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1.
Int J Mol Sci ; 25(13)2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-39000431

RESUMO

Wheat allergy is a major type of food allergy with the potential for life-threatening anaphylactic reactions. Common wheat, Triticum aestivum (hexaploid, AABBDD genome), was developed using tetraploid wheat (AABB genome) and the ancient diploid wheat progenitor (DD genome)-Aegilops tauschii. The potential allergenicity of gluten from ancient diploid wheat is unknown. In this study, using a novel adjuvant-free gluten allergy mouse model, we tested the hypothesis that the glutenin extract from this ancient wheat progenitor will be intrinsically allergenic in this model. The ancient wheat was grown, and wheat berries were used to extract the glutenin for testing. A plant protein-free colony of Balb/c mice was established and used in this study. The intrinsic allergic sensitization potential of the glutenin was determined by measuring IgE response upon transdermal exposure without the use of an adjuvant. Clinical sensitization for eliciting systemic anaphylaxis (SA) was determined by quantifying the hypothermic shock response (HSR) and the mucosal mast cell response (MMCR) upon intraperitoneal injection. Glutenin extract elicited a robust and specific IgE response. Life-threatening SA associated and a significant MMCR were induced by the glutenin challenge. Furthermore, proteomic analysis of the spleen tissue revealed evidence of in vivo Th2 pathway activation. In addition, using a recently published fold-change analysis method, several immune markers positively and negatively associated with SA were identified. These results demonstrate for the first time that the glutenin from the ancient wheat progenitor is intrinsically allergenic, as it has the capacity to elicit clinical sensitization for anaphylaxis via activation of the Th2 pathway in vivo in mice.


Assuntos
Alérgenos , Anafilaxia , Glutens , Camundongos Endogâmicos BALB C , Células Th2 , Triticum , Hipersensibilidade a Trigo , Animais , Anafilaxia/imunologia , Células Th2/imunologia , Células Th2/metabolismo , Camundongos , Triticum/imunologia , Triticum/química , Glutens/imunologia , Hipersensibilidade a Trigo/imunologia , Alérgenos/imunologia , Imunoglobulina E/imunologia , Imunoglobulina E/sangue , Modelos Animais de Doenças , Feminino , Mastócitos/imunologia , Mastócitos/metabolismo , Mastócitos/efeitos dos fármacos , Proteômica/métodos
2.
Int J Mol Sci ; 25(8)2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38673984

RESUMO

Gluten hypersensitivity is characterized by the production of IgE antibodies against specific wheat proteins (allergens) and a myriad of clinical allergic symptoms including life-threatening anaphylaxis. Currently, the only recommended treatment for gluten hypersensitivity is the complete avoidance of gluten. There have been extensive efforts to develop dietary-based novel therapeutics for combating this disorder. There were four objectives for this study: (i) to compile the current understanding of the mechanism of gluten hypersensitivity; (ii) to critically evaluate the outcome from preclinical testing of novel therapeutics in animal models; (iii) to determine the potential of novel dietary-based therapeutic approaches under development in humans; and (iv) to synthesize the outcomes from these studies and identify the gaps in research to inform future translational research. We used Google Scholar and PubMed databases with appropriate keywords to retrieve published papers. All material was thoroughly checked to obtain the relevant data to address the objectives. Our findings collectively demonstrate that there are at least five promising dietary-based therapeutic approaches for mitigating gluten hypersensitivity in development. Of these, two have advanced to a limited human clinical trial, and the others are at the preclinical testing level. Further translational research is expected to offer novel dietary-based therapeutic options for patients with gluten hypersensitivity in the future.


Assuntos
Glutens , Humanos , Glutens/imunologia , Animais , Hipersensibilidade Alimentar/dietoterapia , Hipersensibilidade Alimentar/terapia , Hipersensibilidade Alimentar/imunologia , Alérgenos/imunologia
3.
Int J Mol Sci ; 24(6)2023 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-36982527

RESUMO

Wheat allergies are potentially life-threatening and, therefore, have become a major health concern at the global level. It is largely unknown at present whether genetic variation in allergenicity potential exists among hexaploid, tetraploid and diploid wheat species. Such information is critical in establishing a baseline allergenicity map to inform breeding efforts to identify hyper-, hypo- and non-allergenic varieties. We recently reported a novel mouse model of intrinsic allergenicity using the salt-soluble protein extract (SSPE) from durum, a tetraploid wheat (Triticum durum). Here, we validated the model for three other wheat species [hexaploid common wheat (Triticum aestivum), diploid einkorn wheat (Triticum monococcum), and the ancient diploid wheat progenitor, Aegilops tauschii], and then tested the hypothesis that the SSPEs from wheat species will exhibit differences in relative allergenicities. Balb/c mice were repeatedly exposed to SSPEs via the skin. Allergic sensitization potential was assessed by specific (s) IgE antibody responses. Oral anaphylaxis was quantified by the hypothermic shock response (HSR). The mucosal mast cell response (MMCR) was determined by measuring mast cell protease in the blood. While T. monococcum elicited the least, but significant, sensitization, others were comparable. Whereas Ae. taushcii elicited the least HSR, the other three elicited much higher HSRs. Similarly, while Ae. tauschii elicited the least MMCR, the other wheats elicited much higher MMCR as well. In conclusion, this pre-clinical comparative mapping strategy may be used to identify potentially hyper-, hypo- and non-allergenic wheat varieties via crossbreeding and genetic engineering methods.


Assuntos
Diploide , Triticum , Animais , Camundongos , Triticum/metabolismo , Alérgenos/metabolismo , Tetraploidia , Melhoramento Vegetal , Adjuvantes Imunológicos/metabolismo , Cloreto de Sódio/metabolismo , Cloreto de Sódio na Dieta/metabolismo
4.
Int J Mol Sci ; 24(24)2023 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-38139075

RESUMO

Wheat is a prominent allergenic food that can trigger life-threatening anaphylaxis. Presently, it remains unclear whether wheat glutenin (WG) extract possesses inherent sensitization potential independently, without the use of adjuvants, and whether it can sensitize mice to the extent of inducing life-threatening systemic anaphylaxis. In this study, we tested the hypothesis that repeated skin exposures to WG extract without adjuvant will sensitize mice with the resultant anaphylactic reaction upon systemic WG challenge. Balb/c mice were bred and maintained on a strict plant protein-free diet and were repeatedly exposed to a WG extract or vehicle once a week for 9 weeks. WG-specific (s)IgE and total (t)IgE levels were quantified. Mice were challenged with WG extract to induce anaphylactic reactions as measured by hypothermic shock response (HSR) and mucosal mast cell degranulation response (MMCR). We also conducted proteomic analysis of 120 spleen immune markers. These skin-sensitized mice exhibited exposure-dependent IgE responses and near-fatal anaphylaxis upon challenge. Proteomic analysis identified seven dramatically elevated immune biomarkers in anaphylactic mice. These data reveal that WG is intrinsically allergenic, and that chronic skin exposure to WG extract can prime the mice for potentially fatal anaphylaxis.


Assuntos
Anafilaxia , Camundongos , Animais , Alérgenos , Triticum , Proteômica , Imunoglobulina E , Melhoramento Vegetal , Adjuvantes Imunológicos , Camundongos Endogâmicos BALB C , Adjuvantes Farmacêuticos
5.
Int J Mol Sci ; 23(12)2022 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-35742949

RESUMO

Wheat allergies are potentially life-threatening because of the high risk of anaphylaxis. Wheats belong to four genotypes represented in thousands of lines and varieties. Monitoring changes to wheat allergens is critical to prevent inadvertent ntroduction of hyper-allergenic varieties via breeding. However, validated methods for this purpose are unavailable at present. As a proof-of-concept study, we tested the hypothesis that salt-soluble wheat allergens in our mouse model will be identical to those reported for humans. Groups of Balb/cJ mice were rendered allergic to durum wheat salt-soluble protein extract (SSPE). Using blood from allergic mice, a mini hyper-IgE plasma bank was created and used in optimizing an IgE Western blotting (IEWB) to identify IgE binding allergens. The LC-MS/MS was used to sequence the allergenic bands. An ancient Aegilops tauschii wheat was grown in our greenhouse and extracted SSPE. Using the optimized IEWB method followed by sequencing, the cross-reacting allergens in A. tauschii wheat were identified. Database analysis showed all but 2 of the durum wheat allergens and all A. tauschii wheat allergens identified in this model had been reported as human allergens. Thus, this model may be used to identify and monitor potential changes to salt-soluble wheat allergens caused by breeding.


Assuntos
Panencefalite Esclerosante Subaguda , Triticum , Alérgenos , Animais , Cromatografia Líquida , Hibridização Genética , Imunoglobulina E , Camundongos , Melhoramento Vegetal , Espectrometria de Massas em Tandem , Triticum/genética
6.
Compr Rev Food Sci Food Saf ; 20(6): 6089-6115, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34455695

RESUMO

Wheat allergy is a potentiallylife-threatening disease that affects millions of people around the world. Food processing has been shown to influence the allergenicity of wheat and other major foods. However, a comprehensive review evaluating whether or not food processing can be used to develop hypo-/nonallergenic wheat products is unavailable. There were three objectives for this study: (1) to critically evaluate the evidence on the effect of fermentation, thermal processing, and enzyme or acid hydrolysis on wheat allergenicity so as to identify the potential for and challenges of using these methods to produce hypo-/nonallergenic wheat products; (2) to identify the molecular effects of food processing needed to create such products; and (3) to map the concept questions for future research and development to produce hypo-/nonallergenic wheat products. We performed literature research using PubMed and Google Scholar databases with various combinations of keywords to generate the data to accomplish these objectives. We found that: (1) food processing significantly modulates wheat allergenicity; while some methods can reduce or even abolish the allergenicity, others can create mega allergens; and (2) fermentation and enzymatic hydrolysis hold the most potential to create novel hypo-/nonallergenic wheat products; however, preclinical validation and human clinical trials are currently lacking. We also identify five specific research concepts to advance the research to enable the creation of hypo-/nonallergenic wheat products for application in food, medical, and cosmetic industries.


Assuntos
Hipersensibilidade a Trigo , Alérgenos , Manipulação de Alimentos , Humanos
7.
Int J Mol Sci ; 21(9)2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32369940

RESUMO

Wheat protein is considered a major type of food allergen in many countries including the USA. The mechanisms of allergenicity of wheat proteins are not well understood at present. Both adjuvant-based and adjuvant-free mouse models are reported for this food allergy. However, it is unclear whether the mechanisms underlying wheat allergenicity in these two types of models are similar or different. Therefore, we compared the molecular mechanisms in a novel adjuvant-free (AF) model vs. a conventional alum-adjuvant (AA) model of wheat allergy using salt-soluble wheat protein (SSWP). In the AF model, Balb/cJ mice were sensitized with SSWP via skin exposure. In the AA model, mice were sensitized by an intraperitoneal injection of SSWP with alum. In both models, allergic reactions were elicited using an identical protocol. Robust IgE as well as mucosal mast cell protein-1 responses were elicited similarly in both models. However, an analysis of the spleen immune markers identified strikingly different molecular activation patterns in these two models. Furthermore, a number of immune markers associated with intrinsic allergenicity were also identified in both models. Since the AF model uses skin exposure without an adjuvant, the mechanisms in the AF model may more closely simulate the human wheat allergenicity mechanisms from skin exposure in occupational settings such as in the baking industry.


Assuntos
Adjuvantes Imunológicos , Alérgenos/imunologia , Hipersensibilidade a Trigo/imunologia , Compostos de Alúmen , Animais , Especificidade de Anticorpos/imunologia , Antígenos de Plantas/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Imunização , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Camundongos , Proteínas de Plantas/efeitos adversos , Hipersensibilidade a Trigo/sangue , Hipersensibilidade a Trigo/metabolismo
8.
Molecules ; 24(6)2019 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-30909404

RESUMO

The prevalence of wheat allergy has reached significant levels in many countries. Therefore, wheat is a major global food safety and public health issue. Animal models serve as critical tools to advance the understanding of the mechanisms of wheat allergenicity to develop preventive and control methods. A comprehensive review on the molecular mechanisms of wheat allergenicity using animal models is unavailable at present. There were two major objectives of this study: To identify the lessons that animal models have taught us regarding the molecular mechanisms of wheat allergenicity and to identify the strengths, challenges, and future prospects of animal models in basic and applied wheat allergy research. Using the PubMed and Google Scholar databases, we retrieved and critically analyzed the relevant articles and excluded celiac disease and non-celiac gluten sensitivity. Our analysis shows that animal models can provide insight into the IgE epitope structure of wheat allergens, effects of detergents and other chemicals on wheat allergenicity, and the role of genetics, microbiome, and food processing in wheat allergy. Although animal models have inherent limitations, they are critical to advance knowledge on the molecular mechanisms of wheat allergenicity. They can also serve as highly useful pre-clinical testing tools to develop safer genetically modified wheat, hypoallergenic wheat products, novel pharmaceuticals, and vaccines.


Assuntos
Alérgenos/imunologia , Triticum/efeitos adversos , Hipersensibilidade a Trigo/etiologia , Alérgenos/química , Animais , Modelos Animais de Doenças , Manipulação de Alimentos , Inocuidade dos Alimentos , Humanos , Imunização , Imunoglobulina E/imunologia , Hipersensibilidade a Trigo/diagnóstico , Hipersensibilidade a Trigo/prevenção & controle , Hipersensibilidade a Trigo/terapia
9.
Int Arch Allergy Immunol ; 174(1): 7-16, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28950276

RESUMO

BACKGROUND: Wheat allergy and other immune-mediated disorders triggered by wheat proteins are growing at an alarming rate for reasons not well understood. A mouse model to study hypersensitivity responses to salt-soluble wheat protein (SSWP) extract is currently unavailable. Here we tested the hypothesis that SSWP extract from wheat will induce sensitization as well as allergic disease in mice. METHODS: Female BALB/cJ mice were weaned onto a plant protein-free diet. The mice were injected a total of 4 times with an SSWP (0.01 mg/mouse) fraction extracted from durum wheat along with alum as an adjuvant. Blood was collected biweekly and SSWP-specific IgE (SIgE) and total IgE (TIgE) levels were measured using ELISA. Systemic anaphylaxis upon intraperitoneal injection with SSWP was quantified by hypothermia shock response (HSR). Mucosal mast cell degranulation was measured by the elevation of mMCP-1 in the blood. The mice were monitored for dermatitis. Skin tissues were used in histopathology and for measuring cytokine/chemokine/adhesion molecule levels using a protein microarray system. RESULTS: Injection with SSWP resulted in time-dependent SIgE antibody responses associated with the elevation of TIgE concentration. Challenge with SSWP elicited severe HSR that correlated with a significant elevation of plasma mMCP-1 levels. Sensitized mice developed facial dermatitis associated with mast cell degranulation. Lesions expressed significant elevation of Th2/Th17/Th1 cytokines and chemokines and E-selectin adhesion molecule. CONCLUSION: Here we report a mouse model of anaphylaxis and atopic dermatitis to SSWP extract that may be used for further basic and applied research on wheat allergy.


Assuntos
Anafilaxia/imunologia , Dermatite Atópica/imunologia , Glutens/imunologia , Triticum/imunologia , Hipersensibilidade a Trigo/imunologia , Animais , Anticorpos/sangue , Degranulação Celular/imunologia , Quimases/sangue , Dermatite/imunologia , Modelos Animais de Doenças , Feminino , Imunoglobulina E/sangue , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Células Th1/imunologia , Células Th17/imunologia , Células Th2/imunologia
10.
Int J Food Sci Nutr ; 67(6): 660-9, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27251648

RESUMO

Although food processing can alter food allergenicity, the impact of extrusion processing on in vivo hazelnut allergenicity is unknown. Here, we tested the hypothesis that extrusion processing will alter the immune activation properties of hazelnut protein (HNP) in mice. Soluble extrusion-processed HNP (EHNP) was prepared and evaluated for immune response using an established transdermal sensitization mouse model. Mice were sensitized with identical amounts of EHNP versus raw HNP. After confirming systemic IgE, IgG1 and IgG2a antibody responses, oral hypersensitivity reaction was quantified by hypothermia shock response (HSR). Mechanism was studied by measuring mucosal mast cell (MMC) degranulation. Compared to raw HNP, the EHNP elicited slower but similar IgE antibody (Ab) response, lower IgG1 but higher IgG2a Ab response. The EHNP exhibited significantly lower oral HSR as well as MMC degranulation capacity. These results demonstrate that the extrusion technology can be used to produce soluble HNP with altered immune activation properties.


Assuntos
Corylus/química , Manipulação de Alimentos , Hipersensibilidade a Noz/imunologia , Nozes/química , Proteínas de Plantas/imunologia , Animais , Formação de Anticorpos , Corylus/imunologia , Modelos Animais de Doenças , Feminino , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Hipersensibilidade a Noz/prevenção & controle , Nozes/imunologia , Proteínas de Plantas/isolamento & purificação
11.
J Sci Food Agric ; 94(12): 2562-8, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24497201

RESUMO

BACKGROUND: Acrylamide is a possible carcinogen and known to form in heat-treated carbohydrate-rich foods. This study was designed to investigate the effects of different ingredients (reducing sugars, chemical leavening agents, citric acid), processing conditions (feed moisture content: 22, 24 or 26%, exit die temperature: 110, 150 °C), and extrusion cooking methods (with or without CO2 injection) on acrylamide formation. RESULTS: The type of reducing sugar did not have a considerable effect on acrylamide formation, while increased exit die temperature had a promoting effect. Addition of chemical leavening agents (sodium bicarbonate and ammonium bicarbonate) into formulations increased acrylamide formation levels. The addition of citric acid prevented acrylamide formation, but its effect on textural properties was detrimental. Acrylamide levels of extrudates decreased gradually with increasing feed moisture in all formulations. Acrylamide content of extrudates produced with 22% feed moisture decreased by 61% in the CO2 injection method compared to conventional extrusion. Furthermore, an 82% decrease in acrylamide content was observed with the combined effect of CO2 injection and increasing feed moisture content from 22 to 24% and decreased below the limit of quantification with a further increase in feed moisture. CONCLUSION: A substantial decrease in final acrylamide level is probably due to restriction of two major steps of acrylamide formation: dehydration and decarboxylation.


Assuntos
Acrilamida/análise , Bicarbonatos , Metabolismo dos Carboidratos , Dióxido de Carbono , Ácido Cítrico , Culinária , Zea mays/química , Carboidratos , Descarboxilação , Dessecação , Dieta , Temperatura Alta , Humanos , Reação de Maillard , Bicarbonato de Sódio , Água
12.
Biology (Basel) ; 13(9)2024 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-39336104

RESUMO

Consumption of whole-grain wheat has been associated with positive health outcomes, but it remains unclear whether different types of wheat elicit varying effects on the gut microbiome and intestinal inflammation. The objectives of this research were to investigate the effect of two whole-grain wheat flours versus refined wheat flour on the diversity of the human gut microbiota, as well as on butyrate production capacity and gastrointestinal inflammation, using one-week dietary interventions. For this study, 28 participants were recruited, with ages ranging from 18 to 55 years and a mean BMI of 26.0 kg/m2. For four weeks, participants were provided 80 g daily servings of different wheat crackers: Week A was a run-in period of crackers made from soft white wheat flour, Week B crackers were whole-grain soft white wheat flour, Week C crackers were a wash-out period identical to Week A, and Week D crackers were whole-grain soft red wheat flour. At the end of each week, participants provided fecal samples that were analyzed for markers of intestinal inflammation, including lipocalin and calprotectin, using enzyme-linked immunosorbent assays and quantitative real-time PCR. The primary outcome, gut bacterial community alpha and beta diversity, was similar across timepoints. Three taxa significantly differed in abundance following both whole-grain wheat flour interventions: Escherichia/Shigella and Acidaminococcus were significantly depleted, and Lachnospiraceae NK4A136 group was enriched. Secondary outcomes determined that protein markers of intestinal inflammation and genes related to putative butyrate production capacity were similar throughout the study period, with no significant changes. Lipocalin concentrations ranged from 14.8 to 22.6 ng/mL while calprotectin ranged from 33.2 to 62.5 ng/mL across all 4 weeks. The addition of wheat crackers to the adult human subjects' usual diet had a minimal impact on their gastrointestinal inflammation or the gut microbiota.

13.
Front Allergy ; 4: 1214051, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37841051

RESUMO

Introduction: Gluten allergy is a major public health problem that is growing at an alarming rate. Specific mechanisms underlying sensitization to gluten remain incompletely understood. Currently, it is unclear whether chronic exposure to alcohol-soluble gluten extract via undamaged skin has the capacity to clinically sensitize mice for life-threatening anaphylaxis. Using an adjuvant-free mouse model, here we tested the hypothesis that chronic application of alcohol-soluble durum gluten (ASDG) extract will clinically sensitize mice for life-threatening anaphylaxis. Methods: This study was conducted in a gluten-free Balb/c mouse colony that was established and maintained on a plant protein-free diet. Groups of adult female mice were exposed dermally to ASDG extract or vehicle once a week for 9-weeks. Specific (s) and total (t) IgE levels were quantified. Mice were challenged systemically with ASDG to measure symptoms of systemic anaphylaxis. Hypothermic shock response (HSR) and mucosal mast cell degranulation response (MMCR) were determined upon challenge. Spleen Th1, Th2, and other immune markers were quantified. Results: We found that chronic exposure to ASDG elicited robust elevation of sIgE and tIgE. Systemic challenge with ASDG, but not vehicle, elicited life-threatening anaphylaxis associated with dramatic HSR and MMCR. Correlation analysis demonstrated direct positive inter-relationships among IgE, HSR, and MMCR. Anaphylaxis was associated with significant elevation of prototypic Th2 but not Th1 immune markers in the spleen. Discussion/Conclusion: Our study collectively demonstrates that ASDG is intrinsically allergenic; and chronic exposure to ASDG via undamaged skin can clinically sensitize mice for life-threatening anaphylaxis via activating the systemic Th2 immune responses.

14.
Nat Commun ; 14(1): 1726, 2023 03 28.
Artigo em Inglês | MEDLINE | ID: mdl-36977662

RESUMO

Mis-sense mutations affecting TP53 promote carcinogenesis both by inactivating tumor suppression, and by conferring pro-carcinogenic activities. We report here that p53 DNA-binding domain (DBD) and transactivation domain (TAD) mis-sense mutants unexpectedly activate pro-carcinogenic epidermal growth factor receptor (EGFR) signaling via distinct, previously unrecognized molecular mechanisms. DBD- and TAD-specific TP53 mutants exhibited different cellular localization and induced distinct gene expression profiles. In multiple tissues, EGFR is stabilized by TAD and DBD mutants in the cytosolic and nuclear compartments respectively. TAD mutants promote EGFR-mediated signaling by enhancing EGFR interaction with AKT via DDX31 in the cytosol. Conversely, DBD mutants maintain EGFR activity in the nucleus, by blocking EGFR interaction with the phosphatase SHP1, triggering c-Myc and Cyclin D1 upregulation. Our findings suggest that p53 mutants carrying gain-of-function, mis-sense mutations affecting two different domains form new protein complexes that promote carcinogenesis by enhancing EGFR signaling via distinctive mechanisms, exposing clinically relevant therapeutic vulnerabilities.


Assuntos
Receptores ErbB , Proteína Supressora de Tumor p53 , Proteína Supressora de Tumor p53/metabolismo , Receptores ErbB/genética , Receptores ErbB/metabolismo , Transdução de Sinais , Ativação Transcricional , Fosforilação
15.
Front Allergy ; 3: 926576, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36238931

RESUMO

Wheat is a major food allergen per the regulatory bodies of various nations. Hypersensitivity reactions to wheat have been steadily increasing for reasons that are not completely understood. Wheat-allergy models typically use adjuvants to induce sensitization to wheat proteins followed by an intraperitoneal challenge to elicit anaphylaxis. Although these models are very useful, they lack the ability to reveal the intrinsic allergenicity potential of wheat. To improve the mouse model of wheat allergy, we tested the hypothesis that repeated skin application of salt-soluble protein extract (SSPE) from durum wheat will clinically sensitize the mice to oral anaphylaxis to SSPE. Balb/c mice were bred and maintained on a plant-protein-free diet and used in the experiments. Adult female mice were exposed to SSPE once a week for 9 weeks via a solution on intact skin. Sensitization was measured by SSPE-specific IgE (sIgE) antibody and total IgE (tIgE) levels. Oral anaphylaxis was quantified by hypothermic shock response (HSR), and mucosal mast cell response (MMCR) was quantified by measuring MMCP-1 after oral challenge. Using single mouse data, correlation analyses were performed to determine the relationship among the allergenicity readouts. Spleen cytokines were quantified using a protein microarray method. Our results show that (i) repeated skin exposures to SSPE elicited robust increases in the sIgE and tIgE levels; (ii) skin exposure to SSPE was sufficient to sensitize mice for oral anaphylaxis and MMCR; (iii) both HSR and MMCR showed a strong correlation with each other, as well as with sIgE, and a modest correlation with tIgE levels; (iv) selected Th2/Th17/Th1 cytokines were elevated in skin-sensitized mice; and (v) oral allergen-challenged mice showed selective elevation of IL-6 and a panel of chemokines compared to saline-challenged mice. Together, we report the development and characterization of a novel adjuvant-free wheat-allergy mouse model that uses skin sensitization without tape-stripping followed by oral elicitation of anaphylaxis. Furthermore, validation of quantifiable wheat allergenicity readouts makes this model particularly suitable as a pre-clinical testing tool to assess the intrinsic sensitization/oral-anaphylaxis elicitation potential of novel wheat proteins (e.g., processed wheat) and to develop hypo/non-allergenic wheat products.

16.
JMIR Res Protoc ; 10(10): e29046, 2021 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-34612840

RESUMO

BACKGROUND: Daily fiber intake can increase the diversity of the human gut microbiota as well as the abundance of beneficial microbes and their metabolites. Whole-grain wheat is high in fiber. OBJECTIVE: This manuscript presents a study protocol designed to understand the effects of different types of wheat on gastrointestinal tract microbes. METHODS: Human adults will consume crackers made from three types of wheat flour (refined soft white wheat, whole-grain soft white wheat, and whole-grain soft red wheat). In this study, participants will alternate between crackers made from refined soft white wheat flour to those made from whole-grain soft white wheat and whole-grain soft red wheat flour. Survey and stool sample collection will occur after 7-day treatment periods. We will assess how wheat consumption affects gastrointestinal bacteria by sequencing the V4 region of 16S rRNA gene amplicons and the inflammatory state of participants' intestines using enzyme-linked immunosorbent assays. The butyrate production capacity of the gut microbiota will be determined by targeted quantitative real-time polymerase chain reaction. RESULTS: We will report the treatment effects on alpha and beta diversity of the microbiota and taxa-specific differences. Microbiota results will be analyzed using the vegan package in R. Butyrate production capacity and biomarkers of intestinal inflammation will be analyzed using parametric statistical methods such as analysis of variance or linear regression. We expect whole wheat intake to increase butyrate production capacity, bacterial alpha diversity, and abundance of bacterial taxa responsive to phenolic compounds. Soft red wheat is also expected to decrease the concentration of inflammatory biomarkers in the stool of participants. CONCLUSIONS: This protocol describes the methods to be used in a study on the impact of wheat types on the human gastrointestinal microbiota and biomarkers of intestinal inflammation. The analysis of intestinal responses to the consumption of two types of whole wheat will expand our understanding of how specific foods affect health-associated outcomes. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/29046.

17.
Food Chem ; 302: 125338, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31434027

RESUMO

This study was designed to evaluate the baking performances of 25 edible dry bean (Phaseolus vulgaris L.) varieties and to investigate correlations among cookie features and rapid test indices (i.e., water and lactic acid retention capacities, oil binding capacity and Rapid Visco Analyzer indices). Two bean powder particle sizes (≤0.5 mm, ≤1.0 mm) were investigated. Cookies were evaluated in terms of nutritional, geometrical and textural properties. Bean powders doubled the amount of cookie protein and increased cookie resistant starch content. Baking potential varied according to bean genotype and powder particle size: coarse powders resulted in larger (+26%) and thinner (-19%) cookies characterized by easier breaking texture (fracture strengths of 41-157 vs. 48-226 kPa for fine powders). Water retention and oil binding capacities and pasting properties significantly (p < 0.05) correlated with cookie features. In conclusion, these accumulated findings can be used in designing value-added traditional and gluten-free cookies.


Assuntos
Phaseolus/química , Pós/química , Culinária , Dieta Livre de Glúten , Farinha , Ácido Láctico/análise , Ácido Láctico/química , Valor Nutritivo , Tamanho da Partícula , Amido/análise , Água/análise , Água/química
18.
J Neurosurg ; 111(1): 114-8, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19249924

RESUMO

The authors describe the off-label use of Onyx for embolization of fusiform mycotic and dissecting intracranial aneurysms based on their experience with 3 patients treated at the University of Utah Hospital from 2006 through 2007. Technical success in occluding the parent artery/aneurysm was achieved in all patients. There were no complications. The authors conclude that Onyx can be used to achieve occlusion of fusiform mycotic and dissecting intracranial aneurysms in conjunction with parent artery occlusion.


Assuntos
Dissecção Aórtica/terapia , Dimetil Sulfóxido , Embolização Terapêutica/métodos , Aneurisma Intracraniano/terapia , Polivinil , Adulto , Idoso , Dissecção Aórtica/diagnóstico por imagem , Angiografia Cerebral , Corpo Caloso/irrigação sanguínea , Feminino , Humanos , Aneurisma Intracraniano/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios X
19.
Int J Food Sci Nutr ; 60(6): 449-57, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19212857

RESUMO

Three different commercial starches (Hylon VII, Novelose 330 and CrystaLean) were incorporated into the bread formulation at different levels and their effects on the rheological and baking properties and resistant starch contents of breads were evaluated. Starch-supplemented doughs were weaker and absorbed more water than those of the base flour. Loaf volumes of the breads decreased above the 10% level for Novelose and above the 20% level for Hylon VII and CrystaLean supplementation. Commercial starches did not have a substantial deteriorative effect on crumb color, external appearance and symmetry of breads. Crust color values decreased at a 30% addition level in Novelose-supplemented and CrystaLean-supplemented breads and above a 10% addition level in Hylon VII-supplemented breads. Firmness of breads increased above the 10% level for Novelose and above the 20% level for Hylon VII and CrystaLean supplementation. Resistant starch contents of the breads (after 1 and 7 days storage) increased significantly as the addition level of commercial starches increased.


Assuntos
Pão , Dieta , Carboidratos da Dieta , Fibras na Dieta , Suplementos Nutricionais , Farinha/análise , Amido , Cor , Culinária , Carboidratos da Dieta/análise , Fibras na Dieta/análise , Farinha/normas , Qualidade dos Alimentos , Dureza , Humanos , Reologia , Amido/análise , Água
20.
J Immunol Methods ; 464: 95-104, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30395814

RESUMO

BACKGROUND: Wheat allergy is a major food allergy that has reached significant levels of global public health concern. Potential variation in allergenicity among different wheat genotypes is not well studied at present largely due to the unavailability of validated methods. Here, we developed and validated a novel mouse-based primary screening method for this purpose. METHODS: Groups of Balb/c mice weaned on-to a plant protein-free diet were sensitized with salt-soluble protein (SSP) extracted from AABB genotype of wheat (durum, Carpio variety). After confirming clinical sensitization for anaphylaxis, mice were boosted 7 times over a 6-month period. Using a pooled-plasma mini bank, a wheat-specific IgE-inhibition (II)-ELISA was optimized. Then the relative allergenicity of SSPs from tetraploid (AABB), hexaploid (AABBDD) and diploid (DD) wheat genotypes were determined. The IC50/IC75 values were estimated using IgE inhibition curves. RESULTS: The optimized II-ELISA with an inhibition time of 2.5 h had a co-efficient of variation of <2%. Primary screening for relative allergenicity demonstrated that IgE binding to AABB-SSP was significantly abolished by the other two wheat genotypes. Compared to AABB, the relative allergenicity of SSPs of AABBDD and DD were significantly lower (p < .01). Furthermore, IgE inhibition curves showed significant differences in IC50 and IC75 values among the three wheat genotypes. CONCLUSION: We report a novel mouse-based primary screening method of testing relative allergenicity of wheat proteins from three different wheat genotypes for the first time. This method is expected to have broad applications in wheat allergy research.


Assuntos
Alérgenos , Ensaio de Imunoadsorção Enzimática , Imunoglobulina E/sangue , Proteínas de Plantas/imunologia , Triticum/imunologia , Hipersensibilidade a Trigo/diagnóstico , Alérgenos/genética , Animais , Biomarcadores/sangue , Feminino , Camundongos Endogâmicos BALB C , Proteínas de Plantas/genética , Reprodutibilidade dos Testes , Fatores de Tempo , Triticum/genética , Hipersensibilidade a Trigo/sangue , Hipersensibilidade a Trigo/imunologia
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