Molecular Profile of Sensitization to Dermatophagoides pteronyssinus Dust Mite in Portugal.

BACKGROUND AND OBJECTIVES: To analyze component-resolved diagnosis of sensitization to Dermatophagoides pteronyssinus (Der p) in patients with respiratory allergy and the association between diagnostic findings and clinical severity in different geographical areas. METHODS: The study population comprised 217 patients (mean age, 25.85 [12.7] years; 51.16% female) selected from 13 centers in Portugal (5 from the North, n=65). All had allergic rhinitis with or without asthma and positive skin prick test results to at least 1 dust mite. Specific IgE (sIgE) to Der p, Dermatophagoides farinae, Lepidoglyphus destructor, Der p 1, Der p 2, Der p 10, and Der p 23 was determined using ImmunoCAP. The Mann-Whitney test was applied for the following comparisons: rhinitis vs rhinitis and asthma; mild vs moderate-to-severe rhinitis; North vs South. RESULTS: The prevalence of sensitization was 98.2% for Der p, and 72.4%, 89.4%, 9.7%, and 77% for Der p 1, Der p 2, Der p 10, and Der p 23, respectively. The corresponding median sIgE levels were 8.56, 17.7, 0.01, and 3.95 kUA/L. sIgE to all allergens was higher in patients with moderate-to-severe rhinitis and rhinitis with asthma (nonsignficant). Concentrations of sIgE to Der p 2 were significantly higher in the South than in the North (P=.0496). CONCLUSION: The most common sensitization in Portugal was to Der p. The highest prevalence and median sIgE level were observed for Der p 2. All sIgE values for molecular components were higher in more symptomatic patients (nonsignificant). Concentrations of sIgE to Der p 2 were higher in the South, probably because of the warmer temperature and/or the larger sample size.

Multivariate analysis of biochar-derived carbonaceous nanomaterials for detection of heavy metal ions in aqueous systems.

This article focuses on implementing multivariate analysis to evaluate biochar-derived carbonaceous nanomaterials (BCN) from three different feedstocks for the detection and differentiation of heavy metal ions in aqueous systems. The BCN were produced from dairy manure, rice straw and sorghum straw biochar using our NanoRefinery process. The NanoRefinery process transforms biochar into advanced nanomaterials using depolymerisation/chemical oxidation and purification of nanomaterials using solvent extraction. Dairy manure biochar-derived carbonaceous nanomaterials (DMB-CN), rice straw biochar-derived carbonaceous nanomaterials (RSB-CN) and sorghum straw biochar-derived carbonaceous nanomaterials (SSB-CN) were utilised as probes for the evaluation of their fluorescent properties and the detection of heavy metal ions. The BCN fluorescence quenching and fluorescence recovery was tested with lead (Pb2+), nickel (Ni2+), copper (Cu2+) and mercury (Hg2+). Principal component analysis (PCA) and discriminant analysis were used to differentiate among heavy metal ions in water samples. The BCN from different feedstocks had different characteristics and produced different interactions with heavy metal ions. DMB-CN had the highest quenching for Hg2+ and Ni2+ while SSB-CN and RSB-CN responded best to Cu2+ and Pb2+, respectively. The fluorescence quenching was modelled using linear and empirical functions. PCA and discriminant analysis used the quenching measurements to differentiate heavy metal ions in aqueous system. A key result was that the discriminant analysis had a 100% accuracy to detect Pb2+, 66% for Ni2+ and Cu2+, and 33% for Hg2+. This study has shown that biochar-derived carbonaceous nanomaterials could be used in heavy metal ions sensing applications. This is the first step in the development of a fast and accurate method for the detection of heavy metal ions in waters using environmentally friendly BCN.

NanoRefinery of carbonaceous nanomaterials: Complementing dairy manure gasification and their applications in cellular imaging and heavy metal sensing.

This article describes an efficient method, combining chemical oxidation and acetone extraction, to produce carbonaceous nanomaterials from dairy manure biochar. The optical and mechanical properties are similar to methods previously reported carbonaceous nanomaterials from biomass. Our novel process cuts the processing time in half and drastically reduces the energy input required. The acetone extraction produced 10 fractions with dairy manure biochar-derived carbonaceous nanomaterials (DMB-CNs). The fraction with the carbonaceous nanomaterials, DMB-CN-E1, with highest fluorescence was selected for in-depth characterisation and for initial testing across a range of applications. DMB-CN-E1 was characterised using atomic force microscope, electrophoresis, and spectrophotometric methods. DMB-CN-E1 exhibited a lateral dimension between 11 and 28 nm, a negative charge, and excitation/emission maxima at 337/410 nm, respectively. The bioimaging potential of DMB-CN-E1 evidenced different locations and different interactions with the cellular models evaluated. DMB-CN-E1 was quenched by several heavy metal ions showing a future application of these materials in heavy metal ion detection and/or removal. The demonstrated capabilities in bioimaging and environmental sensing create the opportunity for generating added-value nanomaterials (NanoRefinery) from dairy manure biochar gasification and, thus, increasing the economic viability of gasification plants.

Microalgae biochar-derived carbon dots and their application in heavy metal sensing in aqueous systems.

This research seeks a coupled solution for managing the large amounts of biochar produced by microalgae biofuel production, and the necessity for novel, economic and accurate heavy metal sensing methods. Therefore, this study evaluated the transformation of microalgae biochar (MAB) into carbon dots (Cdots) and their subsequent application as heavy metal ion sensors in aqueous systems. The experimental phase included the transformation of MAB into microalgae biochar-derived carbon dots (MAB-Cdots), MAB-Cdot characterisation and the evaluation of the MAB-Cdots as transducers for the detection of four heavy metal ions (Pb2+, Cu2+, Cd2+, and Ni2+). MAB-Cdot fluorescence was stable over a wide range of pH and resistant to photo-bleaching, making them suitable as fluorescence probes. The MAB-Cdot fluorescence was quenched by all of the metal ions and displayed different quenching levels. Depending upon the ions involved, MAB-Cdots were used to detect the presence of heavy metal ions from concentrations of 0.012 µM up to 2 mM by measuring the reduction in fluorescence intensity. Neutral and slightly alkaline pHs were optimal for Cu2+ Ni2+ and Pb2+ heavy metal quenching. To quantify the concentration of the heavy metal ions, linear and logarithmic functions were used to model the MAB-Cdot fluorescence quenching. The sensing mechanism was determined to be reversible and purely collisional with some fluorophores less accessible than the others. This work demonstrated the ability to produce Cdots from microalgae biochar, examined their application as a transducer for detecting heavy metal ions in aqueous systems and paves the way for novel sensing systems using MAB-Cdots.

Genetic characterization of Guinea-Bissau using a 12 X-chromosomal STR system: Inferences from a multiethnic population.

A male West African sample from Guinea-Bissau (West-African coast) was genetically analyzed using 12 X chromosomal short tandem repeats that are grouped into four haplotype groups. Linkage disequilibrium was tested (p≤0.0008) and association was detected for the majority of markers in three out of the four studied haplotype clusters. The sample of 332 unrelated individuals analyzed in this study belonged to several recognized ethnic groups (n=18) which were used to evaluate the genetic variation of Guinea-Bissau's population. Pairwise genetic distances (FST) did not reveal significant differences among the majority of groups. An additional 110 samples from other countries also belonging to West Africa were as well compared with the sample of Guinea-Bissau. No significant differences were found between these two groups of West African individuals, supporting the genetic homogeneity of this region on the X chromosome level. The generation of over 100 DNA West African sequences provided new insights into the repeat sequence structure of some of the present X-STRs. Parameters for forensic evaluation were also calculated for each X-STR, supporting the potential application of these markers in typical kinship scenarios. Also, the high power of discrimination values for samples of female and male origin observed in this study, confirms the usefulness of the present X-STRs in identification analysis.

Characterisation of systemic reactions to subcutaneous immunotherapy with airborne allergens and classification according to WAO 2010.

BACKGROUND: Systemic reactions (SR) to subcutaneous immunotherapy (SCIT) are rare but potentially severe. The use of different definitions and classifications hampered comparability between studies. AIMS: To determine the frequency of SR to SCIT with airborne allergens, and to characterise and classify them according to the WAO 2010 recommendations. METHODS: Cross-sectional, retrospective study. Data on patients, immunotherapy and SR to SCIT were collected from the SCIT record forms. During the study period, 22,332 SCIT injections were administered (3732 patients). RESULTS: A total of 26 SR (0.1% of administrations) were recorded in 16 (0.6%) patients (median age 22 years, nine males, all with rhinitis and nine with asthma). Twenty-one (81%) SR occurred during the induction phase; eight (31%) in the first hour after administration. According to the WAO 2010 classification, 12 (46%) were grade 1 and 14 (54%) were grade 2. Most grade 2 reactions occurred in asthmatics, presented as mild asthma symptoms and resolved without need for medical observation. Only two individuals without asthma presented grade 2 reactions, both with concurrent cutaneous and low respiratory symptoms; both required medical observation and treatment despite late onset; 82% (n=12) of grade 2 reactions were late. No grade 3-5 reactions were registered and only one patient needed adrenaline treatment. No risk factors for SR to SCIT were identified in this study. CONCLUSIONS: SCIT is a safe treatment when administered by trained staff. The WAO 2010 classification might be useful for retrospectively classifying the severity of reactions, although its usefulness in treatment decision needs further research.

Tetranychus urticae allergy in a population without occupational exposure.

BACKGROUND: Tetranychus urticae is a phytophagus mite found in the leaves of numerous plants. High sensitization rates have been demonstrated, however, provocation tests have only been performed in an occupational setting. OBJECTIVE: To assess accuracy of skin prick tests and clinical relevance of T. urticae sensitization by means of conjunctival provocation tests (CPT) in a population without occupational exposure and to evaluate possible environmental risk factors for T. urticae allergy. METHODS: Patients ≥ 18 years old sensitized to T. urticae (n = 12) and a non-sensitized control group (n = 12) were invited to perform CPT with T. urticae and fulfill a questionnaire including demographic data, questions on environmental exposure to T. urticae and allergy symptoms/diagnosis. A single-blinded placebo-controlled CPT with T. urticae (Leti®) was performed with increasing concentrations (0.002, 0.02, 0.2 and 2 mg/mL) and considered positive if conjunctival hyperemia, palpebral edema or lacrimation were observed in the tested eye. RESULTS: Of T. urticae sensitized patients (mean wheal 4.4 ± 1.5 mm), 9 had a positive CPT, including 3 monosensitized. A good diagnostic accuracy was found for skin prick tests: AUC = 0.952, sensitivity = 100%, specificity = 80%, positive likelihood ratio = 5 and negative likelihood ratio = 0 for a 3 mm wheal. No differences were found between allergic and non-allergic subjects regarding atopy, allergic disease or farming activities. CONCLUSIONS: A high prevalence of allergy to Tetranychus urticae was found in the north of Portugal. Future studies with a larger number of patients are needed to evaluate its relation to clinical symptoms and the impact of environmental factors.

Costs of treatment affect compliance to specific subcutaneous immunotherapy.

BACKGROUND: Specific subcutaneous immunotherapy (SCIT) is cost-effective; however its economical burden can lead to non-adherence. We aimed to identify the reported reasons, patient's personal or socio-demographic characteristics and SCIT-related factors associated with non-adherence to SCIT. METHODS: This is a cross-sectional, observational study held in a Portuguese University Hospital. All records from patients starting SCIT in the last 4 years were reviewed in July 2011. Those without registry of SCIT administration in the previous 3 months were included (n = 181). A telephonic survey was performed and 56 were confirmed as non-adherent; reasons for discontinuation were assessed. Univariate and multiple logistic regression models were developed using characteristics previously described as being associated with non-adhesion. RESULTS: Economical burden of SCIT was the most important factor leading to treatment discontinuation (40%). In the univariate analysis, presence of family history of allergic disease treated with immunotherapy was associated with decreased OR of non-adherence to SCIT (OR [95%CI] = 0.31 [0.11- 0.88]). In the multiple logistic regression model, none of the factors was significantly associated with non-adherence. CONCLUSION: Adherence is influenced by economic factors. More attention should be given to the impact of economic changes in allergic patient's treatment.

Tuberculosis screening in Portuguese healthcare workers using the tuberculin skin test and the interferon-gamma release assay.

The prevalence of latent tuberculosis (TB) infection (LTBI) and the incidence of active tuberculosis in healthcare workers (HCWs) in a Portuguese hospital were examined. This cross-sectional study comprises 4,735 hospital workers screened between May 2005 and September 2008. Tuberculin skin test (TST) and interferon-gamma release assay (IGRA) were used simultaneously in 1,219 HCWs (25.7%). Radiographs were taken in symptomatic HCWs or in test-positive HCWs. The tests were repeated annually or bi-annually depending on risk assessment. IGRA was positive in 32.6% and TST in 74.2% of the HCWs. Years spent in healthcare were a risk factor for a positive IGRA, but not for a positive TST. Repeated bacillus Calmette-Guérin vaccination increased the probability of TST+/IGRA- discordance (35.4% versus 54.4%, respectively). In those tested three times with TST during the study period (n = 59), the mean diameter of TST increased from 5 to 7 to 10 mm. Within 3 yrs, 31 HCWs were diagnosed with active TB (annual incidence rate 191 out of 100,000 people). In eight HCWs with active TB, TST and IGRA were performed at the time of diagnosis and each test was positive. TB burden in HCWs in Portugal is high. With IGRA, the number of radiographs needed to exclude active TB could have been reduced by about half without missing a case of active TB. Therefore IGRA should be introduced into TB screening programmes.

Is ineffective oesophageal motility associated with reflux oesophagitis?

OBJECTIVE: To evaluate the association between ineffective oesophageal motility and reflux oesophagitis controlling for hiatal hernia, hypotensive lower oesophageal sphincter and male sex in patients with gastro-oesophageal reflux disease. METHODS: A total of 387 patients with reflux disease (mean age, 46 years, 42% men) were consecutively selected from a database. All patients underwent upper endoscopy, oesophageal manometry and 24 h oesophageal pH-metry in accordance with a standardized protocol. Reflux disease was confirmed either by endoscopy (oesophagitis grade I-IV according to Savary-Miller) or by pH-metry (increased acid exposure). Hiatal hernia was diagnosed endoscopically, whereas ineffective oesophageal motility and hypotensive lower oesophageal sphincter were characterized during manometry testing. The association between ineffective oesophageal motility and reflux oesophagitis was assessed by logistic regression analysis. RESULTS: A total of 166 patients with oesophagitis (mean age 45 years, 49% men) and 221 without oesophagitis (mean age 46 years, 37% men) were present. Prevalences of ineffective oesophageal motility, hiatal hernia, hypotensive lower oesophageal sphincter and male sex were significantly higher in patients with oesophagitis compared with those without oesophagitis (P<0.05). Ineffective oesophageal motility was independently associated with oesophagitis after multivariate logistic regression analysis (odds ratio=1.68; 95% confidence interval=1.04-2.70). CONCLUSION: Ineffective oesophageal motility is associated with reflux oesophagitis, independently of hiatal hernia, hypotensive lower oesophageal sphincter and male sex.

Indoor mite allergens in patients with respiratory allergy living in Porto, Portugal.

We investigated the levels of mite allergens (Der p 1, Der f 1, Der 2, and Lep d 1) in dust samples from the homes of 59 patients with asthma, 36 sensitized to house-dust mites (HDM) and 23 to grass pollen (controls), living in Porto, northern Portugal. The relationship between exposure and sensitization to HDM and the influence of housing conditions on mite-allergen levels were also evaluated. Der p 1 (median 9.2 micrograms/g) and Der 2 (4.6 micrograms/g) were the main allergens, while Der f 1 and Lep d 1 levels were always < 1 microgram/g dust and undetectable in 11% and 47% of samples, respectively. All HDM-sensitized asthmatics were exposed to Der p 1 levels > 2 micrograms/g and their homes contained significantly higher levels of Der p 1 (median 12.5 vs 6.4 micrograms/g; P = 0.008) and Der 2 (6.2 vs 3.0 micrograms/g; P = 0.004) when compared to the control group. A significant correlation was observed between the exposure to Der p 1 and the wheal area at skin testing with the Dermatophagoides pteronyssinus (Dp) extract (P = 0.01) as well as with serum specific IgE levels to Dp (P = 0.03). Patients with higher levels of serum specific IgE (> or = 17.5 HRU/ml) were also more frequently exposed to Der p 1 levels > or = 10 micrograms/g (P = 0.002). Old homes, presence of carpets, and signs of dampness were conditions associated with significantly higher levels of mite allergens. In conclusion, we found high levels of Der p 1 and Der 2 particularly in the homes of HDM-sensitized patients and we confirm the relationship between exposure and sensitization to HDM, assessed by both in vivo and in vitro methods. In additional to a favorable outdoor climate, we found in our region housing conditions propitious to mite growth, suggesting that specific geographic characteristics must also be taken into account for the correct planning of mite-avoidance measures.

Effects of immunotherapy on symptoms, PEFR, spirometry, and airway responsiveness in patients with allergic asthma to house-dust mites (D. pteronyssinus) on inhaled steroid therapy.

The present study was designed to investigate the effects of immunotherapy (IT) with an extract of Dermatophagoides pteronyssinus (Alergo-Merck Depot) during a 27-month period in patients with allergic asthma to house-dust mites. We included 11 patients (mean age 18 years) treated with a combination of IT and inhaled beclomethasone dipropionate (BDP) in comparison to another 11 (mean age 22 years) treated with BDP alone. We evaluated symptom scores, salbutamol use, peak expiratory flow rates (PEFR), spirometry, and bronchial hyperresponsiveness (BHR) during 18 months of therapy with BDP and in the 9 months after BDP interruption. The two kinds of treatment were efficient and comparable in relation to symptom score, salbutamol use, morning PEFR, FVC, and FEV1, but patients treated with IT and BDP had a faster improvement of BHR and PEFR variability. The interruption of BDP after 18 months of therapy was linked to an impairment of all end points, which were more pronounced in patients previously treated only with BDP. These findings suggest that in selected asthmatic patients allergic to house-dust mites, the association of IT and BDP is more effective than therapy with this inhaled steroid alone due to a faster and more striking improvement during the first months of treatment and to a lower rate of relapse after the interruption of therapy with BDP.

[Cockroach allergy: a study of its prevalence using skin tests with commercial extracts]. / Alergia a la cucaracha: estudio de su prevalencia mediante pruebas cutáneas con extractos comerciales.

Cockroaches have been increasingly recognized as an important source of indoor allergens. In this study we assessed the prevalence of cockroach sensitization among an outpatient population observed at our Department (155 patients with a mean age of 29 +/- 12 years), based on skin "prick" tests with four commercial cockroach extracts along with a common battery of standardized inhalant allergens. We found a positive wheal to at least one of these four extracts in 27 patients: 26 (96.2%) to Blatta orientalis, 10 (37%) both to Blatella germanica and 1 (3.7%) exclusively to Blatella germanica, with no significant concordance between them. We also observed in patients with cockroach positive skin "prick" test an association with atopy (p < 0.001) and with cutaneous reactivity to other indoor allergens, namely house dust mites (p = 0.02), danders (p = 0.01) and fungi (p = 0.01). These data confirm the higher risk of cockroach sensitization among the atopic population sensitized to indoor allergens. However, the heterogeneity of the positive cutaneous responses obtained in this study, possibly reflecting the incomplete standardization of cockroach extracts, questions the real prevalence and clinical significance of this particular sensitization.