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1.
J Biol Chem ; 295(9): 2736-2748, 2020 02 28.
Artículo en Inglés | MEDLINE | ID: mdl-31882546

RESUMEN

Cyclophilins are structurally conserved pan-allergens showing extensive cross-reactivity. So far, no precise information on cross-reactive IgE-epitopes of cyclophilins is available. Here, an 18-kDa IgE-reactive cyclophilin (Rhi o 2) was purified from Rhizopus oryzae, an indoor mold causing allergic sensitization. Based on LC-MS/MS-derived sequences of natural Rhi o 2, the full-length cDNA was cloned, and expressed as recombinant (r) allergen. Purified rRhi o 2 displayed IgE-reactivity and basophil degranulation with sera from all cyclophilin-positive patients. The melting curve of properly folded rRhi o 2 showed partial refolding after heat denaturation. The allergen displayed monomeric functional peptidyl-prolyl cis-trans isomerase (PPIase) activity. In IgE-inhibition assays, rRhi o 2 exhibited extensive cross-reactivity with various other cyclophilins reported as allergens from diverse sources including its homologous human autoantigen. By generating a series of deletion mutants, a conserved 69-residue (Asn81-Asn149) fragment at C terminus of Rhi o 2 was identified as crucial for IgE-recognition and cross-reactivity. Grafting of the Asn81-Asn149 fragment within the primary structure of yeast cyclophilin CPR1 by replacing its homologous sequence resulted in a hybrid molecule with structural folds similar to Rhi o 2. The IgE-reactivity and allergenic activity of the hybrid cyclophilin were greater than that of CPR1. Therefore, the Asn81-Asn149 fragment can be considered as the site of IgE recognition of Rhi o 2. Hence, Rhi o 2 serves as a candidate antigen for the molecular diagnosis of mold allergy, and determination of a major cross-reactive IgE-epitope has clinical potential for the design of next-generation immunotherapeutics against cyclophilin-induced allergies.


Asunto(s)
Alérgenos/inmunología , Reacciones Cruzadas/inmunología , Ciclofilinas/inmunología , Epítopos/análisis , Inmunoglobulina E/inmunología , Rhizopus/inmunología , Alérgenos/genética , Secuencia de Aminoácidos , Secuencia Conservada , Ciclofilinas/genética , Ciclofilinas/aislamiento & purificación , ADN Complementario , Proteínas Fúngicas/inmunología , Humanos , Hipersensibilidad/diagnóstico , Fragmentos de Péptidos/inmunología
3.
Contemp Clin Dent ; 6(3): 313-7, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26321827

RESUMEN

BACKGROUND: Considering an increase in elderly population in recent years, it has become necessary to pay attention to this group so that they can have a higher quality of life (QoL). Oral health is one of the factors affecting the QoL of the elderly. AIMS: The purpose of this study is to evaluate the effect of oral health on the QoL of the elderly population in Babol, Iran. MATERIALS AND METHODS: In this study, we used the multi-stage cluster sampling technique to select 300 elderly subjects, over 65 years of age, living in Babol. Oral health-related QoL was evaluated by the Persian version of oral health impact profile (OHIP-14) questionnaire. Data were collected on gender, age, occupation, education, the date of the last dental visit, use of prosthetic appliances, and dental treatment needs, including the periodontal, prosthetic, and surgical needs, by interview and examination. STATISTICAL ANALYSIS USED: Data were analyzed with SPSS software program using Mann-Whitney and Kruskal-Wallis tests. RESULTS: The mean age of the subjects was 71.4 ± 5.6 years, with 183 males (61%) and 117 females (39%). The mean score of OHIP-14 questionnaire was 22.4 ± 8.2, with a range of 5‒50. The highest score was achieved on the psychological discomfort domain. There was no significant difference in OHIP-14 scores according to gender, with significantly lower scores in subjects with academic education. The subjects wearing prosthetic appliances had lower OHIP-14 scores compared to those who did not wear prosthetic appliances. OHIP-14 scores were lower in the dentate individuals compared to the edentulous individuals. There was no correlation between the age and the OHIP-14 score. CONCLUSION: In general, the results of the present study showed a moderate oral health-related QoL in the elderly living in Babol, who have orodental problems.

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