different solution conditions. Notably, methanol‐based digestions produced fewer missed cleavages while acetonitrile‐based digestions produced the most peptides and the most intense mass spectra. Flow rates through the column were varied from 0.5 to 15 μL/min, corresponding to column residence times of 78 and 2.6 s, respectively. All flow rates produced high sequence coverage of myoglobin, although, at higher flow rates, more missed cleavages were observed. No significant increase in undigested myoglobin was observed with flow rates up to 15 μL/min. The described method was applied to the digestion of human transferrin (hTf), a proteolytically resistant protein. Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometric (MALDI‐TOFMS) analysis detected 42 peptides covering 46% of the hTf sequence. The traditional aqueous method resulted in 12 peptides (8% sequence coverage) only when high concentrations of trypsin were used. Lastly, digestion of low nanomolar myoglobin was shown to produce detectable peptides and resulted in a correct database hit. Thus, we demonstrate a method that is capable of rapid on‐line digestion, thereby lending itself to high‐throughput identification of proteins. Copyright © 2003 John Wiley & Sons, Ltd.
A simplified recursive formulation for the dynamic analysis of spatial mechanisms in point coordinates
Authors: Attia H.A.
Published: April 2003

In the present study, a recursive formulation for generating the equations of motion of spatial mechanisms with revolute and/or spherical joints is presented. The method uses the concepts of linear and angular momentums to generate the rigid body equations of motion in terms of the Cartesian coordinates of a dynamically equivalent constrained system of particles, without introducing any rotational coordinates and the corresponding rotational transformation matrix. It eliminates the necessity of distributing the external forces and couples over the equivalent system of particles. For the open‐chain system, the equations of motion are generated recursively along the serial chains. Closed‐chain system is transformed to open‐chain by cutting suitable kinematic joints and introducing cut‐joint constraints. The method is simple and suitable for computer implementation. An example is chosen to demonstrate the generality and simplicity of the developed formulation.
Air flow influences on local climate: observed United Kingdom climate variations
Authors: Osborn Timothy J. , Jones Philip D.
Published: April 2003

Identifying and removing the influence of atmospheric circulation variability on central England temperature increases the statistical significance of warming trends in spring, autumn and the annual mean over the last 50 years. The trends are more detectable because the circulation changes contribute greatly to the ‘noise’ of interannual to interdecadal variability, but induce only small multi‐decadal trends (the ‘signal’). Factoring out the circulation can thus enhance the signal‐to‐noise ratio. For precipitation, the recent enhancement in the difference between summer rainfall in south‐east England and winter precipitation in northern Scotland can partly be explained by atmospheric circulation variability over the past 40 years (particularly the increase in the North Atlantic Oscillation index from the 1960s to the early 1990s). Copyright © 2000 Royal Meteorological Society.
Extending the ‘stressy’ edge: molecular chaperones flirting with RNA
Authors: Henics Tamás
Published: April 2003

A number of intriguing observations have emerged during the past years indicating that certain classes of the evolutionarily highly conserved heat shock or stress proteins extend their molecular partnerships beyond the originally recognized protein world. In this review, following a brief introduction to the 70‐kDa family of stress proteins, we summarize the main aspects of RNA recognition and binding by this class of molecules. By highlighting some biochemical features of both the protein and RNA partners, we attempt to embed the central parts of this interaction in the context of potential physiological relevance. As perhaps true for many newly recognized molecular interactions, the phenomenon of RNA recognition and binding by molecular chaperones discussed in this review calls for a place of ever‐growing importance in the functional genomic era, where an expanding number of previously unsuspected molecular partnerships are uncovered by virtue of powerful high throughput methodologies. We suggest that integration of this new knowledge into the long‐outlined ‘classical’ network of cellular metabolism at both the biochemical and architectural level is pivotal to the emerging ‘synthesis era’ of today's cell biology.
Offenbarung 1.1–3: Johanneische Autorisierung einer Aufklärungsschrift
Authors: TAEGER JENS-W.
Published: April 2003

This paper addresses certain aspects of the opening passage of Revelation which have received little attention in recent years: the central role ascribed to John and the multiple characterisations of his work. These aspects are analysed in the context of the metatextual character of the passage. Moreover, these verses probably were not composed by the author himself, but rather by an editor/redactor who belonged to the Johannine circle, as can be shown through linguistic data and thematic analogies. The editor authorises John's work and thereby the critical view towards the Empire contained in the book itself – an attitude which stands in opposition to the one commonly adopted by Christians at the time.
Early intervention for relapse in schizophrenia: results of a 12-month randomized controlled trial of cognitive behavioural therapy
Authors: GUMLEY A. , O'GRADY M. , MCNAY L. , REILLY J. , POWER K. , NORRIE J.
Published: April 2003

Background. The paper describes a randomized controlled trial of targeting cognitive behavioural therapy (CBT) during prodromal or early signs of relapse in schizophrenia. We hypothesized that CBT would result in reduced admission and relapse, reduced positive and negative symptoms, and improved social functioning.

Method. A total of 144 participants with schizophrenia or a related disorder were randomized to receive either treatment as usual (TAU) (N=72) or CBT+TAU (N=72). Participants were prospectively followed up between entry and 12 months.

Results. At 12 months, 11 (15·3%) participants in the CBT group were admitted to hospital compared to 19 (26·4%) of the TAU group (hazard ratio=0·53, P=0·10, 95% CI 0·25, 1·10). A total of 13 (18·1%) participants in CBT relapsed compared to 25 (34·7%) in TAU (hazard ratio=0·47, P<0·05, 95% CI 0·24, 0·92). In addition, the CBT group showed significantly greater improvement in positive symptoms, negative symptoms, global psychopathology, performance of independent functions and prosocial activities.

Conclusions. The study provides evidence for the feasibility and effectiveness for targeting CBT on the appearance of early signs of relapse in schizophrenia. The results are discussed in context of the study's methodological limitations.
NEUTRINO CONDENSATES AT CENTER OF GALAXIES AS BACKGROUND FOR THE MSW MECHANISM
Authors: CAPOZZIELLO S. , LAMBIASE G. , IOVANE G.
Published: April 2003

The possibility is explored that neutrino condensates, possible candidates for the explanation of very massive objects in galactic centers, could act as background for the Mikheyev–Smirnov–Wolfeinstein mechanism responsible of neutrino oscillations. Assuming a simple neutrino star model with constant density, the lower limit of the mass squared difference of neutrino oscillations is inferred. Consequences on neutrino asymmetry are discussed.
RECOGNITION OF NUMERAL STRINGS ON MAPS
Authors: CHEN LIANG-HUA , WANG JIING-YUH
Published: April 2003

This paper presents a complete procedure for the extraction and recognition of hand-printed numeral strings on maps. The extraction algorithm can extract individual characters from a map even if the characters touch each other or touch with graphical line. The feature-based recognition algorithm can recognize numeral characters of any size, position and orientation. Our features for discrimination are simple and easily detectable. Experimental results on utility and cadastral maps have shown that the proposed technique is effective in the automatic data capture of geographic information systems.
INVARIANT GEOMETRIC PROPERTIES OF IMAGE CORRESPONDENCE VECTORS AS RIGID CONSTRAINTS TO MOTION ESTIMATION
Authors: LIU Y. , RODRIGUES M. A.
Published: April 2003

Accurate motion estimation algorithms are based on a number of invariant properties that can be inferred from the motion. A large number of calibration algorithms have been proposed over the last two decades mainly based on analytic, perspective, or epipolar geometries. Extending Chasles' screw motion concept to the estimation of motion parameters in computer vision, we have presented an analysis of geometric properties of image correspondence vectors synthesized into a single coordinate frame and developed calibration algorithms using both simulated and real range image data.15,16 In this paper, we extend that work by defining the relevant geometric properties of image correspondence vectors from the point of view of invariants and by developing two calibration algorithms using the Monte Carlo method and median filtering. The algorithms are applied to real and synthetic image data corrupted by noise and outliers. Experimental results demonstrate that the median filter based algorithm is generally more robust and accurate than the Monte Carlo based algorithm and that the geometric analysis of invariant properties of correspondence vectors is a useful framework to motion parameter estimation.
Potential roles of galectins in myeloid differentiation into three different lineages
Authors: Abedin Mohammad J , Kashio Yumiko , Seki Masako , Nakamura Kazuhiro , Hirashima Mitsuomi
Published: April 2003

Little is known about the roles of galectins, a family of β-galactoside-binding lectins, in myeloid cell differentiation. In the present experiments, we used HL-60 cells as a model of myeloid cell differentiation. The HL-60 cells were differentiated into eosinophil-, monocyte-, and neutrophil-like cells by coculture with sodium butyrate under a mild alkaline condition, phorbol 12-myristate 13-acetate, and dimethyl sulfoxide, respectively. Thus, the expression of galectins in HL-60 cells during differentiation into three different lineages was assessed. Reverse transcriptase-polymerase chain reaction analyses revealed that undifferentiated HL-60 cells expressed galectin-1, -3, -8, -9, and -10 (identical to Charcot Leyden crystal) mRNAs, and galectin-2, -4, and -7 were negligible before and after the differentiations. We failed to detect evident changes in the mRNA levels of galectin-1 and -8 during the diferentiations. However, during the eosinophilic differentiation, galectin-9 mRNA expression was gradually decreased, whereas galectin-10 mRNA expression was increased. During the course of monocytic differentiation, galectin-9 mRNA expression was down-regulated, whereas galectin-3 mRNA expression was up-regulated. Moreover, only galectin-10 mRNA expression was enhanced in the process of neutrophilic differentiation. These changes in galectin expressions were confirmed by Western blot and flow cytometry analyses. It is thus suggested that changes in the expressions of galectin-3, -9, and -10 are potentially important for myeloid cell differentiation into specific lineages.
Analysis of mannose receptor regulation by IL-4, IL-10, and proteolytic processing using novel monoclonal antibodies
Authors: Martinez-Pomares Luisa , Reid Delyth M , Brown Gordon D , Taylor Philip R , Stillion Richard J , Linehan Sheena A , Zamze Susanne , Gordon Siamon , Wong Simon Y C
Published: April 2003

The study of the murinemacrophage mannose receptor (MR) has been hampered by the lack of specific reagents. We have generated and characterized novel anti-MR monoclonal antibodies and used them to analyze MR expression in primary mouse macrophages (MØ). In BioGel- and thioglycollate-elicited MØ, interleukin (IL)-4 up-regulated total cell-associated MR (cMR), correlating with enhanced surface expression. We investigated the influence of IL-10, a well-characterized deactivator of MØ function, on MR levels and observed that it had a similar effect to IL-4. In both cases, enhanced cMR levels translated into increased production of the soluble form of the receptor (sMR). We have demonstrated the presence of sMR in cultures of stable non-MØ transductants expressing full-length MR, indicating that the proteolytic activity responsible for cMR cleavage is not MØ-restricted. These data support a role for the MR in T helper cell type 2 cytokine-driven, immune responses and suggest a non-MØ contribution to sMR production in vivo.
Preparation of Nanocrystalline Titanium Oxide by Decomposition of Molecular Precursor α-(NH4)2TiO(SO4)2
Authors: Zhang Qinghong , Gao Lian
Published: April 2003

Hydroxy-free anatase TiO2 nanocrystals (25 nm) with photocatalytic activities higher than that of commercial photocatalyst Degussa P-25, were prepared by nonhydrolytic thermolysis of ammonium titanyl sulfate at 700 °C for 2 h.
Deep Drawing and Impact Extrusion of Magnesium Alloys at Room Temparature
Authors: Wagener H.‐W. , Hosse‐Hartmann J. , Friz R.
Published: April 2003

Magnesium alloys and composites are potentially highly attractive materials for lightweight design in the automotive industry – if common forming technologies, i.e., deep drawing and cold extrusion, are applicable to them; so far, this is seldom the case. The authors have tested some candidate Mg alloys and discuss the results of their measurements in detail. The Figure shows a typical workpiece part obtained by a deep‐drawing technique.
GdAlO3–ZrO2: A Ductile Ceramic Composite
Authors: Panneerselvam M. , Aggarwal N. , Subanna G.N. , Rao K.J.
Published: April 2003

A ceramic with a high fracture toughness KIC that is close to the respective values of cast iron and Mg alloys has been fabricated by microwave sintering of GdAlO3 and ZrO2 into composite materials of different molar fractions; for the 80/20 composite (see Figure for an SEM image) KIC reaches the remarkable value of 17.5 Mpa m1/2. Such composites may become extremely valuable for structural applications in advanced technologies.
Synthesis of Maleic Anhydrides and Maleic Acids by Pd‐Catalyzed Oxidative Dicarbonylation of Alk‐1‐ynes
Authors: Gabriele Bartolo , Veltri Lucia , Salerno Giuseppe , Costa Mirco , Chiusoli Gian Paolo
Published: April 2003

We have found that carbon dioxide effectively promotes the Pd‐catalyzed oxidative carbonylation of terminal alkynes to give maleic anhydrides in fair yields. Reactions were carried out in aqueous dioxane at 60−80 °C in the presence of catalytic amounts of PdI2 in conjunction with KI and under a 4:1:10 mixture of CO/air/CO2 (60 atm total pressure at 25 °C). By working in the presence of a large excess of water, maleic acids were formed selectively with unprecedented catalytic efficiencies for this kind of reaction. In the latter case, the use of an excess of carbon dioxide tended to inhibit, rather than promote, the reaction. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Biochemical properties of the cell wall in the Arabidopsis mutant bor1‐1 in relation to boron nutrition
Authors: Noguchi Kyotaro , Ishii Tadashi , Matsunaga Toshiro , Kakegawa Koichi , Hayashi Hiroaki , Fujiwara Toru
Published: April 2003

We examined concentrations of boron (B) and dimerization of rhamnogalacturonan II (RG‐II), a B‐binding polysaccharide, in the cell wall of a low‐B sensitive mutant of Arabidopsis thaliana, bor1‐1, to investigate possible effects of the bor1‐1 mutation on the biochemical form of pectins in the cell wall. In the bor1‐1 mutant, B concentrations in the cell wall from shoots were lower than those in the wild type at low B supply, whereas they were similar at sufficient B supply. The amount of B present as borate ester of the RG‐II dimer (dRG‐II‐B) in the bor1‐1 mutant was lower than that in the wild type at low B supply. In the wild type, about 90 % of RG‐II was present as dRG‐II‐B, both, at low and sufficient B supply. In the bor1‐1 mutant, about 60 % of RG‐II was in its monomeric form (mRG‐II) at low B supply, whereas more than 85 % of it was present as dRG‐II‐B at sufficient B supply. However, similar as the wild type, mRG‐II derived from the bor1‐1 mutant was able to form dRG‐II‐B in vitro in the presence of borate and lead. Sugar composition of cell wall fractions was similar in both genotypes. These results suggest that the polysaccharide composition in the cell wall was not strongly affected by the bor1‐1 mutation. The observed difference in dimerization of RG‐II at low B supply is most likely due to a reduced B concentration in the shoots of the bor1‐1 mutant.
Professor ARTHUR HUGH BUNTING, CMG
Authors: Betts David
Published: April 2003

A. H. Bunting was born in Johannesburg, South Africa on 7 September 1917 and died peacefully in his sleep at his home in Berkshire, UK on 8 May 2002.
Long-term follow-up of survival of liver transplant recipients with familial amyloid polyneuropathy (Portuguese type)
Authors: Suhr Ole B. , Ericzon Bo-Göran , Friman Styrbjörn
Published: April 2003

Portuguese type familial amyloid polyneuropathy is a dominantly inherited neuropathic amyloidosis caused by a mutant transthyretin (TTR). Because TTR is produced mainly by the liver, liver transplantation (LT) abolishes production of the amyloidogenic variant TTR. To date, the procedure appears to halt the progress of the disease. However, long-term outcome is unknown. The aim of the present study is to evaluate the survival of our initial group of unselected liver transplant recipients with FAP. Seventy patients, 51 transplant recipients and a control group of 19 nontransplantation patients, with disease onset before the age of 55 years were included on the study. Transplant recipients were divided into two categories: (1) early series, with patients followed up for 5 years or longer, and (2) new series, with patients followed up for 1 to 5 years. Nonparametric statistical methods were used. Binary regression analyses were performed by stepwise logistic regression and Cox proportional hazard regression. Survival analysis was performed using Kaplan-Meier analysis, the Cox-Mantel test. Survival analyses and Cox proportional hazard regression analysis were performed from disease onset, not from LT. Significantly decreased survival was noted for transplant recipients with a modified body mass index (mBMI) less than 600 compared with the control group (P < .05). A significant difference in survival also was observed between transplant recipients with an mBMI greater than 600 at the time of LT compared with those with an mBMI less than 600 (P < .02). mBMI and age at LT had a significant impact on survival; whereas late deaths were related to age at LT, early deaths were related to mBMI. The cumulative 10-year survival rate after disease onset was 94% in the new series, with one early death (<6 months) after LT, compared with a 78% survival rate and eight early deaths in the early series (P = .1).
The new liver allocation system: Moving toward evidence-based transplantation policy
Authors: Freeman Richard B. , Wiesner Russell H. , Harper Ann , McDiarmid Sue V. , Lake Jack , Edwards Erick , Merion Robert , Wolfe Robert , Turcotte Jeremiah , Teperman Lewis
Published: April 2003

In 1999, the Institute of Medicine suggested that instituting a continuous disease severity score that de-emphasizes waiting time could improve the allocation of cadaveric livers for transplantation. This report describes the development and initial implementation of this new plan. The goal was to develop a continuous disease severity scale that uses objective, readily available variables to predict mortality risk in patients with end-stage liver disease and reduce the emphasis on waiting time. Mechanisms were also developed for inclusion of good transplant candidates who do not have high risk of death but for whom transplantation may be urgent. The Model for End-Stage Liver Disease (MELD) and Pediatric End-Stage Liver Disease (PELD) scores were selected as the basis for the new allocation policy because of their high degree of accuracy for predicting death in patients having a variety of liver disease etiologies and across a broad spectrum of liver disease severity. Except for the most urgent patients, all patients will be ranked continuously under the new policy by their MELD/PELD score. Waiting time is used only to prioritize patients with identical MELD/PELD scores. Patients who are not well served by the MELD/PELD scores can be prioritized through a regionalized peer review system. This new liver allocation plan is based on more objective, verifiable measures of disease severity with minimal emphasis on waiting time. Application of such risk models provides an evidenced-based approach on which to base further refinements and improve the model.
Orthotopic liver transplantation in patients with human immunodeficiency virus and end-stage liver disease
Authors: Neff Guy W. , Bonham Andrew , Tzakis Andreas G. , Ragni Margaret , Jayaweera Dushyantha , Schiff Eugene R. , Shakil Obaid , Fung John J.
Published: April 2003

Patients with human immunodeficiency virus (HIV) most often have hepatitis C virus (HCV) or hepatitis B (HBV) virus coinfection, or both, as a cause of their liver disease. Recent survival statistics show that patients infected with HIV treated with highly active antiretroviral therapy (HAART) can expect a significant prolongationof life by interfering with the natural progression of HIV to aquired immune deficiency syndrome (AIDS). Therefore, HIV-positive patients experiencing complications of liver failure are at greater immediate risk of dying from their end-stage liver disease (ESLD) rather than their HIV. Many transplant centers still consider HIV infection asa contraindication for orthotopic liver transplantation (OLT). Atour two institutions, we believe that patients with HIV suffering from ESLDshould be considered for OLT. This study evaluates the survival of patients undergoing OLT with HIV under HAART therapy. OLT was performed in 16 patients with HIV suffering from ESLD as a result of chronic HCV, chronic HBV, or fulminant hepatic failure (FHF). Collected data include patient demographics, patient and graft survival, pre-OLT assessments, and postoperative complications (including opportunistic infections). Ten patients at Pittsburgh and 6 patients at Miami received OLT. Of the 16 patients who received OLT, 14 remain alive to date. Thirteen of 16 patients are more than 12 months post-OLT, whereas the last patient is currently 6 months post-OLT. Five patients at Miami and 9 of 10 patients at Pittsburgh received HAART therapy before OLT, although 2 of the Pittsburgh patients had their HAART therapy discontinued before OLT because of significant liver dysfunction. The pre-OLT viral loads were undetectable in 13 of 16 patients. The cluster determinant (CD)4 count was less than 200 in 6 patients and greater than 100 in 2 patients before OLT. In all patients, CD4 counts increased above 200 in the post-OLT period. Tacrolimus toxicity associated with the pharmacologic inhibition of cytochrome p450 metabolism caused by protease inhibitors occurred in 6 patientsafter OLT. Six patients (38%) experienced acute cellular rejection immediately after OLT. Our experience suggests that OLT is effective in selected HIV-positive patients suffering from ESLD. Patient and graftsurvival was similar to non-HIV-positive patients suffering from the same indications for OLT. Acute cellular rejection was no less frequent that seen in non-HIV-positive patients. Given the complex pharmacologic interactions between the protease inhibitors and tacrolimus, careful monitoring, and attention is required to prevent toxicity or underdosing.
A NEW REPRESENTATION FOR QUARTIC CURVES AND COMPLETE SETS OF GEOMETRIC INVARIANTS
Authors: UNEL MUSTAFA , WOLOVICH WILLIAM A.
Published: April 2003

Many free-form object boundaries can be modeled by quartics with bounded zero sets. The fact that any nondegenerate closed-bounded algebraic curve of even degree n=2p can be expressed as the product of p conics, which are real ellipses, plus a remaining polynomial of degree n-2,12 can be utilized to express a nondegenerate quartic as the product of two leading ellipses plus a third conic which might be either a closed curve (an ellipse) or an open curve (a hyperbola). However, it can be shown that the leading ellipses can be modified with appropriate constants by constraining the third conic to be a circle, thus implying a 2-ellipse and 1-circle; i.e. an elliptical-circular(E2C)representation of the quartic. The use of such representations is to simplify the analysis of quartics by exploiting the well-known properties of conics and to develop a set of functionally independent geometric invariants for recognition purposes. Also, it is shown that the underlying Euclidean transformation between two configurations of the same quartic can be determined using the centers of the three conics.
The expression of adhesion molecules in cigarette smoke-induced airways obstruction
Authors: Gonzalez S , Hards J , van Eeden S , Hogg JC
Published: April 2003

Cigarette smoking produces peripheral airway inflammation in all smokers, and chronic airways obstruction in approximately 20% of heavy smokers. The present study was designed to test the hypothesis that airways obstruction is related to changes in the expression of adhesion molecules involved in the recruitment of cells to sites of inflammation in the lung. Freshly resected lungs from heavy smokers with airways obstruction (n = 10) and from heavy smokers with normal lung function (n = 10) were collected in the operating room, inflated with optimal cutting temperature (OCT) medium and frozen over liquid nitrogen. Six micrometres thick cryostat sections cut from random samples of this tissue were stained, using immunohistochemistry, with monoclonal antibodies to the adhesion molecules on leucocytes: L-selectin, very late activation antigen-4 (VLA-4), CD11a/CD18, CD11b/CD18, CD11c/CD18; and on endothelial and epithelial surfaces: E-selectin, P-selectin, vascular cell adhesion molecule (VCAM-1), intercellular adhesion molecule (ICAM)-1 and ICAM-2 using the alkaline phosphatase anti-alkaline phosphatase (APAAP) technique. The slides were coded and the expression of each molecule scored by three observers using a semiquantitative grading system. Two inducible adhesion molecules, E-selectin on endothelium and CD11b on leucocytes, were also evaluated using quantitative morphometric analysis. The results showed a distribution of adhesion molecules that was consistent with the inflammatory response in the airways and parenchyma of all subjects but failed to show any differences between those with or without airways obstruction. We conclude that development of airways obstruction in heavy smokers cannot be explained by differences in the expression of adhesion molecules known to be involved in the control of cell traffic in the lung.
Modulation of glucocorticoid receptor expression in human bronchial epithelial cell lines by IL-1 beta, TNF-alpha and LPS
Authors: Verheggen MM , van Hal PT , Adriaansen-Soeting PW , Goense BJ , Hoogsteden HC , Brinkmann AO , Versnel MA
Published: April 2003

Bronchial epithelium plays a major role in the regulation of inflammatory reactions in the airways. It is thought to be a possible target for glucocorticoid therapy. Glucocorticoid responsiveness requires the presence of specific glucocorticoid receptors (GR). Until now, little was known about the presence of such receptors in the human bronchial epithelium. In this study we demonstrated the expression of GR messenger ribonucleic acid (mRNA) in two simian virus (SV)-40/adenovirus-transformed human bronchial epithelial cell lines, BEAS S6 and BEAS 2B. In a whole cell dexamethasone binding assay, BEAS S6 and BEAS 2B cells were found to possess (mean +/- SEM) 28.9 +/- 4.4 x 10(3) and 32.1 +/- 5.7 x 10(3) binding sites per cell, respectively, with dissociation constant (Kd) values of 8.2 +/- 1.5 and 8.6 +/- 2.4 nM, respectively. Using electrophoretic mobility shift assays we demonstrated the binding of nuclear translocated GR to specific sites on deoxyribonucleic acid (DNA), named glucocorticoid responsive elements (GRE). Lipopolysaccharide (LPS) and interleukin-1 beta (IL-1 beta) significantly increased the number of GR per cell (median = 312% and 171% of control, respectively; p < 0.05), but significantly reduced the ligand affinity of these receptors, i.e. increased the Kd (median = 410% and 145% of control, respectively; p < 0.05) in BEAS 2B cells. These results indicate that the bronchial epithelium may be an actual target for glucocorticoid therapy. Inflammatory mediators, such as IL-1 beta and LPS, modulate the number and ligand affinity of these GR. Therefore, the response of bronchial epithelium to glucocorticoid therapy may be modulated by airway diseases associated with inflammation.
Allergic responses reduce the relaxant effect of beta-agonists but not potassium channel openers in guinea-pig isolated trachea
Authors: Houjou S , Iizuka K , Dobashi K , Nakazawa T
Published: April 2003

This study was conducted to determine whether the relaxant effect of adenosine triphosphate (ATP)-sensitive potassium channel (KATP) openers on airway smooth muscle are reduced during the hyporesponsiveness of beta-adrenergic receptors. Isometric tension was measured and dose-response curves were constructed for levcromakalim (a KATP opener), Y-26763 (another KATP opener), isoprenaline and theophylline in guinea-pig isolated trachea that was challenged with ovalbumin or pretreated in vitro with either isoprenaline or the KATP openers. Antigen challenge in vitro significantly reduced the potency but not the efficacy of isoprenaline in tracheal strips from actively sensitized guinea-pigs. The concentration of drug that produced a half-maximum inhibition (IC50) was 0.014 +/- 0.004 microM in the challenge group versus 0.006 +/- 0.001 microM in the control group (p < 0.05; n = 9). The IC50 of levcromakalim (1.73 +/- 0.17 microM; n = 6) and of theophylline (110 +/- 3.1 microM; n = 6) were unaffected. Exposure to 4 microM isoprenaline for 30 min evoked beta-adrenergic hyporesponsiveness: the IC50 of isoprenaline rose significantly from 0.010 +/- 0.001 to 0.017 +/- 0.002 microM (p < 0.01; n = 6). No effect was observed on the relaxant actions of levcromakalim and theophylline. In contrast, prior incubation with either levcromakalim or Y-26763 (300 microM for 30 min) significantly reduced the subsequent potency and efficacy of levcromakalim, but did not alter the effects of isoprenaline and theophylline. We conclude that the relaxant effect of the adenosine triphosphate-sensitive potassium channel openers was independent of beta-adrenergic hyporesponsiveness in airway smooth muscle of guinea-pigs.
Involvement of PAF in postallergic propranolol-induced bronchoconstriction in guinea-pigs
Authors: Fujimura M , Tsujiura M , Songur N , Myou S , Matsuda T
Published: April 2003

Administration of propranolol can provoke bronchoconstriction in asthmatic patients. Recently, we successfully developed a guinea-pig model for propranolol-induced bronchoconstriction (PIB). We hypothesized that such bronchoconstriction may result from the inflammatory mediators released by an allergic reaction. The purpose of this study was to examine the role of platelet-activating factor (PAF) in the development of PIB after allergic reaction. Propranolol, at a concentration of 10 mg.mL-1 was inhaled 20 min after antigen challenge in passively sensitized, anaesthetized and artificially-ventilated guinea-pigs. The animals were treated intravenously with PAF antagonists, E6123 (1 and 10 micrograms.kg-1) or Y-24180 (1 and 10 mg.kg-1), 10 min before or 15 min after antigen challenge. Propranolol inhaled 20 min after antigen challenge caused bronchoconstriction. E6123 and Y-24180 administered 15 min after antigen challenge as well as 10 min before antigen challenge reduced the PIB in a dose-dependent manner. We conclude that platelet-activating factor may contribute to the development of propranolol-induced bronchoconstriction after allergic reaction in our guinea-pig model.
Autoimmunity and geriatrics: clinical significance of autoimmune manifestations in the elderly
Authors: Ramos-Casals M , García-Carrasco M , Brito M P , López-Soto A , Font J
Published: April 2003

The immune system undergoes continuousmorphologic and functionalchanges throughoutthe years, and it is now believed that the immune response has its peak function in puberty and gradually decreases with age (immunosenescence). Recent studies in healthy octogenarian patients suggest that the immune system, instead of suffering a generalized deterioration, undergoes a remodelling/readjustment of its major functions. Increase in two contrasting phenomena coexist in immunosenescence: on the one hand, a decrease in the capacity of the immune response and, on the other hand autoantibodyproduction.The possibleconsequencesof this progressive‘ageing’ of the immune system are the increasein autoimmune phenomena, incidenceof neoplasiaand predisposition to infections. The study of autoimmune manifestations in elderly populations should be considered a priority for future medical research because of increasing life expectancy, especially in developed countries. This review analyses the main immune disorders associated with immunosenescence, the prevalence and clinical significance of autoantibodiesin the elderly and the clinical expression of the main autoimmune diseases in older patients.
Factor V Leiden mutation in Sneddon syndrome
Authors: Besnier R , Francès C , Ankri A , Aiach M , Piette J C
Published: April 2003

Sneddon syndrome (SNS) is characterizedby the associationof ischaemic cerebrovascularevents and widespread livedo racemosa. Its pathophysiology is still controversial. The aim of this study was to evaluate the prevalence of factor V Leiden mutation in consecutive patients referred for SNS according to antiphospholipid antibodies (aPL) status. Fifty-three Caucasian patients were enrolled from 1996 to 2001. Diagnosis of SNS was based on the presence of a widespread livedo racemosa and at least one clinical neurologic ischaemic event. The following investigations were performed: detection of antithrombin III, protein C and protein S deficiency, lupus anticoagulant, anticardiolipin and anti-b2 glycoprotein I antibodies, biologic false-positive test for syphilis, and factor V Leiden mutation by direct genomic analysis. Fisher’s test and t-test were used for statistics. Detection of aPL on multiple determinationswas negativein 31 patients (group 1) and positivein 22 patients (group 2). Factor V Leiden mutation was detected in six patients (11.3%), heterozygousin all. The frequency of this mutation was statistically higher in group 1 (6/31, 19.3%) than in group 2 (0/22; P 0.035). Within aPL-negative SNS, the comparison of patients with versus without factor V Leiden mutation showed no difference for clinical data or familial history of thrombosis. A high prevalence of heterozygous factor V mutation was found in aPL-negative patients with SNS. This finding adds further arguments to consider SNS as a heterogeneous entity.
Optimization of the rotational moulding process for polyolefins
Authors: Cramez M C , Oliveira M J , Crawford R J
Published: April 2003

Hollow plastic parts can be made by a number of manufacturing methods, but only rotational moulding offers the ability to create one-piece, stress-free parts with attractive economics. However, the fundamental nature of rotational moulding is such that cycle times are long. Consequently, the plastic is subjected to relatively high temperatures, in the presence of air, for excessive periods of time. This can lead to thermal and oxidative degradation at the inner free surface of the plastic, resulting in a deterioration in the performance of the moulded part. The industry relies heavily on the experience of operators to establish the best processing conditions to avoid degradation, but this is problematic and inefficient. Unfortunately, automatic process control is difficult owing to the complex rotation of the mould. Recent developments highlighting the importance of the peak air temperature inside the mould have been an important step forward, but ever-increasing technical demands on moulders make it clear that more sophisticated process control is needed. It is known that the processing conditions that lead to degradation vary with factors that affect the heating rate, such as the type of mould used and the thickness of the end-product. In the work reported here, a method is proposed for predicting the onset of degradation, on the basis that this occurs when the concentration of antioxidant in the polymer reaches zero. Good agreement has been obtained between the experimental and predicted optimum processing temperature for polyethylenes stabilized with different antioxidant systems. A procedure is described for identifying the best rotational moulding conditions so that more efficient manufacturing methods can be achieved.
A high-efficiency rough milling strategy for mould core machining
Authors: Chan K W , Chiu W K , Tan S T , Wong T N
Published: April 2003

Increasing the efficiency of rough machining operations can produce significant productivity improvement in mould and die making because most of the metal is removed in the roughing stage. In this paper, a high-efficiency 2.5-dimensional rough milling strategy for mould core machining is presented. The strategy consists of three different tool paths. The first tool path is generated on the basis of the convex hull boundary of a machining region. Owing to the absence of concave tool path segments, the convex hull based tool path can eliminate the chip load fluctuation problem encountered in corner cutting. The second tool path is an enhanced unidirectional straight-line tool path, which has the virtue of maintaining a steady cutting resistance throughout. The large staircases left behind by these two tool paths are refined by using the third tool path which is a contour-parallel tool path that cuts the mould core layer by layer in an upward manner. After applying these three tool paths, the stock material left on the mould core surface can be post-processed by the subsequent finish milling operation. A case study is illustrated to demonstrate the practicality of the presented rough milling strategy.
Characterization and representation of non-ideal effects in electrochemical machining
Authors: Clifton D , Mount A R , Mill F , Howarth P S
Published: April 2003

The basis of a technique for in-line process characterization of electrochemical machining (ECM) has been established. This technique has been applied to map out parameter interdependence, resulting from non-ideal conditions, in the form of the C-function. This mapping is achieved within a single characterization trial taking only a few minutes to complete, thus avoiding the need for extensive iterative trials, as has previously been the case. This approach has been made possible through the novel use of an ultrasonic time-of-flight system to allow the spatial measurement of erosion parameters. Example C-functions are generated for the case of an Inconel 718/sodium nitrate system. This methodology is shown to apply to the characterization of the machining of both planar and two-dimensional double-cosine profiles under non-ideal conditions. This is an important step towards the implementation of a fully automated process specification procedure for ECM.
Prominence of γδ T Cells in the Elicitation Phase of Dinitrochlorobenzene-induced Contact Hypersensitivity in Lambs
Authors: Jörundsson E. , Press C. M. , Ulvund M. , Landsverk T.
Published: April 2003

Immunohistochemical techniques were used to identify T-cell subpopulations in the ear skin of lambs during the elicitation phase of dinitrochlorobenzene (DCNB)-induced contact hypersensitivity. Thirty lambs (21-26 weeks of age) were divided into groups of 10. The shaved right ear of one group was treated with DNCB. Two weeks later, this group was challenged with DNCB. One group was treated with the vehicle alone, and the remaining group was left untreated. The lambs were killed 48 hours after challenge, and tissue specimens were collected from the ears of the three groups. There was an increase in T-cell populations in the skin of the DNCB-treated lambs 48 hours after challenge. The majority of the T cells were CD8+ and associated predominantly with the blood vessels and adnexa of the superficial dermis. There was also an increased presence of CD4+ cells and γδ T cells in the superficial dermis. In the epidermis, clusters of γδ T cells and CD4+ cells were associated with microlesions. Computer-assisted morphometric analysis was used to estimate the relative presence of the T-cell subpopulations in the superficial and deep dermis and the entire dermis. Statistical analysis of the relative area of immunostaining showed that the significant increases in all T-cell subpopulations (CD4+, CD8+, and γδ T cells) in the entire dermis were accounted for by changes in the superficial dermis. The prominence of γδ T cells in delayed-type hypersensitivity in the skin of domestic ruminants has been the subject of conflicting reports. In the present study, CD4+ cell and γδ T-cell populations were of similar size in the normal and DNCB-treated lambs, suggesting an equal participation in the elicitation phase of contact hypersensitivity.
Generalized Dermal Dysplasia and Perifollicular Mucinosis in a Dog
Authors: Uetsuka K. , Suzuki M. , Suehiro M. , Ejiri N. , Nishimura R. , Sasaki N. , Nakayama H. , Doi K.
Published: April 2003

Multiple nodules 0.5-2 cm in diameter were observed in the dorsal skin of a 1.5-year-old male castrated mongrel dog. Histopathologically, perifollicular mucinosis and clusters of small cells with a dense, round nucleus and scant cytoplasm were seen. The small cell clusters were present in the dermis near the epidermal basal layer or surrounding hair follicles. These cells demonstrated different staining patterns for vimentin and keratin depending on their location; ultrastructurally, they were poor in organelles and connected to each other with cytoplasmic projections. These findings suggested that they were immature epithelial cells. The case was diagnosed as generalized dermal dysplasia and perifollicular mucinosis.
In Situ Hybridization for the Detection and Localization of Swine Chlamydia trachomatis
Authors: Chae C. , Cheon D.-S. , Kwon D. , Kim O. , Kim B. , Suh J. , Rogers D. G. , Everett K. D. E. , Andersen A. A.
Published: April 2003

Gnotobiotic piglets were inoculated intralaryngeally with swine Chlamydia trachomatis strain R33 or orally with swine C. trachmatis strain R27. Archived formalin-fixed, paraffin-embedded tissues from piglets euthanatized 4-7 days postinoculation were examined by in situ hybridization for C. trachomatis nucleic acid using a nonradioactive digoxigenin-labeled DNA probes that targeted specific ribosomal RNA or ompl mRNA molecules of the swine C. trachomatis strains. Positive hybridization signals were detected in bronchial epithelial cells, bronchiolar epithelial cells, pneumocytes, alveolar and interstitial macrophages, and jejunal and ileal enterocytes. Chlamydia-infected cells had a strong signal that was confined to the intracytoplasmic inclusions. Positive hybridization signals were not detected in tissue sections from an uninfected control piglet or in C. psittaci-infected sheep placenta. The morphology of host cells was preserved despite the relatively high temperature required in parts of the incubation procedure. The data indicate that in situ hybridization can be used to detect swine C. trachomatis in formalin-fixed, paraffin-embedded tissue specimens.
Age-related Changes in the Brain of the Dog
Authors: Borràs D. , Ferrer I. , Pumarola M.
Published: April 2003

Although many age-related changes have been described in the nervous system of different species, few authors have specifically studied the topic. Knowledge of such changes is essential to veterinary pathologists, who must distinguish the lesions of specific pathologic processes from those arising as a result of normal aging. The brains of 20 old dogs, ranging in age from 8 to 18 years, were compared with those of 10 young dogs using routine staining techniques (hematoxilin and eosin, periodic acid-Schiff), special staining techniques (periodic acid-methenamine silver stain), and immunohistochemical techniques to detect glial fibrillary acid protein, neurofilaments, ubiquitin, and β-amyloid. Changes affected meninges and choroid plexuses, meningeal and parenchymal vessels, neurons, and glial cells. Of special interest was the presence of polyglucosan bodies, cerebrovascular amyloid deposition, senile plaques, and ubiquitinated bodies. Some of the age-related changes found, particularly lipofuscin, polyglucosan bodies, and β-amyloid protein deposition, may play a role in the pathogenesis of the canine cognitive dysfunction syndrome. The dog could be used as a natural animal model for the study of normal aging and human neurodegenerative diseases.
Actinobacillus pleuropneumoniae Osteomyelitis in Pigs Demonstrated by Fluorescent In Situ Hybridization
Authors: Jensen T. K. , Boye M. , Hagedorn-Olsen T. , Riising H. J. , Angen Ø.
Published: April 2003

Necrotizing osteomyelitis and fibrinopuralent arthritis with isolation of Actinobacillus pleuropneumoniae serotype 2 is reported in two pigs from a herd with lameness and mild coughing problems among 8 to 12-week-old pigs. Application of fluorescent in situ hybridization targeting 16S ribosomal RNA of A. pleuropneumoniae in formalin-fixed tissue was performed to verify the association of A. pleuropneumoniae with the bone and joint lesions. By in situ hybridization A. pleuropneumoniae was demonstrated as multiple microcolonies or single cells dispersed in focal fibrinonecrotizing pleuropneumonia, in joints with arthritis, and in bone necroses including lysis of growth plate and suppurative inflammation in the adjacent trabecular met-aphysis, thus demonstrating that well-known infections manifest new, unusual lesions.