Identification, expression, purification and characterization of new allergens from the Polybia paulista wasp venom

As hipersensibilidades do tipo I são caracterizadas por um grupo heterogêneo de manifestações clínicas que atingem mais de 30% da população mundial. Novas reatividades a alérgenos regionais brasileiros têm sido descritas e muitas fontes ainda não são totalmente conhecidas. Dentre os alérgenos mais prevalentes estão os venenos de insetos. A vespa regional Polybia paulista (Hymenoptera vespidae) é endêmica no sudeste do Brasil e é responsável por acidentes graves, causando reações alérgicas que podem levar ao choque anafilático. Alguns componentes dos venenos de vespas de diferentes espécies apresentam mimetismo molecular ou biológico, podendo gerar reação imunológica cruzada, mas muitas vezes não são os responsáveis pelo desencadeamento da resposta alérgica. Isto ocasiona falha no diagnóstico e consequentemente no tratamento indicado, a imunoterapia alérgeno-específica. Diante desses fatos e do grande número de pacientes que procuram o Serviço de Imunologia Clínica e Alergia do Hospital das Clínicas de São Paulo (HCFMUSP) com manifestações clínicas de alergias a ferroadas de insetos, foi desenvolvida uma sistemática de investigação clínica e laboratorial, com ênfase na abordagem proteômica, para identificar e caracterizar físico-química e imunologicamente novos alérgenos do veneno da vespa Polybia paulista e estudar potenciais reatividades cruzadas com alérgenos já conhecidos. Vinte e um pacientes com história de anafilaxia a venenos de vespa foram selecionados para participar do estudo. Foram realizados testes cutâneos e in vitro com veneno de Polistes spp. disponível comercialmente e com o veneno da Polybia paulista, produzido seguindo o protocolo padronizado anteriormente. Os resultados mostraram que a maioria dos pacientes apresentam IgE específica para os dois venenos com maior reatividade ao veneno de Polybia e que o padrão de proteínas reconhecidas entre os dois venenos é diferente, evidenciando a necessidade de veneno de Polybia paulista na prática clínica nas regiões cuja vespa está presente. Foram identificadas mais de 2000 proteínas no extrato total do veneno de Polybia paulista e algumas proteínas alergênicas ainda não descritas. Dentre elas foi identificada uma nova isoforma ao antígeno 5 da vespa Polybia scutellaris relatada como hipoalergênica. A molécula foi produzida na forma recombinante com conformação adequada, pela primeira vez em E. coli. O alérgeno, registrado na IUIS como Poly p 5, foi reconhecido por IgEs no soro dos pacientes testados e apresenta reatividade cruzada com outros antígenos 5 homólogos. Testes de desgranulação de basófilos em linhagem celular de ratos mostraram que o Poly p 5 induziu pouca desgranulação, indicando seu potencial hipoalergênicoType I hypersensitivity is characterized by heterogeneous clinical manifestations and specialists estimate that today around 30% of the general population suffers from an allergic disease. New allergens are being reported and some sources are not yet identified. Insect venoms are amongst the most prevalent allergen sources. The social wasp Polybia paulista (Hymenoptera vespidae) is endemic in the southeastern of Brazil and is responsible for serious accidents due to their venomous stings, causing allergic reactions that can lead to anaphylactic shock. Several components presenting molecular or biological mimicry can be found in different species of wasps and lead to a cross-immunological reaction but they are not always responsible for the allergic manifestations. Therefore, diagnostic and consequently immunotherapy is unsuccessful, since specific allergen identification is crucial. Considering the high number of patients attended at the \"Serviço de Imunologia Clínica e Alergia do Hospital das Clínicas de São Paulo\" with clinical manifestations of allergies not yet determined or barely studied, an approach involving a systematic clinical, laboratorial and investigative practice through a proteomic analysis was created to identify and characterize new allergens of Polybia paulista venom. Twenty-one patients with clinical history of anaphylaxis to Hymenoptera venoms were selected for this work. Cutaneous and in vitro tests were performed using Polistes venom commercially available as well as Polybia paulista venom, produced following a published protocol. The results shows that the majority of the patients has specific IgE for both venoms with biggest reactivity to Polybia paulista venom and the protein profile recognized in these venoms is different. More than 2000 proteins were identified in the whole venom extract of Polybia paulista and some of the allergenic proteins are not yet described in this venom. Among them, a new isoform that is similar to antigen 5 from Polybia scutellaris, already known as hypoallergenic. The molecule was produced as a recombinant properly folded for the first time in E. coli. The allergen, registered at IUIS as Poly p 5, was recognized by IgEs in the sera of 50% of the patients tested and has cross-reactivity with other homologs of antigen 5. Basophil degranulation tests in rat lineage cells showed that Poly p 5 induced little degranulation, indicating the hypoallergenic potential of this molecul

Manifestação cutânea isolada da COVID-19 conduzida por telemedicina

Objetivos: Este artigo é um relato de caso de uma paciente com diagnóstico confirmado de COVID-19 que teve avaliação médica realizada por telemedicina. Esta paciente apresentou manifestação cutânea isolada e foi avaliada por clínico geral remotamente, com diagnóstico e tratamento orientados. A avaliação correta da manifestação cutânea da COVID-19 é um desafio, e a viabilidade do diagnóstico por telemedicina ainda é incerta. Nosso objetivo é demonstrar a eficácia da teleconsulta em uma manifestação dermatológica durante a pandemia. Material e Métodos: Compilação retrospectiva de dados do prontuário da paciente e assinatura do termo de consentimento livre e esclarecido para o relato. Resultados: A apresentação clínica inicial foi de máculas pruriginosas difusas pelo abdome, que motivou a paciente a procurar atendimento presencial em pronto-socorro no primeiro dia de sintomas. A hipótese inicial foi de reação alérgica não especificada. Devido ao contexto da pandemia foram realizados exames de sangue e RT-PCR (swab nasofaríngeo) para COVID-19. A paciente recebeu alta medicada com antialérgico e com orientação para ser checar resultado dos exames em 24h e ter reavaliação médica se necessário. Ela foi reavaliada por telemedicina no dia seguinte, houve confirmação do diagnóstico de infecção aguda por COVID-19 (swab positivo), não houve manifestação de outros sintomas e foi possível reavaliar com precisão as lesões cutâneas remotamente. A lesão examinada por vídeo era compatível com rash morbiliforme difuso com angioedema local. A paciente recebeu orientações específicas conforme diretriz institucional e manutenção do tratamento sintomático dermatológico. Após 3 semanas, em nova reavaliação por telemedicina, houve confirmação da resolução do quadro cutâneo e ausência de outros sintomas neste período. Conclusão: A avaliação médica por telemedicina permitiu a interpretação da lesão cutânea elementar, tomada de decisão e orientação adequada. A manifestação cutânea isolada pode ser uma manifestação da COVID-19 e é factível de ser diagnosticada por teleconsulta, reforçando a necessidade de expansão do programa de telemedicina para avaliação inicial de pacientes agudos.Objectives: This paper is a case report of a COVID-19 patient with a confirmed diagnosis of COVID-19 who had a medical evaluation by Telemedicine (TM). She presented with an isolated cutaneous manifestation and was examined by a remote general practitioner, having diagnosis and treatment guided. The correct evaluation of COVID-19 cutaneous manifestation is a challenge, and the feasibility of TM diagnosis is still uncertain. We aim to demonstrate the effectiveness of teleconsultation in a dermatological manifestation during a pandemic. Material and Methods: Retrospective data compilation from patient’s medical record and the patient signed the consent form. Results: The initial clinical presentation was a diffuse pruritic macules in the abdomen, which motivated the patient to seek face-to-face care in the emergency department on the first day of symptoms. The initial hypothesis was an unspecified allergic reaction. Due to the context of the pandemic, blood tests and RT-PCR (nasopharyngeal swab) were performed for COVID-19. The patient was discharged medicated with an anti allergy and instructed to check the results of the test within 24 hours and have a medical reassessment if necessary. She was re-evaluated by TM the following day, the diagnosis of acute infection by COVID-19 was confirmed (swab positive), there was no manifestation of other symptoms and it was possible to accurately re-evaluate the skin lesions remotely. The patient had a diffuse morbilliform rash with local angioedema. The patient received specific orientations according to institutional guidelines and maintenance of dermatological symptomatic treatment. After 3 weeks, in a new reassessment by TM, there was confirmation of the resolution of the skin condition and absence of other symptoms during this period. Conclusion: The medical evaluation by TM allowed the interpretation of the elementary skin lesion, decision making and adequate guidance. The isolated cutaneous manifestation can be a manifestation of COVID-19 and it is feasible to be diagnosed by teleconsultation, reinforcing the need to expand the MT program for the initial evaluation of acute patients

In-hospital stroke protocol outcomes before and after the implementation of neurological assessments by telemedicine: an observational case–control study

PurposeStroke is the second leading cause of global adult mortality and the primary cause of disability. A rapid assessment by a neurologist for general and reperfusion treatments in ischemic strokes is linked to decreased mortality and disability. Telestroke assessment is a strategy that allows for neurological consultations with experienced professionals, even in remote emergency contexts. No randomized studies have compared face-to-face neurological care outcomes with telestroke care. Whether neurologists in an institution achieve better results remotely than in person is also unknown. This study aimed to compare mortality and other outcomes commonly measured in stroke protocols for stroke patients assessed by a neurologist via face-to-face evaluations and telestroke assessment.MethodsObservational single-center retrospective study from August/2009 to February/2022, enrolling 2,689 patients with ischemic stroke, subarachnoid hemorrhage, and intracerebral hemorrhage. Group 1 (G1) comprised 2,437 patients with in-person neurological assessments, and Telemedicine Group 2 (G2) included 252 patients.ResultsThe in-person group had higher admission NIHSS scores (G1, 3 (0; 36) vs. G2, 2 (0; 26), p < 0.001). The door-to-groin puncture time was lower in the in-person group than in the telestroke group (G1, 103 (42; 310) vs. G2, 151 (109; 340), p < 0.001). The telestroke group showed superior metrics for door-to-imaging time, symptomatic hemorrhagic transformation rate in ischemic stroke patients treated with intravenous thrombolysis, hospital stay duration, higher rates of intravenous thrombolysis and mechanical thrombectomy, and lower mortality. Symptomatic hemorrhagic transformation rate was smaller in the group evaluated via telestroke (G1, 5.1% vs. G2, 1.1%, p = 0.016). Intravenous thrombolysis and mechanical thrombectomy rates were significantly higher in telestroke group: (G1, 8.6% vs. G2, 18.2%, p < 0.001 and G1, 5.1% vs. G2, 10.4%, p = 0.002, respectively). Mortality was lower in the telestroke group than in the in-person group (G1, 11.1% vs. G2, 6.7%, p = 0.001). The percentage of patients with an mRS score of 0–2 at discharge was similar in both groups when adjusting for NIHSS score and age.ConclusionThe same neurological emergency team may assess stroke patients in-person or by telemedicine, with excellent outcome metrics. This study reaffirms telestroke as a safe tool in acute stroke care