Research needs in allergy: an EAACI position paper, in collaboration with EFA

Abstract In less than half a century, allergy, originally perceived as a rare disease, has become a major public health threat, today affecting the lives of more than 60 million people in Europe, and probably close to one billion worldwide, thereby heavily impacting the budgets of public health systems. More disturbingly, its prevalence and impact are on the rise, a development that has been associated with environmental and lifestyle changes accompanying the continuous process of urbanization and globalization. Therefore, there is an urgent need to prioritize and concert research efforts in the field of allergy, in order to achieve sustainable results on prevention, diagnosis and treatment of this most prevalent chronic disease of the 21 st century. The European Academy of Allergy and Clinical Immunology (EAACI) is the leading professional organization in the field of allergy, promoting excellence in clinical care, education, training and basic and translational research, all with the ultimate goal of improving the health of allergic patients. The European Federation of Allergy and Airways Diseases Patients' Associations (EFA) is a non-profit network of allergy, asthma and Chronic Obstructive Pulmonary Disorder (COPD) patients' organizations. In support of their missions, the present EAACI Position Paper, in collaboration with EFA, highlights the most important research needs in the field of allergy to serve as key recommendations for future research funding at the national and European levels. Although allergies may involve almost every organ of the body and an array of diverse external factors act as triggers, there are several common themes that need to be prioritized in research efforts. As in many other chronic diseases, effective prevention, curative treatment and accurate, rapid diagnosis represent major unmet needs. Detailed phenotyping/endotyping stands out as widely required in order to arrange or re-categorize clinical syndromes into more coherent, uniform and treatment-responsive groups. Research efforts to unveil the basic pathophysiologic pathways and mechanisms, thus leading to the comprehension and resolution of the pathophysiologic complexity of allergies will allow for the design of novel patient-oriented diagnostic and treatment protocols. Several allergic diseases require well-controlled epidemiological description and surveillance, using disease registries, pharmacoeconomic evaluation, as well as large biobanks. Additionally, there is a need for extensive studies to bring promising new biotechnological innovations, such as biological agents, vaccines of modified allergen molecules and engineered components for allergy diagnosis, closer to clinical practice. Finally, particular attention should be paid to the difficult-to-manage, precarious and costly severe disease forms and/or exacerbations. Nonetheless, currently arising treatments, mainly in the fields of immunotherapy and biologicals, hold great promise for targeted and causal management of allergic conditions. Active involvement of all stakeholders, including Patient Organizations and policy makers are necessary to achieve the aims emphasized herein

Polymorphism rs in the PPARG gene in relation to the concentration and composition of fatty acids in Czech adolescents

The prevalence of obesity and type 2 diabetes in the adolescent population has been steadily rising. According to the previous studies, the rs1801282 (Pro12Ala) polymorphism in the PPARG gene is connected to the development of obesity, hypertension, insulin resistance, hyperlipemia, and other markers of type 2 diabetes. Data collection was undertaken as part of the COPAT project (Childhood Obesity Prevalence and Treatment). A sample of 2246 Czech adolescents aged 13-17 was genotyped using RT-PCR. 735 individuals from this sample were subjected to further anthropometric and clinical measurements, and biochemical testing. Data on their nutritional intake and dietary habits were collected as well. Our goal was to 1) determine the effect of the polymorphism on the anthropometric, clinical and biochemical parameters, 2) determine the effect of the genotype-phenotype interaction on the lipid spectra. We measured the frequency of Ala allele as 15%. Ala allele was associated with lower fasting c-peptide (p=0,006) and fasting insulin levels (p=0,035). In obese Ala carriers we detected higher levels of phospholipid and triacylglycerol ω-3 polyunsaturated fatty acid than in lean Ala carriers or obese noncarriers. Keywords: PPARγ2, Pro12Ala, fatty acids, obesity, COPAT projectPrevalence obezity a diabetu 2. typu v adolescentní populaci narůstá. Podle předchozích studií polymorfismus v PPARG rs1801282 (Pro12Ala) se pojí s rozvojem obezity, hypertenze, inzulinové rezistence, hyperlipémie a dalších markerů diabetu 2. typu. Data byla nasbírána v rámci projektu COPAT (Childhood Obesity Prevalence and Treatment). Soubor 2246 českých adolescentů ve věku 13-17 let byl genotypizován pomocí metody RT-PCR. 735 jedinců z tohoto souboru bylo podrobeno dalšímu antropometrickému a klinickému měření a biochemickým testům. Byla také nasbírána data o jejich nutričním příjmu a stravovacím návykům. Naším cílem bylo 1) určit efekt polymorfismu na antropometrické, klinické a biochemické parametry, 2) určit efekt interakce genotyp-fenotyp na lipidové spektrum. Frekvenci alely Ala jsme stanovili 15%. Alela Ala asociovala s nižší hladinou c-peptidu nalačno (p=0,006) a hladinou inzulinu nalačno (p=0,035). U obézních přenašečů alely Ala jsme detekovali vyšší hladiny triacylglycerolů a fosfolipidů ω-3 polynenasycených mastných kyselin než u štíhlých přenašečů Ala či obézních Pro homozygotů. Klíčová slova: PPARγ2, Pro12Ala, mastné kyseliny, obezita, projekt COPATDepartment of Anthropology and Human GeneticsKatedra antropologie a genetiky člověkaPřírodovědecká fakultaFaculty of Scienc

Polymorphism rs180128 in the PPARγ gene in relation to the concentration and composition of fatty acids in Czech adolescents

The prevalence of obesity and type 2 diabetes in the adolescent population has been steadily rising. According to the previous studies, the rs1801282 (Pro12Ala) polymorphism in the PPARγ gene is connected to the development of obesity, hypertension, insulin resistance, hyperlipidaemia, and other metabolic complications of type 2 diabetes. Data collection was undertaken as part of the COPAT project (Childhood Obesity Prevalence and Treatment). A sample of 2246 Czech adolescents aged 13-17 was genotyped using RT-PCR and subjected to further anthropometric and clinical measurements, and biochemical testing. Data on their nutritional intake and dietary habits were collected as well. A randomly selected subgroup of 735 individuals was further tested for serum fatty acids ratios. Our goal was to 1) determine the effect of the rs1801282 polymorphism in the PPARγ gene on the anthropometric, clinical and biochemical parameters, 2) determine the effect of the interaction between the rs1801282 polymorphism in the PPARγ gene and obesity or overweightness on the observed parameters. The frequency of Ala allele in our sample was 15%. Ala allele was associated with lower fasting c-peptide (p=0,006) and fasting insulin levels (p=0,035). In obese Ala boy carriers higher levels of phospholipid and triacylglycerol..

Socialization of Second Generation Armenian Immigrants

A qualitative research focuses on the effects of socialization in Czech cultural environment on the ethnic identity of the second generation Armenian immigrants. The research was conducted with six participants. Their age ranged between 17-28. Two of the participants were male. Author used the method of semi-structured interviews, to find out how the different socialization factors interract with a person's identity. In order to preserve the original ethnic identity, several factors must meet. The first contact of family with the destination country has a significant impact on a child's development of ethnicity. Powered by TCPDF (www.tcpdf.org

Socialization of Second Generation Armenian Immigrants

Kvalitativní výzkum se soustřeďuje na efekty socializace v rámci českého kulturního prostředí na etnickou identitu arménských migrantů druhé generace. Výzkum byl proveden se šesti respondenty. Rozmezí věku se pohybovalo mezi 17-28. Dva z respondentů byli muži. Pro zjištění způsobu, jakým různé socializační faktory interagují s identitou osoby, autorka použila metodu polostrukturovaného rozhovoru. Pro udržení původní etnické identity se musí sejít řada faktorů, největší dopad na vývoj etnické identity má první kontakt rodiny s cílovou zemí. Powered by TCPDF (www.tcpdf.org)A qualitative research focuses on the effects of socialization in Czech cultural environment on the ethnic identity of the second generation Armenian immigrants. The research was conducted with six participants. Their age ranged between 17-28. Two of the participants were male. Author used the method of semi-structured interviews, to find out how the different socialization factors interract with a person's identity. In order to preserve the original ethnic identity, several factors must meet. The first contact of family with the destination country has a significant impact on a child's development of ethnicity. Powered by TCPDF (www.tcpdf.org)Katedra psychologiePedagogická fakultaFaculty of Educatio

Socialization of Second Generation Armenian Immigrants

A qualitative research focuses on the effects of socialization in Czech cultural environment on the ethnic identity of the second generation Armenian immigrants. The research was conducted with six participants. Their age ranged between 17-28. Two of the participants were male. Author used the method of semi-structured interviews, to find out how the different socialization factors interract with a person's identity. In order to preserve the original ethnic identity, several factors must meet. The first contact of family with the destination country has a significant impact on a child's development of ethnicity. Powered by TCPDF (www.tcpdf.org

Characterization of Immune Response towards Generation of Universal Anti-HA-Stalk Antibodies after Immunization of Broiler Hens with Triple H5N1/NA-HA-M1 VLPs

(1) Background: Avian influenza viruses (AIVs) promptly evade preexisting immunity by constantly altering the immunodominant neutralizing antibody epitopes (antigenic drift) or by procuring new envelope serotypes (antigenic shift). As a consequence, the majority of antibodies elicited by infection or vaccination protect only against closely related strains. The immunodominance of the globular head of the main glycoprotein has been shown to mask the immunogenicity of the conserved regions located within the hemagglutinin (HA) protein. It has been shown that the broadly neutralizing universal antibodies recognize the HA2 domain in headless hemagglutinin (HA-stalk). Therefore, the HA-stalk is a highly conserved antigen, which makes it a good candidate to be used in universal vaccine development against AIVs. (2) Methods: Sf9 insect cells were used to produce triple H5N1/NA-HA-M1 influenza virus-like particles (VLPs) via co-expression of neuraminidase, hemagglutinin and matrix proteins from a tricistronic expression cassette. Purified influenza VLPs were used to immunize broiler hens. An in-depth characterization of the immune response was performed with an emphasis on the pool of elicited universal antibodies. (3) Results: Our findings suggest, that after vaccination with triple H5N1/NA-HA-M1 VLPs, hens generate a pool of broad-spectrum universal anti-HA-stalk antibodies. Furthermore, these universal antibodies are able to recognize the mammalian-derived HA-stalk recombinant proteins from homologous H5N1 and heterologous H7N9 AIVs as well as from the heterosubtypic human H1N1 influenza strain. (4) Conclusions: Our findings may suggest that highly pathogenic avian influenza H5 HA protein contain functional epitopes that are attractive targets for the generation of broad-spectrum antibodies against AIVs in their native hosts

The Preliminary Assessment of New Biomaterials Necessitates a Comparison of Direct and Indirect Cytotoxicity Methodological Approaches

Background: Cytotoxicity testing is a primary method to establish the safety of biomaterials, e.g., biocomposites. Biomaterials involve a wide range of medical materials, which are usually solid materials and are used in bone regeneration, cardiology, or dermatology. Current advancements in science and technology provide several standard cytotoxicity testing methods that are sufficiently sensitive to detect various levels of cellular toxicity, i.e., from low to high. The aim was to compare the direct and indirect methodology described in the ISO guidelines UNE-EN ISO 10993-5:2009 Part 5. Methods: Cell proliferation was measured using WST-1 assay, and cytotoxicity was measured using LDH test kit. Results: The results indicate that the molecular surface of biomaterials have impact on the cytotoxicity and proliferation profile. Based on these results, we confirm that the indirect method does not provide a clear picture of the cell condition after the exposure to the surface, and moreover, cannot provide complete results about the effects of the material. Conclusions: Comparison of both methods shows that it is pivotal to investigate biomaterials at the very early stages using both indirect and direct methods to access the influence of the released toxins and surface of the material on the cell condition

Assessment of the Toxicity of Biocompatible Materials Supporting Bone Regeneration: Impact of the Type of Assay and Used Controls

Assessing the toxicity of new biomaterials dedicated to bone regeneration can be difficult. Many reports focus only on a single toxicity parameter, which may be insufficient for a detailed evaluation of the new material. Moreover, published data frequently do not include control cells exposed to the environment without composite or its extract. Here we present the results of two assays used in the toxicological assessment of materials’ extracts (the integrity of the cellular membrane and the mitochondrial activity/proliferation), and the influence of different types of controls used on the obtained results. Results obtained in the cellular membrane integrity assay showed a lack of toxic effects of all tested extracts, and no statistical differences between them were present. Control cells, cells incubated with chitosan extract or chitosan-bioglass extract were used as a reference in proliferation calculations to highlight the impact of controls used on the result of the experiment. The use of different baseline controls caused variability between obtained proliferation results, and influenced the outcome of statistical analysis. Our findings confirm the thesis that the type of control used in an experiment can change the final results, and it may affect the toxicological assessment of biomaterial

Excess filaggrin in keratinocytes is removed by extracellular vesicles to prevent premature death and this mechanism can be hijacked by Staphylococcus aureus in a TLR2‐dependent fashion

Abstract Filaggrin (FLG) protein is indispensable for multiple aspects of the epidermal barrier function but its accumulation in a monomeric filaggrin form may initiate premature keratinocytes death; it is unclear how filaggrin levels are controlled before the formation of storing keratohyalin granules. Here we show that keratinocyte‐secreted small extracellular vesicles (sEVs) may contain filaggrin‐related cargo providing a route of eliminating excess filaggrin from keratinocytes; blocking of sEV release has cytotoxic effects on those cells. Filaggrin‐containing sEVs are found in plasma in both healthy individuals and atopic dermatitis patients. Staphylococcus aureus (S. aureus) enhances packaging and secretion of filaggrin‐relevant products within the sEVs for enhanced export via a TLR2‐mediated mechanism which is also linked to the ubiquitination process. This filaggrin removal system, preventing premature keratinocyte death and epidermal barrier dysfunction, is exploited by S. aureus which promotes filaggrin elimination from the skin that could help safeguard bacterial growth