bluuebunny/crossref_metadata_2025 huggingface dataset:
Five‐year maintenance follow‐up of early‐onset periodontitis patients
Authors: Kamma Joanna J. , Baehni Pierre C.
Published: June 2003

Abstract
Objectives: The purpose of this study was to evaluate the clinical and microbiological status of patients with early‐onset or aggressive periodontitis (EOP) who had received supportive periodontal care (SPC) every 3–6 months for a period of 5 years, following active periodontal treatment.

Material & Methods: The study population consisted of 25 individuals with early‐onset periodontitis. Clinical examination and recordings of probing pocket depth (PPD) and clinical attachment level (CAL) were performed at baseline prior to treatment (T0), 3 months following the termination of active periodontal treatment (T1) and annually at the SPC appointments (T2,T3,T4,T5). Microbiological samples were obtained at the 5‐year SPC (T5). Subgingival plaque samples for each individual were collected from one deep pocket (>5 mm), based on pretreatment measurements, randomly selected in each quadrant. The levels of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola were determined using oligonucleotide probe hybridization.

Results: During the 5‐year period, the mean of SPC/patient was 12.7 sessions. A significant improvement was observed in PPD, CAL, gingival bleeding index and suppuration following treatment. However, between T1 and T5, 134 sites in 20 patients deteriorated with a CAL loss of≥2 mm. Out of these 134 sites showing disease progression, microbial samples were randomly obtained in 13 sites (9.7%) from 8 patients. Among other factors, smoking and stress were found to have significant predictive value on the future attachment loss. P. gingivalis, T. denticola and total bacterial load were statistically significantly higher in patients who experienced disease progression during the 5‐year maintenance period.

Conclusions: For most EOP patients, regular SPC was effective in maintaining clinical and microbiological improvements attained after active periodontal therapy. However, a small percentage of sites was identified as progressive in 20 patients. Variables found to be related to periodontal progression were the presence of as well as the high bacterial counts of P. gingivalis, T. denticola and total bacterial load, number of acute episodes, number of teeth lost, smoking and stress.
Acquisition and loss of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia over a 5‐year period: effect of a triclosan/copolymer dentifrice
Authors: Cullinan M. P. , Hamlet S. M. , Westerman B. , Palmer J. E. , Faddy M. J. , Seymour G. J.
Published: June 2003

Objectives: The present study describes the natural history of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia over a 5‐year period and the effect of a triclosan/copolymer dentifrice on these organisms in a normal adult population.

Material and Methods: Subgingival plaque samples were collected from 504 adult volunteers. Probing pocket depths (PPD) and relative attachment levels were measured using an automated probe. Participants were matched for disease status (CPI), plaque index, age and gender, and allocated to receive either a triclosan/copolymer or placebo dentifrice. Re‐examination and subgingival plaque sampling was repeated after 1, 2, 3, 4 and 5 years. P. gingivalis, A. actinomycetemcomitans and P. intermedia were detected and quantitated using an enzyme linked immunosorbent assay. Logistic regression and generalised linear modelling were used to analyse the data.

Results: This 5‐year longitudinal study showed considerable volatility in acquisition and loss (below the level of detection) of all three organisms in this population. Relatively few subjects had these organisms on multiple occasions. While P. gingivalis was related to loss of attachment and to PPD ≥3.5 mm, there was no relationship between A. actinomycetemcomitans or P. intermedia and disease progression over the 5 years of the study. Smokers with P. gingivalis had more PPD ≥3.5 mm than smokers without this organism. There was no significant effect of the triclosan dentifrice on P. gingivalis or A. actinomycetemcomitans. Subjects using triclosan were more likely to have P. intermedia than those not using the dentifrice; however this did not translate into these subjects having higher levels of P. intermedia and its presence was uniform showing no signs of increasing over the course of the study.

Conclusion: The present 5‐year longitudinal study has shown the transient nature of colonisation with P. gingivalis, A. actinomycetemcomitans and P. intermedia in a normal adult population. The use of a triclosan‐containing dentifrice did not lead to an overgrowth of these organisms. The clinical effect of the dentifrice would appear to be independent of its antimicrobial properties.
Safety and efficiency of novel sonic scaler tips in vitro
Authors: Petersilka Gregor J. , Draenert Miriam , Mehl Albert , Hickel Reinhard , Flemmig Thomas F.
Published: June 2003

Background: The aim of the study was to evaluate a novel sonic scaler tip for subgingival root surface instrumentation combining high efficiency in calculus removal with minimized risk of root damage through subgingival debridement.

Methods: A metal sonic scaler tip with a paddle‐like working end covered with spheroid convexities of 0.8 mm diameter and 0.3 mm height was designed from the aspect of optimized adaptation to the root anatomy (tip‐end dimension: 3.0×1.5×0.6 mm). Using a customary sonic scaler tip as control, instrument efficiency was quantified by measuring the time needed to completely remove calculus on extracted teeth (n=52) under standardized conditions. To quantify the amount of calculus removed, the mean calculus area per tooth was measured on photographs taken before instrumentation. The tips safety was evaluated instrumenting calculus‐free root surfaces in vitro (n=18, lateral forces 0.5, 1, 2 N, 20 s instrumentation time, tip angulation 0°) with subsequent laser‐optical determination of resulting root substance loss.

Results: Debridement efficiency was significantly higher for the novel tip (0.78±0.81 mm2/s completely debrided) than for the conventional tip (0.42±0.33 mm2) (Mann–Whitney test, p<0.05). Concerning instrument safety, the novel tip caused significantly less root substance loss than the conventional tip.

Conclusion: The novel scaler tip appears to be significantly more efficient in calculus removal and less damaging to the root surface than the assessed conventional tip.
Activation of human matrix metalloproteinase 2 by gingival crevicular fluid and Porphyromonas gingivalis
Authors: Grayson R. , Douglas C.W.I. , Heath J. , Rawlinson A. , Evans G.S.
Published: June 2003

Aim: To assess the potential of gingival crevicular fluid (GCF) from adult periodontitis patients and Porphyromonas gingivalis proteases to activate matrix metalloproteinase 2 (MMP‐2) in vitro.

Material and Methods: GCF samples were collected from each of 15 adult periodontitis patients, from a clinically healthy site, a deep (>6 mm) bleeding site, and a deep nonbleeding site. The GCF samples were examined for general proteolytic activity, gelatinolytic activity and ability to activate pro‐MMP‐2 by zymography. Ultrasonic extracts of a range of clinical isolates of P. gingivalis cells and purified arg‐ and lys‐gingipains were also assessed for their ability to activate pro‐MMP‐2.

Results: GCF from deep nonbleeding sites showed higher general proteolytic activity than samples from deep bleeding and healthy sites but this did not reach statistical significance. Pefabloc, a general serine protease inhibitor, inhibited the majority (92%) of the proteolytic activity. GCF samples contained neutrophil MMP‐9 in its latent form in 93% of the samples, and in its activated form in 40% of the samples. In contrast, MMP‐2 was present in only trace amounts in 9% of the samples. When latent MMP‐2 was added to these GCF samples, it was converted to the activated form (59 kDa) in 68% of the samples. Lower molecular weight (55 and 45 kDa) activated forms also appeared in 53% of the samples, particularly those from deep sites. Activation to the 55 and 45 kDa forms was inhibited by MSAAPket (a neutrophil elastase inhibitor), whereas Pefabloc completely inhibited the activation of latent MMP‐2. All ultrasonic extracts of P. gingivalis activated latent MMP‐2 in a concentration‐ and time‐dependent manner. Also, latent MMP‐2 was activated by purified arg‐gingipain but less efficiently by lys‐gingipain.

Conclusion: These findings suggest that P. gingivalis arg‐gingipain and neutrophil elastase present in GCF can activate latent MMP‐2, which may contribute in vivo to local periodontal tissue destruction.
Analysis of the TGF‐β1 promoter polymorphism (C−509T) in patients with chronic periodontitis
Authors: De Souza A. P. , Trevilatto P. C. , Scarel‐Caminaga R. M. , De Brito R. B. , Line S. R. P.
Published: June 2003

Background: A polymorphism in the promoter region of the transforming growth factor beta‐1 (TGF‐β1) gene was described at position −509. This polymorphism represents a C‐to‐T base exchange, which creates a YY1 consensus sequence in an area involved with down transcription regulation. This polymorphism has been associated with risk for asthma and allergies. In this study we investigated the association between this polymorphism and chronic periodontitis severity.

Methods: Genomic DNA from oral mucosa of 87 Caucasian subjects was amplified by PCR, and digested with Eco81I restriction endonuclease. The alleles were separated by polyacrylamide gel electrophoresis. The differences in genotype distribution from those expected by Hardy–Weinberg equilibrium, and the significance of the differences in observed frequencies of the polymorphism in moderate and severe disease and healthy groups were assessed by the χ2 test.

Results: There was a difference in the presence of the different alleles and genotypes among the healthy, moderate and severe periodontitis groups. The allele T was seen at 57.7% in the group with severe periodontitis and 37.8% and 35.4% in the healthy group and moderate periodontitis group, respectively (p=0.0387). The genotype T/T was found at 38.5% in the group with severe periodontitis, and at a frequency of 8% in the healthy group (p=0.0258).

Conclusion: These results demonstrate that the polymorphism at bp −509 in the TGF‐β1 promoter may have a small effect on the modulation of the inflammatory process during periodontitis.
Post‐transfusion purpura (PTP) and disseminated intravascular coagulation (DIC)
Authors: Welling Karen‐Lise , Taaning Ellen , Lund Birgit V. , Rosenkvist Jørgen , Heslet Lars
Published: June 2003

A 47‐yr‐old female with acute pancreatitis received four units of fresh frozen plasma because of subtle signs of disseminated intravascular coagulation (DIC). Seven days later, she developed severe thrombocytopenia. Serological studies demonstrated antibodies against HPA‐1a together with pan‐reactive antibodies against platelet glycoproteins (GPIIb‐IIIa, GPIb‐IX and GPIa‐IIa), which was consistent with the diagnosis of PTP. The patient was treated with platelet transfusions, corticosteroids and intravenous immunoglobulin (IVIG) without permanent beneficial effect. After treatment with plasma exchange the platelet count increased to normal values.
Variation in resistance and virulence among demes of a plant host–pathogen metapopulation
Authors: Thrall Peter H. , Burdon Jeremy J. , Young Andrew
Published: June 2003

Despite the recognition that spatial structure can have major consequences for the ecological and evolutionary dynamics of host–pathogen interactions, few empirical studies have examined how regional processes affect within‐population co‐evolutionary patterns.

 Considerable variation was detected within and among demes of a metapopulation in the resistance ofLinum marginaleto a range of local isolates of the rustMelampsora lini.Only four resistance phenotypes were common (occurring at an overall frequency of > 5%) and pathogen populations were similarly variable, with five out of 44 virulence phenotypes classified as common.

 Resistance and virulence structure were related to the occurrence of two distinct ecotypes of the host. Hill populations were consistently more resistant than those in bogs and their pathogen populations more virulent. Allozyme analysis showed a high degree of genetic differentiation between host ecotypes, with fixed allelic differences. Host and pathogen phenotypes were non‐randomly distributed between bog and hill sites.

 When ecotypic differences were factored out, there was clear evidence of a non‐random spatial distribution of resistance, with nearby populations being more likely to share resistance phenotypes, suggesting at least weak isolation by distance. While still apparent, this effect was much less noticeable in the pathogen populations, indicating significant differences in the spatial scale of dispersal of host and pathogen.
Ozone‐induced bronchial epithelial cytokine expression differs between healthy and asthmatic subjects
Authors: Bosson J. , Stenfors N. , Bucht A. , Helleday R. , Pourazar J. , Holgate S. T. , Kelly F. J. , Sandström T. , Wilson S. , Frew A. J. , Blomberg A.
Published: June 2003

Background Ozone (O3) is a common air pollutant associated with adverse health effects. Asthmatics have been suggested to be a particularly sensitive group.

Objective This study evaluated whether bronchial epithelial cytokine expression would differ between healthy and allergic asthmatics after ozone exposure, representing an explanatory model for differences in susceptibility.

Methods Healthy and mild allergic asthmatic subjects (using only inhaled β2‐agonists prn) were exposed for 2 h in blinded and randomized sequence to 0.2 ppm of O3 and filtered air. Bronchoscopy with bronchial mucosal biopsies was performed 6 h after exposure. Biopsies were embedded in GMA and stained with mAbs for epithelial expression of IL‐4, IL‐5, IL‐6, IL‐8, IL‐10, TNF‐α, GRO‐α, granulocyte–macrophage colony‐stimulating factor (GM–CSF), fractalkine and ENA‐78.

Results When comparing the two groups at baseline, the asthmatic subjects showed a significantly higher expression of IL‐4 and IL‐5. After O3 exposure the epithelial expression of IL‐5, GM–CSF, ENA‐78 and IL‐8 increased significantly in asthmatics, as compared to healthy subjects.

Conclusion The present study confirms a difference in epithelial cytokine expression between mild atopic asthmatics and healthy controls, as well as a differential epithelial cytokine response to O3. This O3‐induced upregulation of T helper type 2 (Th2)‐related cytokines and neutrophil chemoattractants shown in the asthmatic group may contribute to a subsequent worsening of the airway inflammation, and help to explain their differential sensitivity to O3 pollution episodes.
Early endotoxin exposure and atopy development in infants: results of a birth cohort study
Authors: Bolte G. , Bischof W. , Borte M. , Lehmann I. , Wichmann H.‐E. , Heinrich J. ,   for the LISA Study Group
Published: June 2003

Background Exposure to endotoxin in childhood is currently discussed to protect from the development of allergic diseases.

Objective To study the effect of early endotoxin exposure on incidence of atopic sensitization, atopic dermatitis and wheezing until the age of 2 years in infants with different risk status in terms of parental atopy.

Methods Data of 1942 infants of an ongoing birth cohort study were analysed by logistic regression. Endotoxin was measured in settled dust of the mothers' mattresses at infants' age of 3 months. Data on allergic symptoms and physicians' diagnoses were gathered by questionnaire. Sensitization to common food and inhalant allergens was assessed by specific serum IgE.

Results High endotoxin levels increased the risk of repeated wheeze [adjusted odds ratio (OR) for 4th exposure quartile (Q4) 1.52, 95% confidence interval (CI) 1.08–2.14], but were associated with neither sensitization to food allergens nor atopic dermatitis. Stratification by parental atopy showed that there was an association of endotoxin exposure with incidence of repeated wheeze as well as with sensitization to inhalant allergens (P for trend = 0.008 and 0.044, respectively) only in infants with parental atopy, with the highest risk in the 4th exposure quartile (repeated wheeze: ORQ4 1.77, 95% CI 1.14–2.73; sensitization to inhalant allergens: ORQ4 1.69, 95% CI 0.70–4.11).

Conclusion Early endotoxin exposure in terms of mattress dust endotoxin levels seemed to increase the risk of atopic reactions to inhalant allergens at the age of 2 years, especially in infants at risk due to parental atopy. Our data disagree with an early protective effect of endotoxin on atopy development until the age of 2 years.
Normative data for total serum immunoglobulin E measurements in children of three ethnicities
Authors: Chan E. Y. , McKenzie S. A.
Published: June 2003

Background It is about 20 years since IgE measurements were published for children without atopic disease. It is possible that the recent increase in atopic disease is reflected in altered measurements in subjects who have no clinical expression of atopy. If the measurement of IgE is to be used as a marker for atopy to characterize disease categories, contemporary normative data must be available.

Objective To measure total serum IgE in healthy children of three ethnicities born and living in an inner city environment.

Methods Subjects were aged 1 to ≤ 12 years, of Afro‐Caribbean, Bangladeshi and white British ethnicities, with no personal history of current atopic disease. An extra 1 mL of blood for the measurement of total serum IgE was collected when blood was taken for other purposes or when a surgical procedure was being undertaken.

Results Measurements were taken from 151 boys (median age 5.4 years) and 106 girls (median age 6.0 years), who included 127 Bangladeshis, 58 Afro‐Caribbeans and 72 white British children. Measurements increased with age but were not related to gender or ethnicity. The data were significantly higher than previous measurements by sixfold.

Conclusion These contemporary normative data allow the generation of z scores for total IgE measurements for clinical or epidemiological use.
Relationships of allergic sensitization, total immunoglobulin E and blood eosinophils to asthma severity in children of the EGEA Study
Authors: Siroux V. , Oryszczyn M.‐P. , Paty E. , Kauffmann F. , Pison C. , Vervloet D. , Pin I.
Published: June 2003

Background Although allergy is highly associated with childhood asthma, it is not well known if there is a relationship between the intensity of allergic sensitization and asthma severity.

Objective The objectives of the study were to examine the relationships between several markers of allergy and asthma severity in asthmatic children included in the Epidemiological study on the Genetics and Environment of Asthma, bronchial hyper‐responsiveness and atopy (EGEA).

Methods The population comprised 216 asthmatic children below 16 years of age. Total IgE and blood eosinophil counts were measured and skin prick tests to 11 aeroallergens were performed. The intensity of the allergic sensitization was assessed by the number of positive skin prick tests and by skin weal sizes. Asthma severity was measured with four criteria: a clinical severity score, history of hospitalization for asthma, FEV1% predicted and inhaled steroid use in the last 12 months.

Results Most of the children were sensitized to at least one aeroallergen (88.2%). Atopy was not related to the severity of asthma, except for a tendency for a more severe clinical score in non‐atopic children. The type and intensity of the allergic sensitization were not associated with any criteria of asthma severity. Total IgE was significantly increased in children treated with inhaled corticosteroids and in children ever hospitalized for asthma (P‐values 0.009 and 0.04, respectively). Eosinophil counts were not related to asthma severity.

Conclusion Our results suggest that severe childhood asthma may be related to a high level of total IgE but not to blood eosinophil counts. The lack of positive relationships between both atopy and the intensity of allergic sensitization with asthma severity supports the hypothesis of different risk factors being associated with asthma and with the severity of asthma.
Bronchial responsiveness to adenosine‐5′‐monophosphate and methacholine as predictors for nasal symptoms due to newly introduced allergens. A follow‐up study among laboratory animal workers and bakery apprentices
Authors: De Meer G. , Postma D. S. , Heederik D.
Published: June 2003

Background In asthma patients, bronchial hyper‐responsiveness (BHR) to adenosine‐5′‐monophosphate (AMP) reflects bronchial inflammation more closely than BHR to methacholine. In this follow‐up study we studied bronchial responsiveness to both stimuli as predictors of new‐onset airway symptoms.

Methods We included 118 laboratory animal workers and bakery apprentices with a work experience of maximally 1 year. The baseline survey comprised a questionnaire, skin prick tests (SPTs) to common and work allergens, blood eosinophil counting, and bronchial challenge with methacholine and AMP. At follow‐up, questionnaire and SPTs to work allergens were repeated. Airway symptoms to common allergens and work allergens were defined as nasal symptoms, chest tightness or asthma attack during or after contact with either common or work allergen. Bronchial challenge tests were analysed by BHR at a 15% fall in forced expiratory volume of 1 s (FEV1), and by dose–response‐slope (DRS).

Results Fourteen subjects (12%) developed airway symptoms to work allergens, of whom 12 had nasal symptoms. A positive SPT to work allergens occurred in 64%, and was the strongest predictor of airway symptoms [relative risk (RR) 7.5, 95% confidence interval (CI) 2.0–28.6]. Other predictors were airway symptoms to common allergens (RR 4.3, 95% CI 1.4–12.8), blood hypereosinophilia (RR 4.4, 95% CI 1.2–15.4) and BHR, with a slightly higher risk estimate for AMP than for methacholine (RRAMP 3.7, 95% CI 1.1–12.5 and RRmeth 2.8, 95% CI 1.0–8.5). The difference was more distinct analysing airway responsiveness by DRS, for which AMP predicted symptoms better than methacholine (P < 0.05).

Conclusions Pre‐existent bronchial inflammation or a preinflammatory state marked by AMP (hyper)responsiveness increases the vulnerability to develop nasal symptoms.
Air filtration units in homes with cats: can they reduce personal exposure to cat allergen?
Authors: Gore R. B. , Bishop S. , Durrell B. , Curbishley L. , Woodcock A. , Custovic A.
Published: June 2003

Background Domestic air filtration units have previously been shown to cause a dramatic fall in airborne pet allergen levels in homes with pets. Clinical trials of air filtration units, however, have failed to reveal a significant beneficial effect. Personal pet allergen exposure during air filtration unit use has never been measured.

Objective To determine the effect of air filtration on inhaled cat allergen exposure in homes with cats.

Methods Nasal air samplers were worn to measure personal cat allergen exposure. The study was carried out in five homes with cats on 4 separate days examining four experimental conditions (cat absent or present, air filtration off or on). The two operators collected four baseline samples and two 15‐min samples/h over three consecutive hours. Cat allergen‐bearing particles were detected by immunoblotting and allergen concentrations measured by amplified enzyme‐linked immunosorbent assay (ELISA).

Results There was a significant reduction in the quantity of the inhaled Fel d 1 when the air cleaner was used with the cat in the room. Fel d 1 halo counts (detransformed means) were 29.3 at baseline, 11.8 after 1 h, 10.0 after 2 h and 14.1 after 3 h, with no change on control days (P = 1.00). With the cat elsewhere in the house, a marginal, but statistically significant reduction was observed only after 3 h with the use of air cleaner (Fel d 1 halo count: baseline 12.4; 3 h 5.5; P = 0.01).

Conclusions The use of air filtration units appears to result in a much smaller reduction of inhaled cat allergen exposure than suggested by previous studies using standard air samplers. Cat removal remains the best advice to cat‐allergic patients who experience symptoms upon exposure.
Identification of Cucumisin (Cuc m 1), a subtilisin‐like endopeptidase, as the major allergen of melon fruit
Authors: Cuesta‐Herranz J. , Pastor C. , Figueredo E. , Vidarte L. , De Las Heras M. , Durán C. , Fernández‐Caldas E. , De Miguel J. , Vivanco F.
Published: June 2003

Background Allergenic components in melon extracts have not been described in spite of the fact that melon (Cucumis melo) is a frequent allergy‐eliciting fruit. The aim of this study was to evaluate allergenic components in melon extract and to report the identification of cucumisin as a major melon allergen.

Materials and methods Sera from 35 patients allergic to melon were selected on the basis of clinical symptoms, skin prick tests and oral challenge test. Allergenic components were detected by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting. Molecular characterization of IgE‐binding bands was performed by N‐terminal amino acid sequencing.

Results More than 10 IgE‐binding bands, between 10 and 80 kDa, were identified in melon extract. Out of them, four IgE‐binding bands were major allergens: 14 kDa, 36 kDa, 54 kDa and 67 kDa. These major allergens, except 14 kDa band, showed the same N‐terminal sequence: T‐T‐R‐S‐W‐D‐F‐L. Research conducted with protein databases identified this N‐terminal sequence as cucumisin, an alkaline serine protease, which shares structural homology with microbial subtilisin. The molecular mass of the identified bands corresponds with different molecular forms of cucumisin produced during the processing or degradation of the enzyme: 67 kDa native cucumisin, 54 kDa mature cucumisin and 36 kDa NH2‐terminal cucumisin fragment.

Conclusion Cucumisin (Cuc m 1) and several N‐terminal cucumisin fragments are the major allergens of melon. The ubiquitous distribution of this protein family (cucumisin‐like proteases) in many plant species and its high structural similarity suggest its potential role as a new panallergen in plant foods.
Inhibitory effects of interferon‐γ on the heterologous desensitization of β‐adrenoceptors by transforming growth factor‐β1 in tracheal smooth muscle
Authors: Ishikawa T. , Kume H. , Kondo M. , Ito Y. , Yamaki K. , Shimokata K.
Published: June 2003

Background Transforming growth factor‐β1 (TGF‐β1) is generally considered to play an important role in the pathogenesis of chronic inflammation and fibrosis.

Objective and methods This study was designed to determine mechanisms of reduced responsiveness of guinea‐pig tracheal smooth muscle to β‐adrenoceptor agonists by TGF‐β1, using isometric tension records and tissue cAMP measurement. Moreover, we examined the involvement of the signal transduction processes of TGF‐β superfamily in the desensitization of β‐adrenoceptors.

Results After exposure to 0.2–2000 pm TGF‐β1 for 4–8 h, the inhibitory effects of 1 µm isoprenaline (ISO) and 10 µm forskolin on 1 µm MCh‐induced contraction were markedly reduced in a concentration‐dependent fashion. The desensitization by TGF‐β1 was greater against ISO than for forskolin. The values of EC75 for the curves for ISO after exposure to the normal bathing solution and TGF‐β1 were 0.039 ± 0.02 and 0.38 ± 0.28 µm, respectively. The values of EC50 for the curves for forskolin under these conditions were 0.50 ± 0.12 and 0.89 ± 0.21 µm, respectively. On the other hand, the inhibitory effects of phosphodiesterase inhibitors such as theophylline and rolipram were not attenuated after exposure to TGF‐β1. Concentration–inhibition curve for ISO was shifted to the right after exposure to 2000 pm TGF‐β1 for 8 h more than that curve for forskolin. In contrast, the curve for theophylline was not shifted to the right by TGF‐β1. When the tissues were incubated with TGF‐β1 in the presence of IFN‐γ, an intracellular antagonist of TGF‐β signalling, IFN‐γ inhibited the reduced response to ISO and forskolin after exposure to TGF‐β1 in a concentration‐dependent fashion. After exposure to TGF‐β1, the effects of cAMP accumulation of ISO was significantly reduced, however, neither forskolin‐nor theophylline‐induced cAMP accumulation was affected. IFN‐γ had no significant effect on cAMP accumulation either to ISO or forskolin.

Conclusions Impairment of the β‐adrenoceptors/adenylyl cyclase pathway are involved in heterologous desensitization of β‐adrenoceptors induced by TGF‐β1 in airway smooth muscle. IFN‐γ functionally suppresses this phenomenon via cAMP‐independent processes. Phosphodiesterase is still intact under this condition.
Effects of mediator antagonism on mannitol and adenosine monophosphate challenges
Authors: Currie G. P. , Haggart K. , Lee D. K. C. , Fowler S. J. , Wilson A. M. , Brannan J. D. , Anderson S. D. , Lipworth B. J.
Published: June 2003

Background Airway hyper‐responsiveness (AHR) to indirect stimuli is a useful non‐invasive surrogate inflammatory marker in the evaluation of asthma, while histamine and cysteinyl leukotrienes are important inflammatory mediators.

Objective To evaluate AHR to indirect bronchoconstrictor stimuli and time taken to recover following single doses of montelukast 10 mg and desloratadine 5 mg in combination, montelukast 10 mg alone and placebo.

Methods Fifteen mild‐to‐moderate persistent asthmatics completed a randomized, double‐blind, cross‐over study. Patients received encapsulated montelukast 10 mg/desloratadine 5 mg combination, montelukast 10 mg alone and placebo, 10–14 h prior to challenge on two separate occasions. The mannitol threshold dose, AMP threshold concentration and recovery times after challenge were measured along with lung function.

Results Compared to placebo, montelukast/desloratadine conferred improvements (P < 0.05) in adenosine monophosphate (AMP) threshold concentration and mannitol threshold dose: a 3.2‐fold (95% CI 2.2–4.6) and 2.4‐fold (95% CI 1.7–3.3) difference, respectively, while compared to montelukast this amounted to a 2.0‐fold (95% CI 1.2–3.4) and 1.5‐fold (95% CI 1.1–2.4) improvement, respectively. Montelukast was not significantly different from placebo. Both montelukast/desloratadine and montelukast compared to placebo, shortened recovery following both challenges (P < 0.05): a 27‐min (95% CI 17–37) and 29‐min (95% CI 20–36) reduction, respectively, for AMP, and a 27‐min (95% CI 17–37) and 26‐min (95% CI 17–35) reduction, respectively for mannitol.

Conclusion The dissociated effects of single doses of montelukast alone but not montelukast/desloratadine combination on AHR and recovery time, highlights the relative roles of histamine in initiating the bronchoconstrictor response and cysteinyl leukotrienes in sustaining it. Similar improvements in AHR and recovery time were observed following both indirect bronchoconstrictor stimuli.
Montelukast as an adjuvant to mainstay therapies in patients with seasonal allergic rhinitis
Authors: Topuz B. , Öǧmen G. G.
Published: June 2003

Background Drug selection for optimal treatment of allergic rhinitis may be difficult and involve many diverse factors.

Objective To evaluate montelukast, in the treatment of patients with seasonal allergic rhinitis, as an adjuvant to mainstay therapies, i.e., antihistamines and corticosteroids.

Methods The study was a prospective, three‐phased (per lasted 2 weeks) clinical trial. In phase I, patients were separated into two groups, based on the predominating symptoms: (1) runners (patients with moderate to severe sneezing/itchy, watery nose/itchy, watery eyes), and (2) blockers (patients with moderate to severe nasal congestion). The runners received antihistamine loratidine 10 mg daily, and the blockers received intranasal corticosteroid mometasone furoate 200 µg. In phase II, if patients were dissatisfied with the initial treatment, they were assigned to receive another study drug additionally. In phase III, for the patients unsatisfied with the treatment of loratidine plus mometasone furoate, montelukast 10 mg once daily was added.

Results Of the 169 patients who entered phase I, 150 could be evaluated for treatment efficacy and safety. There were 108 runners and 42 blockers. Physicians' and patients' assessments indicated that in phase I 58 runners (60.0% of 50 runners) and 36 blockers (33.3% of six blockers) were satisfied with their therapy. In phase II, 30 runners (27.7%) and two blockers (4.7%) were satisfied with the addition of other study drug. Totally 62.6% of patients were satisfied at the end of phase I, and 84.0% at the end of phase II. Sixteen patients (66.6% of 24 patients) were satisfied with the addition of montelukast to the previous two drugs (in total, 10.6% of patients). Fourteen patients (12.2%) treated with loratadine, and three patients (0.3%) treated with mometasone, reported side‐effect.

Conclusion The results of this trial indicate that 10% of patients with seasonal allergic rhinitis may be treated with the supplement of montelukast to mainstay therapy.
Suppression of fructokinase encoded by LeFRK2 in tomato stem inhibits growth and causes wilting of young leaves
Authors: German Marcelo A. , Dai Nir , Matsevitz Tanya , Hanael Rana , Petreikov Marina , Bernstein Nirit , Ioffe Marina , Shahak Yosepha , A. Schaffer Arthur ,   David Granot
Published: June 2003

Fructokinases catalyze the key step of fructose phosphorylation in plants. LeFRK2, the major fructokinase‐encoding gene in tomato plants, is abundantly expressed in roots, stems, and fruits. To analyze the role of LeFRK2 in plant development, we analyzed transgenic tomato plants with sense and antisense expression of StFRK, the potato homolog of LeFRK2. Increased fructokinase activity had no effect. However, plants in which LeFRK2 was specifically suppressed, either via antisense suppression or via co‐suppression, exhibited growth inhibition and wilting of young leaves at daytime. Grafting experiments indicated that a stem interstock of antisense plants was sufficient to inhibit growth and cause leaf wilting. Stem secondary xylem exhibited particular suppression of LeFRK2 and the area of active xylem, estimated by eosin uptake, was significantly smaller in antisense stem compared to that of wild‐type plants. These results suggest that LeFRK2 might be required for proper development of xylem that affected growth and wilting.
The GA octodinucleotide repeat binding factor BBR participates in the transcriptional regulation of the homeobox gene Bkn3
Authors: Santi Luca , Wang Yamei , Stile Maria Rosaria , Berendzen Kenneth , Wanke Dierk , Roig Cristina , Pozzi Carlo , Müller Kai , Müller Judith , Rohde Wolfgang , Salamini Francesco
Published: June 2003

In the dominant mutant Hooded (K), the barley gene BKn3 is overexpressed as a result of a duplication of 305 bp in intron IV. When fused to a cauliflower mosaic virus 35S minimal promoter, the 305 bp element activates gene expression in tobacco, as does a 655 bp BKn3 promoter sequence. Both DNA fragments contain a (GA)8 repeat (GA/TC)8. A one‐hybrid screen using the 305 bp element as the DNA target led to the cloning of the barley b recombinant (BBR) protein, which binds specifically to the (GA/TC)8 repeat. BBR is nuclear targeted and is a characterized nuclear localization signal (NLS) sequence, a DNA‐binding domain extended up to 90 aa at the C‐terminus and a putative N‐terminal activation domain. The corresponding gene has no introns and is ubiquitously expressed in barley tissues. In co‐transfection experiments, BBR activates (GA/TC)8‐containing promoters, and its overexpression in tobacco leads to a pronounced leaf shape modification. BBR has properties of a GAGA‐binding factor, but the corresponding gene has no sequence homology to Trl and Psq of Drosophila, which encode functionally analogous proteins. In Arabidopsis, (GA/TC)8 repeats occur particularly within 1500 bp upstream of gene start codons included in some homeodomain genes of different classes. The data presented suggest that expression of the barley BKn3 is regulated, at least in part, by the binding of the transcription factor BBR to GA/TC repeats.
Protein interaction analysis of SCF ubiquitin E3 ligase subunits from Arabidopsis
Authors: Risseeuw Eddy P. , Daskalchuk Timothy E. , Banks Travis W. , Liu Enwu , Cotelesage Julian , Hellmann Hanjo , Estelle Mark , Somers David E. , Crosby William L.
Published: June 2003

Ubiquitin E3 ligases are a diverse family of protein complexes that mediate the ubiquitination and subsequent proteolytic turnover of proteins in a highly specific manner. Among the several classes of ubiquitin E3 ligases, the Skp1‐Cullin‐F‐box (SCF) class is generally comprised of three ‘core’ subunits: Skp1 and Cullin, plus at least one F‐box protein (FBP) subunit that imparts specificity for the ubiquitination of selected target proteins. Recent genetic and biochemical evidence in Arabidopsis thaliana suggests that post‐translational turnover of proteins mediated by SCF complexes is important for the regulation of diverse developmental and environmental response pathways. In this report, we extend upon a previous annotation of the Arabidopsis Skp1‐like (ASK) and FBP gene families to include the Cullin family of proteins. Analysis of the protein interaction profiles involving the products of all three gene families suggests a functional distinction between ASK proteins in that selected members of the protein family interact generally while others interact more specifically with members of the F‐box protein family. Analysis of the interaction of Cullins with FBPs indicates that CUL1 and CUL2, but not CUL3A, persist as components of selected SCF complexes, suggesting some degree of functional specialization for these proteins. Yeast two‐hybrid analyses also revealed binary protein interactions between selected members of the FBP family in Arabidopsis. These and related results are discussed in terms of their implications for subunit composition, stoichiometry and functional diversity of SCF complexes in Arabidopsis.
The Arabidopsis thaliana CUTA gene encodes an evolutionarily conserved copper binding chloroplast protein
Authors: Burkhead Jason L. , Abdel‐Ghany Salah E. , Morrill Jonathan M. , Pilon‐Smits Elizabeth A. H. , Pilon Marinus
Published: June 2003

The Arabidopsis thaliana CUTA gene encodes a 182‐amino‐acid‐long putative precursor of a chloroplast protein with high sequence similarity to evolutionarily conserved prokaryotic proteins implicated in copper tolerance. Northern analysis indicates that AtCUTA mRNA is expressed in all major tissue types. Analysis of cDNA clones and RT‐PCR with total mRNA revealed alternative splicing of AtCUTA by retention of an intron. The intron‐containing mRNA encodes a truncated 156‐amino‐acid protein as a result of stop codons in the included intron. The sequence of AtCutAp encoded by the fully spliced transcript suggests that the precursor consists of three domains: an N‐terminal chloroplast transit sequence of 70 residues, followed by a domain with prokaryotic signal‐sequence‐like characteristics and finally the most conserved C‐terminal domain. The N‐terminal chloroplast transit sequence was functional to route a passenger protein into isolated pea chloroplasts with possible sorting to the envelope. Chloroplast localization was confirmed by Western blot analysis of isolated and fractionated chloroplasts. Recombinant AtCutA protein was expressed in Escherichia coli without the N‐terminal 70‐amino‐acid chloroplast transit sequence. This recombinant AtCutAp was routed to the bacterial periplasm of E. coli. Purified recombinant AtCutAp is tetrameric and selectively binds Cu(II) ions with an affinity comparable to that reported for mammalian prion proteins.
A pea antisense gene for the chloroplast stromal processing peptidase yields seedling lethals in Arabidopsis: survivors show defective GFP import in vivo
Authors: Zhong Rong , Wan Jiangxin , Jin Rongguan , Lamppa Gayle
Published: June 2003

The stromal processing peptidase (SPP) of chloroplasts is a metalloendopeptidase that cleaves in vitro a broad range of precursor substrates. Here, we have investigated SPP's role in vivo. Two pea cDNA antisense constructs encoding either full‐length SPP (AS4.0) or its N‐terminal half (AS2.2) are introduced into Arabidopsis, which contains one gene for SPP that codes for one isoform. Our analyses show that AS4.0 produces a strong mutant phenotype, with a large percentage of the plants dying as seedling lethals. Surviving plants exhibited slower shoot and root growth, and grossly aberrant leaf morphology. Green and white sectoring, and purple pigmentation was observed. In cells where chloroplasts could be identified, they were fewer in number by at least 40%, thylakoids were not fully developed, and starch granules accumulated. The phenotype produced by AS2.2 was less severe. Using green fluorescent protein (GFP) fused to a transit peptide as a reporter, we examined import into chloroplasts in vivo. In the Arabidopsis antisense lines, GFP was located primarily in the cytosol, indicating that an early step in the import pathway was impeded. In a tobacco AS14 line expressing AS2.2, GFP was located in the cytosol, on the envelope, and in the stroma. The three patterns were observed in different cells, suggesting that the import capacity of individual cells was not the same. Our in vivo studies demonstrate that SPP is essential for chloroplast biogenesis and plant survival. SPP does not act independently in the stroma, but its activity influences earlier steps in the import pathway.
Monitoring expression profiles of Arabidopsis gene expression during rehydration process after dehydration using ca. 7000 full‐length cDNA microarray
Authors: Oono Youko , Seki Motoaki , Nanjo Tokihiko , Narusaka Mari , Fujita Miki , Satoh Rie , Satou Masakazu , Sakurai Tetsuya , Ishida Junko , Akiyama Kenji , Iida Kei , Maruyama Kyonoshin , Satoh Shinobu , Yamaguchi‐Shinozaki Kazuko , Shinozaki Kazuo
Published: June 2003

Plants respond and adapt to drought stress in order to survive under stress conditions. Several genes that respond to drought at the transcriptional level have been described, but there are few reports on genes involved in the recovery from dehydration. Analysis of rehydration‐inducible genes should help not only to understand the molecular mechanisms of stress responses in higher plants, but also to improve the stress tolerance of crops by gene manipulation. We used a full‐length cDNA microarray containing ca. 7000 Arabidopsis full‐length cDNAs and identified 152 rehydration‐inducible genes. Venn diagram analysis showed relationship of the rehydration‐inducible genes to proline‐inducible and water‐treatment‐inducible genes. Among the 152 rehydration‐inducible genes, 58 genes contained the ACTCAT sequence involved in proline‐ and hypoosmolarity‐inducible gene expression in their promoter regions, suggesting that ACTCAT sequence is a major cis‐acting element involved in rehydration‐inducible gene expression, and that some novel cis‐acting elements are involved in rehydration‐inducible gene expression. Functional analysis of rehydration‐inducible and rehydration‐repressed genes revealed their functions not only in the release from a stressed status but also in the recovery of growth in plants.
Distribution of the oriental fruit moth Grapholita molesta Busck (Lep., Tortricidae) infestation on newly planted peaches before and during 2 years of mating disruption
Authors: Il'ichev A. L. , Williams D. G. , Drago A.
Published: June 2003

Abstract:  Oriental fruit moth (OFM) Grapholita molesta Busck (Lep., Tortricidae) is one of the most important pests of commercial stone fruit orchards in the Goulburn‐Murray Valley region of Victoria, Australia. OFM populations have been successfully controlled by the use of the mating disruption (MD) technique for many years, but damage to shoot tips and fruit has now started to increase. The most severe damage under MD is found at the edge of peach blocks, adjacent to the pear blocks under insecticide treatment. In 1997–98, OFM infestation levels were examined in a newly planted peach block surrounded by older peaches, pears, apples and pasture. The infestation distribution was followed up for four consecutive years. No treatments were used against OFM for the first 2 years in the newly planted peaches, but in years 3 and 4 the whole block was treated with MD. At the end of year 2, shoot tip damage was randomly distributed throughout the newly planted peach block with no ‘edge effect’. After MD was applied in year 3, the damage was confined to the edges of the block adjacent to insecticide‐sprayed apples and pears. No ‘edge effect’ was detected along the border with an older peach block treated with MD or on the border with pasture. Extending the MD treated area for 25–30 m into the neighbouring apples and pears in year 4 reduced the ‘edge effect’.
Partial characterization of cotton plants expressing two toxin proteins from Bacillus thuringiensis: relative toxin contribution, toxin interaction, and resistance management
Authors: Greenplate J. T. , Mullins J. W. , Penn S. R. , Dahm A. , Reich B. J. , Osborn J. A. , Rahn P. R. , Ruschke L. , Shappley Z. W.
Published: June 2003

Abstract:  Laboratory studies were performed to characterize the lepidopteran toxicity of cotton plants expressing two different toxin proteins from Bacillus thuringiensis (Bt), in order to assess insect resistance management implications of a commercial, two‐toxin transgenic cotton. An independent and additive interactive effect of two Bt δ‐endotoxins expressed by the transgenic cotton variety 15985 was demonstrated by examining the responses of Heliothis virescens (F.), Helicoverpa zea (Boddie), and Spodoptera frugiperda (J.E. Smith) larvae to field‐ or greenhouse‐grown tissue from genetic near‐isolines, which expressed Cry1A only, Cry2Ab only, or both toxins. In all cases, the Cry2Ab component was the larger contributor to total toxicity in the two‐toxin isoline. Toxin‐specific, quantitative enzyme‐linked immunosorbent assay (ELISA) tests confirmed that the levels of each toxin in tissues of the two‐toxin isoline were not statistically different (P > 0.05) from the levels found in the corresponding tissues of the respective single‐toxin isoline. Resistance management considerations were discussed. Considering the additive interaction of toxins, a relatively simple insect resistance‐monitoring procedure was proposed for the monitoring of commercial cotton varieties expressing both toxins.
The impact of regular hospitalization of children living with thalassaemia on their parents in Sri Lanka: A phenomenological study
Authors: Nahalla Chitra Kumari , FitzGerald Mary
Published: June 2003

This paper reports on a small phenomenological study that set out to describe what it means to have a child with thalassaemia regularly attending hospital for blood transfusion. Ten parents were invited to participate from the thalassaemia unit of a large hospital in a north‐west province of Sri Lanka. Data were collected by in‐depth interviews with parents. The participants’ experiences are described as they reveal, in their own words, what it means to support children with this disease. With startling reality, this research portrays the problems associated with caring for these children and their regular admission to hospital. Three themes are used to present the participant's experience: worries, medical services, and helping and being helped. The findings of the study show that there have been improvements in the service and treatment of these children in Sri Lanka. However, the lives of the parents are still dire. This research should be useful locally in terms of increasing awareness of thalassaemia and raising the consciousness of Sri Lankan nurses and other health care workers, so that they are more aware of the parents’ needs. Although phenomenological research is not generalizable, it is likely that the experience of the parents in this study has similarities throughout the developing world.