An Unusual IgE-Mediated Hypersensitivity: Two Case Reports of Paracetamol Allergy.

Paracetamol is one of the most commonly used analgesic and antipyretic agents worldwide, attributed in part to its excellent safety profile when administered at recommended doses. Paracetamol allergy is not common, and the majority of the reactions are related to the pharmacological action of cyclooxygenase 1 inhibition. Selective and Immunoglobulin E (IgE)-mediated hypersensitivity reactions are rare. In this article, the authors report two cases of paracetamol allergy in which the mechanism of IgE-mediated hypersensitivity was demonstrated by positive skin tests and basophil activation tests. We highlight the relevance of identifying the mechanism underlying the reaction since patients with IgE-mediated paracetamol allergies will be able to tolerate non-steroidal anti-inflammatory drugs.

Diaporthe species associated with twig blight and dieback of Vaccinium corymbosum in Portugal, with description of four new species.

Blueberry, an increasingly cultivated fruit crop in Portugal, is known to be susceptible to twig blight and dieback caused by species of Diaporthe. The diversity of Diaporthe species associated with symptomatic and asymptomatic Vaccinium corymbosum plants in Portugal was assessed. A multilocus sequence analysis of the rDNA internal transcribed spacer (ITS) region and the translation elongation factor 1-alpha (tef1-α), ß-tubulin (tub2), calmodulin (cal), and histone 3 (his3) genes revealed the presence of Diaporthe foeniculina, Diaporthe rudis, and four new species, which are described as Diaporthe crousii, Diaporthe phillipsii, Diaporthe rossmaniae, and Diaporthe vacuae. These new species were characterized in terms of their morphology, mating strategies, and temperature growth requirements. In artificial inoculation trials of V. corymbosum cv. Bluecrop plants, all Diaporthe species caused minor symptoms. Further, no differences in aggressiveness were apparent between species. This study provides the first survey of Diaporthe species associated with blueberry twig blight and dieback in Portugal. It disclosed the occurrence of a diverse assemblage of Diaporthe species, whose status and impact as pathogens of blueberry is not yet fully understood.

Contribution of molecular diagnosis to bee venom allergic patients with systemic reactions during the build-up phase of bee venom immunotherapy / Contribuição do diagnóstico molecular em doentes alérgicos ao veneno de abelha com reações sistêmicas durante o ultra-rush

Introduction: Bee venom (BV) allergy is one of the most common causes of severe anaphylaxis. Venom immunotherapy (VIT) is considered the most effective treatment, but systemic reactions may occur. This study aimed to characterize the sensitization profile by molecular components of patients with BV anaphylaxis under VIT and to evaluate whether systemic reactions during the build-up phase of VIT protocol are related to different sensitization patterns. Methods: A retrospective study of 30 patients under VIT for 1 year. The group of patients who reacted during the build-up phase (group A) was compared with the group with no reactions (group B). Specific IgE (sIgE) and IgG4 (sIgG4) for BV and recombinants (rApi m1, rApi m2, rApi m3, rApi m5, and rApi m10) were evaluated before and 1 year after VIT. Statistical analysis was performed using GraphPad Prism v5.01. Results: Men accounted for 80% of the sample, and mean age was 47 years (14-74 years). Group A consisted of 10 patients, and group B of 20 patients. Before VIT, sIgE to rApi m1 was detected in 86.7% of patients, rApi m2 in 46.7%, rApi m3 in 16.7%, rApi m5 in 43.3%, and rApi m10 in 70%. Positive results to at least 1 BV allergen were detected in 100%; 73% of patients were sensitized to >1 allergen, and 13.3% to all allergens. The profile of the two groups did not differ significantly before VIT, but group B showed a significant decrease in whole BV extract (p=0.045), rApi m 3 (p=0.017), and rApi m 10 (p=0.021) 1 year after VIT. Regarding sIgG4, there was a significant increase in rApi m1, which was not observed in other allergens, such as rApi m3 and rApi m10. Conclusion: The analysis of a panel of BV recombinants can improve diagnostic sensitivity, when compared to rApi m1 alone. There was no association between systemic reactions during the build-up phase of VIT and molecular sensitization profile. Nevertheless, it is important to study a greater number of patients.

Introdução: A alergia ao veneno de abelha (VA) é uma das causas mais comuns de anafilaxia grave. A imunoterapia com veneno de abelha (VIT) é considerada o tratamento mais eficaz, mas reações sistêmicas podem ocorrer. O objetivo deste estudo foi caracterizar o perfil de sensibilização por componentes moleculares de doentes com anafilaxia a VA e avaliar se reações sistêmicas durante o ultrarush estão relacionadas com diferentes padrões de sensibilização. Métodos: Estudo retrospectivo incluindo 30 doentes submetidos a VIT durante 1 ano. Considerou-se dois grupos: grupo de doentes que reagiu durante o ultra-rush (Grupo A), que foi comparado com o grupo sem reação (Grupo B). Foram avaliadas as IgE (sIgE) e IgG4 (sIgG4) específicas para VA(i1) e componentes moleculares: rApi m1, rApi m2, rApi m3, rApi m5 e rApi m10 antes e 1 ano após VIT. Os testes estatísticos foram realizados com Graph-PadPrism v5.01. Resultados: 80% sexo masculino, média de idade 47 anos (14-74). Grupo A com 10 doentes, Grupo B com 20 doentes. Previamente à VIT, sIgE para rApi m1 foi detectada em 86,7%; rApi m2 em 46,7%; rApi m3 em 16,7%; rApi m5 em 43,3%; e rApi m10 em 70%. Resultados positivos para pelo menos um alergênio de VA foram detectados em 100%. 73% dos doentes eram sensibilizados a mais de um alergênio, e 13,3% a todos os alergênios. Não houve diferenças estatisticamente significativas no perfil dos dois grupos antes da VIT, porém verificouse uma diminuição significativa: p = 0,045; p = 0,017 e p = 0,021 de i1, rApi m3 e rApi m10, respectivamente, no grupo B um ano após VIT. Relativamente à sIgG4, observou-se um aumento significativo de rApi m1, não observado nos restantes alergênios como rApi m3 e rApi m10. Conclusão: A análise de um painel de recombinantes de VA pode melhorar a sensibilidade diagnóstica, quando comparado com rApi m1 isolado. Não se verificou associação entre a ocorrência de reações sistêmicas durante o ultra-rush e o perfil de sensibilização molecular. No entanto, é importante para estudar um maior número de doentes.

Chlorhexidine: a hidden life-threatening allergen.

Chlorhexidine is a commonly used antiseptic and disinfectant in the health-care setting. Anaphylaxis to chlorhexidine is a rare but potentially life-threatening complication. Epidemiologic data suggest that the cases of chlorhexidine allergy appears to be increasing. In this article we report a life-threatening anaphylactic shock with cardiorespiratory arrest, during urethral catheterization due to chlorhexidine. The authors also performed a literature review of PubMed library of anaphylactic cases reports due to this antiseptic between 2014 and 2018, demonstrating the increase in the number of cases occurring worldwide and the importance of detailed anamnesis and appropriate diagnostic workup of allergic reactions to disinfectants.

Mating type gene analyses in the genus Diplodia: From cryptic sex to cryptic species.

Cryptic species are common in Diplodia, a genus that includes some well-known and economically important plant pathogens. Thus, species delimitation has been based on the phylogenetic species recognition approach using multigene genealogies. We assessed the potential of mating type (MAT) genes sequences as phylogenetic markers for species delimitation in the genus Diplodia. A PCR-based mating type diagnostic assay was developed that allowed amplification and sequencing of the MAT1-1-1 and MAT1-2-1 genes, and determination of the mating strategies used by different species. All species tested were shown to be heterothallic. Phylogenetic analyses were performed on both MAT genes and also, for comparative purposes, on concatenated sequences of the ribosomal internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1-α) and beta-tubulin (tub2). Individual phylogenies based on MAT genes clearly differentiated all species analysed and agree with the results obtained with the commonly used multilocus phylogenetic analysis approach. However, MAT genes genealogies were superior to multigene genealogies in resolving closely related cryptic species. The phylogenetic informativeness of each locus was evaluated revealing that MAT genes were the most informative loci followed by tef1-α. Hence, MAT genes can be successfully used to establish species boundaries in the genus Diplodia.

Mating type genes in the genus Neofusicoccum: Mating strategies and usefulness in species delimitation.

The genus Neofusicoccum includes species with wide geographical and plant host distribution, some of them of economic importance. The genus currently comprises 27 species that are difficult to identify based on morphological features alone. Thus, species differentiation is based on phylogenetic species recognition using multigene genealogies. In this study, we characterised the mating type genes of Neofusicoccum species. Specific primers were designed to amplify and sequence MAT genes in several species and a PCR-based mating type diagnostic assay was developed. Homothallism was the predominant mating strategy among the species tested. Furthermore, the potential of mating type gene sequences for species delimitation was evaluated. Phylogenetic analyses were performed on both MAT genes and compared with multigene genealogies using sequences of the ribosomal internal transcribed spacer region, translation elongation factor 1-alpha and beta-tubulin. Phylogenies based on mating type genes could discriminate between the species analysed and are in concordance with the results obtained with the more conventional multilocus phylogenetic analysis approach. Thus, MAT genes represent a powerful tool to delimit cryptic species in the genus Neofusicoccum.

Influence of household cooking methods on amino acids and minerals of Barrosã-PDO veal.

The effect of commonly household cooking methods (boiling, microwaving and grilling) on amino acid and mineral (Fe, Mg, K and Zn) contents was investigated in the longissimus lumborum muscle of Barrosã-PDO veal. Fifteen Barrosã purebred calves at 7-8 months of age and an average weight of 177±37 kg were slaughtered. Cooking had a strong effect (P<0.05) on yield, being higher (67.5%) in boiling compared to microwave and grilling (64.0% and 64.5%, respectively). Grilling increased most of the percentage retention of individual amino acids (>100%), in particular for leucine. No significant differences (P>0.05) were observed for iron and zinc retentions among the cooking methods, while the retention of magnesium and potassium was strongly affected, mainly after boiling. Our findings indicate that the different cooking methods clearly affect the chemical composition and nutritional value of meat, which may have a strong impact on the intake of essential nutrients.

Biomineralization studies on cellulose membrane exposed to biological fluids of Anodonta cygnea.

The present work proposes to analyse the results obtained under in vitro conditions where cellulose artificial membranes were incubated with biological fluids from the freshwater bivalve Anodonta cygnea. The membranes were mounted between two half 'Ussing chambers' with different composition solutions in order to simulate epithelial surfaces separating organic fluid compartments. The membrane surfaces were submitted to two synthetic calcium and phosphate solutions on opposite sides, at pH 6.0, 7.0 or 9.0 during a period of 6 hours. Additional assays were accomplished mixing these solutions with haemolymph or extrapallial fluid from A. cygnea, only on the calcium side. A selective ion movement, mainly dependent on the membrane pore size and/or cationic affinity, occurred with higher permeability for calcium ions to the opposite phosphate chamber supported by calcium diffusion forces across the cellulose membrane. In general, this promoted a more intense mineral precipitation on the phosphate membrane surface. A strong deposition of calcium phosphate mineral was observed at pH 9.0 as a primary layer with a homogeneous microstructure, being totally absent at pH 6.0. The membrane showed an additional crystal phase at pH 7.0 exhibiting a very particular hexagonal or cuttlebone shape, mainly on the phosphate surface. When organic fluids of A. cygnea were included, these crystal forms presented a high tendency to aggregate under rosaceous shapes, also predominantly in the phosphate side. The cellulose membrane was permeable to small organic molecules that diffused from the calcium towards the phosphate side. In the calcium side, very few similar crystals were observed. The presence of organic matrix from A. cygnea fluids induced a preliminary apatite-brushite crystal polymorphism. So, the present results suggest that cellulose membranes can be used as surrogates of biological epithelia with preferential ionic diffusion from the calcium to the phosphate side where the main mineral precipitation events occurred. Additionally, the organic fluids from freshwater bivalves should be also thoroughly researched in the applied biomedical field, as mineral nucleators and crystal modulators on biosynthetic systems.

Reproductive cycle and strategy of Anodonta anatina (L., 1758): notes on hermaphroditism.

Freshwater mussels have decreased dramatically in Iberia over the last decades. These animals are responsible for important ecosystem services such as recycling nutrients and improving water clarity. Under this view a better knowledge on the biological features of these animals is extremely important for future conservation and management actions. In this study the reproductive and gametogenic cycle of Anodonta anatina were studied during 2 years in one population as well as the sex ratio and hermaphroditism in six distinct populations, using standard histology. Gametogenesis was continuous in both sexes and germinal epithelium in early stages of development. Gametes were present throughout the reproductive cycle. Oogenesis and spermatogenesis occurred mainly between January and May. Larvae brooding occurred between September and March and main glochidia discharge occurred over a short period (2-3 weeks) in March. For the sex-ratio and hermaphroditism assessments a variable number of individuals were collected from several populations from lakes and rivers. Previous studies described A. anatina as mainly dioecious with only a few populations presenting occasional hermaphroditism. However, the present study indicates that A. anatina sexual behavior is influenced by environmental conditions, being mainly dioecious in rivers with increased hermaphroditism in standing waters. Although self-fertilization was not confirmed, additional studies with molecular characterization of larvae using fast evolving markers should be used in future studies to enlighten this process. Overall, this study indicates that for more efficient conservation actions and management plans, freshwater mussel reproductive biology should be studied at the population level mainly in the subfamily Anodontinae.

Cuidador informal: sobrecarga e atitudes face à morte / Informal caregiver: burden and attitudes towards death

A sobrecarga do cuidador informal (CI) tem-se tornado numa importante preocupação para a enfermagem de reabilitação, não só pela sua influência na qualidade de vida destas pessoas e independência funcional, como também pelos elevados custos sociais associados (Goodhead e McDonald, 2007; Petronilho, 2010, Cruz et al., 2010). Apesar de as questões relacionadas com a morte terem adquirido nas últimas décadas estatuto relevante nas ciências sociais e humanas (Loureiro, 2010), estas tem merecido pouca atenção quando se estuda a sobrecarga do CI (Reimer, 2007). Realizou-se um estudo quantitativo descritivo-correlacional com os objetivos principais de: analisar a influência das variáveis sociodemográficas e contextuais na sobrecarga do CI; analisar a influência da sobrecarga do CI nas suas atitudes perante a morte. Utilizou-se para o efeito o Questionário de Avaliação da Sobrecarga do Cuidador Informal (QASCI) e a Escala de Avaliação do Perfil de Atitudes Acerca da Morte (EAPAM). Foram inquiridas através de questionário 32 cuidadores informais. Os resultados evidenciaram correlações significativas entre : "idade" e atitude de "Evitamento"; "anos de cuidado" e atitude de "Neutralidade"; "estado civil" e as dimensões "Reações a Exigências" e "Perceção de Mecanismos Eficácia e Controlo"; "motivo da dependência" e atitude de "Medo", bem como com as dimensões "Sobrecarga Financeira" e "Implicações na Vida Pessoal"; "Implicações Vida Pessoal" e "Neutralidade"; "Sobrecarga Emocional" e "Medo". Estes resultados são sustentados pela literatura, nomeadamente relacionada com as atitudes perante a morte (Reimer, 2007), afirmando-as um importante foco de atenção dos enfermeiros.

Informal Caregiver: Overburden and Attitudes Towards Death Informal caregiver's (IC) overburden has become a major concern to rehabilitation nursing, not only because of their influence on these people's quality of life and functional independency, as well as the associated high social costs (Goodhead & McDonald, 2007; Petronilho, 2010, Cruz et al., 2010). While issues related to death have acquired relevant status in recent decades in what social sciences and humanities concerns (Loureiro, 2010), these have received little attention when studying IC's overburden (Reimer, 2007). A descriptive-correlational quantitative study has been conducted with the main objetives: to analyze the influence of sociodemographic and contextual variables in IC's overburden; analyze the influence of IC's overburden in their attitudes towards death. For this purpose, the Assessment Questionnaire of Informal Caregiver Overburden (QASCI) and the Profile Rating Scale of Attitudes About Death (EAPAM) were used. 32 informal caregivers were surveyed by questionnaire. Results showed significant correlations between "Age" and "Avoidance" attitude; "Years of care" and "Neutrality" attitude; "marital status" and the dimensions "Reactions to Demandings" and "Efficacy and Control Perception Mechanisms "; "cause of dependency" and "Fear's" attitude, as well as with the dimensions "Financial Burden" and "Implications for Personal Life", "Implications for Personal Life" and "Neutrality, "Emotional Overload" and "Fear". Literature supports these results, particularly regarding the attitudes towards death (Reimer, 2007), affirming them as an important focus of nurses attention.

Effect of cooking methods on fatty acids, conjugated isomers of linoleic acid and nutritional quality of beef intramuscular fat.

The effect of boiling, microwaving and grilling on the composition and nutritional quality of beef intramuscular fat from cattle fed with two diets was investigated. Longissimus lumborum muscle from 15 Alentejano young bulls fed on concentrate or pasture was analyzed. Cooking losses and, consequently, total lipids, increased directly with the cooking time and internal temperature reached by meat (microwaving>boiling>grilling). The major changes in fatty acid composition, which implicated 16 out of 34 fatty acids, resulted in higher percentages in cooked beef of SFA and MUFA and lower proportions of PUFA, relative to raw meat, while conjugated linoleic acid (CLA) isomers revealed a great stability to thermal processes. Heating decreased the PUFA/SFA ratio of meat but did not change its n-6/n-3 index. Thermal procedures induced only slight oxidative changes in meat immediately after treatment but hardly affected the true retention values of its individual fatty acids (72-168%), including CLA isomers (81-128%).

Seasonal variations of pH, pCO2, pO2, HCO3- and Ca2+ in the haemolymph: implications on the calcification physiology in Anodonta cygnea.

A study about the relationship between the physical-chemical parameters and the calcium carbonate balance between the haemolymph fluid and mantle calcareous structures was carried out in Anodonta cygnea. An intense peak of HCO(3) (-) and a highest pH in December-January months may be understood as a preparation period for creating alkaline conditions. An intense pH decrease from January to February in parallel with the HCO(3) (-) reduction seems to indicate the beginning process of carbonate precipitation. On the other hand, the following calcium and HCO(3) (-) increases in February-May associated with a continuous and gradual pH fall profile may infer two combined aspects: calcium and HCO(3) (-) absorption from external environment and a simultaneous intense calcium carbonate deposition in the haemolymph. So, the pCO(2) peak in this period reflects a subsequent result on equilibrium balance between HCO(3) (-) absorption and deposition. The only significant pO(2) increase in the next period, from February to June, is related with an energetic increase to support the metabolic activity favouring the posterior intense pCO(2) peaks. The extended time of CO(2) production in the haemolymph from May to November should induce an increased metabolic acidosis with subsequent intense formation of both HCO(3) (-) and Ca(2+) ions in the same period. This seems to result from CaCO(3) deposits dissolution in the haemolymph, the most direct calcareous source. Additionally, the later increase of metabolic succinic acid during autumn may greatly potentiate this acidosis increasing the dissolution process. Consequently, the pH profile present two simultaneous alkaline peaks in July and October, probably due to a strong HCO(3) (-) release from the CaCO(3) dissolution. So, the present seasonal results indicate that in the freshwater bivalve A. cygnea, the low metabolism with higher pH from the early winter is the main cause which may favour a calcareous precipitation, while the high metabolism with lower pH from the early summer may function as an inductor of calcareous dissolution in the haemolymph.