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Pollen from common ragweed is an important allergen source worldwide and especially in western and southern Romania. More than 100 million patients suffer from symptoms of respiratory allergy (e.g., rhinitis, asthma) to ragweed pollen. Among the eleven characterized allergens, Amb a 6 is a non-specific lipid transfer protein (nsLTP). nsLTPs are structurally stable proteins in pollen and food from different unrelated plants capable of inducing severe reactions. The goal of this study was to produce Amb a 6 as a recombinant and structurally folded protein (rAmb a 6) and to characterize its physicochemical and immunological features. rAmb a 6 was expressed in Spodoptera frugiperda Sf9 cells as a secreted protein and characterized by mass spectrometry and circular dichroism (CD) spectroscopy regarding molecular mass and fold, respectively. The IgE-binding frequency towards the purified protein was evaluated using sera from 150 clinically well-characterized ragweed-allergic patients. The allergenic activities of rAmb a 6 and the nsLTP from the weed Parietaria judaica (Par j 2) were evaluated in basophil activation assays. rAmb a 6-specific IgE reactivity was associated with clinical features. Pure rAmb a 6 was obtained by insect cell expression. Its deduced molecular weight corresponded to that determined by mass spectrometry (i.e., 10,963 Da). rAmb a 6 formed oligomers as determined by SDS-PAGE under non-reducing conditions. According to multiple sequence comparisons, Amb a 6 was a distinct nsLTP with less than 40% sequence identity to currently known plant nsLTP allergens, except for nsLTP from Helianthus (i.e., 52%). rAmb a 6 is an important ragweed allergen recognized by 30% of ragweed pollen allergic patients. For certain patients, rAmb a 6-specific IgE levels were higher than those specific for the major ragweed allergen Amb a 1 and analysis also showed a higher allergenic activity in the basophil activation test. rAmb a 6-positive patients suffered mainly from respiratory symptoms. The assumption that Amb a 6 is a source-specific ragweed allergen is supported by the finding that none of the patients showing rAmb a 6-induced basophil activation reacted with Par j 2 and only one rAmb a 6-sensitized patient had a history of plant food allergy. Immunization of rabbits with rAmb a 6 induced IgG antibodies which strongly inhibited IgE binding to rAmb a 6. Our results demonstrate that Amb a 6 is an important source-specific ragweed pollen allergen that should be considered for diagnosis and allergen-specific immunotherapy of ragweed pollen allergy.
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Alérgenos , Antígenos de Plantas , Proteínas de Transporte , Imunoglobulina E , Humanos , Alérgenos/imunologia , Imunoglobulina E/imunologia , Antígenos de Plantas/imunologia , Antígenos de Plantas/química , Animais , Proteínas de Transporte/imunologia , Proteínas de Transporte/metabolismo , Proteínas de Plantas/imunologia , Proteínas de Plantas/química , Feminino , Rinite Alérgica Sazonal/imunologia , Masculino , Adulto , Ambrosia/imunologia , Spodoptera/imunologia , Proteínas Recombinantes/imunologia , Sequência de Aminoácidos , Células Sf9 , Pessoa de Meia-Idade , Extratos VegetaisRESUMO
Ragweed pollen allergy is the most common seasonal allergy in western Romania. Prolonged exposure to ragweed pollen may induce sensitization to pan-allergens such as calcium-binding proteins (polcalcins) and progression to more severe symptoms. We aimed to detect IgE sensitization to recombinant Amb a 9 and Amb a 10 in a Romanian population, to assess their potential clinical relevance and cross-reactivity, as well as to investigate the relation with clinical symptoms. rAmb a 9 and rAmb a 10 produced in Escherichia coli were used to detect specific IgE in sera from 87 clinically characterized ragweed-allergic patients in ELISA, for basophil activation experiments and rabbit immunization. Rabbit rAmb a 9- and rAmb a 10-specific sera were used to detect possible cross-reactivity with rArt v 5 and reactivity towards ragweed and mugwort pollen extracts. The results showed an IgE reactivity of 25% to rAmb a 9 and 35% to rAmb a 10. rAmb a 10 induced basophil degranulation in three out of four patients tested. Moreover, polcalcin-negative patients reported significantly more skin symptoms, whereas polcalcin-positive patients tended to report more respiratory symptoms. Furthermore, both rabbit antisera showed low reactivity towards extracts and showed high reactivity to rArt v 5, suggesting strong cross-reactivity. Our study indicated that recombinant ragweed polcalcins might be considered for molecular diagnosis.
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Proteínas de Ligação ao Cálcio , Reações Cruzadas , Imunoglobulina E , Rinite Alérgica Sazonal , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Reações Cruzadas/imunologia , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/sangue , Romênia , Proteínas de Ligação ao Cálcio/imunologia , Antígenos de Plantas/imunologia , Alérgenos/imunologia , Feminino , Masculino , Ambrosia/imunologia , Coelhos , Adulto , Extratos VegetaisRESUMO
Currently, allergen-specific immunotherapy (AIT) for ragweed allergy is still based on natural allergen extracts. This study aimed to analyse the ability of four commercially available AIT vaccines (CLUSTOID, TYRO-SIT, POLLINEX Quattro Plus and Diater Depot) regarding their ability to induce IgG antibodies against ragweed pollen allergens in rabbits. Accordingly, the IgG reactivity of AIT-induced rabbit sera was tested for ten different ragweed pollen allergens (Amb a 1, 3, 4, 5, 6, 8, 9, 10, 11 and 12) by an ELISA. Furthermore, the ability of rabbit AIT-specific sera to block allergic patients' IgE binding to relevant ragweed allergens (Amb a 1, 4, 6, 8 and 11) and to inhibit allergen-induced basophil activation was evaluated by an IgE inhibition ELISA and a mediator release assay. Only two AIT vaccines (Diater Depot > CLUSTOID) induced relevant IgG antibody levels to the major ragweed allergen Amb a 1. The IgG responses induced by the AIT vaccines against the other ragweed allergens were low and highly heterogeneous. Interestingly, the kinetics of IgG responses were different among the AIT vaccines and even within one AIT vaccine (Diater Depot) for Amb a 1 (long-lasting) versus Amb a 8 and Amb a 11 (short-lived). This could be due to variations in allergen contents, the immunogenicity of the allergens, and different immunization protocols. The IgE inhibition experiments showed that rabbit AIT-specific sera containing high allergen-specific IgG levels were able to inhibit patients' IgE binding and prevent the mediator release with Diater Depot. The high levels of allergen-specific IgG levels were associated with their ability to prevent the recognition of allergens by patients' IgE and allergen-induced basophil activation, indicating that the measurement of allergen-induced IgG could be a useful surrogate marker for the immunological efficacy of vaccines. Accordingly, the results of our study may be helpful for the selection of personalized AIT vaccination strategies for ragweed-allergic patients.
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Common ragweed pollen allergy has become a health burden worldwide. One of the major allergens in ragweed allergy is Amb a 1, which is responsible for over 90% of the IgE response in ragweed-allergic patients. The major allergen isoform Amb a 1.01 is the most allergenic isoform in ragweed pollen. So far, no recombinant Amb a 1.01 with similar allergenic properties to its natural counterpart (nAmb a 1.01) has been produced. Hence, this study aimed to produce a recombinant Amb a 1.01 with similar properties to the natural isoform for improved ragweed allergy management. Amb a 1.01 was expressed in insect cells using a codon-optimized DNA construct with a removable N-terminal His-Tag (rAmb a 1.01). The recombinant protein was purified by affinity chromatography and physicochemically characterized. The rAmb a 1.01 was compared to nAmb a 1.01 in terms of the IgE binding (enzyme-linked immunosorbent assay (ELISA), immunoblot) and allergenic activity (mediator release assay) in well-characterized ragweed-allergic patients. The rAmb a 1.01 exhibited similar IgE reactivity to nAmb a 1.01 in different IgE-binding assays (i.e., IgE immunoblot, ELISA, quantitative ImmunoCAP inhibition measurements). Furthermore, the rAmb a 1.01 showed comparable dose-dependent allergenic activity to nAmb a 1.01 regarding basophil activation. Overall, the results showed the successful expression of an rAmb a 1.01 with comparable characteristics to the corresponding natural isoform. Our findings provide the basis for an improvement in ragweed allergy research, diagnosis, and immunotherapy.
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Alérgenos , Ambrosia , Antígenos de Plantas , Imunoglobulina E , Proteínas Recombinantes , Humanos , Antígenos de Plantas/imunologia , Antígenos de Plantas/genética , Antígenos de Plantas/química , Imunoglobulina E/imunologia , Animais , Alérgenos/imunologia , Alérgenos/genética , Ambrosia/imunologia , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/genética , Feminino , Adulto , Proteínas de Plantas/imunologia , Proteínas de Plantas/genética , Proteínas de Plantas/química , Rinite Alérgica Sazonal/imunologia , Masculino , Pessoa de Meia-Idade , Extratos Vegetais/químicaRESUMO
Patients with schizophrenia tend to have deficits in emotion recognition (ER) that affect their social function. However, the commonly-used ER measures appear incomprehensive, unreliable and invalid, making it difficult to comprehensively evaluate ER. The purposes of this study were to develop the Computerized Emotion Recognition Video Test (CERVT) evaluating ER ability in patients with schizophrenia. This study was divided into two phases. First, we selected candidate CERVT items/videos of 8 basic emotion domains from a published database. Second, we validated the selected CERVT items using Rasch analysis. Finally, the 269 patients and 177 healthy adults were recruited to ensure the participants had diverse abilities. After the removal of 21 misfit (infit or outfit mean square > 1.4) items and adjustment of the item difficulties of the 26 items with severe differential item functioning, the remaining 217 items were finalized as the CERVT items. All the CERVT items showed good model fits with small eigenvalues (≤ 2) based on the residual-based principal components analysis for each domain, supporting the unidimensionality of these items. The 8 domains of the CERVT had good to excellent reliabilities (average Rasch reliabilities = 0.84-0.93). The CERVT contains items of the 8 basic emotions with individualized scores. Moreover, the CERVT showed acceptable reliability and validity, and the scores were not affected by examinees' gender. Thus, the CERVT has the potential to provide a comprehensive, reliable, valid, and gender-unbiased assessment of ER for patients with schizophrenia.
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Emoções , Psicometria , Esquizofrenia , Humanos , Masculino , Feminino , Adulto , Emoções/fisiologia , Esquizofrenia/diagnóstico , Psicometria/normas , Psicometria/instrumentação , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Psicologia do Esquizofrênico , Reconhecimento Psicológico/fisiologia , Testes Neuropsicológicos/normas , Adulto JovemRESUMO
BACKGROUND: Ragweed is an invasive plant in Europe, causing hay fever and asthma in allergic patients. Climate change is predicted to increase expansion and allergenicity. Elevated NO2 induced upregulation of a new allergen in ragweed pollen, an enolase, Amb a 12. OBJECTIVE: of this study was producing ragweed enolase as a recombinant protein and characterizing its physicochemical and immunological features. METHODS: Amb a 12 was designed for E. coli and insect cell expression. Physicochemical features were determined by mass spectrometry, circular dichroism measurements and enzymatic activity assay. Immunological characteristics were determined in ELISA, in a mediator release assay and by investigation of association with clinical symptoms. Common allergen sources were screened for similar proteins. RESULTS: Ragweed enolase was produced as a 48 kDa protein forming oligomers in both expression systems, showing differences in secondary structure content and enzymatic activity depending on expression system. IgE frequency and allergenicity were low regardless of expression system. Enolase-specific serum bound to similar sized molecules in mugwort, timothy grass and birch pollen, as well as food allergen sources, while highest IgE inhibition was achieved with peach pulp extract. CONCLUSIONS: Amb a 12 had high sequence similarity and comparable IgE frequency to enolase allergens from different sources. 50 kDa proteins were found in other pollen and food allergen sources, suggesting that enolases might be pan-allergens in pollen and plant foods.
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Ambrosia , Proteínas de Plantas , Humanos , Escherichia coli , Imunoglobulina E , Alérgenos , Antígenos de Plantas , Pólen , Fosfopiruvato Hidratase/análiseRESUMO
Ragweed (Ambrosia artemisiifolia) pollen is a major endemic allergen source responsible for severe allergic manifestations in IgE-sensitized allergic patients. It contains the major allergen Amb a 1 and cross-reactive allergen molecules, such as the cytoskeletal protein profilin, Amb a 8 and calcium-binding allergens Amb a 9 and Amb a 10. To assess the importance of Amb a 1, profilin and calcium-binding allergen, the IgE reactivity profiles of clinically well-characterized 150 ragweed pollen-allergic patients were analysed regarding specific IgE levels for Amb a 1 and cross-reactive allergen molecules by quantitative ImmunoCAP measurements, IgE ELISA and by basophil activation experiments. By quantifying allergen-specific IgE levels we found that Amb a 1-specific IgE levels accounted for more than 50% of ragweed pollen-specific IgE in the majority of ragweed pollen-allergic patients. However, approximately 20% of patients were sensitized to profilin and the calcium-binding allergens, Amb a 9 and Amb a 10, respectively. As shown by IgE inhibition experiments, Amb a 8 showed extensive cross-reactivity with profilins from birch (Bet v 2), timothy grass (Phl p 12) and mugwort pollen (Art v 4) and was identified as a highly allergenic molecule by basophil activation testing. Our study indicates that molecular diagnosis performed by the quantification of specific IgE to Amb a 1, Amb a 8, Amb a 9 and Amb a 10 is useful to diagnose genuine sensitization to ragweed pollen and to identify patients who are sensitized to highly cross-reactive allergen molecules present in pollen from unrelated plants, in order to enable precision medicine-based approaches for the treatment and prevention of pollen allergy in areas with complex pollen sensitization.
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Alérgenos , Hipersensibilidade , Humanos , Alérgenos/química , Profilinas , Cálcio , Proteínas de Plantas , Antígenos de Plantas , Extratos Vegetais , Reações Cruzadas , Imunoglobulina E/metabolismo , Ambrosia/metabolismoRESUMO
Wastewater-based epidemiology (WBE) represents an efficient approach for public pathogen surveillance as it provides early warning of disease outbreaks; however, it has not yet been applied to dengue virus (DENV), which might cause endemics via mosquito spread. In this study, a working platform was established to provide direct virus recovery and qPCR quantification from wastewater samples that were artificially loaded with target DENV serotypes I to IV and noncognate spike control viral particles. The results showed qPCR efficiencies of 91.2 %, 94.8 %, 92.6 % and 88.7 % for DENV I, II, III, and IV, respectively, and a broad working range over 6 orders of magnitude using the preferred primer sets. Next, the results revealed that the ultrafiltration method was superior to the skimmed milk flocculation method for recovering either DENV or control viral particles from wastewater. Finally, DENV-2 was loaded simultaneously with the noncognate spike control and could be recovered at comparable levels either in PBS or in wastewater, indicating the applicability of noncognate spike control particles to reflect the efficiency of experimental steps. In conclusion, our data suggest that DENV particles in wastewater could be recovered and quantitatively detected in absolute amounts, indicating the feasibility of DENV surveillance using the WBE approach.
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Culicidae , Vírus da Dengue , Dengue , Animais , Vírus da Dengue/genética , Dengue/diagnóstico , Águas Residuárias , Surtos de Doenças , SorogrupoRESUMO
Ragweed pollen is highly allergenic and elicits type I hypersensitivity reactions in the exposed populations. Amb a 11 is a recently discovered component of this pollen, and its biological role in allergy is still being researched. In our study, ragweed allergy patients were recruited prospectively over a three-year period; a comprehensive questionnaire was administered, and sera were collected and stored. The production of recombinant Amb a 11 was achieved in parallel with patients' recruitment. The gene coding for mature protein was inserted in E. coli and in Sf9 Spodoptera frugiperda cells. The recombinant allergens (designated eAmb a 11 and iAmb a 11) were tested for His-tag presence in Western blot. IgE reactivity was evaluated in 150 patients' sera for both recombinant allergen forms in ELISA, with 5 positive sera being tested further by hRBL (humanized rat basophilic leukemia) hexosaminidase release assay. Both allergen forms were proven to be IgE-reactive His-tagged proteins, with an extensive overlap of positive sera (92 toward the former recombinant allergen, 100 toward the latter) and an overall Amb a 11 sensitization prevalence estimated at 68.67%. The hRBL mediator release assay revealed a significant, slightly weaker effect of recombinant allergens when compared with nAmb a 1. Sensitization to this major allergen appears to be associated with more severe asthma symptoms (OR = 4.71, 95% CI = 1.81-12.21). In conclusion, recombinant Amb a 11 is a bona fide allergen, which is IgE-reactive and an inducer of hRBL degranulation. It is an important IgE-reactive component from ragweed pollen, with high IgE sensitization prevalence in the sample population and allergenicity of the recombinant allergen comparable to Amb a 1.
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Asma , Hipersensibilidade , Ratos , Animais , Alérgenos/genética , Ambrosia/metabolismo , Proteínas de Plantas/metabolismo , Escherichia coli/metabolismo , Imunoglobulina E , Proteínas Recombinantes/genéticaRESUMO
Background: Ragweed (Ambrosia artemisiifolia) is one of the most important allergen sources, worldwide, causing severe respiratory allergic reactions in late summer and fall, in sensitized patients. Amb a 1 has been considered as the most important allergen in ragweed but 12 ragweed pollen allergens are known. The aim of our study was to investigate IgE reactivity profiles of ragweed allergic patients and to associate them with clinical symptoms. Methods: IgE sensitization profiles from clinically well-characterized ragweed allergic patients (n = 150) were analyzed using immunoblotted ragweed pollen extract. Immunoblot inhibition experiments were performed with two Amb a 1 isoforms and CCD markers and basophil activation experiments were performed with IgE serum before and after depletion of Amb a 1-specific IgE. Results: By IgE-immunoblotting 19 different IgE reactivity patterns with and without Amb a 1-sensitization were found. The majority of patients (>95%) suffered from rhino-conjunctivitis, around 60% reported asthma-like symptoms and about 25% had skin reactions. Patients with complex IgE sensitization profiles tended to have more clinical symptoms. Serum with and without Amb a 1-specific IgE induced basophil activation. Conclusions: Ragweed pollen allergic patients exhibit complex IgE reactivity profiles to ragweed allergens including Amb a 1 isoforms and cross-reactive carbohydrates indicating the importance of Amb a 1 isoforms and additional allergens for diagnosis and allergen-specific immunotherapy of ragweed allergy.
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IMPORTANCE: Patients with schizophrenia tend to have severe deficits in theory of mind, which may limit their interpretation of others' behaviors and thereby hamper social participation. Commonly used measures of theory of mind assess the ability to understand various social situations (e.g., implied meaning or hinting, faux pas), but these measures do not yield valid, reliable, and gender unbiased results to inform interventions for managing theory-of-mind deficits. We used understanding of implied meaning, which appears to be a unidimensional construct highly correlated with social competence, as a promising starting point to develop a theory-of-mind assessment. OBJECTIVE: To develop a Rasch-calibrated computerized test of implied meaning. DESIGN: Cross-sectional design. SETTING: Psychiatric hospitals and community. PARTICIPANTS: 344 participants (240 patients with schizophrenia and 104 healthy adults). RESULTS: We initially developed 27 items for the Computerized Implied Meaning Test. After inappropriate items (12 misfit items and 1 gender-biased item) were removed, the remaining 14 items showed acceptable model fit to the Rasch model (infit = 0.84-1.16; outfit = 0.65-1.34) and the one-factor model (comparative fit index = .91, standardized root mean square residual = .05, root-mean-square error of approximation = .08). Most patients (81.7%) achieved individual Rasch reliability of ≥.90. Healthy participants performed significantly better on the test than patients with schizophrenia (Cohen's d = 2.5, p < .001). CONCLUSIONS AND RELEVANCE: Our preliminary findings suggest that the Computerized Implied Meaning Test may provide reliable, valid, and gender-unbiased results for patients with schizophrenia. What This Article Adds: We developed a new measure for assessing theory-of-mind ability in patients with schizophrenia that consists of items targeting the understanding of implied meaning. Preliminary findings suggest that the Computerized Implied Meaning Test is reliable, valid, and gender unbiased and may be used in evaluating patients' theory-of-mind deficits and relevant factors.
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Esquizofrenia , Adulto , Estudos Transversais , Humanos , Psicometria , Reprodutibilidade dos Testes , Inquéritos e QuestionáriosRESUMO
BACKGROUND: Worldwide, pollen of the weed mugwort (Artemisiavulgaris) is a major cause of severe respiratory allergy, with its major allergen, Art v 1, being the key pathogenic molecule for millions of patients. Humanized mice transgenic for a human T-cell receptor specific for the major Art v 1 T-cell epitope and the corresponding HLA have been made. OBJECTIVE: We sought to characterize IgE epitopes of Art v 1-sensitized patients and humanized mice for molecular immunotherapy of mugwort allergy. METHODS: Four overlapping peptides incorporating surface-exposed amino acids representing the full-length Art v 1 sequence were synthesized and used to search for IgE reactivity to sequential epitopes. For indirect mapping, peptide-specific rabbit antibodies were raised to block IgE against surface-exposed epitopes on folded Art v 1. IgE reactivity and basophil activation studies were performed in clinically defined mugwort-allergic patients. Secondary structure of recombinant (r) Art v 1 and peptides was determined by circular dichroism spectroscopy. RESULTS: Mugwort-allergic patients and humanized mice sensitized by allergen inhalation showed IgE reactivity and/or basophil activation mainly to folded, complete Art v 1 but not to unfolded, sequential peptide epitopes. Blocking of allergic patients' IgE with peptide-specific rabbit antisera identified a hitherto unknown major conformational IgE binding site in the C-terminal Art v 1 domain. CONCLUSIONS: Identification of the new major conformational IgE binding site on Art v 1, which can be blocked with IgG raised against non-IgE reactive Art v 1 peptides, is an important basis for the development of a hypoallergenic peptide vaccine for mugwort allergy.
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Artemisia , Hipersensibilidade , Alérgenos , Aminoácidos , Animais , Antígenos de Plantas , Artemisia/química , Epitopos de Linfócito T , Humanos , Soros Imunes , Imunoglobulina E , Imunoglobulina G , Camundongos , Peptídeos , Proteínas de Plantas , CoelhosRESUMO
OBJECTIVE: To compare the effects of botulinum toxin and steroid septal injections in treating allergic rhinitis (AR) by evaluating improvements in the rhinitis control assessment test (RCAT), visual analog scale (VAS), nasal obstruction symptom evaluation (NOSE) scores, and active anterior rhinomanometry (RMM) measurements. METHODS: This prospective, single-blinded cohort study was conducted at the Department of Otolaryngology, Taipei City Hospital between January 2017 and December 2018. Ninety-five patients were randomized to receive botulinum toxin, dexamethasone, or normal saline (group A, group B, and placebo, respectively). The main outcome measures were pretreatment subjective nasal symptoms (RCAT, VAS, and NOSE) and active anterior RMM measurements. All measurements were repeated during posttreatment 1, 2, and 3 months. RESULTS: No significant difference was observed in pretreatment questionnaire scores and RMM values between the study and placebo groups. The mean posttreatment RCAT, VAS, and NOSE scores after 1 and 2 months significantly improved in the treatment groups compared to placebo. The VAS and NOSE at posttreatment 2 months and RCAT, VAS, and NOSE at posttreatment 3 months were significantly different comparing group A to group B. All RMM parameters showed better values in group A than in group B at 1, 2, and 3 months posttreatment, with significant differences in four parameters in posttreatment 3 months. CONCLUSIONS: Botulinum toxin septal injection is a safe treatment option for AR and improves subjective nasal symptoms for 3 months. Botulinum toxin A injection tended to be more effective than steroid septal injection in terms of duration and degree.Level of Evidence: 2b, individual cohort study.
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Cirrhosis refers to irreversible liver damage where healthy tissue is replaced by scar tissue, resulting in impaired liver function. There is no cure and current treatments only prevent further liver damage; thus, novel therapeutic options are urgently needed. Here, we report a new approach that enables the formation of self-assembled 3D spheroids of adipose-derived stem cells (ADSCs) and murine hepatocytes (AML12) via reconstituted collagen fibers. Compared with the spheroids formed in the commercially available EZSHERE dish, the collagen fiber-based ADSC/hepatocyte spheroids offer a notable benefit in structure formation and paracrine factor secretion. To test the regenerative capability of the collagen fiber-based 3D ADSC/hepatocyte spheroids, a rat model of thioacetamide (TAA)-induced liver cirrhosis was employed. The transplantation of the collagen fiber-based 3D ADSC/hepatocyte spheroids show an improvement in liver function and ameliorates pathological liver cirrhosis in TAA-treated rats. In summary, our data show collagen fiber-based self-assembled 3D ADSC/hepatocyte spheroids to possess the excellent regenerative capacity in response to TAA-induced liver injury, promising an alternative therapeutic strategy for liver cirrhosis.
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Hepatócitos/transplante , Cirrose Hepática/terapia , Esferoides Celulares/transplante , Animais , Colágeno/metabolismo , Modelos Animais de Doenças , Humanos , Cirrose Hepática/induzido quimicamente , Masculino , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Ratos Sprague-Dawley , TioacetamidaRESUMO
Error patterns of facial emotion recognition (FER) indicate how individuals misinterpret others' facial expressions, which helps clinicians to manage related deficits. However, previous investigations are limited and may have been biased due to methodological issues (e.g., no consideration of response bias). This study aimed to propose a detectability index (d') for adjusting response bias and examine the error patterns of FER in patients with schizophrenia. Responses to 168 photos showing seven basic emotions, obtained from 351 patients with schizophrenia and 101 healthy adults, were extracted from a previous study. The differences in the d's between the two groups (Δd') were calculated to examine the error patterns of FER among the seven emotions. The findings were generally overlapped with those identified by the traditional confusion matrix. Four error patterns were found. First, the patients were insensitive to some negative emotions (i.e., sadness [Δd' = 0.83] and fear [Δd' = 0.72]). Second, they misrecognized happy faces as showing negative emotions (e.g., disgust [Δd' = 0.43] and sadness [Δd' = 0.37]). Third, they misinterpreted surprised faces as all the other emotions (Δd' = 0.41-0.87), except neutral. Fourth, they confused some negative emotions (e.g., misrecognizing fear as anger [Δd' = 0.87]). Our findings suggest that patients with schizophrenia show four error patterns of FER compared to healthy adults. Accordingly, interventions could be selected to improve their sensitivity to faces with negative emotions, differentiation of faces among positive and negative emotions, understanding of surprised faces, and discrimination of faces with negative emotions.
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Reconhecimento Facial , Esquizofrenia , Adulto , Emoções/fisiologia , Expressão Facial , Felicidade , HumanosRESUMO
BACKGROUND: House dust mite (HDM) allergens are major elicitors of allergic reactions worldwide. OBJECTIVE: Identification, characterization, and evaluation of diagnostic utility of a new important HDM allergen was performed. METHODS: A cDNA coding for a new Dermatophagoides pteronyssinus (Dp) allergen, Der p 37, was isolated from a Dp expression library with allergic patients' IgE antibodies. Recombinant Der p 37 (rDer p 37) expressed in Escherichia coli was purified, then characterized by mass spectrometry, circular dichroism, and IgE reactivity by ImmunoCAP ISAC technology with sera from 111 clinically defined HDM-allergic patients. The allergenic activity of rDer p 37 was studied by basophil activation and CD4+ T-cell responses by carboxyfluorescein diacetate succinimidyl ester dilution assays. Specific antibodies raised against rDer p 37 were used for the ultrastructural localization of Der p 37 in mites by immunogold transmission electron microscopy. RESULTS: Der p 37, a 26 kDa allergen with homology to chitin-binding proteins, is immunologically distinct from Der p 15, 18, and 23. It is located in the peritrophic membrane of fecal pellets. Der p 37 reacted with IgE antibodies from a third of HDM-allergic patients and induced specific basophil- and CD4+ T-cell activation. Der p 37 IgE-positive patients had significantly higher IgE levels to major HDM allergens, reacted with more HDM allergens, and had a higher risk (odds ratio = 3.1) of asthma compared to Der p 37-negative patients. CONCLUSIONS: Der p 37, a new Dp allergen recognized by a third of HDM-allergic patients, may serve as a surrogate marker for severe HDM sensitization and asthma.
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Asma , Hipersensibilidade , Alérgenos , Animais , Antígenos de Dermatophagoides , Proteínas de Artrópodes , Asma/diagnóstico , Poeira , Escherichia coli/genética , Humanos , Imunoglobulina E , PyroglyphidaeRESUMO
Background: Several studies indicate that Der p 7 is an important and clinically relevant allergen of Dermatophagoides pteronyssinus which should be included in vaccines for treatment of house dust mite (HDM) allergy. Aim of this study was to characterize the IgE epitopes of Der p 7. Methods: Recombinant Der p 7 was expressed and purified, analyzed for fold by circular dichroism and tested for its allergenic activity by basophil activation. Seven overlapping, surface-exposed peptides (P1-P7) with a length of 27 to 37 amino acids, which spanned the Der p 7 sequence, were synthesized and tested for IgE reactivity and allergenic activity by basophil activation assay. Carrier-bound peptides were studied for their ability to induce allergen-specific IgG antibodies in rabbits. Peptide-specific antibodies were used to inhibit allergic patients` IgE binding to Der p 7 by ELISA for mapping of IgE epitopes. Results: rDer p 7 showed high allergenic activity comparable with Der p 5, Der p 21, and Der p 23. None of the seven tested peptides showed any IgE reactivity or allergenic activity when tested with HDM- allergic patients indicating lack of sequential IgE epitopes on Der p 7. IgE inhibition experiments using anti-peptide specific IgGs and molecular modeling enabled us to identify discontinuous, conformational IgE epitopes of Der p 7. Conclusion and Clinical Relevance: IgE epitopes of Der p 7 belong to the conformational and discontinuous type whereas sequential Der p 7 peptides lack IgE reactivity. It should thus be possible to construct hypoallergenic vaccines for Der p 7 based on carrier-bound allergen peptides.
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Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Epitopos Imunodominantes , Imunoglobulina E/sangue , Pyroglyphidae/imunologia , Hipersensibilidade Respiratória/imunologia , Alérgenos/química , Alérgenos/genética , Animais , Antígenos de Dermatophagoides/química , Antígenos de Dermatophagoides/genética , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Estudos de Casos e Controles , Linhagem Celular Tumoral , Mapeamento de Epitopos , Humanos , Modelos Moleculares , Conformação Proteica , Dobramento de Proteína , Pyroglyphidae/genética , Coelhos , Ratos , Hipersensibilidade Respiratória/sangueRESUMO
Background Facial emotion recognition deficit (FERD) seems to be an obvious feature of patients with schizophrenia and has great potential for classifying patients and non-patients. The FERD screener was previously developed to classify patients from healthy adults. However, an obvious drawback of this screener is that the recommended cut-off scores could enhance either sensitivity or specificity (about 0.92) only, while the other one is at an only acceptable level (about 0.66). Machine learning (ML) algorithms are famous for their feature extraction and data classification abilities, which are promising for improving the discriminative power of screeners. This study aimed to improve the discriminative power of the FERD screener using an ML algorithm. Methods The data were extracted from a previous study. Artificial neural networks were generated to estimate the probability of being a patient with schizophrenia or a healthy adult based on the examinee's responses on the FERD screener (168 items). The performances of the ML-FERD screener were examined using a stratified five-fold cross-validation method. Results Across the five subsets of data, the ML-FERD screener showed extremely high areas under the receiver operating characteristic curve of 0.97-0.99. With the optimized cut-off scores, the average sensitivity and specificity of the ML-FERD screener were 0.90 (0.85-0.93) and 0.93 (0.86-1.00), respectively. Limitations The characteristics of patients were not representative, and the age was mismatched to control group. Conclusion The ML-FERD screener appears to have a better discriminative power to classify patients with schizophrenia and healthy adults than does the FERD screener.
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Reconhecimento Facial , Esquizofrenia , Adulto , Algoritmos , Humanos , Aprendizado de Máquina , Esquizofrenia/diagnóstico , Sensibilidade e EspecificidadeRESUMO
We investigate oscillations in coupled systems. The methodology is based on the Hopf bifurcation theorem and a condition extended from the Routh-Hurwitz criterion. Such a condition leads to locating the bifurcation values of the parameters. With such an approach, we analyze a single-cell system modeling the minimal genetic negative feedback loop and the coupled-cell system composed by these single-cell systems. We study the oscillatory properties for these systems and compare these properties between the model with Hill-type repression and the one with protein-sequestration-based repression. As the parameters move from the Hopf bifurcation value for single cells to the one for coupled cells, we compute the eigenvalues of the linearized systems to obtain the magnitude of the collective frequency when the periodic solution of the coupled-cell system is generated. Extending from this information on the parameter values, we further compute and compare the collective frequency for the coupled-cell system and the average frequency of the decoupled individual cells. To compare these scenarios with other biological oscillators, we perform parallel analysis and computations on a segmentation clock model.
Assuntos
Relógios Biológicos , Conceitos Matemáticos , RetroalimentaçãoRESUMO
The aim of the present study was to determine the pattern of symptoms of ragweed pollen-induced allergic disease in sensitized patients from Romania and to compare the molecular diagnosis of allergy with the skin prick test, in order to better characterize allergic patients and to guide therapy. A total of 97 subjects, including patients with ragweed pollen-induced allergic rhinoconjunctivitis with/without asthma, as well as healthy controls, were recruited prospectively in one ragweed pollen season, submitted to allergy questionnaires, skin prick tests and multiplex specific IgE (immunoglobulin E) measurement by ImmunoCAP ISAC (ImmunoCAP Immuno-Solid phase Allergy Chip) assay. A total of 83 patients were sensitized to ragweed pollen. Most patients (73%) were diagnosed with moderate-severe intermittent allergic rhinoconjunctivitis and 25% of the patients also had allergic asthma. The most common symptoms were watery rhinorrhea (91.57%), nasal obstruction (86.75%), and sneezing (85.54%). Most patients were polysensitized (62.65%), especially to other pollens, house dust mites and animal danders. Only 90% of the patients with positive skin prick test to ragweed pollen extract also had increased specific serum IgE to Amb a 1. Current options for specific molecular diagnosis of ragweed allergy are limited, as they only contain one or few of the sensitizing allergens present in ragweed pollen. An improved component-resolved diagnosis, using several ragweed pollen allergens, is required for better patient characterization and subsequent selection of an appropriate allergen immunotherapy product, thereby enabling a more personalized approach to the management of the ragweed-allergic patient.