Identification and characterization of a novel IL-4 receptor α chain (IL-4Rα) antagonist to inhibit IL-4 signalling

BACKGROUND/AIMS: In recent times, allergy has become a financial, physical and psychological burden to the society as a whole. In allergic cascades, cytokine IL-4 binds to IL-4 receptor (IL-4R), consequently producing allergen-specific IgE antibodies by B cells. In addition, among other functions, IL-4 is also responsible for B and T cell proliferation and differentiation. Hence, characterization of novel antagonists that inhibit IL-4 signalling forms the overall aim of this study. METHODS: Phage display was used to screen a random 12-mer synthetic peptide library with a human IL-4Rα to identify peptide candidates. Once identified, the peptides were commercially synthesized and used for in vitro immunoassays. RESULTS: We have successfully used phage display to identify M13 phage clones that demonstrated specific binding to IL-4Rα. The peptide N1 was synthesized for use in ELISA, demonstrating significant binding to IL-4Rα and inhibiting interaction with cytokine IL-4. Furthermore, the peptide was tested in a transfected HEK-Blue IL-4 reporter cell line model, which produces alkaline phosphatase (AP). QUANTI-Blue, a substrate, breaks down in the presence of AP producing a blue coloration. Using this colorimetric analysis, >50% inhibition of IL-4 signalling was achieved. CONCLUSION: We have successfully identified and characterised a synthetic peptide antagonist against IL-4Rα, which effectively inhibits IL-4 interaction with the IL-4Rα in vitro. Since IL-4 interaction with IL-4Rα is a common pathway for many allergies, a prophylactic treatment can be devised by inhibiting this interaction for future treatment of allergies.

VITALITY trial: protocol for a randomised controlled trial to establish the role of postnatal vitamin D supplementation in infant immune health

Introduction Postnatal vitamin D supplementation may be associated with a reduction in IgE-mediated food allergy, lower respiratory tract infections and improved bone health. Countries in the Northern hemisphere recommend universal infant vitamin D supplementation to optimise early vitamin D levels, despite the absence of large trials proving safety or efficacy for any disease outcome. With the aim of determining the clinical and cost-effectiveness of daily vitamin D supplementation in breastfed infants from age 6–8 weeks to 12 months of age, we have started a double-blind, randomised, placebo-controlled trial of daily 400 IU vitamin D supplementation during the first year of life, VITALITY. Methods nd analysis Infants (n=3012) who are fully breastfed and not receiving vitamin D supplementation will be recruited at the time of their first immunisation, from council-led immunisation clinics throughout metropolitan Melbourne, Australia. The primary outcome is challenge-proven food allergy at 12 months of age. Secondary outcomes are food sensitisation (positive skin prick test), number of lower respiratory infections (through hospital linkage), moderately-severe and persistent eczema (by history and examination) and vitamin D deficiency (serum vitamin D <50 nmol/L) at age 12 months. The trial is underway and the first 130 participants have been recruited.

Molecular and immunological analysis of hen's egg yolk allergens with a focus on YGP42 (Gal d 6)

Allergy to hen's (Gallus domesticus) egg white is one of the most common forms of food allergy. Allergy to hen's yolk also exists however, to a lesser extent when compared to egg white allergy. Two minor allergens from the hen's egg yolk known as α-livetin (Gal d 5) and YGP42 (Gal d 6) were discovered recently. In this study, we investigated whether sensitization to egg white is associated with reactivity to egg yolk as well. Sera obtained from 25 patients with allergy to egg white were tested for specific IgE binding for egg yolk proteins through western immunoblotting. 36% of patients were found with true IgE-sensitization against egg yolk proteins. It was found that most of the IgE reactive yolk proteins were fragments of major precursor proteins of hen; vitellogenin-1 (VTG-1), vitellogenin-2 (VTG-2) and apolipoprotein B (Apo B). The egg yolk allergen Gal d 6 is the C-terminal part of VTG-1 and was found to be allergenic in significant percentage of egg white allergy patients. These results highlight the significance of Gal d 6 as an important allergen of egg yolk. Therefore, the secondary aim of this study involved developing a recombinant version of YGP42 in an Escherichia coli expression system. Recombinant Gal d 6 (rGal d6) was expressed as a fusion peptide with a 6 × His tag and purified using metal chelating resin. The inhibition ELISA results showed that rYGP42 was IgE reactive and was able to inhibit IgE binding to crude egg yolk (CEY) by up to 30%. Traditionally, it was thought that allergy to egg yolk occurred independently from sensitization to egg white. This study underlies the importance of concomitant sensitization to egg yolk proteins in patients allergic to egg white. Evidence reported in this study strongly suggests that egg yolk has potentially undiscovered allergens and therefore warrants further investigation. Furthermore, IgE reactive Gal d 6 presented in this study has the potential to be used in diagnosis and immunotherapy to treat egg allergy.

Exploring the effects of omega-3 and omega-6 fatty Acids on allergy using a HEK-blue cell line.

BACKGROUND: Allergic reactions can result in life-threatening situations resulting in high economic costs and morbidity. Therefore, more effective reagents are needed for allergy treatment. A causal relationship has been suggested to exist between the intake of omega-3/6 fatty acids, such as docosahexanoic acid (DHA), eicosapentanoic acid (EPA), docosapentanoic acid (DPA) and arachidonic acid (AA), and atopic individuals suffering from allergies. In allergic cascades, the hallmark cytokine IL-4 bind to IL-4 receptor (IL-4R) and IL-13 binds to IL-13 receptor (IL-13R), this activates the STAT6 phosphorylation pathway leading to gene activation of allergen-specific IgE antibody production by B cells. The overall aim of this study was to characterize omega-3/6 fatty acids and their effects on STAT6 signaling pathway that results in IgE production in allergic individuals. METHODS: The fatty acids were tested in vitro with a HEK-Blue IL-4/IL-13 reporter cell line model, transfected with a reporter gene that produces an enzyme, secreted embryonic alkaline phosphatase (SEAP). SEAP acts as a substitute to IgE when cells are stimulated with bioactive cytokines IL-4 and/or IL-13. RESULTS: We have successfully used DHA, EPA and DPA in our studies that demonstrated a decrease in SEAP secretion, as opposed to an increase in SEAP secretion with AA treatment. A statistical Student's t-test revealed the significance of the results, confirming our initial hypothesis. CONCLUSION: We have successfully identified and characterised DHA, EPA, DPA and AA in our allergy model. While AA was a potent stimulator, DHA, EPA and DPA were potential inhibitors of IL-4R/IL-13R signalling, which regulates the STAT6 induced pathway in allergic cascades. Such findings are significant in the future design of dietary therapeutics for the treatment of allergies.

Políticas europeias para a educação em Ciências: O caso das Geociências na educação não superior em Portugal

Apesar da contribuição específica das Geociências para a prossecução da cidadania democrática, não existem referências a esta área do saber nos documentos emanados da União Europeia (UE), ou pelo menos os autores não as conhecem. Este artigo reune breves notas e preocupações acerca da educação em ciências e analisa o lugar que as Geociências assumem na educação não superior no sistema educativo português, tema que interessa ao Brasil. A Carta dos Direitos Fundamentais da União Europeia (cf. www.europarl.europa.eu/charter/pdf/text_pt.pdf) tenciona dar maior visibilidade aos três grandes pilares – liberdade, igualdade, solidariedade –, e reforçar sua proteção à luz da evolução da sociedade, do progresso social e da evolução científica e tecnológica. Com o objetivo de desenvolver valores comuns e, simultaneamente, respeitar a diversidade das identidades nacionais, destaca-se aqui o artigo 14º: “1. Todas as pessoas têm direito à educação, bem como o acesso à formação profissional e contínua. 2. Este direito inclui a possibilidade de frequentar gratuitamente o ensino obrigatório.” É reconhecida, nos documentos da União Europeia (UE), a necessidade de os cidadãos intervirem na vida politica e social não só para assegurar o desenvolvimento de valores democráticos fundamentais mas, também, para fomentar a coesão social em período de diversidade de identidades. Neles a educação é vista como via facilitadora de promoção da equidade, constituindo um dos grandes objetivos do atual quadro estratégico para cooperação europeia, em vigor até 2020. A formulação de estratégias eficazes para atingir a equidade coloca desafios nos níveis político, científico, económico e social. É, assim, compreensível que, na sequência da Estratégia Europa 2010, a educação seja tida como centralidade da Estratégia Europa 2020, prevendo-se, por exemplo, reduções de abandono escolar para níveis inferiores a 10%, enquanto se aponta o patamar de 40% para que cidadãos entre os 30 e 34 anos concluam o ensino superior, tudo isto até 2020. Seria bom que, ao mesmo tempo em que se ressalta a importância da Educação dos cidadãos, sejam significativamente ampliados “os níveis de investimento em recursos humanos, a fim de dar prioridade ao mais importante trunfo da Europa (os seus cidadãos)” (cf. http://eur-lex.europa.eu/ legal-content/EN/TXT/HTML/?uri=URISERV:c11047&from=EN) – não se dê preferência a uma perspetiva de “educação contábil”, isto é, segundo Licínio Lima (cf. http://repositorium.sdum. uminho.pt/bitstream/1822/11788/1/Artigo%20RBE.pdf), orientação centrada na preparação do cidadão para a grande finalidade da competição, assente na ideia do mercado global adaptado à chamada racionalidade económica. Seria muito desejável que nas orientações de Bruxelas estivessem presentes as preocupações de Fraser & Gordon (1994 citadas por Licínio Lima, p. 74 da URL ‘RBE.pdf’ acima): “hoje, quando a retórica acerca do ‘triunfo da democracia’ acompanha a devastação económica, é tempo de insistir que não pode existir cidadania democrática sem direitos sociais”.

Allergological evaluation of a dog population in a veterinary immuno-allergology consultation: What correlates in a canine model

Introduction: Allergic dermatitis (AD) is the most common canine pruritic condition in veterinary dermatology. Allergic dermatitis to flea bites presents the highest prevalence, followed by atopic dermatitis and food AD. This study aimed to identify possible correlation between data from clinical signs, intradermal tests (IDT) and specific IgE levels, which are used in dog AD assessment. Methods: Fifty five dogs from the Veterinary Hospital of the University of Évora (Portugal) and Rof Codina University Hospital (Lugo, Spain) outpatient consultations were studied by means of clinical inquiry, IDT and specific IgE determination. Thirty five of the patients belonged to predisposed breeds, 30 were females and 25 males. Forty one (74%) were indoor. Results: In 82% of cases first clinical signs appeared before the age of 3 years and 24% even before 1 year old. In 70% of the individuals clinical signs included itching, which was generalized in 66%, with 78% of paw licking and chewing. Clinical profile showed seasonal worsening in 64% of cases. From the 69.1% of dogs already presenting with dermatitis, 50% also presented external otitis and 28.9% self-inflicted alopecia. "Intense itching" was found in 10.5%, "medium itching" in 81.6% and “mild itching” in 5.26% of the patients. Prevalence of positive IDT was 37.3 % to Lep d, 29.41% to Der f, 27.5% to Der p, 25.5% to Dac g and 21.6% to Malassezia sp. From the 37 dogs submitted to food IDT, 40.5% revealed positive to beef, 27% to chicken, 27% to porc and 5.4% to lamb. Specific IgE > 150 EAU was found in 84% of dogs to indoor allergen sources and in 68% to pollens. A negative correlation was found between an outdoor life and the intensity (p = 0.033) and precocity (p = 0.026) of clinical signs. Sensitization to pollens was found positively correlated with the seasonality of clinical signs (p = 0.001) and the positivity for Dac g (p = 0.007). The prevalence of chronic otitis correlated positively with alopecia and reactivity to Lep d (p = 0.008), Plantago lanceolata (p = 0.026) and Platanus acerifolia (p = 0.017). There was no correlation between the results of ITD and specific IgE. Conclusion: We can conclude that correlation between different clinical signs and positive testing for some allergenic sources may occur, as well as between sensitization to pollens and the beginning, the intensity and the seasonality of dog patient clinical signs.

Avaliação de híbridos de milho na Região Sul do Brasil, na safra 2015/2016.

2016

EDUCAÇÃO GEOGRÁFICA E ENSINO DE SOLOS: UMA EXPERIÊNCIA EM SALA DE AULA

Neste trabalho temos como objetivo analisar a importância de abordar a temática solo na perspectiva da educação geográfica apresentando trabalhos realizados em sala de aula. O homem ao apropriar-se do espaço modifica-o e, na tentativa de “dominar” essas modificações os professores de geografia têm um papel importante, de pensar em conteúdos que favoreçam a compreensão dos alunos, dos processos que ocorrem no espaço vivido. Os conteúdos com o tema Solos são sempre deixados de lado ou pouco discutidos no ensino. Ao analisar os currículos de Geografia, percebemos pouca discussão mais aprofundada conceitual e temática sobre o significado do uso do solo atualmente, dos seus componentes, suas características e de seus diversos usos. Mas como as discussões do que ocorre no espaço vivido estão sendo abordadas em sala de aula? Qual a preocupação com o uso do solo? Desmatamentos resultantes da expansão das fronteiras agrícolas, das manchas urbanas degradam o solo provocando disfunções, que comprometem a manutenção da biosfera. Os professores devem preocupar-se com essas questões, buscando trabalhar de maneira diferenciada sobre o tema para que os alunos tenham uma compreensão do que significa o solo com um dos componentes que estão no espaço e às vezes, não percebidos por eles.