Functional Outcome of Fixation of Complex Intra-articular Distal Radius Fractures with a Variable-Angle Distal Radius Volar Rim Plate.

Aim To evaluate the outcome of these complex fractures using a volar approach and the DePuy Synthes variable-angle 2.4-mm distal radius rim plate. This plate is precontoured to the volar rim for placement distal to the watershed line allowing purchase of the rim fragment of the lunate facet. Its low profile and smooth edges are designed to minimize flexor tendon irritation. Method We report on a consecutive series of far distal AO-23B3 and AO-23C3 fractures treated using this plate in a tertiary hand center between November 2011 and May 2014. Range of motion, grip strength, and complications were assessed at the final clinical review. Disabilities of the arm, shoulder, and hand (DASH) and patient evaluation measure (PEM) scores were assessed at 12 months after surgery. Results Twenty-six patients were included in this review. Six patients were lost to follow-up at 3 months. This plate was used in isolation in 17 cases, and in combination with a dorsal plate, in cases of dorsal instability after volar plating, in 10 patients. DASH and PEM scores 1 year after surgery were 17.6 and 27%, respectively. Visual analog scores for patient treatment satisfaction and severity of pain showed good satisfaction with treatment and mild intermittent pain on activity. Postoperative range of motion was variable and grip strength was of 71% of the uninjured contralateral side. There were no cases of flexor or extensor tendon rupture. Tendon irritation was noted in two patients. Removal of metal was performed in four patients. Loss of reduction occurred in one case and neurologic complications in two cases. Conclusion This implant is specifically designed for the management of far distal complex intra-articular fractures of the distal radius. Despite the complexity of these fracture patterns and the challenge they pose to accurate reduction and stable fixation, outcomes were satisfactory in this small series. There were no cases of tendon rupture. Removal of metal is not necessary in all cases, but prompt removal should be performed in cases of tendon irritation in view of the risk of tendon rupture.

Reimagining the General Health Questionnaire as a measure of emotional wellbeing: a study of postpartum women in Malta.

BACKGROUND: Postpartum health has been subject to a focus on psychological morbidity, despite positive associations between postpartum recovery and maternal emotional wellbeing. There are currently many validated tools to measure wellbeing and related concepts, including non-psychiatric morbidity. The General Health Questionnaire, 12 items (GHQ-12) is one such instrument, widely used and validated in several languages. Its use in postpartum settings has been documented with disagreement about the instrument's utility in this population, particularly in relation to scoring method and threshold. The GHQ-12 has never been translated into Maltese. This study explored the psychometric properties of the GHQ-12 in a Maltese postpartum population to consider if the use of a different scoring method (visual analogue scale) in the GHQ-12 can determine postpartum wellbeing. METHODS: One hundred and twenty-four postpartum women recruited from one hospital in Malta completed the translated and adapted GHQ-12 as a wellbeing measure (GHQ-12(WB)) at four postpartum time points. The psychometric properties of the GHQ-12(WB) were explored using confirmatory factor analysis, discriminant and divergent validity and reliability analysis. RESULTS: The GHQ-12(WB) demonstrated good divergent and known-groups validity and internal consistency. No models offered a good fit to the data. The overall consistent best-fit to the data was an eight item, two factor model (GHQ-8). Model fit improved across all models in terms of CFI at 13 weeks. CONCLUSION: Findings generally support the reliability and validity of the Maltese version of the GHQ-12(WB). Model fit changes over time reflect the dynamic nature of postpartum recovery. Further evaluation of the GHQ-8(WB) is recommended.

Improvement of the nutritional quality of foods as a public health tool.

OBJECTIVES: To assess the potential contribution of improving the nutritional quality of processed foods on individuals' nutritional intake and food supply. This paper also discusses the means to encourage firms to implement these reformulations, particularly in public/private partnerships. STUDY DESIGN: The French Observatory of Food Quality was created by the Government for the quantification and follow-up of food reformulation by the food industry. This nutritional composition database on branded products was matched with two consumption databases: TNS Kantar Worldpanel, which provides details on quantities bought and food expenditures; and INCA 2, an individuals' food consumption survey completed by the French Food Safety Agency. Three food groups were considered: breakfast cereals (355 items in 2008), biscuits and pastries (1805 items in 2008), and bread-based products (620 items in 2009). METHODS: First, the variability in nutrient composition within food categories was determined, which made it possible to consider several food composition modification scenarios within each category. The formulation of the food items with the lowest nutritional quality was modified to three different levels to improve the overall level of quality in a given category. Second, the quantities of sugars, fat, fibre and sodium delivered to the French market through breakfast cereals, biscuits, pastries and bread-based products were calculated for each scenario. Finally, the distribution of individuals' nutrient consumption from the three food groups among the French population was assessed. RESULTS: These scenarios generated important improvements of 1-22% (increase in the amount of fibre or decrease in the amounts of sugars, fat and sodium delivered to the market), depending on the scenario, the food group and the nutrient considered. Improvement of the products with the lowest nutritional quality would also lead to significant variation in individuals' nutrient consumption for the average adult and child consumers of the three groups (range 4.2-18.8%, depending on the scenario, the food group and the nutrient considered). CONCLUSION: Encouraging the reformulation of foods, especially for products with the lowest nutritional quality in each category of processed foods, is a worthy target for health policy makers. The methodology presented in this paper provides information for negotiations between policy makers and firms to quantify commitments in terms of their potential impacts on individuals' nutrient intake, and to check that the firms' commitments are actually met.

Rectal distension: the effect on bladder function.

AIMS: To assess how rectal distension affects urodynamics parameters and diagnosis. METHODS: Thirty women underwent filling cystometry with a rectal balloon inserted and filled with 150 ml of normal saline and repeated without the balloon distended. The volume at which first desire, strong desire and bladder capacity were reported by the women was recorded as well as urodynamics diagnosis. Women were randomized, using the closed envelope method, into having the rectal balloon distended during the first or during the second filling phase. Women with any bowel disease, history of bleeding per rectum were excluded, or women with any contraindication to undergoing urodynamics, or insertion of a device per rectum. All women of a reproductive age underwent pregnancy test and excluded if found to be pregnant. RESULTS: Thirty patients were recruited, 16 reported mixed urinary incontinence (53%), 5 (17%) had isolated overactive bladder (OAB) symptoms and 9 (30%) reported isolated stress urinary incontinence. Patients with distended rectum had statistically significant lower bladder volumes at which first (46% reduction) and strong desire (33% reduction) was felt and reduced maximum bladder capacity (26% reduction) when compared to the rectum being undistended. In four patients (13%) with a history of OAB a diagnosis of detrusor overactivity was found with the rectum was distended but not when the rectum was empty. CONCLUSION: Rectal distension alters bladder sensation and in some cases urodynamics diagnosis.

Nutritional quality and labelling of ready-to-eat breakfast cereals: the contribution of the French observatory of food quality.

BACKGROUND/OBJECTIVES: To assess developments in the nutritional quality of food products in various food groups in France, an Observatory of Food Quality (Oqali) was created in 2008. To achieve its aims, Oqali built up a new database to describe each specific food item at the most detailed level, and also included economic parameters (market share and mean prices). The objective of this paper is to give a detailed analysis of the monitoring of the ready-to-eat breakfast cereals (RTEBCs) sector in order to show the benefits of the Oqali database. SUBJECTS/METHODS: Analysis was limited to products with nutritional information on labels. Packaging was provided by manufacturers or retailers, or obtained by buying products in regular stores. Economic parameters were obtained from surveys on French food consumption and data from consumer purchase panels. The breakfast cereal sector was divided into 10 categories and 5 types of brand. Oqali has developed anonymous indicators to describe product characteristics for each category of RTEBC and each type of brand by cross-referencing nutritional values with economic data. Packaging-related data were also analysed. The major nutritional parameters studied were energy, protein, fat, saturated fat, carbohydrates, sugars, fibre and sodium. Analysis was performed on the basis of descriptive statistics, multivariate statistics and a Kruskal-Wallis test. RESULTS: For the RTEBC, there is large variability in nutrient content throughout the sector, both within and between product categories. There is no systematic relation between brand type and nutritional quality within each product category, and the proportion of brand type within each product category is different. Nutritional labels, claims and pictograms are widespread on packages but vary according to the type of brand. CONCLUSIONS: These findings form the basis for monitoring developments in the nutritional composition and packaging-related data for breakfast cereals in the future. The final objective is to expand the approach illustrated here to all food sectors progressively.

FTIR spectroscopic analysis of sputum: preliminary findings on a potential novel diagnostic marker for COPD.

COPD is a common, progressively disabling disease and a major health burden worldwide. Fourier transform infrared (FTIR) spectroscopy provides for sensitive analysis of complex biological samples. COPD pathogenesis involves quantitative and qualitative changes in sputum biosynthesis. This first study explores whether FTIR can produce distinct spectral profiles of human sputum, and capture differences between COPD and health. Sputum obtained from 15 COPD patients and 15 healthy volunteers was analysed using FTIR spectroscopy; differences in peak positions, height and configuration were identified and measured. All samples gave reproducible characteristic IR absorption spectra. The most relevant regions identified were the amide and glycogen rich regions, showing crucial spectral differences between health and COPD relating to peak position shifts or intensity alteration. These novel preliminary findings support further exploration of FTIR sputum profiling in a clinical study to determine its potential as a practical method for monitoring COPD.

RhoA signaling modulates cyclin D1 expression in human lung fibroblasts; implications for idiopathic pulmonary fibrosis.

BACKGROUND: Idiopathic Pulmonary Fibrosis (IPF) is a debilitating disease characterized by exaggerated extracellular matrix deposition and aggressive lung structural remodeling. Disease pathogenesis is driven by fibroblastic foci formation, consequent on growth factor overexpression and myofibroblast proliferation. We have previously shown that both CTGF overexpression and myofibroblast formation in IPF cell lines are dependent on RhoA signaling. As RhoA-mediated regulation is also involved in cell cycle progression, we hypothesise that this pathway is key to lung fibroblast turnover through modulation of cyclin D1 kinetic expression. METHODS: Cyclin D1 expression was compared in primary IPF patient-derived fibroblasts and equivalent normal control cells. Quantitative real time PCR was employed to examine relative expression levels of cyclin D1 mRNA; protein expression was confirmed by western blotting. Effects of Rho signaling were investigated using transient transfection of constitutively active and dominant negative RhoA constructs as well as pharmacological inhibitors. Cellular proliferation of lung fibroblasts was determined by BrdU incorporation ELISA. To further explore RhoA regulation of cyclin D1 in lung fibroblasts and associated cell cycle progression, an established Rho inhibitor, Simvastatin, was incorporated in our studies. RESULTS: Cyclin D1 expression was upregulated in IPF compared to normal lung fibroblasts under exponential growth conditions (p < 0.05). Serum deprivation inhibited cyclin D1 expression, which was restored following treatment with fibrogenic growth factors (TGF-beta1 and CTGF). RhoA inhibition, using a dominant negative mutant and a pharmacological inhibitor (C3 exotoxin), suppressed levels of cyclin D1 mRNA and protein in IPF fibroblasts, with significant abrogation of cell turnover (p < 0.05). Furthermore, Simvastatin dose-dependently inhibited fibroblast cyclin D1 gene and protein expression, inducing G1 cell cycle arrest. Similar trends were observed in control experiments using normal lung fibroblasts, though exhibited responses were lower in magnitude. CONCLUSION: These findings report for the first time that cyclin D1 expression is deregulated in IPF through a RhoA dependent mechanism that influences lung fibroblast proliferation. This potentially unravels new molecular targets for future anti-IPF strategies; accordingly, Simvastatin inhibition of Rho-mediated cyclin D1 expression in IPF fibroblasts merits further exploitation.

Inhaled ethanolic and aqueous solutions via Respimat Soft Mist Inhaler are well-tolerated in asthma patients.

BACKGROUND: Respimat Soft Mist Inhaler (SMI) is a new-generation inhaler offering improved lung deposition compared with other devices. Bronchodilators administered via Respimat SMI are preserved and stabilized with benzalkonium chloride (BAC) and ethylene diamine tetra-acetic acid (EDTA); both have been reported to cause paradoxical bronchoconstriction if a threshold dose is exceeded. OBJECTIVE: The aim of this randomized, double-blind, three-period, crossover study was to establish that the safety of inhaled ethanolic and aqueous placebo solutions (containing BAC and EDTA) is equivalent to that of inhaled normal saline solution when administered to asthma patients via Respimat SMI. METHODS: Thirty-seven asthma patients with airway hyper-reactivity were randomized to receive four actuations of each of the following three treatments via Respimat SMI, one on each of 3 study days: ethanolic placebo (12 microl 96% ethanol + 0.13 mug EDTA/actuation), aqueous placebo (12 microl water + 5.5 microg EDTA + 1.1 mug BAC/actuation), and normal saline (12 microl 0.9% sodium chloride/actuation). Pulmonary function tests were performed at baseline and at 5, 15, 30, 60, 120 and 180 min after inhalation; the primary endpoint was the lowest FEV(1) recorded between 0 and 30 min. RESULTS: The mean lowest FEV(1) recorded between 0 and 30 min after inhalation minus the study day baseline was -0.090 litres for ethanolic placebo, -0.121 litres for aqueous placebo and -0.094 litres for normal saline (SEM 0.034 litres for all). The mean treatment differences were: ethanolic placebo versus normal saline 0.004 litres (90% CI -0.075-0.083 litres, p = 0.002), and aqueous placebo versus normal saline -0.028 litres (90% CI -0.107-0.052 litres, p = 0.006). Since both 90% CIs fell within the pre-determined equivalence region of +/-0.15 litres, both treatments were considered equivalent to normal saline. CONCLUSION: Ethanolic and aqueous solutions administered via Respimat SMI are safe with regard to paradoxical bronchoconstriction in asthma patients with airway hyper-reactivity.

An association study between the Clara cell secretory protein CC16 A38G polymorphism and asthma phenotypes.

BACKGROUND: Previously, an association has been reported between an increased risk of asthma and a polymorphism in the Clara cell secretory protein (CC16) gene [namely, an adenine to guanine substitution in the CC16 gene at position 38 (A38G) downstream from the transcription initiation site within the noncoding region of exon 1]. Homozygous individuals for the polymorphic sequence (AA genotype) were reported to have a significant (6.9 fold) increased risk of developing asthma. This finding has not been confirmed independently. OBJECTIVE: To validate the association of CC16 A38G polymorphism to asthma in a separate well-characterized population through a case-control study. METHODS: We conducted an association study using a sample of 217 unrelated Northern European Caucasians. Individuals were clinically characterized by a validated respiratory questionnaire, spirometry and bronchial reactivity measurement, and genotyped for the A38G polymorphism using PCR and restriction digestion. Association analysis was performed using the nonparametric Chi-squared tests. RESULTS: In the unselected population, 43.3% participants were homozygous for the CC16*G allele and 45.4% were heterozygous (AG). We observed no significant difference in the distribution of positive bronchial reactivity to methacholine (at FEV1 PC20 of

Virus-induced asthma.

Clinical and experimental investigations indicate that respiratory viral infections are important triggers for asthma attacks. Viral upper respiratory infections have been associated with 80% of asthma exacerbations in children and 50% of all asthma episodes in adults. Human Rhinovirus (HRV) has been implicated as the most common virus associated with asthma episodes. The observation that the great majority of wheezing lower respiratory tract illnesses in early life are associated with acute viral infections suggests that viruses may also alter the development of the lungs or of the immune system, acting as co-factors for the inception of asthma. Whilst there is no doubt that viruses are important asthma exacerbation factors, the role of viral infections in the development of asthma still remains controversial.

Simultaneous identification of GSTP1 Ile105-->Val105 and Ala114-->Val114 substitutions using an amplification refractory mutation system polymerase chain reaction assay: studies in patients with asthma.

BACKGROUND: The glutathione S-transferase (GST) enzyme GSTP1 utilizes byproducts of oxidative stress. We previously showed that alleles of GSTP1 that encode the Ile105-->Val105 substitution are associated with the asthma phenotypes of atopy and bronchial hyperresponsiveness (BHR). However, a further polymorphic site (Ala114-->Val114) has been identified that results in the following alleles: GSTP1*A (wild-type Ile105-->Ala114), GSTP1*B (Val105-->Ala114), GSTP1*C (Val105-->Val114) and GSTP1*D (Ile105-->Val114). METHODS: Because full identification of GSTP1 alleles may identify stronger links with asthma phenotypes, we describe an amplification refractory mutation system (ARMS) assay that allows identification of all genotypes. We explored whether the GSTP1 substitutions influence susceptibility to asthma, atopy and BHR. RESULTS: Among 191 atopic nonasthmatic, atopic asthmatic and nonatopic nonasthmatic individuals, none had the BD, CD, or DD genotypes. GSTP1 BC was significantly associated with reduced risk for atopy (P = 0.031). Compared with AA, trend test analysis identified a significant decrease in the frequency of GSTP1 BC with increasing severity of BHR (P = 0.031). Similarly, the frequency of GSTP1 AA increased with increasing BHR. CONCLUSION: These data suggest that GSTP1*B and possibly GSTP1*C are protective against asthma and related phenotypes.

Glutathione-S-transferase family of enzymes.

The loci encoding the glutathione-S-transferase (GST) enzymes comprise a large supergene family located on at least seven chromosomes. The function of the GST enzymes has traditionally been considered to be the detoxication of electrophiles by glutathione conjugation. A wide variety of endogenous (e.g. by-products of reactive oxygen species activity) and exogenous (e.g. polycyclic aromatic hydrocarbons) electrophilic substrates have been identified. Interestingly, recent data has suggested a role, at least for the pi class gene product, in jun kinase inhibition. Since many GST genes are polymorphic, there has been considerable interest in determining whether particular allelic variants are associated with altered risk (or outcome) of a variety of diseases. We describe recent studies in patients with asthma and cutaneous basal cell carcinoma that demonstrate associations between GSTP1 and GSTT1 genotypes and disease phenotypes. Thus, GSTP1val(105)/val(105) was protective against asthma symptoms and GSTT1 null was associated with a subgroup of basal cell carcinoma patients who develop large numbers of primary tumours in clusters. Importantly, these associations were characterised by relatively large odds ratios (0.11 and 7.4, respectively) implying that the allelic variants exert a substantial biological effect. These and other data indicate the importance of GST polymorphism in determining disease phenotype.

Differential mRNA expression of insulin-like growth factor-1 splice variants in patients with idiopathic pulmonary fibrosis and pulmonary sarcoidosis.

Insulin-like growth factor-1 (IGF-1) is a highly mitogenic polypeptide detectable in human lung. Using competitive reverse transcriptase/polymerase chain reaction (RT-PCR), expression of four IGF-1 transcripts was examined in bronchoalveolar lavage cells (BALC) from normal subjects, idiopathic pulmonary fibrosis (IPF), stage I/II (no fibrosis), and stage III/IV (confirmed fibrosis) pulmonary sarcoidosis patients, and fibroblast strains isolated from normal and IPF lungs. Transcripts studied were Class 1 and Class 2 (exons 1 or 2, respectively) with IGF-1Eb or IGF-1Ea (exons 5 or 6, respectively). Total IGF-1 expression was downregulated in BALC of both patients with IPF (p < 0.01) and patients with sarcoidosis (p < 0.04) compared with healthy subjects. In contrast, both constitutive (p < 0.003) and transforming growth factor-beta (TGF-beta)- induced (p < 0.02) IGF-1 expression was higher in fibrotic, compared with normal, fibroblasts. These changes were associated with differential expression of IGF-1 splice variants. Healthy subjects and sarcoidosis patients without fibrosis showed similar expression of Class 1/Class 2 and IGF-1Ea/IGF-1Eb. However, patients with fibrosis demonstrated discordant, increased relative abundance of Class 1 transcripts (p < 0.01). In parallel, all fibrosis patients failed to express Class 2, IGF-1Eb forms and sarcoidosis patients with fibrosis did not express the Class 1, IGF-1Eb variant either. Fibrotic fibroblasts expressed higher constitutive levels of Class 1, IGF-1Ea transcripts compared with normal fibroblasts. Class 2, IGF-1Eb forms were moderately expressed by fibroblasts only after stimulation with TGF-beta, which also further increased levels of Class 1, IGF-1Ea transcripts. Our findings suggest that transition from a healthy to a fibrotic phenotype occurs in association with a changing pattern of IGF-1 mRNA heterogeneity and leads us to hypothesize a potential role for specific IGF-1 variants in fibrogenesis.

Expression of growth hormone-releasing factor, growth hormone, insulin-like growth factor-1 and its binding proteins in human lung.

Reverse transcription PCR showed that mRNA encoding the neurohormones growth hormone-releasing factor (GRF) and GH, and its receptor GH-R, together with IGF-1 splice variants and IGFBPs are expressed by inflammatory cells found in the normal human airway. Unfractionated BALC moderately express GRF, GH and GH-R, IGFBP-2 to IGFBP-6, and IGFBP-rPl. In addition, BALC preferentially express the class 1 IGF-1Ea splice variant of the IGF-1 gene. A similar pattern of expression occurs in purified AM, except they do not appear to express GH-R. In marked contrast, AM precursor peripheral blood monocytes, do not express neuropeptides or IGF-1 and only express IGFBP-1, -4 and -6 and IGFBP-rP1. These data suggest that normal human inflammatory airway cells possess a powerful array of neurohormones and IGFBPs that are available for modulating local IGF-1 bioavailability in the lung.

Polymorphisms at the glutathione S-transferase, GSTP1 locus: a novel mechanism for susceptibility and development of atopic airway inflammation.

A common feature of environmental irritants is their ability to cause local inflammation which could alter airway function. The principal targets of such injury are the epithelial cells lining the airway passages and the lower respiratory gas-exchange areas. While host atopy is a recognized risk factor for airway inflammation, atopy alone cannot cause asthma. We hypothesize that susceptibility to persistent airway inflammation in atopic individuals is characterized by an inherited deficiency in the effectiveness of detoxification of inhaled irritants and products of oxidative stress such as reactive oxygen species (ROS). Our case-control studies show that polymorphisms at the glutathione S-transferase, GSTP1, locus on chromosome 11q13 may account for variation in host response to oxidative stress, a key component of airway inflammation. Frequency of the GSTP1 Val/Val genotype is reduced in atopic subjects compared with nonatopic subjects. Trend analysis also shows a significant decrease of GSTP1 Val/Val (with parallel increase of GSTP1 Ile/Ile) genotype frequency with increasing severity of airflow obstruction/bronchial hyperresponsiveness. The implication of specific polymorphisms at the GSTP1 locus in airway inflammation is entirely novel: however, GST are recognized as a supergene family of enzymes critical in 1) cell protection from the toxic products of ROS-mediated reactions, 2) modulation of eicosanoid synthesis.

Expression of intercellular adhesion molecule-1 (ICAM-1) in nasal epithelial cells of atopic subjects: a mechanism for increased rhinovirus infection?

Since clinical experimental studies indicate that upper respiratory tract viral infections may exacerbate acute asthma symptoms in atopic/asthmatic individuals, we have investigated the expression and modulation of ICAM-1 on human nasal epithelial cells (HNEC) from normal and atopic subjects. ICAM-1 is the attachment molecule for the majority of serotypes of human rhinovirus (HRV), including HRV-14, and is also critical for the migration and activation of immune effector cells. Basal ICAM-1 expression was significantly higher in HNEC obtained by brushings from atopic compared with non-atopic subjects (P = 0.031), and was also significantly increased on atopic HNEC harvested in season compared with out of season (P < 0.05). Atopic HNEC showed further up-regulation in ICAM-1 expression when cultured with clinically relevant allergen (P = 0.032). ICAM-1 levels on normal HNEC were also increased by infection with HRV-14 (P < 0.05). Basal expression of ICAM-1 on atopic nasal polyp epithelial cells (EC) was significantly higher than on both normal and atopic nasal HNEC. This elevated nasal polyp ICAM-1 level was not increased further by allergen, although HRV infection resulted in a small significant increase. Recovered viral titres from HRV-infected nasal polyp EC were 1.5-fold higher than from infected normal nasal HNEC. The data are consistent with the hypothesis that allergen, by enhancing expression of the HRV attachment target on host cells, facilitates viral infection in atopic subjects; simultaneously HRV-induced increases in ICAM-1 levels would favour migration and activation of immune effector cells to the airway, resulting in enhanced atopic inflammation.

Polymorphism at the glutathione S-transferase GSTP1 locus. A new marker for bronchial hyperresponsiveness and asthma.

Most genetic studies of asthma have concentrated on genes on chromosomes 11q and 5q and their association with the key asthma-related phenotypes of bronchial hyperresponsiveness (BHR) and atopy. Although asthma is characterized by airway inflammation, a critical component of which is oxidative stress, few data exist on genes involved in protecting against this insult. We describe an association study designed to examine whether allelic variation at the glutathione-S-transferase GSTP1 locus influences expression of the BHR and atopy phenotypes in asthma. The enzyme encoded by GSTP1 utilizes a variety of lipid and DNA products of oxidative stress, and polymorphic variants of this gene are associated with altered catalytic function of this enzyme. We found that the frequency of GSTP1 Val(105)/Val(105) was significantly lower in asthmatic than in control subjects. Indeed, the presence of this genotype conferred a sixfold lower risk of asthma than did GSTP1 Ile(105)/Ile(105). Remarkably, asthma risk in Val(105) homozygotes was further reduced (by ninefold) after correction for atopic indices, age, and gender. Trend analysis after stratification according to the degree of bronchial reactivity/obstruction showed that the frequency of GSTP1 Val(105)/Val(105) correlates with decreasing severity of airway dysfunction. Furthermore, subjects with GSTP1 Val(105)/Val(105) have four- and 10-fold lower risks, respectively, of exhibiting atopy defined by skin test positivity and IgE level. These data show that GSTP1 polymorphism is strongly associated with asthma and related phenotypes, and provide an alternative explanation for the linkage of chromosome 11q13 with BHR and atopy.

The -403 G-->A promoter polymorphism in the RANTES gene is associated with atopy and asthma.

Asthma is a complex inflammatory condition often associated with bronchial hyperreactivity and atopy. Genetic and environmental factors are implicated and several candidate genes have been implicated. Of these, the chemokine RANTES is responsible for the recruitment of inflammatory cells such as eosinophils and T-lymphocytes. We have recently identified a polymorphism within the RANTES promoter (-403 G-->A) and have examined its role, using a PCR-RFLP assay, in the development of atopy and asthma in 201 Caucasian subjects. Atopic status was determined using skin prick testing and serum IgE levels. Severity of airway dysfunction was assessed using spirometric measurement (FEV1) and methacholine challenge (PC20). The -403 A allele was associated with an increased susceptibility to both atopy and asthma. Thus, the proportion of subjects carrying this allele was higher in each of atopic non-asthmatics, non-atopic asthmatics and atopic asthmatics compared with non-atopic, non-asthmatic controls. In particular, this allele was associated with skin test positivity but not IgE level. Homozygosity for the -403 A allele conferred a 6.5-fold increased risk of moderate/severe airway obstruction (FEV1 < or = 80% predicted), a marker for established asthma. Our data, whilst preliminary, indicate that the association of RANTES genotype with both atopy and asthma reflect independent effects, suggesting different mechanisms for the role of this chemokine in atopy and development of airway obstruction.

Early detection of ozone-induced hydroperoxides in epithelial cells by a novel infrared spectroscopic method.

In the lower atmosphere ozone is a toxic and an unwanted oxidising pollutant causing injury to the airway epithelial cells by lipid peroxidation to yield products such as phospholipid hydroperoxides (PLHP). Measurements of PLHP, which are primary oxidation products, may reflect an early susceptibility of the target cell to oxidative stress. Biphasic cultures of bronchial epithelial cells (BEAS-2B) were exposed to ozone at environmentally relevant concentrations (0.1-1.0 ppm) for 4 and 12 h. Detection of PLHP was made using a novel technique based on fourier transform infrared spectroscopy (FTIR) in combination with high performance thin-layer chromatography (HPTLC). Six phospholipids were identified on the HPTLC plate; lysophosphatidylcholine (LPC), sphingomyelin (SM), phosphatidylcholine (PC), lysophosphatidylethanolamine (LPE), phosphatidylinositol (PI), and phosphatidylethanolamine (PE). From the FTIR spectra, O-O stretching of hydroperoxides was identified in the range 890-820cm(-1). Multivariate data analysis revealed a positive correlation (r = 0.99 for 4 h exposure and r = 0.98 for 12h exposure) between ozone exposure levels and the region of the FTIR-spectrum comprising the main wavelengths for hydroperoxides. These data support this alternative, versatile and novel spectroscopic approach for the early detection of ozone-mediated damage in human airway epithelial cells.