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1.
J Family Med Prim Care ; 11(6): 2499-2502, 2022 Jun.
Article in English | MEDLINE | ID: mdl-36119155

ABSTRACT

Background: Though, smoking is the leading cause of chronic obstructive pulmonary disease worldwide, the household air pollution due to use of solid biomass fuel is considered as a major risk factor for the development of obstructive lung disease. The aim of the study was to assess the effect of solid biomass fuel exposure on lung functions in non-smoking female population. Methods: A hospital based, descriptive cross sectional study was carried out among 140 non-smoking female patients aged 40 or more and who had been exposed to solid biomass fuel. These patients underwent spirometry to assess their lung function and were classified as obstructive, restrictive or mixed. Modified medical research council (mMRC) dyspnoea scale for symptom assessment, 6-minute walk test (6 MWT) to determine the exercise capacity and Cumulative exposure index to assess the duration of exposure were also done. Results: All 140 (100%) patients having abnormal lung function, 4 (2.86%) had restrictive pattern, 5 (3.57%) had mixed pattern and 131 (93.57%) had obstructive pattern. Of 131 patients having obstructive pattern, 11 had mild obstruction, 49 had moderate obstruction, 39 had severe obstruction and 32 had very severe obstruction. Most commonly used biomass fuel was wood (43.57%). All the patients had shortness of breath, whereas cough was present in only 35.71% cases. 77 (55%) patients presented with a dyspnoea of mMRC grade 3 and above. Conclusion: Cumulative exposure index for solid biomass fuel is directly proportional to the severity of lung impairment as well as the symptom severity.

2.
Pneumonol Alergol Pol ; 83(4): 275-82, 2015.
Article in English | MEDLINE | ID: mdl-26166789

ABSTRACT

INTRODUCTION: Several studies in developed countries have shown association between indoor air pollution and asthma in children. The present research was undertaken to study this association at Delhi, India. MATERIAL AND METHODS: This study took place at Delhi, capital of India. Eight locations based on the source of pollution such as industrial, residential and villages were included. Recording of the demographic profile and clinical examination of each child was conducted at their residence. Indoor SO2, NO2 and SPM (suspended particulate matter) levels were measured by using Handy Air Sampler (Low Volume Sampler). RESULTS: A total of 3104 children were examined of which 60.3% were male and 39.7% were female. 32.4% children were exposed to environmental tobacco smoke. 31.5 % children's families were using biomass fuels for cooking. History of respiratory symptoms included cough (43.9%), phlegm production (21.9%), shortness of breath (19.3%) and wheezing (14.0%). 7.9% children were diagnosed as having asthma, which was highest in industrial areas (11.8%), followed by residential (7.5%) and village areas (3.9%). The mean indoor SO2, NO2 and SPM levels were 4.28 ± 4.61 mg/m³, 26.70 ± 17.72 mg/m³ and 722.0 ± 457.6 mg/m³ respectively. Indoor SPM was the highest in industrial area followed by residential area and urban village area. Indoor SPM level was significantly (p < 0.001) higher in the asthmatic children's houses. CONCLUSION: This study suggests that industry plays an important role in increasing the concentration of indoor suspended particulate matter and occurrence of asthma in children in developing countries like India.


Subject(s)
Air Pollution, Indoor , Asthma/epidemiology , Asthma/etiology , Housing , Industry , Air Pollutants , Biomass , Child , Cooking , Cough/etiology , Environmental Monitoring , Female , Humans , India/epidemiology , Male , Nitric Oxide/adverse effects , Nitric Oxide/toxicity , Particulate Matter/toxicity , Respiratory Sounds/etiology , Sulfur Dioxide/toxicity , Tobacco Smoke Pollution/adverse effects
4.
Antimicrob Agents Chemother ; 57(6): 2845-8, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23507274

ABSTRACT

Schizophyllum commune (n = 30) showed lowest geometric mean MICs of isavuconazole (0.19 µg/ml), itraconazole (0.2 µg/ml), voriconazole (0.24 µg/ml), and amphotericin B (0.29 µg/ml) and high geometric mean MICs of fluconazole (19.39 µg/ml) and flucytosine (17.28 µg/ml). Five cases (of 8) of allergic bronchopulmonary mycosis that were treated with itraconazole had no recrudescence after 6 to 24 months of follow-up. One case each of invasive pulmonary mycosis and fungal ball were treated successfully with voriconazole and itraconazole.


Subject(s)
Antifungal Agents , Invasive Pulmonary Aspergillosis/drug therapy , Mycoses/drug therapy , Schizophyllum/drug effects , Amphotericin B/pharmacology , Amphotericin B/therapeutic use , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Humans , Invasive Pulmonary Aspergillosis/microbiology , Itraconazole/pharmacology , Itraconazole/therapeutic use , Microbial Sensitivity Tests , Molecular Sequence Data , Mycoses/microbiology , Pyrimidines/pharmacology , Pyrimidines/therapeutic use , Schizophyllum/classification , Schizophyllum/genetics , Sequence Analysis, DNA , Treatment Outcome , Triazoles/pharmacology , Triazoles/therapeutic use , Voriconazole
5.
PLoS One ; 7(12): e52871, 2012.
Article in English | MEDLINE | ID: mdl-23285210

ABSTRACT

Azole resistance is an emerging problem in Aspergillus which impacts the management of aspergillosis. Here in we report the emergence and clonal spread of resistance to triazoles in environmental Aspergillus fumigatus isolates in India. A total of 44 (7%) A. fumigatus isolates from 24 environmental samples were found to be triazole resistant. The isolation rate of resistant A. fumigatus was highest (33%) from soil of tea gardens followed by soil from flower pots of the hospital garden (20%), soil beneath cotton trees (20%), rice paddy fields (12.3%), air samples of hospital wards (7.6%) and from soil admixed with bird droppings (3.8%). These strains showed cross-resistance to voriconazole, posaconazole, itraconazole and to six triazole fungicides used extensively in agriculture. Our analyses identified that all triazole-resistant strains from India shared the same TR(34)/L98H mutation in the cyp51 gene. In contrast to the genetic uniformity of azole-resistant strains the azole-susceptible isolates from patients and environments in India were genetically very diverse. All nine loci were highly polymorphic in populations of azole-susceptible isolates from both clinical and environmental samples. Furthermore, all Indian environmental and clinical azole resistant isolates shared the same multilocus microsatellite genotype not found in any other analyzed samples, either from within India or from the Netherlands, France, Germany or China. Our population genetic analyses suggest that the Indian azole-resistant A. fumigatus genotype was likely an extremely adaptive recombinant progeny derived from a cross between an azole-resistant strain migrated from outside of India and a native azole-susceptible strain from within India, followed by mutation and then rapid dispersal through many parts of India. Our results are consistent with the hypothesis that exposure of A. fumigatus to azole fungicides in the environment causes cross-resistance to medical triazoles. The study emphasises the need of continued surveillance of resistance in environmental and clinical A. fumigatus strains.


Subject(s)
Antifungal Agents/pharmacology , Aspergillus fumigatus/drug effects , Aspergillus fumigatus/genetics , Cytochrome P-450 Enzyme System/genetics , Drug Resistance, Multiple, Fungal/genetics , Fungal Proteins/genetics , Triazoles/pharmacology , Amino Acid Substitution/physiology , Aspergillus fumigatus/isolation & purification , Clone Cells , Colony Count, Microbial , Environment , Fungicides, Industrial/pharmacology , Histidine/genetics , Humans , India/epidemiology , Leucine/genetics , Mutation, Missense/physiology , Tandem Repeat Sequences/genetics
6.
J Antimicrob Chemother ; 67(2): 362-6, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22028200

ABSTRACT

OBJECTIVES: Azole resistance in Aspergillus fumigatus isolates impacts on the management of aspergillosis since azoles are primary agents used for prophylaxis and therapy. We report the emergence of resistance to triazoles in two A. fumigatus isolates from patients in Delhi, India. METHODS: One hundred and three A. fumigatus isolates, collected from 85 patients suspected of bronchopulmonary aspergillosis during 2005-10, were investigated for susceptibility to itraconazole, voriconazole, posaconazole and isavuconazole. We undertook a mixed-format real-time PCR assay for the detection of mutations leading to triazole resistance in A. fumigatus. The resistant isolates were compared with 25 Dutch TR/L98H-positive isolates by microsatellite analysis. RESULTS: Of the 103 A. fumigatus isolates tested, only 2 had high MIC values of itraconazole (>16 mg/L), voriconazole (2 mg/L), posaconazole (2 mg/L) and isavuconazole (8 mg/L). The resistant A. fumigatus isolates exhibited the TR/L98H genotype and showed identical patterns by microsatellite typing, but were different from 25 Dutch TR/L98H isolates. CONCLUSIONS: We report for the first time from India the occurrence of TR/L98H mutations in the cyp51A gene (responsible for reduced azole susceptibility) in two A. fumigatus isolates from patients with chronic respiratory disease who had not previously been exposed to azoles. The presence of TR/L98H is consistent with a route of resistance development through exposure to azole compounds in the environment. Given the emergence of azole resistance in environmental strains, continued surveillance of resistance in clinical A. fumigatus strains is desirable for successful therapy of aspergillosis.


Subject(s)
Antifungal Agents/pharmacology , Aspergillus fumigatus/drug effects , Aspergillus fumigatus/genetics , Cytochrome P-450 Enzyme System/genetics , Drug Resistance, Fungal , Fungal Proteins/genetics , Mutation, Missense , Triazoles/pharmacology , Aspergillus fumigatus/isolation & purification , Cluster Analysis , DNA, Fungal/genetics , Humans , India , Male , Microbial Sensitivity Tests , Middle Aged , Pulmonary Aspergillosis/microbiology , Real-Time Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology
7.
Med Mycol ; 50(3): 281-90, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22103346

ABSTRACT

Indoor fungi are potential sensitizing agents in children and their detection and quantification in indoor air are important in the diagnosis and environmental management of fungal allergies. The objective of this investigation was to assess the prevalence of fungal allergies in children in Delhi and to study the association between mold counts in the homes of children and their sensitization to respective fungal extracts. Fungal concentrations and seasonality were studied at two-week intervals for one year using Andersen Volumetric and Burkard Slide samplers. Sensitization to fungi frequently encountered in patients' homes was assessed by Skin Prick Tests (SPTs). Total fungal specific IgE was measured by ELISA in the sera of patients positive to fungal extracts. Skin Prick Tests revealed that 39.3% (33/84) of patients were markedly positive (2 + and above) to one or the other fungal allergens. Raised serum IgE to predominant indoor fungal species was observed in patients with marked SPT results. Highest marked skin reactivity (2 + and above) was obtained with Alternaria alternata allergens in 17.9% of the children, which was followed by the response to fungal antigens of Aspergillus fumigatus and Penicillium citrinum (15.5%). Exposure to high fungal counts of some dominant fungi (Penicillium, A. nidulans and A. fumigatus) was found associated with increased fungal sensitization in the patients. Total serum IgE level was revealed to be significantly linked with the intensity of skin reactions, as well as with skin index (r(2) = 0.052; P < 0.05). We concluded that children in Delhi are exposed to high concentrations of fungi in the indoor environment and that respiratory allergies were connected with higher prevalence of skin sensitization.


Subject(s)
Air Microbiology , Fungi/isolation & purification , Housing , Hypersensitivity/epidemiology , Adolescent , Antibodies, Fungal/blood , Child , Child, Preschool , Colony Count, Microbial , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin E/blood , India , Male , Prevalence , Seasons
8.
Allergy Rhinol (Providence) ; 2(1): 21-32, 2011 Jan.
Article in English | MEDLINE | ID: mdl-22852111

ABSTRACT

Allergy to fungi has been linked to a wide range of illnesses, including rhinitis and asthma. Therefore, exposure to fungi in home environment is an important factor for fungal allergy. The present study was aimed to investigate types of airborne fungi inside and outside the homes of asthmatic children and control subjects (nonasthmatic children). The dominant fungi were evaluated for their quantitative distribution and seasonal variation. The air samples were collected from indoors and immediate outdoors of 77 selected homes of children suffering from bronchial asthma/allergic rhinitis using Andersen volumetric air sampler. The isolated fungal genera/species were identified using reference literature, and statistical analysis of the dominant fungi was performed to study the difference in fungal concentration between indoor and immediate outdoor sites as well as in between different seasons. A total of 4423 air samples were collected from two indoor and immediate outdoor sites in a 1-year survey of 77 homes. This resulted in the isolation of an average of 110,091 and 107,070 fungal colonies per metric cube of air from indoor and outdoor sites, respectively. A total of 68 different molds were identified. Different species of Aspergillus, Alternaria, Cladosporium, and Penicillium were found to be the most prevalent fungi in Delhi homes, which constituted 88.6% of the total colonies indoors. Highest concentration was registered in autumn and winter months. Total as well as dominant fungi displayed statistically significant differences among the four seasons (p < 0.001). The largest number of isolations were the species of Aspergillus (>40% to total colony-forming units in indoors as well as outdoors) followed by Cladosporium spp. Annual concentration of Aspergillus spp. was significantly higher (p < 0.05) inside the homes when compared with outdoors. Most of the fungi also occurred at a significantly higher (p < 0.001) rate inside the homes when compared with immediate outdoors. Asthmatic children in Delhi are exposed to a substantial concentration of mold inside their homes as well as immediate outdoor air. The considerable seasonal distributions of fungi provide valuable data for investigation of the role of fungal exposure as a risk for respiratory disorders among patients suffering from allergy or asthma in Delhi.

9.
Immunobiology ; 215(7): 527-34, 2010 Jul.
Article in English | MEDLINE | ID: mdl-19897276

ABSTRACT

Asthma is a chronic immune inflammatory disease characterized by variable airflow obstruction and increased bronchial hyperreactivity (BHR). Therapeutic interventions reduce airway inflammation and relieve symptoms but associated with potential side effects that limit their usefulness. The present study was undertaken to assess the effect of choline on immune inflammation and BHR in asthma subjects. The patients of asthma (n=76) were recruited and treated with choline supplement (1500 mg twice) or standard pharmacotherapy for 6 months in two groups. The patients were evaluated by clinical, immunologic and biochemical parameters. The treatment with choline showed significant reduction in symptom/drug score and improvement in PC(20) FEV1 compared to baseline or standard pharmacotherapy (p<0.01). Choline therapy significantly reduced IL-4, IL-5 and TNF-alpha level as compared to baseline or standard pharmacotherapy after 6 months (p<0.01). Blood eosinophil count and total IgE levels were reduced in both the treatment groups. Cysteinyl leukotriene and leukotriene B4 were suppressed significantly by choline treatment (p<0.01). This was accompanied by decreased 8-isoprostanes, a biomarker for oxidative stress after choline treatment (p<0.01). Choline therapy modulates immune inflammation and suppresses oxidative stress in asthma patients. It can be used as an adjunct therapy for asthma patients.


Subject(s)
Asthma/immunology , Choline/pharmacology , Eosinophils/drug effects , Adolescent , Adult , Asthma/diagnosis , Asthma/physiopathology , Bronchial Hyperreactivity , Cells, Cultured , Cytokines/metabolism , Eosinophils/pathology , Female , Follow-Up Studies , Forced Expiratory Volume , Humans , Immunosuppression Therapy , Inflammation , Leukotrienes/metabolism , Male , Middle Aged , Oxidative Stress/drug effects , Oxidative Stress/immunology , Skin Tests , Th2 Cells/drug effects , Th2 Cells/immunology
10.
J Proteome Res ; 8(6): 2650-5, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19290623

ABSTRACT

The knowledge on epitopes of proteins can help in devising new therapeutic modalities for allergic disorders. In the present study, five B (P1-P5) and five T cell (P6-P10) epitopes were predicted in silico based on sequence homology model of Cur l 3, a major allergen of Curvularia lunata. Peptides (epitopes) were synthesized and assessed for biological activity by ELISA, competitive ELISA, lymphoproliferation and cytokine profiling using Curvularia allergic patients' sera. B cell peptides showed higher IgE binding by ELISA than T cell epitopes except P6. Peptides P1-P6 achieved EC(50) at 100 ng, whereas P7-P10 required 10 mug in inhibition assays. Peripheral blood mononuclear cells from Curvularia allergic patients (n = 20) showed higher lymphoproliferation for T cell epitopes than B cell epitopes except P6 confirming the properties of B and T cell prediction. The supernatant from these patients show highest interleukin-4 release on stimulation with P6 followed by B cell peptides. P4 and P6 together identified 35/37 of Curvularia positive patients by skin tests. In summary, experimental analysis confirmed in silico predicted epitopes containing important antigenic regions of Cur l 3. P6, a predicted T cell epitope, showed the presence of a cryptic B cell epitope. Peptides P4 and P6 have potential for clinical application. The approach used here is relevant and may be used to delineate epitopes of other proteins.


Subject(s)
Allergens/immunology , Ascomycota/immunology , Epitopes, B-Lymphocyte/immunology , Epitopes, T-Lymphocyte/immunology , Fungal Proteins/immunology , Allergens/chemistry , Allergens/genetics , Amino Acid Motifs/immunology , Antibodies, Fungal/blood , Ascomycota/genetics , Cell Proliferation , Cells, Cultured , Computational Biology , Computer Simulation , Cytochromes c/chemistry , Cytochromes c/immunology , Cytokines/metabolism , Enzyme-Linked Immunosorbent Assay , Epitope Mapping , Epitopes, B-Lymphocyte/chemistry , Epitopes, B-Lymphocyte/genetics , Epitopes, T-Lymphocyte/chemistry , Epitopes, T-Lymphocyte/genetics , Fungal Proteins/chemistry , Humans , Hypersensitivity/immunology , Immunoglobulin E/metabolism , Lymphocytes/metabolism , Peptide Fragments/chemistry , Peptide Fragments/immunology , Protein Binding
11.
J Clin Immunol ; 29(1): 63-70, 2009 Jan.
Article in English | MEDLINE | ID: mdl-18683033

ABSTRACT

RATIONALE: Epitopes were delineated for allergenic proteins, but studies are required to identify residues mediating IgE binding. In the present study, the in silico approach was used to identify IgE-binding residues of Alt a 13(1-50) fragment and confirmed by experimental approach. METHOD AND RESULTS: IgE-binding epitopes of Alt a 13 mapped computationally were cloned, expressed, purified, and characterized using various immunochemical and biophysical methods. Among four fragments of Alt a 13, Alt a 13(1-50) demonstrated maximum IgE binding with two immunodominant regions and was mutated at these regions. The mutation in first region, Alt a 13(1-50)-K4A_S6F, did not show any change in immunological and biophysical properties of protein. However, mutations in the second region, Alt a 13(1-50)-T21F_N27I, caused reduced IgE binding, histamine release, and low IL-4 release on stimulation of Alternaria alternata positive patients peripheral blood mononuclear cells in vitro. CONCLUSION: This suggests that residues T21 and N27 are important for the secondary structure. In conclusion, Alt a 13(1-50)-T21F_N27I with reduced Th 2 response and intact T-cell proliferation capacity has potential for clinical use.


Subject(s)
Alternaria/immunology , Antigens, Fungal/immunology , Epitopes/immunology , Hypersensitivity/immunology , Immunoglobulin E/immunology , T-Lymphocytes/immunology , Adolescent , Adult , Antibodies, Fungal/blood , Antigens, Fungal/chemistry , Antigens, Fungal/genetics , Computational Biology , Epitope Mapping , Epitopes/chemistry , Epitopes/genetics , Female , Histamine Release/immunology , Humans , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Male , Middle Aged , Mutant Proteins/genetics , Mutant Proteins/immunology , Mutant Proteins/metabolism , Mutation/genetics , Peptide Fragments/immunology , Peptide Fragments/metabolism , Protein Engineering , Protein Structure, Secondary , Sequence Analysis, Protein , Skin Tests , Young Adult
12.
J Agric Food Chem ; 56(24): 12099-104, 2008 Dec 24.
Article in English | MEDLINE | ID: mdl-19035641

ABSTRACT

Genetically modified crops have resistance to abiotic stress by introduction of choline oxidase protein. In the present study, the safety of choline oxidase protein derived from Arthrobacter globiformis was assessed for toxicity and allergenicity. The protein was stable at 90 degrees C for 1 h. Toxicity studies of choline oxidase in mice showed no significant difference (p > 0.05) from control in terms of growth, body weight, food consumption, and blood biochemical indices. Histology of gut tissue of mice fed protein showed normal gastric mucosal lining and villi in jejunum and ileum sections. Specific IgE in serum and IL-4 release in splenic culture supernatant were low in choline oxidase treated mice, comparable to control. Intravenous challenge with choline oxidase did not induce any adverse reaction, unlike ovalbumin group mice. Histology of lung tissues from choline oxidase sensitized mice showed normal airways, whereas ovalbumin-sensitized mice showed inflamed airways with eosinophilic infiltration and bronchoconstriction. ELISA carried out with food allergic patients' sera revealed no significant IgE affinity with choline oxidase. Also, choline oxidase did not show any symptoms of toxicity and allergenicity in mice.


Subject(s)
Alcohol Oxidoreductases/immunology , Alcohol Oxidoreductases/toxicity , Arthrobacter/enzymology , Bacterial Proteins/immunology , Bacterial Proteins/toxicity , Plants, Genetically Modified/immunology , Plants, Genetically Modified/toxicity , Adult , Alcohol Oxidoreductases/chemistry , Alcohol Oxidoreductases/genetics , Animals , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Cells, Cultured , Enzyme Stability , Female , Food Hypersensitivity/immunology , Food, Genetically Modified/standards , Humans , Male , Mice , Mice, Inbred BALB C , Plants, Genetically Modified/physiology , Random Allocation , Spleen/immunology , Young Adult
13.
Ann Allergy Asthma Immunol ; 99(3): 273-80, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17910332

ABSTRACT

BACKGROUND: Mosquito allergy is well established, but mosquito immunotherapy requires validation using clinical and immunologic variables. OBJECTIVE: To evaluate the tolerability and efficacy of specific immunotherapy with Culex quinquefasciatus (mosquito) extract. METHODS: We performed a randomized, double-blind, placebo-controlled trial of immunotherapy for 1 year in 40 patients with asthma, rhinitis, or both. Patients were evaluated by means of intradermal testing, symptom and drug scores, and histamine provocation testing before and after 1 year of immunotherapy. Mosquito specific IgE and IgG subclass antibody responses were evaluated at the basal level and after 1 year. RESULTS: Patients receiving allergen immunotherapy for 1 year showed a significant improvement compared with baseline and patients receiving placebo regarding skin reactions, symptom scores (rhinitis and asthma), and forced expiratory volume in 1 second. Provocation concentration of histamine that caused a decrease in forced expiratory volume in 1 second of 20% by inhalation was elevated in the group receiving immunotherapy. In the active group serologic analysis showed a slight reduction in IgE levels (P = .02) but a significant elevation in IgG4 levels (P = .001), with a significant decrease in the IgE/IgG4 ratio (P = .001). All these changes in the placebo group were nonsignificant. CONCLUSIONS: Allergen immunotherapy with mosquito extract was well tolerated, with improvement in symptoms and airway reactivity. Good clinical outcome was associated with increased IgG4 antibody levels.


Subject(s)
Asthma/therapy , Complex Mixtures/therapeutic use , Culex/immunology , Desensitization, Immunologic/methods , Rhinitis, Allergic, Perennial/therapy , Adolescent , Adult , Allergens/immunology , Allergens/therapeutic use , Animals , Asthma/physiopathology , Bronchial Provocation Tests , Complex Mixtures/immunology , Culex/chemistry , Double-Blind Method , Female , Forced Expiratory Volume , Humans , Immunoglobulin E/blood , Immunoglobulin G/blood , Male , Rhinitis, Allergic, Perennial/physiopathology , Skin Tests , Treatment Outcome
14.
Biologicals ; 35(2): 131-7, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17071100

ABSTRACT

Commercial cockroach extracts for diagnosis and therapy show batch-to-batch variation. This study aimed to standardize Periplaneta americana extract based on major IgE binding components using hypersensitive patients' sera. Extracts were prepared in phosphate buffered saline (PBS) or NH(4)HCO(3), from freeze-dried or 37 degrees C dried material and compared with commercial extracts by immunobiochemical methods. Cockroach positive patients' sera were collected after intradermal tests and specific IgE enzyme linked immunosorbent assay (ELISA). Allergenic proteins were identified by western-blotting and potency of extracts determined by ELISA-inhibition. Adult P. americana extract from freeze dried source material in PBS (PA extract) resolved into 45 protein bands and showed 22 IgE binding components with pooled patients' sera. It required 9-12 ng self-proteins for 50% ELISA-inhibition. Individual patients' sera identified 23, 28, 35, 38, 40, 49, 72, 78 and 97 kDa as major IgE binding components in PA extract. Nymph extract exhibited similar potency and protein profile to PA extract with 72 and 78 kDa proteins present in high intensities. Commercial extracts exhibited only 6-11 IgE reactive bands compared to PA extract and required 40 folds or more protein for 50% ELISA-inhibition. PA extract from freeze-dried source material seems a potent allergen preparation with 9-major IgE binding components. It can be referred to upgrade the quality of commercial extracts exhibiting low potencies due to poor quality source material, inadequate extraction procedures and improper storage.


Subject(s)
Allergens/isolation & purification , Hypersensitivity/diagnosis , Periplaneta/chemistry , Periplaneta/immunology , Allergens/chemistry , Animals , Enzyme-Linked Immunosorbent Assay , Female , Humans , Hypersensitivity/blood , Immunoglobulin E/blood , Male , Skin Tests , Tissue Extracts/chemistry , Tissue Extracts/isolation & purification
15.
Immunobiology ; 211(9): 733-40, 2006.
Article in English | MEDLINE | ID: mdl-17015148

ABSTRACT

Pollen from the mesquite tree, Prosopis juliflora, is an important source of respiratory allergy in tropical countries. Our aim was to partially characterize the IgE binding proteins of P. juliflora pollen extract and study cross-reactivity with prevalent tree pollen allergens. Intradermal tests with P. juliflora and five other tree pollen extracts were performed on respiratory allergy patients from Bikaner (arid) and Delhi (semi arid). Prosopis extract elicited positive skin reactions in 71/220 of the patients. Sera were collected from 38 of these 71 patients and all demonstrated elevated specific IgE to P. juliflora. Immunoblotting with pooled patients' sera demonstrated 16 IgE binding components, with components of 24, 26, 29, 31, 35, 52, 58, 66 and 95 kDa recognized by more than 80% of individual patients' sera. P. juliflora extract is allergenically potent requiring 73 ng of self-protein for 50% inhibition of IgE binding in ELISA inhibition. Cross-inhibition assays showed close relationship among P. juliflora, Ailanthus excelsa, Cassia siamea and Salvadora persica. IgE binding components of 14, 41, 52 and 66 kDa were shared allergens whereas 26 and 29 kDa were specific to P. juliflora. The findings suggest that purification of cross-reactive allergens will be helpful for diagnosis and immunotherapy of tree pollen allergic patients.


Subject(s)
Antigens, Plant/chemistry , Immunoglobulin E/blood , Plant Extracts/chemistry , Pollen/chemistry , Prosopis/chemistry , Adolescent , Adult , Ailanthus/immunology , Antigens, Plant/immunology , Blotting, Western , Cinnamomum aromaticum/immunology , Cross Reactions , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Humans , Hypersensitivity, Immediate/diagnosis , Middle Aged , Plant Extracts/immunology , Pollen/immunology , Prosopis/immunology , Salvadoraceae/immunology , Skin Tests , Ulmus/immunology
16.
Mol Immunol ; 43(12): 1927-32, 2006 May.
Article in English | MEDLINE | ID: mdl-16430961

ABSTRACT

Recombinant proteins are used for vaccines, therapy and diagnosis of many diseases. Biological activity of these may differ from native counterpart and needs investigation. The present study aimed to compare recombinant (r) and native (n) glutathione-S-transferase (GST) from Alternaria alternata. Glutathione-S-transferase sequence showed an ORF of 696bp encoding 26-kDa protein with N-terminus conserved domain. Secondary structure of both forms was comparable with melting temperature of 57 and 59 degrees C, respectively. rGST and nGST showed similar enzymatic activity, allergenicity and potency by ELISA inhibition. Histamine release was comparable in 14/17 patients for both the GSTs. rGST and nGST induced proliferation in PBMC at different concentration. Cell supernatant revealed higher IL-4 and IL-5 levels with low levels of IFN-gamma. In summary, recombinant and native GST demonstrated similar physio-chemical, biological and immunological properties and induced comparable cell mediated and humoral response to be used for diagnosis and specific immunotherapy for the fungal allergy cases.


Subject(s)
Allergens/immunology , Alternaria/enzymology , Fungal Proteins/immunology , Glutathione Transferase/immunology , Rhinitis, Allergic, Seasonal/immunology , Adolescent , Adult , Allergens/chemistry , Allergens/genetics , Alternaria/genetics , Alternaria/immunology , Case-Control Studies , Female , Fungal Proteins/chemistry , Fungal Proteins/genetics , Glutathione Transferase/genetics , Glutathione Transferase/isolation & purification , Humans , Intradermal Tests , Male , Middle Aged , Recombinant Proteins/immunology , Recombinant Proteins/isolation & purification
17.
Asian Pac J Allergy Immunol ; 24(4): 191-9, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17348241

ABSTRACT

Studies suggest the importance of serum total and specific IgE in clinical evaluation of allergic manifestations. Such studies are lacking in Indian subcontinent, though a large population suffer from bronchial asthma. Here relevance of serum total and specific IgE was investigated in asthmatics with food sensitization. A total of 216 consecutive patients (mean age 31.9 years, S.D. 11.8) were screened by various diagnostic testing. Out of 216 patients, 172 were with elevated serum total IgE (201 to > 800 IU/ml). Rice elicited marked positive skin prick test reactions (SPT) in 24 (11%) asthma patients followed by black gram 22 (10%), lentil 21 (9.7%) and citrus fruits 20 (9.2%). Serum total IgE and specific IgE showed significant correlation, p = 0.005 and p = 0.001, respectively, with positive skin tests. Blinded food challenges (DBPCFC) with rice and or black gram confirmed food sensitization in 28-37% of cases. In summary, serum total IgE of 265 IU/ml or more with marked positive SPT (4 mm or more) can serve as marker for atopy and food sensitization. Specific IgE, three times of normal controls correlates well with positive DBPCFC and offers evidence for the cases of food allergy.


Subject(s)
Allergens , Food Hypersensitivity/blood , Immunoglobulin E/blood , Oryza , Adolescent , Adult , Aged , Asthma/complications , Asthma/diagnosis , Biomarkers/blood , Child , Female , Food Hypersensitivity/complications , Food Hypersensitivity/diagnosis , Humans , India , Male , Middle Aged , Predictive Value of Tests , Skin Tests
18.
Saudi Med J ; 26(3): 421-4, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15806211

ABSTRACT

OBJECTIVE: A single blind parallel group study was conducted to evaluate the effects of oral choline [given as tricholine citrate (TRI)] in patients with allergic rhinitis, and compare its efficacy with intranasal budesonide (BUD). METHOD: The study was conducted at the Department of Respiratory Medicine, Vallabhbhai Patel Chest Institute, Delhi, India from February 2001 to April 2002. Sixty patients were randomized into 2 groups after a run-in period of 2 weeks. Group A received intranasal BUD 200 microg twice daily and group B received TRI 500 mg thrice daily. The patients were reviewed every 2 weeks up to 8 weeks. The mean individual symptom score, total symptom score and drug score were significantly reduced in both groups (p<0.05) compared to baseline values, with maximum effect occurring within 4 weeks of therapy. RESULTS: Budesonide showed statistically significant reduction (p<0.05) in all the outcome parameters, when compared to TRI. Crossover study between the 2 treatment groups also showed similar results. Seventy-six percent of patients with BUD and 43% of patients with TRI found the drug to be effective. CONCLUSION: Both intranasal BUD and oral TRI are effective in relieving symptoms of allergic rhinitis. Budesonide was found to be the statistically superior drug.


Subject(s)
Anti-Inflammatory Agents/therapeutic use , Budesonide/therapeutic use , Choline/analogs & derivatives , Choline/therapeutic use , Rhinitis, Allergic, Perennial/drug therapy , Administration, Inhalation , Administration, Oral , Adolescent , Adult , Cross-Over Studies , Humans , Middle Aged , Single-Blind Method
20.
Int Arch Allergy Immunol ; 127(1): 38-46, 2002 Jan.
Article in English | MEDLINE | ID: mdl-11893852

ABSTRACT

BACKGROUND: Curvularia lunata is an important inhalant allergen. The present study was undertaken to investigate the shared IgG- and IgE-binding components among seven Curvularia species prevalent in the aerospora. METHODS: Seven different Curvularia species were grown in a semisynthetic medium for 13 days. The extracts were analyzed by SDS-PAGE, immunoblot and ELISA/immunoblot inhibition using sera from C. lunata-positive patients and anti-C. lunata rabbit serum. RESULTS: Different Curvularia species showed 11-19 protein bands on SDS-PAGE. Proteins of 12, 20, 31, 45, 53, 78 and 97 kD were present in all the species. Eight out of 98 nasobronchial patients exhibited positive skin tests to C. lunata and to at least five Curvularia species. ELISA using these sera showed IgE binding with Curvularia species. Immunoblot using pooled anti-C. lunata sera from patients showed 5-12 allergenic proteins. Proteins of 12, 31, 45, 53 and 78 kD showed IgE binding in Curvularia species. Antibodies against C. lunata detected 6-14 antigenic proteins on immunoblot. Proteins of 31, 45 and 53 kD showed IgG binding in all the species. Proteins of 31 and 53 kD showed complete IgE/IgG binding inhibition. IgE/IgG ELISA inhibition showed dose-dependent inhibition in Curvularia species. C. lunata extract required 0.17 and 0.11 microg of protein for 50% IgE and IgG inhibition, respectively. C. clavata and C. pallescens required 10 times more protein to exhibit the same inhibition and other species required similar protein levels as those required by C. lunata. CONCLUSIONS: A high degree of cross-reactivity was observed between C. lunata and the six other Curvularia species tested. C. lunata and C. senegalensis shared maximum allergenic and antigenic components.


Subject(s)
Allergens/immunology , Ascomycota/immunology , Fungal Proteins/immunology , Fungal Proteins/metabolism , Immunoglobulin E/metabolism , Immunoglobulin G/metabolism , Allergens/adverse effects , Allergens/metabolism , Animals , Ascomycota/classification , Cross Reactions , Enzyme-Linked Immunosorbent Assay , Fungal Proteins/adverse effects , Humans , Immunoblotting , Immunoglobulin E/immunology , Immunoglobulin G/immunology , Rabbits , Respiratory Hypersensitivity/etiology , Skin Tests
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