Isolation and identification of allergens and biogenic amines of Prosopis juliflora genotypes

Background: Prosopis, or mesquite (Prosopis juliflora (Sw.) DC.), was introduced in Saudi Arabia several decades ago and is heavily used in street, roadside, and park plantations. It shows great adaptation to the prevailing climatic conditions such as high temperature, severe drought, and salinity and spreads naturally in many parts of the Kingdom. This research was conducted to isolate allergen proteins and biogenic amines from the pollen grains of P. juliflora genotypes in Saudi Arabia from two regions, namely Al-Qassim and Eastern regions. Results: The results showed that 18 different allergen proteins were detected in P. juliflora genotypes, with molecular weight ranging from 14 to 97 kDa. Moreover, P. juliflora genotypes from the two studied regions contained eight biogenic amines, namely histamine, tyramine, tryptamine, ß-phenylethylamine, butricine, codapherine, spermidine, and spermine. All genotypes from the Al-Qassim region were found to contain all eight amines, while in the Eastern region, histamine was absent in three genotypes, spermine was absent in six genotypes, and spermidine was absent in three genotypes. Genotypes B23, E20, and E21 had the lowest biogenic amine quantity. Conclusions: All identified proteins from mesquite trees from both regions (Eastern and Al-Qassim) cause allergies in patients who are sensitive to pollen grains. Bioamines, except histamine and tyramine, were recorded at varying concentrations in different genotypes.

Identification of IgE- binding pollen protein from Cannabis sativa in pollen-hypersensitive patients from north Pakistan

Cannabis sativa [C.sativa] is well-known for its medicinal, industrial and recreational use. However, allergies in relation to Cannabis sativa [C.sativa] are rarely reported. C. sativa is one of the common weeds found in Pakistan and its pollen grains are common in spring and fall season. Although categorized as an aeroallergen, there are limited number of reports regarding allergenic potential in C. sativa. Therefore, the current study is aimed at exploring the IgE- binding potential among the C.sativa pollen in local pollen allergic patients. Initial screening of C. sativa sensitized individuals was carried out by dot blot from the sera of pollen allergic patients. Proteins from the pollen grains were extracted and resolved on 10% gel. Eight bands were visible on gel however only one protein fragment i.e. of 14KDa size was found to bind to IgE as analyzed through protein gel blot analysis. Strong IgE affinity of a 14 kDa protein fragment from C. sativa pollen extract suggests its allergenic potential. Further study is required to find the exact nature of this protein fragment

Identification of IgE-mediated food allergy and allergens in older children and adults with asthma and allergic rhinitis.

Background and objective. Prevalence of immunoglobulin (Ig) E-mediated food allergy is primarily reported for certain pediatric populations and adults. The present study was aimed to investigate the relative prevalence of food allergy and allergens in older children and adults with asthma and allergic rhinitis. Methods. Patients (12-62 years) were screened using standard questionnaire and skin prick-test (SPT) with common foods and aeroallergens. Specific IgE level was determined by enzyme linked immunosorbent assay (ELISA) and allergy was established by blinded food challenges. Results. Of 1860 patients screened, 1097 (58.9%) gave history of food allergy. Of the history positive patients skin tested (n=470), 138 (29.3%) showed a marked positive reaction to food extracts. Rice elicited positive SPT reaction in maximum number of cases 29 (6.2%) followed by blackgram 28 (5.9%), lentil 26 (5.5%), citrus fruits 25 (5.3%), pea 18 (3.8%), maize 18 (3.8%) and banana 17 (3.6%). The SPT positive patients showed elevated specific IgE levels (range: 0.8-79 IU/mL) against respective food allergens than normal controls (0.73 IU/mL, mean±2SD). Food allergy was confirmed in 21/45 (46.6%) of the patients by blinded controlled food challenges. The prevalence of food allergy was estimated to be 4.5% (2.6%-6.34%) at 95% confidence interval (95% CI) in test population (n=470). Sensitisation to food was significantly associated with asthma (p=0.0065) while aeroallergens were strongly related to rhinitis (p<0.01). Conclusions. Food allergy is estimated to be 4.5% in adolescents and adults with asthma, rhinitis or both. Rice, citrus fruits, blackgram and banana are identified as major allergens for inducing allergic symptoms.

Evaluation of skin sensitivity in dogs bearing allergic dermatitis to standardized allergenic extract of house dust and storage mites

The objective of the study was to evaluate whether allergenic extracts of five house dust and storage mite species standardized for humans might be used for the diagnosis of canine atopic dermatitis (CAD). Extracts of Dermatophagoides pteronyssinus (Pyroglyphidae), D. farinae (Pyroglyphidae), Blomia tropicalis (Glycyphagidae), Lepidoglyphus destructor (Glycyphagidae) and Tyrophagus putrescentiae (Acaridae) were evaluated by intradermal testing in 20 healthy dogs (control) and 25 dogs with allergic dermatitis. A significant difference in the response was observed between the two groups (p<0.05). Only one dog (5 percent) in the control group reacted to the intradermal test, whereas 14 dogs (56 percent) in the allergic group were positive for at least one extract (odds ratio = 24.2). Most of the positive reactions observed in the allergic group occurred against the extracts of T. putrescentiae or L. destructor, each inducing reactions in 10 dogs (40 percent). D. farinae, D. pteronyssinus e B. tropicalis extracts induced reactions in 7 (28 percent), 3 (12 percent) and 3 (12 percent) dogs, respectively. The allergenic extracts standardized for humans evaluated in the present study may be used as a tool to complement the diagnosis of the disease, as well as to select potential allergen candidates for allergen-specific immunotherapy.

O presente trabalho teve como objetivo avaliar se extratos alergênicos de cinco espécies de ácaros da poeira domiciliar e produtos armazenados, padronizados para humanos, podem ser utilizados no diagnóstico da dermatite atópica canina. Extratos de Dermatophagoides pteronyssinus (Pyroglyphidae), D. farinae (Pyroglyphidae), Blomia tropicalis (Glycyphagidae), Lepidoglyphus destructor (Glycyphagidae) e Tyrophagus putrescentiae (Acaridae) foram avaliados através de testes intradérmicos em 45 cães, dos quais 20 normais e 25 com dermatite alérgica. Uma diferença significativa foi observada no padrão de respostas obtidas dos dois grupos (p<0.05). Apenas um animal (5 por cento) do grupo controle reagiu ao teste cutâneo, enquanto que no grupo dos alérgicos 14 cães (56 por cento) apresentaram pelo menos uma reação positiva (odds ratio = 24.2). As maiores freqüências de reações positivas observadas no grupo dos alérgicos foram aos extratos de T. putres-centiae ou L. destructor, cada um induzindo reações em 10(40 por cento) cães. Os extratos de D. farinae, D. pteronyssinus e B. tropicalis foram responsáveis por reações positivas em 7(28 por cento), 3(12 por cento) e 3(12 por cento) cães, respectivamente. Os extratos padronizados para humanos avaliados no presente estudo podem ser utilizados como complemento no diagnóstico da doença, assim como na seleção de alérgenos para a imunoterapia alérgeno-específica.

Production and characterization of a monoclonal antibody against an Ascaris suum allergenic component

Ascaris suum allergenic components (PIII) separated by gel filtration chromatography of an adult worm extract were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells using polyethylene glycol (MW 1450) as fusogen. The hybridomas were cultured in HAT-containing medium and cloned at limiting dilutions. Supernatants from the growing hybrids were screened by ELISA using plates coated with PIII or the A. suum crude extract. The monoclonal antibody obtained, named MAC-3 (mouse anti-A. suum allergenic component), is an IgG1 kappa mouse immunoglobulin that specifically recognizes a 29,000 molecular weight protein (called allergenic protein) with an affinity constant of 1.7 x 10(9) M-1. The A. suum components recognized by MAC-3 induce specific IgE antibody production in immunized BALB/c mice. Ascitic fluid induced in Swiss mice by injecting ip the hybridoma cells and incomplete Freund's adjuvant was purified by affinity chromatography using a protein A-Sepharose column. The purified monoclonal antibody was then coupled to activated Sepharose beads in order to isolate the A. suum allergenic component from the whole extract by affinity chromatography

Alergenos frecuentes del Valle de México en niños / Frequent allergens in children of Mexico

Se evaluó la incidencia de positividad en las pruebas cutáneas para 48 alergenos en niños con asma bronquial. Se estudiaron 347 tarjetas del expediente clínico de niños alérgicos durante 32 meses. Los alergenos más frecuentes fueron los Dermatophagoides pteronyssinus seguido de gato y los otros 24 alergenos fueron menores del 10 por ciento. Estos alergenos se encuentran en el Valle de México y la incidencia a éstos debe considerarse como la más frecuente en niños alérgicos a quienes se les deben reducir las pruebas cutáneas para el diagnóstico de enfermedades alérgicas

Variación anual de la microflora en la ciudad de San Juan y en hogares de pacientes con patología respiratoria IgE dependiente / Annual mycoareological variation in San Juan city, Argentine Republic, and in houses of IgE dependent respiratory pathology patients

El objetivo del estudio fue establecer la variación anual de la microflora en la ciudad de San Juan y en hogares de pacientes con patología respiratoria IgE dependiente. San Juan es una provincia del oeste de la República Argentina, con escasísimas precipitaciones pluviales anuales (menos de 10 mm), con un promedio de humedad relativa ambiente de 46 por ciento (min.: 36 por ciento y máx.: 55 por ciento) y una temperatura promedio de 17,6ºC (min.: 7,8ºC y máx.: 26,5ºC). Para poder conocer el desarrollo de hongos y definir mejor la correlación existente entre ellos y las enfermedades alérgicas respiratorias, estudiamos durante un período anual (julio 1994-junio 1995) la presencia de seis géneros de hongos: Alternaria, Cladosporium, Penicillium, Aspergillus, Mucor y Rhizopus, en el ambiente exterior de la ciudad de San Juan, y en el domicilio de nueve (9) niños con enfermedad respiratoria (rinitis y/o asma), con Prick Test positivo para todos los hongos en estudio. Se controló la reactividad sanguínea de estos pacientes a los mismos géneros de hongos por IgE RAST. La flora micológica fue estudiada por el método gravimétrico, exponiendo mensualmente cápsulas de Petri (diámetro: 10 cm) en medio ambiente externo y en los domicilios (4 cápsulas por domicilio), con medios de cultivo de Sabouraud y Czapek-Dox (Lab. Merck). Los Prick Test se realizaron con antígenos Allergon AB (Lab. Welt), y las IgE RAST (Allerex Labs. Inc - USA). El hongo que se desarrolló con más frecuencia en los domicilios estudiados fue el Penicillium (40,63 por ciento), seguido por Rhizopus (17,24 por ciento), Aspergillus (14,09 por ciento), Mucor (11,28 por ciento), Cladosporium (4,81 por ciento), Alternaria (4,75 por ciento), y otros (7,0 por ciento). Aunque en distinto porcentaje, la distribución de hongos en ambientes externos sigue una curva muy similar a las halladas en los domicilios: Penicillium (17,86 por ciento), Rhizopus (12,76 por ciento), Aspergillus (11,13 por ciento), Mucor (9,97 por ciento), Alternaria (8,12 por ciento), Cladosporium (6,03 por ciento) y otros (1,39 por ciento). La mayor concentración de hongos se alcanzó en octubre para domicilios y en noviembre-diciembre para ambiente exterior. La mejor correlación entre el Prick Test y el RAST se obtuvo para Alternaria y Aspergillus (87 por ciento), seguidos por Cladosporium (75 por ciento), Penicillium (62 por ciento), Rhizopus (50 por ciento), y Mucor (37 por ciento). No encontramos correlación directa entre la respuestas de los pacientes y la concentración de hongos en el período de estudio, confirmando lo esperado de acuerdo con las diferentes potencias alergénicas de cada género

Influencia de la contaminación atmosférica en la calidad de aire de interiores / Influence of outdoor pollution on indoor air quality

The influence of outdoor pollution on indoor air quality was studied in downtown Santiago (Bandera street). Carbon monoxide (CO), nicotine, particulate matter, respirable fraction (PM5) and total and carcinogenic polyaromatic hydrocarbons (PAHs) were simultaneously monitores indoors and outdoors and indoors in restaurants, offices and other places. The levels of CO changed simultaneously outdoors and indoors (r=0.89) specially during traffic rush hours, demonstrating the importance of outdoor infiltration into the indoor air quality and masking the contribution of other CO indoor sources. The maximun CO concentrations were over 800 percent and over 1000 percent higher indoors and outdoors respectively than the 9 ppm CO National Ambient Air Quality. The PM5 concentrations were very high and showed no significant differences (p>0.05) from indoor to outdoors, or between indoor levels in restaurants, offices and other places. Total ans carcinogenic PAHs levels were also very high, outdoors and indoors, outodoor levels being generally higher than those indoors and no significant; differences (p>0.05) were found for the indoor levels between restaurants, offices and other places. Nicotine levels showed significant differences (p<0.05) between indoor and outdoor levels. In addition, great differences (p<0.05) in indoor levels, were found between offices and restaurants, and offices and other places. Among indoor sources cigarrete smoke seems to be a minor source since nicotine concentrations, bieng 2.3 times higher in restaurants and other places than in offices, do not contribute to enhance significantly PM5 and total and carcinogenic HAPs in the first ones. These results suggest that in downtown Santiago, infiltration might be the main source of indoor pollution. This is supported by 2 evidences: a) coronene, a tracer of vehicle emissions was found in high concentraion indoors and b) in restaurants (in wich PAHs emissions might be higher indoor) a correlation coefficient of 0.987 for the indoor and outdoor concentrations of carcinogenic PAHs was found. Furthermore a survey asking for different symptoms and effects probably related to air pollution was made to people working in Bandera and in a rural area located 40 Km from Santiago. The results showed that excluding smoking as a confounded factor, people working in Bandera showed a significantly greater (p<0.05) risk of ill effects on their health than people working in the rural area

Cladosporium as an airborne allergen in Saudi Arabia

Airborne mold monitoring in three cities in Saudi Arabia has documented the presence of Cladosporium in high concentrations. The spore counts reached threshold levels for sensitization at Jeddah and Al-Khobar with prominent seasonal variations. The concentration in Riyadh [central region] showed levels with minor fluctuations and a minor peak towards the month of November. Indoor monitoring also revealed the presence of Cladosporium. Ten species of Cladosporium were identified with C. sphaerospermum, C. herbarum, C. cladosporoides and C. macrocarpum being most prevalent. The species are relevant in relation to extracts for diagnosis. Skin testing of asthmatic children in Riyadh and Makkah revealed reactivities to Cladosporium extract of 5.8% and 31.3%, respectively. This indicates that Cladosporium may be an important allergen in the region that necessitates further biochemical and immunochemical studies to confirm the allergenic impact of the species of Cladosporium prevalent in the region

Sensibilidad cutánea a ácaros domésticos y de deposito en niños y adultos asmáticos / Cutaneous sensibility to domestic and of deposit acarus in asthmatics children aqnd adults

La alergia a los ácaros del polvillo doméstico en asociación al asma es, actualmente considerado un problema de diseminación mundial. Entre el 70 al 90% de los pacientes asmáticos tienen test cutáneo positivos a extractos de ácaros, particularmente a D. pteronyssinus y D. farinae. Numerosos estudios han encontrado sensibilidad cutánea a otros Piroglífidos, tal el caso de E. maynei. En los depósitos de granos, alimentos y maderas, entre otros, se han hallado ácaros que se desiganaron "de almacenamiento" y que ahora son llamados "domésticos" ya que también se pueden encontrar en las viviendas. No conocíamos la prevalencia de sensibilidad a extractos de otros ácaros del sub-orden astigmata. En este estudio se presentan la frecuencia de sensibilización cutánea en niños asmáticos de Córdoba y niños y adultos asmáticos de Santa Fe a extractos de D. pteronyssinus, D. farinae, B. tropicalis, A. ovatus, C. arcuatus y L. destructor. Tanto en los pacientes asmáticos de Córdoba como de Santa Fe, la mayor frecuencia de sensibilización cutánea fue a D. pteronyssinus, 63.3% y 72.7% respectivamente. B. tropicalis fue el segundo alergeno hallado en Córdoba (58.3%) y D. farinae en Santa Fe (66.6%). Los extractos alergénicos de los otros ácaros estudiados arrojaron positividades entre el 6 y el 57.5% de los casos estudiados. Los ácaros del sub-orden Astigmata, no Piroglífidos, son capaces de inducir sensibilidad cutánea en niños y adultos asmáticos de nuestro país

Structural studies on allergen RC-13 from Ricinus communis L.: isolation and characterization of a major glycopeptide

A glycoprotein, RC-13, isolated from Ricinus communis seeds was reduced, S-alkylated and cleaved by trypsin. The tryptic digest was fractionated by ion-exchange chromatography and a glycopeptide was isolated and purified by high-voltage paper electrophoresis. When submitted to amino acid and carbohydrate analyses this major glycopeptide showed the following chemical composition: Lys1, Asp1, Thr2, Ser4, Glu1, Pro2, Gly2, Ala2, Val2, GlcN6, Man6 and Gal8. Hydrazynolysis positioned Ser as the C-terminal residue. It is postulated that this glycopeptide belongs to the C-terminal region of the allergen