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1.
Neuroophthalmology ; 48(4): 249-256, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38933751

RESUMO

To translate the the Utah Photophobia Symptom Impact Scale-12 questionnaire into Persian and assess the psychometric aspects to check its validity and reliability based on the Rasch modelling method. Translation and cultural adjustment of the English language UPSIS-12 questionnaire to Persian was undertaken. A total of 61 patients with complaints of photophobia participated in evaluating validity and reliability aspects. All the participants were asked to complete the Persian translation of the UPSIS-12 questionnaire. Rasch analyses of the survey items were conducted using WINSTEPS. All items fit the Rasch model. Point-measure correlation values varied from 0.41 to 0.77, providing a preliminary indication of adequate construct validity. All factor loadings were found more than 0.4. All items obtained infit and outfit mean square (MnSq) values of < 2.0. All participants except 5 had normal outfit values. Patients' abilities relative to the items' difficulty were analysed. Item difficulty was estimated and item characteristic curves were included. Sufficient unidimensionality, hierarchical order, and equal interval scoring were obtained. In conclusion, the Persian UPSIS-10 questionnaire has excellent psychometric properties and it will be valuable in both clinical practice and research. It will help Persian practitioners to assess their patients' photophobia.

2.
Diagnostics (Basel) ; 13(18)2023 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-37761339

RESUMO

AIM: This study sought to compare two common risk stratification systems in terms of their diagnostic performance for the evaluation of thyroid malignancy. METHODS: The American College of Radiology (ACR) Thyroid Imaging, Reporting and Data System (TI-RADS) and the American Thyroid Association (ATA) guidelines were compared among 571 thyroid nodules with definitive fine needle aspiration (FNA) cytology or postoperative histopathology. Ultrasound characteristics such as composition, echogenicity, shape, margin, size, and vascularity were assessed for each thyroid nodule. Diagnostic performance measures were determined and compared through receiver operating characteristic (ROC) curves, and decision curve analysis (DCA). RESULTS: Of 571 nodules, 65 (11.4%) were malignant. The AUC, sensitivity, specificity, positive predictive value, and negative predictive value were 0.691, 49.2%, 84.9%, 29.6%, and 92.8% for ATA guideline, and 0.776, 72.3%, 79.2%, 30.9%, and 95.7%, for ACR TI-RADS, respectively. ACR TI-RADS was more sensitive (p = 0.003), while the ATA guideline was more specific (p < 0.001). DCA demonstrated that the ACR TI-RADS provided a greater net benefit than the ATA guideline. In addition, the net reduction in unnecessary biopsies is higher for ACR TI-RADS than ATA guidelines. The total number of indicated biopsies and unnecessary FNA rates were lower in ACR TI-RADS compared to ATA guideline (293 vs. 527 and 80.2 vs. 87.8). ACR TI-RADS presented no biopsy indication in seven malignant nodules (all categorized as TR2), whereas ATA guideline missed one. Hypoechogenicity was the most significant predictor of malignancy (OR = 8.34, 95% CI: 3.75-19.45), followed by a taller-than-wide shape (OR = 6.73, 95% CI: 3.07-14.77). CONCLUSIONS: Our findings suggest that each system has particular advantages in the evaluation of thyroid nodules. ACR TI-RADS reduces unnecessary FNA rates, however, malignant nodules categorized as TR2 might be missed using this system. Further evaluation of this group of nodules using Doppler and other ultrasound modalities is recommended.

3.
Curr J Neurol ; 21(3): 156-161, 2022 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-38011354

RESUMO

Background: The accuracy of current laboratory and imaging studies for diagnosis and monitoring of Parkinson's disease (PD) severity is low and diagnosis is mainly dependent on clinical examination. Proton magnetic resonance spectroscopy (MRS) is a non-invasive technique that can assess the chemical profile of the brain. In this study, we evaluated the utility of proton MRS in diagnosis of PD and determination of its severity. Methods: Patients with PD and healthy age-matched controls were studied using proton MRS. The level of N-acetylaspartate (NAA), total creatine (Cr), and total choline (Cho), and their ratios were calculated in substantia nigra (SN), putamen (Pu), and motor cortex. PD severity was assessed by the Unified Parkinson's Disease Rating Scale (UPDRS) and the Hoehn and Yahr scale. Results: Compared to 25 healthy controls (18 men, age: 59.00 ± 8.39 years), our 30 patients with PD (24 men, age: 63.80 ± 12.00 years, 29 under treatment) showed no significant difference in the metabolite ratios in SN, Pu, and motor cortex. Nigral level of NAA/Cr was significantly correlated with total UPDRS score in patients with PD (r = -0.35, P = 0.08). Moreover, patients with PD with Hoehn and Yahr scale score ≥ 2 had a lower NAA/Cr level in SN compared to patients with a lower stage. Conclusion: This study shows that 1.5 tesla proton MRS is unable to detect metabolite abnormalities in patients with PD who are under treatment. However, the NAA/Cr ratio in the SN might be a useful imaging biomarker for evaluation of disease severity in these patients.

4.
Curr Rheumatol Rev ; 18(2): 160-167, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34548002

RESUMO

BACKGROUND: Neutrophil to lymphocyte ratio (NLR) is a marker for many inflammatory diseases. Ankylosing spondylitis (AS) is among these inflammatory diseases, and many studies have compared the NLR ratio between patients with AS and healthy controls. AIM: This study aims to systematically review and analyze the available evidence about the significance of NLR values in AS. METHOD: Based on Meta-analysis of Observational Studies in Epidemiology (MOOSE) guidelines, we searched Embase, Pubmed, ISI Web of Science, and Scopus databases from inception to August 2020 using ("Ankylosing spondyl* " OR "Bechterew Disease" OR "Rheumatoid Spondylitis") AND ((neutrophil* AND lymphocyte*) OR NLR) as key terms of the search strategy. Data selection and extraction were conducted separately by two authors. We appraised the included articles according to the Joanna Briggs checklist. Comprehensive Meta-analysis Version 2 was used for analysis and assessment of publication bias. I2 > 75% and p-value < 0.05 were considered significant. RESULT: In total, 182 studies resulted from a search in all databases. Duplicate removal, title, abstract, and full-text screening yielded 12 related studies, with 11 included in the meta-analysis. Quality assessment was satisfying in all studies. Pooled difference in NLR means value between patients and controls was 0.38 (95% CI: 0.24-0.52, p-value <0.0001). An I2 of 51% and a Cochran Q test p-value of <0.05 indicated moderate heterogeneity; thus, subgroup analysis had no indication. Publication bias was not significant (Funnel plot with an Egger's intercept of -0.07; p-value=0.95). CONCLUSION: Significant higher amounts of NLR may be strongly indicative of underlying inflammation in AS.


Assuntos
Artrite Reumatoide , Espondilite Anquilosante , Biomarcadores , Humanos , Linfócitos , Neutrófilos
5.
Int Immunopharmacol ; 86: 106690, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32585607

RESUMO

BACKGROUND: Sublingual immunotherapy (SLIT) was introduced to deliver allergens in an effective and non-invasive route, which can be considered as an alternative for allergen-specific subcutaneous immunotherapy (SCIT). On the other hand, the use of gold nanoparticles (AuNPs) in allergen delivery has beneficial effects on sublingual immunotherapy. In addition, the molecular targeting agents like aptamers (Apt), have been widely applied for targeted drug delivery. Therefore, the current study aimed to evaluate the effects of dendritic cells (DCs)-specific Aptamer-modified AuNPs coated with ovalbumin (OVA) on the improvement of the SLIT outcome in the mouse model of allergy. MATERIAL AND METHODS: AuNPs with approximately 15 nm diameter were prepared by citrate reduction of HAuCl4. Afterward, Apt-modified AuNP complex was prepared and OVA was then loaded onto this complex. Following sensitization of Balb/c mice to OVA, SLIT was performed with Apt-AuNPs containing 5 µg OVA twice a week for a 2-month period. Allergen-specific IgE in serum, as well as cytokines secretion of spleen cells, were analyzed using ELISA. Also, nasopharyngeal lavage Fluid (NALF) was collected for total and eosinophil counts. Moreover, the lungs were removed for histopathological examination. RESULTS: SLIT with Apt-modified AuNPs complex containing 5 µg OVA, decreased the IgE levels compared to the other groups. Also, IL-4 production has significantly decreased in spleen cells, while TGF-ß and IFN-γ have significantly increased. The assessment of NALF in the group treated by this complex showed a decrease in total cell as well as in eosinophil count. Also, the examination of lung tissues revealed that, in the group treated by this complex, inflammation and perivascular infiltration were lesser than the other groups, which were observed in only one vessel of tissue. CONCLUSION: It was shown that, Sublingual immunotherapy with DC specific Apt-modified AuNPs containing 5 µg OVA can improve the Th1 and Treg immunomodulatory responses.


Assuntos
Materiais Biocompatíveis/administração & dosagem , Células Dendríticas/metabolismo , Portadores de Fármacos/administração & dosagem , Ouro/administração & dosagem , Hipersensibilidade/tratamento farmacológico , Nanopartículas Metálicas/administração & dosagem , Imunoterapia Sublingual/métodos , Alérgenos/administração & dosagem , Alérgenos/química , Alérgenos/imunologia , Animais , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/uso terapêutico , Materiais Biocompatíveis/química , Citocinas/metabolismo , Células Dendríticas/imunologia , Modelos Animais de Doenças , Portadores de Fármacos/química , Eosinófilos/metabolismo , Feminino , Ouro/química , Imunoglobulina E/sangue , Pulmão/patologia , Nanopartículas Metálicas/química , Camundongos Endogâmicos BALB C , Líquido da Lavagem Nasal/imunologia , Ovalbumina/administração & dosagem , Ovalbumina/química , Ovalbumina/imunologia , Baço/imunologia , Linfócitos T Reguladores/metabolismo , Células Th1/metabolismo
6.
Protein Pept Lett ; 27(7): 604-613, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31721685

RESUMO

BACKGROUND: The engineered chimeric peptides including functional multi-epitope structures fused by various peptide linkers are widely applied in biotechnological research to improve the expression level and biological activity of chimera. OBJECTIVE: The aim of our study was to evaluate the effect of helical and flexible linkers on solubility, expression level and folding of multi-epitope chimera containing four epitopes of Human T Lymphotropic Virus Type 1 (HTLV-1). METHODS: For this purpose, the chimera sequences connected by the helical or flexible linker were inserted into different plasmid vectors and expressed in E. coli strains. The expressed products were analyzed using SDS-PAGE and Western blot techniques. Additionally, the molecular modeling study of the chimera with helical or flexible linker was performed using iterative threading assembly refinement (I-TASSER) to attain their three-dimensional structures. RESULTS: Comparison of the chimera expression indicated that the insertion of a flexible (GGGGS)3 linker among chimera epitopes could significantly enhance the level of expression, whereas, the low-level of chimera expression was observed for chimera containing the contiguous helical (EAAAK)5 linker. According to the results of sequence alignment and plasmid stability test, the structure and function of a consecutive helical linker among chimera epitopes were similar to porins as the outer-membrane pore-forming proteins. The molecular modeling results confirmed our experimental study. CONCLUSION: This investigation illustrated the key role of linker design in determining the expression level of multi-epitope chimera and conformational folding.


Assuntos
Epitopos , Vírus Linfotrópico T Tipo 1 Humano/genética , Engenharia de Proteínas , Proteínas Recombinantes de Fusão , Proteínas Virais , Epitopos/biossíntese , Epitopos/química , Epitopos/genética , Humanos , Estrutura Secundária de Proteína , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Virais/biossíntese , Proteínas Virais/química , Proteínas Virais/genética
7.
Arch Sex Behav ; 47(8): 2287-2298, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30128981

RESUMO

Disorders of sex development (DSD) are congenital conditions in which the typical genetic and hormonal profiles are affected and thereby the usual process of sexual differentiation. Most of these studies, however, have been conducted in Western countries. In the present study, preschool sex-typed activities of Iranian individuals with DSD and their age-matched non-affected male and female relatives were assessed using the Pre-School Activities Inventory (PSAI) modified for retrospective self-report. A total of 192 individuals participated in our study, including 33 46,XX individuals with congenital adrenal hyperplasia (CAH; M age = 10.36, SD = 5.52), 15 46,XY individuals with complete androgen insensitivity syndrome (CAIS; M age = 19.8, SD = 7.14), and 16 46,XY individuals with 5-alpha reductase deficiency type-2 (5α-RD-2; M age = 17.31, SD = 7.28), as well as one age-matched non-affected male and female relative for each patient. With regard to PSAI scores, male-identifying participants with 5α-RD-2 and male controls reported similar levels of male-typical childhood play. Female-identifying participants with 5α-RD-2 and CAH showed comparable scores: significantly less masculine and more feminine than male controls, but significantly more masculine and less feminine than females with CAIS and female controls. These findings support the role of androgens in the development of sex-typical childhood play behavior, with those being exposed to higher levels of fetal functional androgens expressing more masculine behavior at preschool ages.


Assuntos
Comportamento Infantil , Identidade de Gênero , Caracteres Sexuais , Desenvolvimento Sexual , 3-Oxo-5-alfa-Esteroide 4-Desidrogenase/deficiência , 3-Oxo-5-alfa-Esteroide 4-Desidrogenase/genética , 3-Oxo-5-alfa-Esteroide 4-Desidrogenase/metabolismo , Adolescente , Hiperplasia Suprarrenal Congênita/genética , Hiperplasia Suprarrenal Congênita/metabolismo , Hiperplasia Suprarrenal Congênita/fisiopatologia , Adulto , Síndrome de Resistência a Andrógenos/genética , Síndrome de Resistência a Andrógenos/metabolismo , Síndrome de Resistência a Andrógenos/fisiopatologia , Androgênios/metabolismo , Criança , Pré-Escolar , Transtorno 46,XY do Desenvolvimento Sexual/genética , Transtorno 46,XY do Desenvolvimento Sexual/metabolismo , Transtorno 46,XY do Desenvolvimento Sexual/fisiopatologia , Feminino , Humanos , Hipospadia/genética , Hipospadia/metabolismo , Hipospadia/fisiopatologia , Irã (Geográfico) , Masculino , Estudos Retrospectivos , Autorrelato , Diferenciação Sexual , Erros Inatos do Metabolismo de Esteroides/genética , Erros Inatos do Metabolismo de Esteroides/metabolismo , Erros Inatos do Metabolismo de Esteroides/fisiopatologia
8.
Ann Vasc Surg ; 49: 219-228, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29421414

RESUMO

To date, there is still no treatment protocol for patients with thromboangiitis obliterans (TAO) who are also afflicted with critical limb ischemia (CLI). Smoking cessation on its own cannot be considered a treatment for the purposes of salvaging a limb of a TAO patient with CLI. The aim of this review was to evaluate different studies of various treatment protocols for avoiding amputation in TAO patients. A systematic search for relevant studies dating from 1990 to the end of 2016 was performed on the PubMed, SCOPUS, and Science Direct databases. Only 24 studies fulfilled the inclusion criteria, of which only one was a randomized controlled trial (RCT). The remaining studies were quasi-experimental with various treatments and follow-up durations. Therefore, meta-analysis was not performed. Judging from the major amputation rates after the suggested treatments were performed, no treatment was particularly effective. This review demonstrated that more standard RCTs are needed to resolve this treatment issue involved in TAO. In addition, because health insurance coverage for TAO patients differs by country, regional cost-benefit and cost-efficacy studies of the suggested treatments for TAO are highly recommended.


Assuntos
Fármacos Cardiovasculares/uso terapêutico , Salvamento de Membro/métodos , Estimulação da Medula Espinal , Transplante de Células-Tronco , Simpatectomia , Tromboangiite Obliterante/terapia , Procedimentos Cirúrgicos Vasculares/métodos , Adulto , Amputação Cirúrgica , Fármacos Cardiovasculares/efeitos adversos , Humanos , Salvamento de Membro/efeitos adversos , Pessoa de Meia-Idade , Fatores de Risco , Estimulação da Medula Espinal/efeitos adversos , Transplante de Células-Tronco/efeitos adversos , Simpatectomia/efeitos adversos , Tromboangiite Obliterante/diagnóstico , Tromboangiite Obliterante/cirurgia , Resultado do Tratamento , Procedimentos Cirúrgicos Vasculares/efeitos adversos
9.
Int J Biometeorol ; 61(1): 1-9, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27255304

RESUMO

Air pollutants and their interaction with environmental allergens have been considered as an important reason for the recent increase in the prevalence of allergic diseases. The aim of this study was to investigate the traffic pollution effect, as a stressor, on Platanus orientalis pollen allergens messenger RNA (mRNA) and protein expression. P. orientalis pollen grains were collected along main streets of heavy traffic and from unpolluted sites in Mashhad city, in northeast Iran. The pollen samples were examined by scanning electron microscopy. To assess the abundance of pollen allergens (Pla or 1, Pla or 2, and Pla or 3) from polluted and unpolluted sites, immunoblotting was performed. Moreover, the sequences encoding P. orientalis allergens were amplified using real-time PCR. Scanning electron microscopy showed a number of particles of 150-550 nm on the surface of pollen from polluted sites. Also, protein and gene expression levels of Pla or 1 and Pla or 3 were considerably greater in pollen samples from highly polluted areas than in pollen from unpolluted areas (p < 0.05). In contrast, no statically significant difference in Pla or 2 protein and mRNA expression level was found between samples from the two areas. We found greater expression of allergens involved in plant defense mechanisms (Pla or 1 and Pla or 3) in polluted sites than in unpolluted ones. The high expression of these proteins can lead to an increase in the prevalence of allergic diseases. These findings suggest the necessity of supporting public policies aimed at controlling traffic pollution to improve air quality and prevent the subsequent clinical outcomes and new cases of asthma.


Assuntos
Poluentes Atmosféricos/análise , Alérgenos , Magnoliopsida , Proteínas de Plantas , Pólen , Poluição do Ar/análise , Alérgenos/genética , Alérgenos/imunologia , Alérgenos/ultraestrutura , Animais , Anticorpos/imunologia , Cidades , DNA de Plantas/genética , Escherichia coli/genética , Feminino , Irã (Geográfico) , Microscopia Eletrônica de Varredura , Veículos Automotores , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Pólen/genética , Pólen/imunologia , Pólen/ultraestrutura , Coelhos , Proteínas Recombinantes/genética
10.
Med Arch ; 70(5): 389-391, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27994303

RESUMO

AIM: An accessory spleen is an additional tissue of the spleen that may be found near the spleen. It is a congenital anomaly of the spleen that its incidence has been reported 10-30% of the population. Hence, the objective of this study was evaluation the incidence of accessory spleen in Iranian cadavers. METHOD: Sixty hundred and ninety three spleens (541 males, 152 females) were excised from cadavers in the dissection hall of Mashhad Forensic Medicine Organization cadavers. Inclusion criteria were as follows: Fresh Iranian cadavers with no history of alcohol, poisoning or drug abuse, and no evidence of pathologic abnormality or injury to the spleen. The presence of accessory spleens, its dimension and weight investigated in cadavers. RESULTS: During routine postmortem examination, five cases with an accessory spleen were found in the autopsy laboratory of Mashhad legal Medicine Organization between June 2014 and July 2015. Of the cases, 3 were male and 2 were female. The accessory spleens were observed at the splenic hilum. The length of the accessory spleens ranged from 2-3.5 cm, while the range of width was between 0.5 and 2.5 cm. The accessory spleens were confirmed by histological examination. CONCLUSION: An accessory spleen has clinical importance in some locations. When an accessory spleen is situated in another site, it may mimic some tumors such as pancreatic tumor and adrenal tumor. In addition, accessory spleen may cause hyperplasia after splenectomy and be responsible for a recurrence of the hematological disorders.


Assuntos
Baço/anormalidades , Adulto , Autopsia , Feminino , Humanos , Irã (Geográfico) , Masculino , Pessoa de Meia-Idade , Adulto Jovem
11.
Rep Biochem Mol Biol ; 5(1): 40-45, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-28070533

RESUMO

BACKGROUND: Platanus species are widely cultured around the world and considered an important cause of allergic reactions. In the present study, we developed a sandwich ELISA to quantify Pla or 3 allergen in P. orientalis pollen extracts grown near high-traffic roads and compared it to pollen extracts collected from rural areas as control. METHODS: Pollen samples were collected from three polluted and two unpolluted sites in Mashhad, northeast Iran. Recombinant Pla or 3 was expressed and used for polyclonal antibody production in rabbit. A sandwich ELISA was developed and validated to quantify Pla or 3 levels in pollen extracts from the different sites. RESULTS: The coefficients of variation (CVs) for the intra- and inter- day assays were less than 5 and 18%, respectively. The working range of the standard curve was between 0.1 and 25 ng/ml, with the detection limit being 0.037 ng/ml. The recovery percentage was 88-106.4% at working concentrations from 0.31 to 26.5 ng/ml. Pla or 3 levels were significantly greater in pollens grown near high-traffic roads than in those grown in rural regions (p < 0.0001). CONCLUSION: A sandwich ELISA was developed and validated to quantify Pla or 3 in pollen extracts. Using this validated ELISA, we showed a substantial difference between the amounts of Pla or 3 in pollens grown in different environments. This finding should be considered in developing public policies to reduce traffic pollution, which leads to reduced allergic reactions in atopic subjects.

13.
Int J High Risk Behav Addict ; 2(2): 87-91, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24971281

RESUMO

BACKGROUND: Quality of life (QoL) stands for to the general well-being of the populations in the societies. Quality of life should not be confused with the concept of standard of living, which is based primarily on income. Instead, standard indicators of the QOL include not only wealth and employment, but also built environment, physical and mental health, education, recreation and leisure time and social belonging. OBJECTIVES: This study attempted to evaluate the QoL of the young addicted women compared to non-addict controls. MATERIALS AND METHODS: This case-control study was conducted on young addict women aged between 16-25 years old using with BREF-WHOQOL questionnaire. The subgroups included vulnerable addicts, non-vulnerable addicts and healthy controls. RESULTS: The findings of the current study demonstrated that drug abuse led to reduce health-related QoL in all aspects of health compared with control group. CONCLUSIONS: Health-related QoL was lower in all aspects of health compared with control group.

14.
Mol Biol Rep ; 39(3): 3169-78, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21713408

RESUMO

Chenopodium album pollen is one of the main sources of pollen allergy in desert and semi-desert areas and contains three identified allergens, so the aim of this study is comparison of the diagnostic potential of C. album recombinant allergens in an allergenic cocktail and C. album pollen extract. Diagnostic potential of the allergenic cocktail was investigated in 32 individuals using skin prick test and obtained results were compared with the acquired results from C. album pollen extract. Specific IgE reactivity against the pollen extract and allergenic cocktail was determined by ELISA and western blotting tests. Inhibition assays were performed for the allergenic cocktail characterization. The exact sensitization profile of all patients was identified which showed that 72, 81 and 46% of allergic patients had IgE reactivity to rChe a 1, rChe a 2 and rChe a 3, respectively. Almost all of C. album allergic patients (30/32) had specific IgE against the allergenic cocktail. In addition, there was a high correlation between IgE levels against the allergenic cocktail and IgE levels against the pollen extract. The allergenic cocktail was able to completely inhibit IgE binding to natural Che a 1, Che a 2 and Che a 3 in C. album extract. In addition, positive skin test reactions were seen in allergic patients that tested by the allergenic cocktail. The reliable results obtained from this study confirmed that the allergenic cocktail with high diagnostic potential could be replaced with natural C. album allergen extracts in skin prick test and serologic tests.


Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Proteínas de Ligação ao Cálcio/imunologia , Chenopodium album/imunologia , Pólen/imunologia , Proteínas Recombinantes/imunologia , Rinite Alérgica Sazonal/diagnóstico , Adulto , Alérgenos/genética , Antígenos de Plantas/genética , Western Blotting , Proteínas de Ligação ao Cálcio/genética , Primers do DNA/genética , DNA Complementar/genética , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina E/imunologia , Irã (Geográfico) , Masculino , Pessoa de Meia-Idade , Pólen/química , Rinite Alérgica Sazonal/imunologia , Testes Cutâneos/métodos
15.
Allergol Int ; 60(3): 309-16, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21430432

RESUMO

BACKGROUND: Pollinosis from Amaranthus retroflexus pollen is a common cause of respiratory allergy in Iran with a high positive rate (68.8%) among Iranian allergic patients. The aim of the present study was to evaluate the allergenicity of the A. retroflexus pollen profilin. METHODS: Using sera from twelve patients allergic to A. retroflexus pollen, IgE-binding proteins from the A. retroflexus pollen extract was identified by immunoblotting. The cDNA of A. retroflexus pollen profilin was amplified, then cloned into the pET-21b (+) vector, expressed in Escherichia coli, and finally purified by metal affinity chromatography. The IgE-binding capacity of the recombinant protein was then analyzed by the ELISA, immunoblotting, and inhibition assays, as well as by the skin prick test (SPT). RESULTS: Immunoblotting results indicated a 14.6kDa protein with IgE-reactivity to 33% (4/12) among A. retroflexus pollen-allergic patients. Nucleotide sequencing of the cDNA revealed an open reading frame of 399 bp encoding for 133 amino acid residues which was belonged to the profilin family and designated as Ama r 2. A recombinant Ama r 2 (rAma r 2) was then produced in E. coli as a soluble protein which showed a strong IgE-reactivity via ELISA confirmed by the SPT. Inhibition experiments revealed high IgE cross-reactivities with the profilins from other plants. CONCLUSIONS: The profilin from the A. retroflexus pollen, Ama r 2, was firstly identified as an allergen. Moreover, rAma r 2 was produced in E. coli as a soluble immunoreactive protein with an IgE-reactivity similar to that of its natural counterpart.


Assuntos
Alérgenos/imunologia , Amaranthus/imunologia , Pólen/imunologia , Profilinas/imunologia , Adulto , Alérgenos/genética , Alérgenos/metabolismo , Amaranthus/genética , Sequência de Aminoácidos , Reações Antígeno-Anticorpo/imunologia , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Humanos , Hipersensibilidade/etiologia , Hipersensibilidade/imunologia , Hipersensibilidade/metabolismo , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Pólen/genética , Pólen/metabolismo , Profilinas/genética , Profilinas/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Adulto Jovem
16.
Allergol Int ; 60(1): 61-7, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21173569

RESUMO

BACKGROUND: Oral allergy syndrome resulted from plant-derived foods is frequent among adults. Allergy to melon (cucumis melo) is one of the most frequent fruit allergies in Iran. Three different major allergens have been found in Cucumis melo that Cuc m 1 (cucumisin) has been identified as the major allergen of melon. Cucumisin is an alkaline serine protease that it is found as a 78kDa protein in precursor form. The aim of this study was production of recombinant Cuc m 1 in Escherichia coli (E. coli) cells and characterization of its allergenicity property. METHODS: Production of recombinant Cuc m 1 was carried out by cDNA cloning technique into the pET32b(+) vector using specific primers designed based on cucumisin nucleotide sequence available in Genebank database, cucumisin encoding gene and directional cloning method. Cloned plasmid into E. coli TOP10 was transformed into E. coli BL21 and expression of the protein was induced by IPTG. The recombinant protein was purified via Ni-NTA affinity chromatography using histidine tag in recombinant protein. IgE binding of this protein was assessed by IgE-immunoblotting, ELISA and inhibition ELISA. RESULTS: The directional cloning was resulted in expression of a fusion Cuc m 1. Immunoblotting with sera of patients allergic to melon showed strong reactivity with purified protein band. Inhibition assays demonstrated that purified rCuc m 1 could be the same with natural form of Cuc m 1 in total extract. CONCLUSIONS: In the present study, we have provided a functional recombinant cucumisin allergen, rCuc m 1 with 86kDa, which may be used as a standard allergen for clinical diagnosis and study of allergy to melon.


Assuntos
Cucumis melo/enzimologia , Escherichia coli/genética , Escherichia coli/metabolismo , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Subtilisinas , Clonagem Molecular , Cucumis melo/genética , Hipersensibilidade Alimentar/imunologia , Humanos , Imunoglobulina E/imunologia , Subtilisinas/metabolismo
17.
Allergol Int ; 58(3): 429-35, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19628979

RESUMO

BACKGROUND: Profilin is a panallergen that is recognized by IgE in allergic patients. Allergy to saffron (Crocus sativus) pollen has been described in people exposed to its pollen. Saffron contains a profilin that may cause allergic reactions in atopic subjects. The aim of this study was to describe the cloning, expression and purification of saffron profilin from pollen. METHODS: Cloning of saffron profilin was performed by polymerase chain reaction using specific primers from saffron pollen RNA. Expression was carried out in Escherichia coli BL21 (DE3) using a vector pET-102- TOPO. A recombinant fusion protein was expressed and the recombinant profilin was purified by metal precipitation. Immunological characterization was performed by immunoblotting experiments. RESULTS: The 34kDa- recombinant saffron profilin, Cro s 2, as a fusion protein was purified. Immunoblotting tested with the sera of allergic patients showed a specific reaction with the recombinant Cro s 2 band. CONCLUSIONS: The sequence of Cro s 2 showed a high degree of identity and similarity to other plant profilins and the recombinant saffron profilin, Cro s 2, may be used for target-specific diagnosis and structural analyses and investigation of cross reactivity of Cro s 2 with other plant profilins.


Assuntos
Antígenos de Plantas/biossíntese , Crocus/imunologia , Proteínas de Plantas/genética , Profilinas/biossíntese , Sequência de Aminoácidos , Antígenos de Plantas/química , Antígenos de Plantas/genética , Clonagem Molecular , Crocus/genética , Escherichia coli/metabolismo , Dados de Sequência Molecular , Peso Molecular , Proteínas de Plantas/biossíntese , Proteínas de Plantas/química , Pólen/imunologia , Profilinas/química , Profilinas/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Alinhamento de Sequência
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