bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthetic peptides derived from the β2−β3 loop of Raphanus sativus antifungal protein 2 that mimic the active site
Authors: Schaaper W. M. M. , Posthuma G.A. , Meloen R.H. , Plasman H.H. , Sijtsma L. , Van Amerongen A. , Fant F. , Borremans F. A. M. , Thevissen K. , Broekaert W.F.
Published: March 2003

Abstract: Rs‐AFPs are antifungal proteins, isolated from radish (Raphanus sativus) seed or leaves, which consist of 50 or 51 amino acids and belong to the plant defensin family of proteins. Four highly homologous Rs‐AFPs have been isolated (Rs‐AFP1–4). The structure of Rs‐AFP1 consists of three β‐strands and an α‐helix, and is stabilized by four cystine bridges. Small peptides deduced from the native sequence, still having biological activity, are not only important tools to study structure−function relationships, but may also constitute a commercially interesting target. In an earlier study, we showed that the antifungal activity of Rs‐AFP2 is concentrated mainly in the β2−β3 loop. In this study, we synthesized linear 19‐mer peptides, spanning the entire β2−β3 loop, that were found to be almost as potent as Rs‐AFP2. Cysteines, highly conserved in the native protein, are essential for maintaining the secondary structure of the protein. Surprisingly, in the 19‐mer loop peptides, cysteines can be replaced by α‐aminobutyric acid, which even improves the antifungal potency of the peptides. Analogous cyclic 19‐mer peptides, forced to adopt a hairpin structure by the introduction of one or two non‐native disulfide bridges, were also found to possess high antifungal activity. The synthetic 19‐mer peptides, like Rs‐AFP2 itself, cause increased Ca2+ influx in pregerminated fungal hyphae.
Amphipathic control of the 310‐/α‐helix equilibrium in synthetic peptides
Authors: Hammarström L. G. J. , Gauthier T.J. , Hammer R.P. , McLaughlin M.L.
Published: March 2003

Abstract: A series of short, amphipathic peptides incorporating 80% Cα,Cα‐disubstituted glycines has been prepared to investigate amphipathicity as a helix‐stabilizing effect. The peptides were designed to adopt 310‐ or α‐helices based on amphipathic design of the primary sequence. Characterization by circular dichroism spectroscopy in various media (1 : 1 acetonitrile/water; 9 : 1 acetonitrile/water; 9 : 1 acetonitrile/TFE; 25 mm SDS micelles in water) indicates that the peptides selectively adopt their designed conformation in micellar environments. We speculate that steric effects from ith and ith + 3 residues interactions may destabilize the 310‐helix in peptides containing amino acids with large side‐chains, as with 1‐aminocyclohexane‐1‐carboxylic acid (Ac6c). This problem may be overcome by alternating large and small amino acids in the ith and ith + 3 residues, which are staggered in the 310‐helix.
Structural and biosensor analyses of a synthetic biotinylated peptide probe for the isolation of adenomatous polyposis coli tumor suppressor protein complexes
Authors: Wade J.D. , Catimel B. , Faux M.C. , Burgess A.W. , Nice E. , Otvos L.
Published: March 2003

Abstract: Large numbers of colon tumors stem from mutations in the gene coding for the production of the adenomatous polyposis coli (APC) tumor suppressor protein. This protein contains a coiled‐coil N‐terminal domain that is known to be responsible for homodimerization. Previous work by others has led to the design of a specific 54‐residue anti‐APC peptide (anti‐APCp1) that dimerizes preferentially with this domain. We have undertaken the chemical synthesis of a modified form of this peptide (anti‐APCp2) that bears a biotin moiety at its N‐terminus for use in subsequent ligand‐binding analysis studies. The peptide was subjected to comprehensive chemical characterization to confirm its purity. Secondary structural analysis by circular dichroism spectroscopy and Fourier transform infrared spectroscopy indicated that the peptide could assume a wide range of potential conformations, depending upon the precise microenvironment. Significantly, a stable α‐helical structure was generated when the solvent conditions supported intramolecular salt‐bridge formation along the helix barrel. The biotinylated anti‐APCp2 was immobilized onto a streptavidin sensor surface, in a specific orientation leaving all amino acids available to form a coiled structure. In one experiment, injection of colonic cell lysate extracts (LIM1215) onto a size‐exclusion column resulted in the isolation of a high molecular mass protein peak (> 600 kDa) that reacted specifically with the immobilized anti‐APCp2 on the biosensor surface. In another experiment, a high molecular mass protein (Mr > 250 kDa on SDS−PAGE) could be specifically immunoprecipitated from this peak using either the anti‐APCp2 peptide or an anti‐APC polyclonal antibody. This demonstrates the specific interaction between the anti‐APCp2 peptide and native APC and highlights the potential use of the former peptide in a multidimensional micropreparative chromatographic/biosensor/proteomic protocol for the purification of APC alone and APC complexed with different biopolymers in various cell lines, and stages of tumor development.
A hydrophilic residue at position 2 can improve specific biological responses in fMLP‐OMe analogs
Authors: Cavicchioni G. , Spisani S.
Published: March 2003

Abstract: The peptides for‐Met‐Ser‐Phe‐OMe 1, for‐Met‐Cys‐Phe‐OMe 2, for‐Met‐Lys‐Phe‐OMe 3, and for‐Met‐Tyr‐Phe‐OMe 4 were synthesized in order to investigate the importance of a hydrophilic side‐chain on the residue at position 2 on biological activities of human neutrophils. Our results seem to highlight that this type of substitution does not facilitate good chemotaxis, although it elicits both superoxide anion production and particularly lysozyme release, in some cases even more potent than the parent fMLP‐OMe, if the hydrophilicity is associated with steric hindrance.
Relative hydrophobicity of amino acid residues in homooligopeptides as measured by aqueous two‐phase partitioning
Authors: Gulyaeva N. , Zaslavsky A. , Chait A. , Zaslavsky B.
Published: March 2003

Abstract: Partitioning of 17 amino acids and their homooligopeptides of different lengths in an aqueous dextran–PEG two‐phase system containing 0.15 m NaCl in 0.01 M sodium phosphate buffer, pH 7.4 and 0.11 m sodium phosphate buffer, pH 7.4 was examined. The relative hydrophobicity of the amino acid residues was estimated and expressed in equivalent numbers of methylene units. Analysis of the data shows that the additivity principle does hold for the hydrophobicity of homooligopeptides. The relative hydrophobicity of essentially all amino acid residues is noticeably affected by the ionic composition of aqueous media.
NMR structure of a minimum activity domain of human parathyroid peptide hormone: structural origin of receptor activation
Authors: Jung J. , Lee W. , Lim S.‐K. , Kim Y.
Published: March 2003

Abstract: Parathyroid hormone (PTH) which increases osteoblast numbers and bone formation by activating bone‐lining cells to osteoblasts plays an important role in calcium and phosphate homeostasis and bone remodeling by activating PTH receptors. To determine the structural origin of a minimum activity domain of hPTH, we initiated a detailed structural determination of the hPTHH14 in aqueous solution using NMR spectroscopy. Circular dichroism and NMR data demonstrated that hPTHH14 maintains a typical helical conformation in both membrane‐mimicking environments and 30% TFE solution. The solution structure clearly showed that the residues from Ser3 to Leu11 of hPTHH14 formed a stable helical structure, especially having charged side‐chains oriented in opposite directions relative to one another for optimum interaction with the receptor molecule.
Structural characterization of peptide fragments from hCD81–LEL
Authors: Dhanasekaran M. , Baures P. W. , Prakash O. , VanCompernolle S. , Todd S.
Published: March 2003

Abstract: The solution conformation of two peptides [1: PSGSNIISNLFKED; 2: GSSTLTALTTSVLKNNL] from human CD81 (hCD81) large extra‐cellular loop (LEL) with known importance in the hepatitis C virus glycoprotein E2 (HCV‐E2) binding interaction was characterized using circular dichroism spectroscopy. In addition, the solution structure of peptide 1 that contains a phenylalanine residue (F186 in hCD81) known to be critical in the binding interaction with HCV‐E2 was determined using 1D and 2D 1H NMR spectroscopy. Both peptides are unstructured in water but begin forming significant helical conformation following the addition of 20% or more trifluoroethanol (v/v), a result consistent with their α‐helical conformation found in the native protein. The CD data recorded as a function of pH and NaCl concentration are consistent with stabilization of the helical structure from electrostatic forces for both peptides. Peptide 1 is able to block the binding interaction of recombinant HCV‐E2 (rHCV‐E2) to hCD81 expressed on Molt‐4 T cells at high concentrations (3.5 mm), a low affinity that we attributed to the random coil structure in water.
Practical assay and molecular mechanism of aggregation inhibitors of β‐amyloid
Authors: Akikusa S. , Watanabe K‐I. , Horikawa E. , Nakamura K. , Kodaka M. , Okuno H. , Konakahara T.
Published: March 2003

Abstract: β‐Amyloid peptide (Aβ) is the main protein component of neuritic plaques in the brain of patients of Alzheimer's disease (AD), and its neurotoxicity would be exposed by the formation of aggregates. The aggregation inhibitors composed of an Aβ recognition element (KLVFF) and a hydrophilic moiety are evaluated by a novel fluorescence assay. These compounds inhibit growth of the model aggregates on the KLVFF immobilized surface. In addition, some compounds also possess disrupting activities of preformed aggregates. These compounds could be a key candidate for therapeutic drugs for AD by their novel molecular mechanisms.
Immunomagnetic recovery of human neutrophil defensins from the human gingival crevice
Authors: McKay M. S. , Olson E. , Hesla M. A. , Panyutich A. , Ganz T. , Perkins S. , Rossomando E. F.
Published: March 2003

The human neutrophilic protein defensins are cationic, antimicrobial, cytotoxic, corticostatic chemotactic, opsonic peptides found in azurophilic granules and constitute about 5% of the total protein in human neutrophils. In the present study, defensins were recovered from the human gingival crevice using paramagnetic microspheres (beads), coated with anti‐defensin antibodies. The bead‐bound defensins were assayed using an enzyme‐linked immunosorbent assay developed in this laboratory. Twenty sites were sampled; defensin was found in 100% of the sites and ranged in amount from 270–2000 ng/site. The large local concentrations of defensins, estimated in the mg/ml range, may have major effects on the microbiology of the gingival crevice.
Membrane locus and pH sensitivity of paraben inhibition of alkali production by oral streptococci
Authors: Ma Y. , Rutherford G. C. , Curran T. M. , Reidmiller J. S. , Marquis R. E.
Published: March 2003

Parabens were found to be potent inhibitors of alkali production from arginine

by oral streptococci such as Streptococcus rattus, Streptococcus sanguis and Streptococcus gordonii. For example, 2 μmol butylparaben per ml completely and irreversibly inhibited arginolysis by intact cells of S. rattus FA‐1 and was lethal for the organism. In contrast, butylparaben was not a very effective inhibitor of ureolysis by intact cells of Streptococcus salivarius 57.I, although it did kill the cells. Butylparaben irreversibly inhibited the cytoplasmic enzymes arginine deiminase, carbamate kinase and urease in permeabilized cells or isolated form. However, inhibition of arginolysis by intact cells appeared to be due primarily to irreversible inhibition of transport systems for arginine uptake, because butylparaben added to intact cells did not reduce levels of arginine deiminase when the cells were subsequently permeabilized after washing. The insensitivity of ureolysis by intact cells to butylparaben can be related to the known high permeability of cell membranes to urea and the cytoplasmic location of urease. The potency of butylparaben as an inhibitior of arginolysis or glycolysis and as a lethal agent was found to be greater at acid pH that at neutral or alkaline pH.
In vitro susceptibility of Candida albicans isolates from apical and marginal periodontitis to common antifungal agents
Authors: Waltimo T. M. T. , Ørstavik D. , Meurman J. H. , Samaranayake L. P. , Haapasalo M. P. P.
Published: March 2003

The susceptibility of a total of 70 Candida albicans strains to five common antifungal agents was determined. Thirty‐five of the strains were isolated from persistent cases of apical periodontitis and 35 from cases of marginal periodontitis. The susceptibility of the strains to amphotericin B, 5‐fluorocytosine and three azoles: fluconazole, miconazole and clotrimazole, was tested. The antifungal agents and yeast inoculums were prepared according to the NCCLS (National Committee for Clinical Laboratory Standards) recommendations. The yeasts were incubated with ten different concentrations of antifungal agents at 35°C for 48 h. Yeast growth was measured spectrophotometrically. All strains from both isolation sources were susceptible to low concentrations of amphotericin B and 5‐fluorocytosine, whereas the susceptibility to the three azoles varied, and three of the strains showed azole cross‐resistance. These findings are in agreement with recent reports of increased azole resistance in Candida species in general and suggest the possibility that the oral cavity may act as a reservoir of resistant yeast isolates in systemic infections.
Serum and salivary antibodies to a mycobacterial 65‐kDa stress protein are elevated in HIV‐positive patients and modified by oral candidiasis
Authors: Coogan M. M. , Challacombe S. J.
Published: March 2003

Serum immunoglobulin G (IgG) and IgA, and salivary IgA antibodies to a mycobacterial stress protein (mSP65) were determined in human immunodeficiency virus (HIV)–positive patients, acquired immunodeficiency syndrome (AIDS) patients and HIV‐negative controls with or without oral candidiasis. Serum IgG antibodies were elevated in patients with HIV infection and AIDS and especially in subjects with candidiasis compared with controls (P<0.02, P<0.005). This was not apparent with serum IgA. In the absence of candidiasis, salivary IgA antibodies were elevated in HIV‐positive patients compared with AIDS (P<0.005) patients and healthy controls (P=0.001). The relative avidity of serum IgG antibodies to mSP65 in controls with candidiasis was lower than healthy controls (P<0.0001). In saliva there was a decrease in the relative avidity of IgA antibodies in AIDS patients with candidiasis compared with HIV patients (P< 0.03). In patients without candidiasis, the relative avidity was higher in HIV patients than healthy controls (P=0.02). The results suggest that HIV infection leads to raised serum and salivary antibodies to heat shock proteins. Concurrent Candida infection may modify both the titer and relative avidity differently for serum and saliva.
Analysis of gene expression in Treponema denticola with differential display polymerase chain reaction
Authors: Tsai J. P. , Shi W.
Published: March 2003

Treponema denticola is an oral spirochete associated with the periodontal diseases. A great deal of the molecular components of T. denticola will be learned soon since its genome sequence project is on the way. One of the most important works after genome sequence is to analyze the function of these genes and their regulation. However, like many other oral pathogens, there are currently a very limited number of molecular and genetic tools available to study gene expression in T. denticola. In this article, we describe a method of adapting differential display polymerase chain reaction (ddPCR) for use in the T. denticola system. To test for effectiveness of this protocol, we used three different temperature conditions, 4°C, 25°C and 42°C, to test for differential gene expression. With various ddPCR conditions, we found a number of genes that were expressed differentially. Some of these differentially expressed genes were cloned and sequenced and found to be homologous with the known temperature‐regulated genes, including HtrA. The study indicates that the ddPCR method can be effectively used in T. denticola for analyzing gene expression under various conditions.
The effect of monoclonal antibody and route of immunization on the humoral immune response against Porphyromonas gingivalis
Authors: Van Tilburg M. L. J. A. , Kozarov E. V. , Progulske‐Fox A. , Brady L. J.
Published: March 2003

Immunomodulation mediated by exogenous antibodies has been proposed as a vaccine strategy to improve immune protection against pathogenic microorganisms and suggested to contribute to protection following passive immunization. To test whether a monoclonal antibody directed against an adhesion epitope of the periodontal pathogen Porphyromonas gingivalis could influence the humoral immune response following mucosal immunization, BALB/c mice were immunized orally or intranasally with P. gingivalis alone or P. gingivalis coated with monoclonal antibody 61BG1.3. Differences in antigenic specificity of anti–P. gingivalis serum immunoglobulin G (IgG) were demonstrated between groups of mice that received monoclonal antibody–coated P. gingivalis versus those that received P. gingivalis alone by either route of immunization. Binding of monoclonal antibody 61BG1.3 to P. gingivalis prior to immunization did not influence the serum IgG subclass distribution. However, minor differences in subclass distribution were observed between the various routes of mucosal immunization. These results support the hypothesis that specific monoclonal antibody bound to a bacterial vaccine can alter the quality of the humoral immune response to that organism.
Amine and tin fluoride inhibition of Streptococcus sanguis adhesion under continuous flow
Authors: Embleton J. V. , Newman H. N. , Wilson M.
Published: March 2003

This study evaluated the ability of topically applied amine fluoride (AmF) and AmF‐tin fluoride to inhibit the adhesion of Streptococcus sanguis within a parallel plate flow cell system. One of three AmF compounds and two tin fluoride preparations significantly reduced the net bacterial adhesion to conditioned glass over a 1‐h period. Tin(IV) fluoride inhibited S. sanguis adhesion to the greatest extent, and this was shown to be dependent on the formation of the conditioning film prior to agent application.
Supragingival and subgingival microbiota of adult patients with Down’s syndrome. Changes after periodontal treatment
Authors: Sakellari D. , Belibasakis G. , Chadjipadelis T. , Arapostathis K. , Konstantinidis A.
Published: March 2003

In this longitudinal study, five adult Down’s syndrome patients with periodontitis were placed on a frequent recall visit schedule (every 6 weeks) after treatment, in order to investigate: 1) the microbiological status, both supragingivally and subgingivally, and the changes that occurred after treatment and 2) the effect of frequent professional supragingival plaque control on the subgingival microbiota and clinical variables in these patients. The clinical variables recorded were probing pocket depth, probing attachment level, bleeding on probing and presence of plaque (full mouth, six surfaces per tooth). Microbiological examination was performed separately for supragingival and subgingival samples from the same site for 14 species, using whole genomic DNA probes and the “checkerboard” DNA‐DNA hybridization technique. The findings indicate that, although a reduction of periodontal indices was noticed, plaque levels remained high (60%) even at the end of the experimental period. Periodontal pathogens including Porphyromonas gingivalis, Bacteroides forsythus and Actinobacillus actinomycetemcomitans were frequently detected both supragingivally and subgingivally (>30%). The presence of a species supragingivally and the presence at the same time points subgingivally were correlated. This finding suggested that supragingival plaque acts as a reservoir for reinfection of treated sites. A reduction of the percentages of detection of these species was noticed 1 month after an oral hygiene period as well as at 3 and 6 months after treatment. Inadequate oral hygiene as performed by these patients probably affected supragingival, and consequently subgingival, plaque composition.
Molecular mediators of Porphyromonas gingivalis‐induced T‐cell apoptosis
Authors: Harris J. I. , Russell R. R. B. , Curtis M. A. , Aduse‐Opoku J. , Taylor J. J.
Published: March 2003

Porphyromonas gingivalis produces virulence factors which can modify the molecular and cellular components of the host immune response. In the present work we investigated the role of specific virulence factors from P. gingivalis in the induction of apoptosis in Jurkat T cells. P. gingivalis culture supernatants mimicked the effect of butyric acid on T‐cell apoptosis and this effect was associated with an increase in histone H4 acetylation. A role for proteases was excluded in experiments which demonstrated that neither protease inhibitors nor use of P. gingivalis mutants defective in protease synthesis had any effect on the stimulation of T‐cell apoptosis in this system.
gbpC and pac gene mutations detected in Streptococcus mutans strain GS‐5
Authors: Sato Y. , Okamoto K. , Kizaki H.
Published: March 2003

The Streptococcus mutans gbpC gene encoding cell wall‐anchoring glucan‐binding protein C is involved in the dextran(alpha‐1,6 glucan)‐dependent aggregation (ddag) of this organism. Unlike cells of other strains of S. mutans, strain GS‐5 cells did not exhibit dextran(alpha‐1,6 glucan)‐dependent aggregation under any conditions. We therefore hypothesized that the gbpC gene may be mutated in strain GS‐5. Sequencing analysis of the 1752‐nucleotide GS‐5 gbpC gene revealed a point mutation that switched codon 65 to a TAA termination codon. Strain GS‐5 was previously reported also to have a mutation in the pac gene encoding the cell wall‐anchored major protein antigen. The laboratory‐maintained strain GS‐5 is regarded as having lower cariogenicity than the original isolate. The decreased cariogenicity developed during the laboratory culture of strain GS‐5 may have been caused by mutations in an environment lacking appropriate selective pressures.
Effects of tobacco smoke on the secretion of interleukin‐1β, tumor necrosis factor‐α, and transforming growth factor‐β from peripheral blood mononuclear cells
Authors: Ryder M. I. , Saghizadeh M. , Ding Y. , Nguyen N. , Soskolne A.
Published: March 2003

Alterations of the host response caused by short‐term exposure to high levels of smoke during the act of smoking (acute smoke exposure) as well as long‐term exposure to lower levels of tobacco substances in the bloodstream of smokers (chronic smoke exposure) may play a role in the pathogenesis of periodontal diseases in smokers. In this study, we examined the secretion of three cytokines [interleukin (IL)‐1β, tumor necrosis factor (TNF)‐α, and transforming growth factor (TGF)‐β] from mononuclear blood cells from current smokers and non‐smokers exposed to in vitro tobacco smoke (which may be comparable to in vivo acute smoke exposure) and mononuclear blood cells from current smokers not exposed to further in vitro smoke (which may be comparable to chronic smoke exposure). Peripheral blood mononuclear cells were isolated from eight healthy current smokers and eight healthy non‐smokers, plated in culture wells, exposed in vitro for 1–5 min to cigarette smoke in a smoke box system or not exposed (baseline controls), and then incubated without further smoke exposure for another 24 h. Supernatants from each well were then collected and assayed for the concentrations of the three cytokines by enzyme‐linked immunosorbent assay (ELISA). At baseline, mean IL‐1β levels were higher in smokers than in non‐smokers (mean: 10.6 vs. 5.9 pg/ml, anova: P < 0.05). In both smokers and non‐smokers, secreted levels of IL‐1β increased from 0 to 5 min of in vitro smoke exposure (mean: 5.9–9.9 pg/ml, t‐test: P < 0.05 for non‐smokers only) with levels in smokers higher than in non‐smokers (P > 0.05). Mean TNF‐α levels increased from 0 to 2 min of smoke exposure and decreased from 2 to 5 min in smokers and non‐smokers, with higher levels in non‐smokers than smokers at all time‐points (P > 0.05). Mean TGF‐β levels were higher in smokers than in non‐smokers at all time‐points (mean: 180.5 vs. 132.0 pg/ml, P < 0.05 at 5 min only) with no significant alteration of the pattern of secretion with cigarette smoke exposure. These observed alterations in the secretion of cytokines from mononuclear blood cells in smokers, relative to non‐smokers, and with in vitro smoke exposure may play a role in the pathogenesis of periodontal diseases in smokers.
The relative importance of socio‐economic status, parental smoking and air pollution (SO2) on asthma symptoms, spirometry and bronchodilator response in 11‐year‐old children
Authors: García‐Marcos Luis , Guillén José J. , Dinwiddie Robert , Guillén Agustín , Barbero Paloma
Published: March 2003

The aim of this study was to evaluate the relative contribution of several risk factors to the prevalence of allergic respiratory symptoms, and the positivity of the bronchodilator test with fenoterol, and to establish the relative importance of these factors on the variability of FVC, FEV1, PEF, MEF25, MEF50 and MEF75. A total of 340 11‐year‐old children attending school in polluted and non‐polluted areas of the city of Cartagena, Spain, were studied. The polluted area had had an annual mean of 75 μg/m3 of SO2 over the last 10 years and the non‐polluted area had < 20 μg/m3 during this period. A questionnaire about allergic respiratory symptoms was completed by the parents. Specific questions about parental smoking habits and socio‐economic level were included. Each child’s performance in spirometry before and after administration of 0.2 mg of inhaled fenoterol was evaluated. The only significant predictive variables in the logistic regression (for suffering any symptom or a positive bronchodilator response) were male sex for nasal symptoms (RR 1.37; p = 0.04) and housing near heavy traffic for eye symptoms (RR 1.45; p = 0.01). Living in the polluted area reduced the risk of a positive bronchodilator response (RR 0.61; p = 0.004). Maternal smoking, even though not statistically significant, tended to increased the risk of suffering any symptom (RR 1.26; p = 0.07) or of having a positive bronchodilator response (RR 1.23; p = 0.1). None of the risk factors studied was of significant importance in explaining the variability of spirometry results. Although none of the risk factors were specifically determinant to the symptom questions, bronchodilator test or spirometric measurements, having a mother who smokes seems more important than living in a polluted area if statistically non‐significant trends are considered.
Treatment preferences of adolescent patients with asthma
Authors: Weinberg Eugene G. , Naya Ian
Published: March 2003

The study objective was to assess whether asthmatic adolescents who were regular users of inhaled corticosteroids preferred treatment with zafirlukast tablets or inhaled beclomethasone dipropionate (BDP), and, secondarily, to assess adolescents’ inhaler technique and their opinions about treatment. An open‐label, randomized, two‐period, cross‐over study was conducted in 18 centres (primary care to specialist asthma centres) in South Africa, the UK, Finland and the Czech Republic. One hundred and thirty‐two adolescents aged 12–17 years with asthma for at least 1 year and FEV1 ≥ 75% of predicted, treated with short‐acting bronchodilators and inhaled corticosteroids, entered the study. Patients received oral zafirlukast tablets (Accolate™) 20 mg bd or inhaled BDP 100 or 200 µg bd, provided by a standard pressurized metered‐dose inhaler, for 4 weeks each. One questionnaire was used to determine preference (the primary outcome measure) and a second questionnaire was used to determine patients’ likes and dislikes of treatment. Investigators also scored inhaler technique. Of 113 adolescents, 79 (70%) preferred zafirlukast compared with 31 (27%) who preferred the BDP inhaler (p < 0.001); three had no preference. Only 35 (29%) of 122 adolescents could use their inhaler correctly at study entry. Seventy‐six patients (65%) rated zafirlukast tablets as ‘very easy’ to use, compared with 35 (30%) for the BDP inhaler. Both treatments were well tolerated. This study shows that asthmatic adolescents prefer zafirlukast tablets by a ratio of 2.6:1 over inhaled BDP, and these results may have implications for improving adolescent patient compliance with asthma therapy.
Pre‐natal sensitization in humans
Authors: Warner J.O. , Jones C.A. , Kilburn S.A. , Vance G.H.S. , Warner J.A.
Published: March 2003

Allergy has a very strong hereditary component but even in identical twins, concordance for the development of allergic disease can be as low as 50%. This suggests that there is a very strong environmental influence on manifestations of sensitization. To what extent environment might have an influence on the ontogeny of sensitization antenatally has hitherto not been a focus of much research. However, circumstantial evidence suggests that this may be important.
Interleukin‐15 enhances CD4+ CD45RA+ expression on umbilical cord blood mononuclear cells
Authors: Lin Syh‐Jae , Chao Hsun‐Chin
Published: March 2003

The reduced incidence of graft‐vs.‐host disease following umbilical cord blood (CB) transplantation may be related to the functional immaturity of newborn T cells expressing mainly the naive CD45RA phenotype. Expansion of CD4+ CD45RA+ T cells using cytokines may benefit neonates and infants with human immunodeficiency virus (HIV) infection, as a preferential decline in CD4+ CD45RA+ cells has been noted as HIV disease progresses. The aim of the study was to investigate the effect of interleukin (IL)‐15, a novel cytokine similar to IL‐2 in biological activities, on CD45RA/RO expression and apoptosis in umbilical cord blood (CB) and adult peripheral blood (APB) mononuclear cells (MNCs). Prior to culture, CB MNCs contained a greater number of CD4+ CD45RA+ cells and fewer CD4+ CD45RO+ cells than did APB MNCs. When incubated with RPMI‐1640 containing 10% fetal calf serum for 7 days, the percentage of CD45RA+ cells within CD4+ T cells (%CD45RA+/CD4+) significantly decreased compared to that of fresh CB MNCs. IL‐15 exerted a dose‐dependent increase of %CD45RA+/CD4+ and a corresponding decrease of %CD45RO+/CD4+ in CB MNCs, an effect not observed with APB MNCs treated with IL‐15. The percentages of CD45RA+ and CD45RO+ expression within CD8+ cells, however, were not influenced by IL‐15, in either CB or APB MNCs. A greater number of CB MNCs underwent apoptosis than did APB MNCs after 7 days of culture in RPMI‐1640 containing 10% fetal calf serum. IL‐15 did not inhibit apoptosis but induced proliferation comparable to that achieved in APB MNCs. The ability of IL‐15 to preferentially enhance the proliferation of CD4+ CD45RA+ cells in CB MNCs suggests a role for immunomodulative therapy in HIV‐infected newborns and infants.
Allergic rhinoconjunctivitis, eczema, and sensitization in two areas with differing climates
Authors: Hesselmar Bill , Åberg Birgitta , Eriksson Bo , Åberg Nils
Published: March 2003

In this 5‐year follow‐up study we compared the prevalence of allergic rhinoconjunctivitis, eczema, and sensitization, in relation to several background factors, in two Swedish regions (Göteborg and Kiruna). In Göteborg, a city on the southwest coast, the climate is mild and humid. Kiruna is a town north of the Arctic Circle. Questionnaire replies and results of interviews were collected from all 412 7–8‐year‐old children of a population‐based sample (203 in Göteborg and 209 in Kiruna); in addition, 192 children from Göteborg and 205 from Kiruna were skin‐prick tested for sensitization to common aero‐allergens. After 5 years, at 12–13 years of age, almost all of the initial study cohort were re‐investigated. At follow‐up the prevalence of allergic rhinoconjunctivitis was 17%, eczema 23%, and sensitization 32%. Allergic rhinoconjunctivitis and eczema were as common in Göteborg as in Kiruna, whereas sensitization was far more common in Kiruna. Children born during the pollen season had allergic rhinoconjunctivitis less often – and were sensitized to pollen and animal protein less often – than those born during the rest of the year. Sensitization to birch pollen, cat protein, and horse protein was less common in children living in Göteborg, the region with the highest frequency of cat ownership and horseback riding, and with the longest birch‐pollen season. The girls were more commonly horseback riders but the boys were more often sensitized to horses. The results reinforce our previous findings: indoor climate may affect the development of sensitization and allergic diseases, to some extent independently; and if exposure to antigen is unavoidable, high doses might be better than low doses.
Early life factors related to clinical manifestations of atopic disease but not to skin‐prick test positivity in young children
Authors: Stazi Maria‐Antonietta , Sampogna Francesca , Montagano GIuseppe , Grandolfo Michele E. , Couilliot Marie‐France , Annesi‐Maesano Isabella
Published: March 2003

The relative influence of early life events in the development of IgE‐mediated allergy is still undetermined. We investigated early life factors in relation to skin‐prick test positivity (SPT) and clinical manifestations of atopic disease in a population‐based sample of 201 Italian children (3 months−5 years), after considering their interactions with known determinants of allergy. Among them, 143 children had SPT performed to common allergens. Threatened abortions, general anesthesia at delivery, prematurity, birthweight < 2500 g, maternal smoking, dampness and gas heating exposure were all significantly related to an increased risk of frequent rhinitis in the absence of cold (18%). In utero smoking, threatened abortions, fetal health complications, infantile colic, maternal smoking in childhood (satisfactorily correlated with maternal expired CO during the survey) and respiratory infections were all independent determinants of frequent wheezing (23%). Doctor's diagnosis of asthma (3%) was related to in utero smoking, being born in spring, infantile colic and respiratory infections. A simultaneous exposure to in utero smoking and infantile colic put the infants to a fourfold higher risk of frequent wheezing and to a ninefold risk of asthma, respectively. Having a pet and washing blankets at < 60°C were inversely related to frequent wheezing. Data confirmed also that maternal phenotype influences the inheritance of atopic disease. No event, except a low intake of fruit (< 3/week), was significantly associated with positive SPT (20%) or eczema. Besides allergic sensitization, other events, which occur early in life, seem critical to the development of IgE‐mediated allergy.
Glucocorticoids enhance interleukin‐4 production to neo‐antigen (hyaluronidase) in children immunocompromised with cytostatic drugs
Authors: Edelbauer Monika , Gerstmayr Marianne , Loibichler Christine , Jost Eva , Huemer Martina , Urbanek Radvan , Szépfalusi Zsolt
Published: March 2003

Immunoglobulin E (IgE)‐mediated immediate‐type allergic reactions to hyaluronidase have been observed in children with central nervous system (CNS) tumors. Glucocorticoids, used as therapy for brain edema, are discussed controversially as T helper 2 (Th2) stimulatory factors. In this study we investigated the role of glucocorticoids on a Th2 cytokine‐promoting effect in children with CNS tumors. Peripheral blood mononuclear cells (PBMCs) from: 29 children suffering from malignant brain tumors, of whom 23 received short‐term glucocorticoid treatment (for 3–4 days) during the course of chemotherapy; 18 children with nephrotic syndrome or renal transplantation receiving long‐term glucocorticoid treatment; and 13 healthy children, were incubated with phytohemagglutinin (PHA) and/or anti‐CD28 monoclonal antibody (mAb) and, in a second approach, with hyaluronidase. The concentrations of Th cell‐mediated cytokines – interleukin (IL)‐4, IL‐10, and interferon‐γ (IFN‐γ) – were measured in supernatants. The IL‐4 production of PBMCs incubated with PHA/anti‐CD28 mAb from children with repeated co‐administration of glucocorticoids, hyaluronidase, and cytostatic drugs (median: 249.9 pg/ml; range: 234.4–261.7) was significantly higher (p < 0.0001) than IL‐4 production of PBMC from children of all the other groups (median: 86.18; range: 16.0–212.5). There was no significant difference in the levels of IL‐10 and IFN‐γ within the groups. PBMCs stimulated only with hyaluronidase failed to produce detectable levels of cytokines. The results of this study indicate that repeated co‐administration of glucocorticoids and hyaluronidase (a neo‐antigen) enhance IL‐4 production in vitro and thus may induce the production of specific IgE antibodies in children immunocompromised with cytostatic drugs. Hyaluronidase itself does not stimulate in vitro IL‐4 synthesis in PBMCs of children receiving cytostatic drugs.
What is the role for mycophenolate mofetil in pediatric renal transplantation?
Authors: Neu A. M. , Benfield M.
Published: March 2003

Abstract:Mycophenolate mofetil (MMF) has gained considerable popularity in pediatric renal transplantation. This popularity is largely a result of data from three large trials of MMF in adult cadaveric transplant patients who demonstrated a decreased rate of acute rejection episodes when treated with cyclosporin A (CsA), prednisone, and MMF compared with those receiving CsA, prednisone, and azathioprine (AZA) or placebo. However, the ability of MMF to reduce acute rejection appears to be limited to the first month post‐transplant, and its effectiveness with microemulsion CsA or tacrolimus‐based regimens has not been proven. In addition, there are currently no data that convincingly demonstrate that this agent improves graft survival, patient survival, graft function or protects against chronic rejection. Finally, there may be an increased risk for severe cytomegalovirus (CMV) disease and lymphoproliferative disorder with central nervous system involvement in patients treated with MMF. These data call into question the role of MMF in current immunosuppressive regimens.
Acute lymphoblastic leukaemia: Whom and when should we transplant?
Authors: Eden O. B.
Published: March 2003

Abstract: To date, the evidence upon which a decision is made to transplant a child with acute lymphoblastic leukaemia either in first or subsequent remission has rarely been based on randomized trial data. Modern era management of infection and graft‐versus‐host disease lessens risks but procedure‐related deaths still remain higher than with chemotherapy alone. However, disease control appears superior with successful bone marrow transplantation (BMT). The critical need is for international consensus on who is at such great risk of recurrent disease that BMT is required (from no mattter what donor source) and conversely those for whom transplantation is not needed to achieve long‐term cure.
The impact of acute rejection on chronic rejection: A report of the North American Pediatric Renal Transplant Cooperative Study
Authors: Tejani A. , Sullivan E. K.
Published: March 2003

Abstract: The North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) group has analyzed its database from January 1987 to October 1998. During this time we enrolled 6395 transplants: of these, 5323 were primary and 1072 were repeat transplants. Overall, 30.8% (483/1566) of the grafts failed as a result of chronic rejection. For living donor (LD) grafts, the failure rate as a result of chronic rejection was 32% (175/553), and it was 30% (308/1013) for cadaveric donor (CD) transplants. A proportional hazards model identified first acute rejection, multiple rejections, and a late acute rejection as risk factors for the development of chronic rejection. Additional risk factors for the development of chronic rejection were African–American race, a repeat transplant, and a cyclosporin A (CsA) dose of < 5 mg/kg/day. Our analysis found that one acute rejection episode increases the risk of chronic rejection graft failure three‐fold. Patients with two or more acute rejections have a 12‐fold increased relative risk (RR) of chronic rejection graft loss (CRGL). A late acute rejection (> 365 days post‐transplant) increases the RR by six‐fold. Two or more acute rejections, when the first is a late initial rejection, increases the RR 26‐fold. Based on this information we have initiated a multicenter trial of intervention in patients with one or more acute rejections.
Impact of acute rejection on development of chronic rejection in pediatric renal transplant recipients
Authors: Matas Arthur J.
Published: March 2003

Abstract: For pediatric kidney transplant recipients, chronic rejection has become the predominant cause of graft loss. This article reviews risk factors for chronic rejection and what can be done to lower the risk of chronic rejection for future transplant recipients.
Modified temporary end‐to‐side portocaval shunt in liver and small bowel transplantation
Authors: Molmenti Ernesto P. , Marsh J. Wallis , Molmenti Hebe , Reyes Jorge , Fung John J.
Published: March 2003

Abstract: Piggyback orthotopic liver transplantation (LTx) has permitted the elimination of extra‐corporeal venovenous bypass. In some instances, an internal temporary portocaval shunt has to be constructed in order to prevent hemodynamic instability. We describe a technique in which a donor iliac vein graft is used to bridge the distance between the portal vein and vena cava in cases where a direct shunt cannot be constructed. This technique can be applied to liver Tx as well as to liver and small bowel Tx.
Heart transplantation in children and infants: Short‐term outcome and long‐term follow‐up
Authors: Bauer J. , Thul J. , Krämer U. , Hagel K. J. , Akintürk H. , Valeske K. , Schindler E. , Bohle R. M. , Schranz D.
Published: March 2003

Abstract: Since 1988, 82 heart transplants have been performed in 80 infants and children. Diagnoses pretransplant were: hypoplastic left heart syndrome (HLHS) (n = 43); cardiomyopathy (n = 19); endocardial fibroelastosis (n = 6); and other complex congenital heart diseases (n = 12). Age at transplantation was < 1 yr in 61 patients. Overall survival rate was 79% at 1 yr and 73% at 5 and 10 yr. To date, 20 patients have died after transplantation. Causes of death were: rejection (eight patients); right ventricular failure (four patients); transplant coronary artery disease (TCAD) (two patients); and other causes (six patients). In the majority of patients somatic growth is not impaired, and renal function is reduced (but stable) in all patients. Two patients developed post‐transplant lymphoproliferative disease, which was treated successfully. Major long‐term morbidity is neurologic deficit – severe in three patients and minor in six. TCAD was present or suspected in six surviving patients. We conclude that heart transplantation in infants and children can be performed with good early and late results. Quality of life is excellent in most patients. TCAD, however, will become an increasing problem in the long term.
Drug‐induced lupus as a cause of relapsing inflammatory disease after renal transplantation
Authors: Pape Lars , Strehlau Juergen , Latta Kay , Ehrich Jochen Hh , Offner Gisela
Published: March 2003

Abstract: We discuss the case of an 18‐year‐old‐boy presenting with relapsing fever, arthralgia, myalgia and renal failure, 7 yr after renal transplantation. A thorough diagnostic work‐up for infectious and inflammatory diseases revealed a mastoiditis and atypical mycobacteria, but symptoms persisted after treatment. Persistent antinuclear antibodies in combination with cardiolipin and myeloperoxidase antibodies, despite negative dsDNA antibodies, suggested a drug induced lupus‐like syndrome. Six months after withdrawal of dihydralazine, all symptoms had disappeared. Drug‐induced lupus should be considered as an important differential diagnosis in transplanted patients with recurrent inflammatory disease in conjunction with lupus‐like symptoms and negative dsDNA antibodies. It may prevent a potentially hazardous reduction of immunosuppression in persistent inflammation.
Treatment of leukemia relapse with recombinant granulocyte‐macrophage colony stimulating factor (rhGM‐CSF) following unrelated umbilical cord blood transplant: Induction of graft‐vs.‐leukemia
Authors: Worth Laura L. , Mullen Craig A. , Choroszy Mary , Koontz Susannah , Chan KaWah
Published: March 2003

Abstract: An infant with congenital leukemia in complete remission (CR1) received an unrelated donor umbilical cord blood cell transplant from a one‐HLA disparate donor. The conditioning regimen consisted of thiotepa, busulfan and cyclophosphamide. GVHD prophylaxis consisted of tacrolimus and mini‐methotrexate. Engraftment occurred and a bone marrow aspirate obtained on day 28 showed 100% donor cells. The post‐transplant course was complicated by skin and liver GVHD, grade III, that responded to therapy with methylprednisolone, anti‐thymocyte globulin and daclizumab (Zenapax), in addition to tacrolimus. A bone marrow aspirate obtained on day 187 showed relapse, with 17% blasts. The patient was then treated for 30 days with recombinant human granulocyte‐macrophage colony‐stimulating factor treatment (rhGM‐CSF). A bone marrow aspirate obtained 17 days after the initiation of rhGM‐CSF treatment showed 2% blasts. Ascites was the predominant side‐effect of the rhGM‐CSF treatment. The patient remains in complete remission 24 months after relapse and 30 months after transplantation. This case documents that rhGM‐CSF and withdrawal of immunosuppression can induce a durable complete remission after relapse following an unrelated donor cord blood transplant.
Early heart transplant in a child with advanced lymphoma
Authors: Morgan Elaine , Pahl Elfriede
Published: March 2003

Abstract: A child with advanced stage large cell lymphoma in remission developed overwhelming, irreversible anthracycline cardiac toxicity incompatible with life early in her treatment course. The medical complexities and ethical issues concerning a decision to proceed with orthotopic heart transplantation are discussed.
Megakaryocyte chimerism after allogeneic stem cell transplantation in children
Authors: Eklund Ola , Dalianis Tina , Wester Daniela , Winiarski Jacek
Published: March 2003

The aim of this study was to investigate the extent and the clinical implications of mixed chimerism in megakaryocytes after stem cell transplantation (SCT). Polymerase chain reaction analyzing allele length polymorphisms was used to determine the origin of immunomagnetically isolated megakaryocytes and leukocyte subpopulations after SCT in 13 children. Eleven were unselected consecutive cases while two were included due to known leukocyte mixed chimerism. Recipient DNA was detected in the megakaryocytes in six out of the 11 cases at levels between 1 and 100%. Coinciding mixed chimerism in the leukocyte populations was detected in two of the 11 cases. Of the two selected cases with known leukocyte mixed chimerism, two boys with aplastic anemia and Wiskott–Aldrich syndrome had 1–5 and 70% recipient megakaryocytes, respectively. Although the four relapses or deaths, within the 13 months of observation, were restricted to patients with multilineage or isolated megakaryocyte (n = 1) mixed chimerism, it was not possible to link any other apparent clinical problems, except a prolonged thrombocytopenia in one case, to the mixed chimerism in this limited study group.