Alterações oculares decorrentes de picada de abelha na córnea / Ocular changes due to corneal bee sting

RESUMO O propósito deste estudo foi reportar as alterações oculares observadas após picada de abelha com ferrão retido na córnea. Destacamos o tratamento e o desfecho de uma lesão de córnea incomum e sua patogênese. Trata-se de relato de caso e revisão da literatura de lesões oculares por picada de abelha. Paciente do sexo feminino, 63 anos, procurou atendimento oftalmológico de urgência devido à picada de abelha na córnea do olho direito há 6 dias. Queixava-se de embaçamento visual, dor e hiperemia ocular. Apresentou acuidade visual de vultos no olho afetado. Ao exame, notaram-se hiperemia moderada de conjuntiva bulbar, edema corneano com dobras de Descemet e presença do ferrão alojado na região temporal, no estroma profundo da córnea. A paciente foi internada para ser abordada no centro cirúrgico sob anestesia geral. Durante a cirurgia, o ferrão teve que ser retirado via câmara anterior, mediante a realização de uma paracentese e uma lavagem da câmara anterior, com dupla via e solução salina balanceada. Ainda não existe na literatura um tratamento padrão na abordagem de pacientes com lesões oculares por picada de abelha, sendo importantes a identificação e o reconhecimento precoce de possíveis complicações que ameacem a visão.

ABSTRACT The purpose of this study was to report the ocular changes observed after a bee sting with a stinger retained in the cornea. We show the treatment and outcome of an unusual corneal injury and its pathogenesis. This is a case report and literature review of ocular injuries caused by bee stings. A 63-year-old female patient sought emergency ophthalmic care because of a bee sting on the cornea of her right eye six days before. She complained of blurred vision, pain, and ocular hyperemia. She had glare sensitivity on visual acuity in the affected eye. Examination revealed moderate hyperemia of the bulbar conjunctiva, corneal edema with Descemet's folds and a stinger lodged in the temporal region, in the deep stroma of the cornea. The patient was admitted to the operating room under general anesthesia. During surgery, the stinger had to be removed via the anterior chamber, by performing a paracentesis and washing the anterior chamber with a double flushing and balanced saline solution. There is still no standard treatment in the literature for patients with eye injuries caused by bee stings, and early identification and recognition of possible sight-threatening complications is important.

Epidemiological study of bee stings in the state of Paraiba, Northeastern Brazil, from 2015 to 2019

INTRODUCTION: In Brazil, envenomation by bee sting is a public health problem due to its incidence in all regions of the country, as well as the severity of cases. Despite the medical and sanitary importance, the literature on this topic in Brazil is scarce, in the Northeast region. OBJECTIVE: To describe the epidemiological characteristics of bee stings in Paraíba, in the Northeast region of Brazil, from 2015 to 2019. METHODS: The current study is a descriptive and retrospective epidemiological investigation conducted through the recovery of secondary data made available by the database from the Ministry of Health Notification Information System. RESULTS: A total of 1,151 cases were analyzed, and they were reported during all the months of the period of the study and were more frequent in the regions Agreste and Sertão of the Paraíba. The victims were men aged 20 to 59 years. Most individuals received medical assistance within 3 hours of the incident. The cases were classified as mild and progressed to cure. CONCLUSION: In Paraíba, bee stings have increased in recent years, especially in regions with a semi-arid climate. In addition, this study provides data that may be used in the development of educational health actions to promote the prevention and control of bee stings in this region.

Anaphylactic shock with pulmonary eosinophilic infiltration due to honeybee attack in a donkey: case report / [Choque anafilático com infiltração eosinófila pulmonar devido a ataque de abelhas em um burro: relato de caso]

A case of a donkey attacked by Africanized honeybee is reported here with clinical signs of agitation, dehydration, congestion of the ocular mucous membranes, tongue edema, tachycardia and inspiratory dyspnea, and progression to death. At necropsy, diffuse, severe subcutaneous edema at face and cervical regions and severe diffuse pulmonary hyperemia with abundant edema without parenchymal collapse were observed. Microscopically, marked, diffuse deep dermis and panniculus carnosus edema and marked diffuse alveolar edema, with moderate population of eosinophils predominantly around larger caliber vessels were noted. The final diagnosis of anaphylactic shock was supported by history, clinical signs, and anatomic pathology findings. This is the first report of a honeybee attack with pulmonary eosinophilic infiltration in a mammal.(AU)

Descreve-se um caso de ataque de abelha africanizada em um burro, com sinais clínicos de agitação, desidratação, mucosas oculares congestas, edema de língua, taquicardia e dispneia inspiratória, com progressão e morte. Na necropsia, foram verificados edema subcutâneo difuso grave nas regiões de face e cervical, hiperemia pulmonar difusa grave com edema abundante e sem colapso do parênquima. Microscopicamente, foram observados edema marcado difuso na derme profunda e panículo carnoso e edema alveolar difuso acentuado, com população moderada de eosinófilos predominantemente em torno de vasos de maior calibre. O diagnóstico de choque anafilático foi baseado no histórico, em sinais clínicos e em achados anatomopatológicos. Este é o primeiro relato de ataque de abelhas com infiltração eosinofílica pulmonar em um mamífero.(AU)

Venenos animais como possíveis ferramentas terapéuticas contra o estresse oxidativo em diversas doengas: Revisao da Literatura / Animal venoms as possible therapeutic tools against oxidative stress in several diseases: Literature Review

Resumo Venenos sao uma substancia tóxica (composta por uma ou mais toxinas) que podem causando lesao fisiológica dependente da dose. As toxinas sao moléculas bioativas formadas principalmente por compostos enzimáticos e nao enzimático que porque provocam consequéncias indesejáveis nas presas, além disso, exibem atividades biológicas únicas, diversas e específicas que perturbam os processos fisiológicos normais. Entretanto, muitas toxinas, de diferentes animais, tém sido isoladas e muitas delas sao consideradas ótimas ferramentas para pesquisa básica e alvos terapéuticos. Foi relatado que o estresse oxidativo desempenha um papel fundamental na patogénese de várias doengas, como distúrbios neurodegenerativos, distúrbios cardiovasculares e cáncer. O mecanismo pelo qual as toxinas animais atuam nos parametros de estresse oxidativo em várias doengas, ainda nao está estabelecido. O foco principal desta revisao é destacar os principais estudos com toxinas animais como ferramenta terapéutica em diversas doengas, atuando no balango redox do organismo.

Abstract Venoms are a toxic substance (comprised of one or more toxins) that can cause dose-dependent physiological injury. Toxins are bioactive molecules formed primarily by enzymatic and non-enzymatic compounds that cause undesirable conse-quences in prey, in addition, exhibit unique, diverse and specific biological activities that disrupt normal physiological processes. However, many toxins, from different animals, have been isolated and many of them are considered great tools for basic research and therapeutic targets. Oxidative stress has been reported to play a key role in the pathogenesis of various diseases such as neurodegenerative disorders, cardiovascular disorders and cancer. How animal toxins act on oxidative stress parameters in several diseases is not yet established. The main focus of this review is to highlight the main studies with animal toxins as a therapeutic tool in several diseases, acting on the organism's redox balance.

Resumen Los venenos son sustancias tóxicas (compuestas por una o más toxinas) que pueden causar daño fisiológico dependiente de la dosis. Las toxinas son moléculas bioactivas formadas principalmente por compuestos enzimáticos y no enzimáticos que debido a que causan consecuencias indeseables en las presas, además, exhiben actividades biológicas únicas, diversas y específicas que alteran los procesos fisiológicos normales. Sin embargo, se han aislado muchas toxinas de diferentes animales, y muchos de ellos se consideran grandes herramientas para la investigación básica y dianas terapéuticas. Se ha informado que el estrés oxidativo juega un papel clave en la patogenia de diversas enfermedades, como los trastornos neurodegenerativos, enfermedades cardiovasculares y cáncer. El mecanismo por el cual las toxinas animales actúan sobre los parámetros de estrés oxidativo en vários enfermedades, aún no está establecido. El enfoque principal de esta revisión es resaltar los principales estudios con toxinas animales como herramienta terapéutica en diversas enfermedades, actuando en el equilibrio redox del organismo.

Melittin-induced metabolic changes on the Madin-Darby Bovine Kidney cell line / [Mudanças metabólicas induzidas por melitina em células da linhagem Madin-Darby Bovine Kidney]

In this study, the toxic effects of melittin on Madin-Darby Bovine Kidney cells (MDBK) were analyzed with respect to mitochondrial functionality by reduction of MTT and flow cytometry, apoptosis potential, necrosis, oxygen reactive species (ROS) production, lipid peroxidation, and DNA fragmentation using flow cytometry and cell membrane destabilization by confocal microscopy. The toxicity presented dose-dependent characteristics and mitochondrial activity was inhibited by up to 78.24 ±3.59% (P<0.01, n = 6) in MDBK cells exposed to melittin (10µg/mL). Flow cytometry analysis revealed that melittin at 2µg/mL had the highest necrosis rate (P<0.05) for the cells. The lipoperoxidation of the membranes was also higher at 2µg/mL of melittin (P<0.05), which was further confirmed by the microphotographs obtained by confocal microscopy. The highest ROS production occurred when the cells were exposed to 2.5µg/mL melittin (P<0.05), and this concentration also increased DNA fragmentation (P<0.05). There was a significative and positive correlation between the lipoperoxidation of membranes with ROS (R=0.4158), mitochondrial functionality (R=0.4149), and apoptosis (R=0.4978). Thus, the oxidative stress generated by melittin culminates in the elevation of intracellular ROS that initiates a cascade of toxic events in MDBK cells.(AU)

Neste estudo, os efeitos tóxicos da melitina em células Madin-Darby Bovine Kidney (MDBK) foram analisados quanto à funcionalidade mitocondrial, por redução de MTT e citometria de fluxo, potencial de apoptose, necrose, produção de espécies reativas de oxigênio (ROS), peroxidação lipídica e fragmentação de DNA, utilizando-se citometria de fluxo e desestabilização da membrana celular, por microscopia confocal. A toxicidade apresentou características dose-dependentes e a atividade mitocondrial foi inibida até 78,24±3,59% (P<0,01, n = 6) em células MDBK expostas à melitina (10µg/mL). Análises por citometria de fluxo revelaram que a melitina a 2µg/mL apresentou o maior índice necrótico celular (P<0,05). A maior lipoperoxidação de membranas também foi na concentração de 2µg/mL de melitina (P<0,05), o que foi posteriormente confirmado por microscopia confocal. A maior produção de ROS aconteceu quando as células foram expostas a 2,5µg/mL de melitina (P<0,05), e essa concentração também aumentou a fragmentação de DNA (P<0,05). Houve uma significativa correlação positiva entre a lipoperoxidação de membranas e a produção de ROS (R=0,4158), funcionalidade mitocondrial (R=0,4149) e apoptose (R=0,4978). Portanto, o estresse oxidativo gerado pela melitina culminou na elevação de ROS intracelular, que inicia uma cascata de eventos tóxicos nas células MDBK.(AU)

Bee sting envenomation severe cases in Manaus, Brazilian Amazon: clinical characteristics and immune markers of case reports

Abstract Bee venom is a natural toxin composed of several peptides. Massive envenoming causes severe local and systemic reactions. We report two cases of severe bee envenomation, of which one was fatal. We also describe clinical characteristics and immune markers. Both victims suffered from respiratory distress, renal failure, rhabdomyolysis, and shock. They required invasive mechanical ventilation, vasoactive drugs, and renal replacement therapy. Moreover, serum levels of chemokines, cytokines, and cell-free circulating nucleic acids demonstrated an intense inflammatory process. Massive envenoming produced systemic injury in the victims, with an uncontrolled inflammatory response, and a more significant chemotactic response in the fatal case.

Characterization of the sensitization profile of bee venom allergic patients ­ Association with the severity of reaction? / Caracterização do perfil de sensibilização molecular de doentes alérgicos à picada de abelha - Associação com a gravidade da reação?

Introduction: Bee venom (BV) allergy, a common cause of anaphylaxis in adults, is often associated with severe reactions. The use of component-resolved diagnostics (CRD) increases diagnostic accuracy. Objectives: To characterize the sensitization profile of BV allergic patients and a possible correlation with the severity of reaction. Materials and methods: We selected patients with a clinical history of BV allergy, positive skin tests, and specific IgE (sIgE) for BV. The allergenic profile was analyzed by both CRD and Western blot using a well-defined and properly characterized BV extract. Results: Forty-four patients were included, 30 (68.2%) were men. Mean age was 48.9 (SD 17.9) years. Eleven (25%) had large local reactions (LLRs) and 33 (75%) had systemic sting reactions (SSRs). One patient with negative sIgE for BV had positive sIgE for Api m 1, Api m 5, and Api m 10. The sensitization frequency for BV, Api m 1, Api m 2, Api m 3, Api m 5, and Api m 10 was 97.7%, 75%, 47.7%, 20.5%, 40.9%, and 61.4%, respectively. Five patients (11.4%) were sensitized to all BV components. CRD association showed that 5 patients (11.4%) were sensitized only to Api m 1, 8 (18.2%) to Api m 1/Api m 3/Api m 10, and 16 (36.6%) to Api m 1/ Api m 10. Twenty-eight patients (84.8%) with SSRs were sensitized to Api m 1, and concomitant sensitization to Api m 1/Api m 10 was detected in 20 (60.6%). There was a significant difference in Api m 1 between patients with LLRs and SSRs (p = 0.0104). Similar profiles were identified by Western blot analysis, with relevance for the detection of Api m 6 in 28 (64%) and Api m 4 in 16 (36%) patients. Conclusion: The analysis of the sensitization profile using CRD and the association of several of these components can increase diagnostic accuracy in BV allergy. Our data showed that concomitant sensitization to Api m 1 and Api m 10, detected by both CRD and electrophoretic profile, may be associated with SSRs. We emphasize the identification of sensitization to Api m 6 in > 50% of patients, which may be considered a major allergen, and to Api m 4, which may be related to reactions during BV immunotherapy.

Introdução: A alergia ao veneno de abelha (VA) é uma causa frequente de anafilaxia em adultos e está muitas vezes associada a reações graves. O diagnóstico por componentes moleculares (CRD) contribui para uma melhor caracterização desta alergia. Objetivos: Caracterização do perfil de sensibilização molecular de doentes alérgicos ao veneno de abelha e possível correlação com a gravidade da reação. Material e métodos: Selecionaram-se doentes com história de alergia a VA, testes cutâneos e IgE específica (sIgE) positivos para VA. Avaliou-se o perfil alergênico por CRD e por Western Blot, utilizando extrato de VA bem caracterizado. Resultados: 44 doentes, 30 (68,2%) sexo masculino. Média de idades 48,9 ± 17,9 anos, 11 (25%) com reacções locais exuberantes e 33 (75%) com reações sistêmicas à picada (SSR). Um doente tinha sIgE negativa para VA, mas Api m 1, Api m 5 e Api m 10 positivas. A frequência de sensibilização para VA, Api m 1, Api m 2, Api m 3, Api m 5 e Api m 10 foi 97,7%; 75%; 47,7%; 20,5%; 40,9% e 61,4%, respectivamente. Cinco (11,4%) doentes estavam sensibilizados a todos os componentes. Por associação de CRD, detectaram-se 5 (11,4%) doentes sensibilizados apenas a Api m 1, 8 (18,2%) a Api m 1/Api m 3/Api m 10, e 16 (36,6%) a Api m 1/Api m 10. Vinte e oito (84,8%) doentes com SSR tinham Api m 1 positiva e 20 (60,6%) tinham Api m 1/Api m 10 simultaneamente positivas. Observou-se uma diferença estatisticamente significativa para a Api m 1 entre doentes com reações locais exuberantes e sistêmicas (p = 0,0104). Os perfis detectados por Western Blot foram semelhantes, de referir, à detecção de Api m 6 em 28 (64%) e Api m 4 em 16 (36%) dos doentes. Conclusão: A análise do perfil de sensibilização através de CRD e a sua associação aumentam a precisão do diagnóstico de alergia a VA. Sensibilização simultânea a Api m 1 e Api m 10 identificados tanto por CRD como por perfil eletroforético, pode estar associada à ocorrência de SSR. Destaca-se a sensibilização a Api m 6 em > 50% dos doentes, podendo ser considerado um alergênio major, e a Api m 4, possivelmente associado a reações durante a imunoterapia com VA.

Caracterización de pacientes alérgicos a picadura de abeja que reciben inmunoterapia / Characterization of allergic patients to bee sting that receive immunotherapy

La alergia al veneno de abejas provoca reacciones de leves a severas con compromiso para la vida. La inmunoterapia con veneno de himenópteros es un tratamiento eficaz y protege a los pacientes alérgicos de sufrir reacciones sistémicas ante nuevas picaduras. Nos propusimos caracterizar los pacientes alérgicos a picaduras de abeja que reciben inmunoterapia. Se realizó un estudio observacional descriptivo de corte longitudinal en pacientes alérgicos a las picaduras de abeja tratados con inmunoterapia de extracto de abeja en el Hospital Universitario General Calixto García de La Habana, Cuba. La muestra fue de 17 pacientes que cumplieron los criterios de inclusión. Usamos técnicas de estadística descriptiva: promedio, probabilidad y puntaje estandarizado, así como técnicas de estadística inferencial tales como Chi cuadrado, verificando asociación significativa entre las variables; el nivel de significación empleado fue del 5 por ciento (p˂0,05). La tercera década de la vida fue la edad promedio de los pacientes. Se observó predominio del sexo masculino y residencia en zona urbana. Alrededor de la mitad de los pacientes tenían rinitis y antecedentes familiares de asma. Todos los pacientes tuvieron reacciones locales, la mayoría se re-expusieron a la picadura; de ellos, solo el 20 por ciento presentaron reacciones alérgicas sistémicas después de la inmunoterapia. Se concluye que la reactividad cutánea al extracto de abeja se redujo con el tratamiento de inmunoterapia(AU)

Allergy to bee venom may cause from mild to severe reactions threatening the patient´s life. Immunotherapy with hymenopter venom is an effective treatment that can protect allergic patients from suffering systemic reactions to new stings. The aim of this study was to characterize allergic patients to bee sting that receive immunotherapy. A descriptive longitudinal observational study was carried out in allergic patients to bee sting receiving immunotherapy with bee extracts in the University Hospital General Calixto García, Havana, Cuba. A sample of 17 patients with inclusion criteria was analyzed. Descriptive statistical techniques were used: mean, probability, standardized score, as well as, inferential statistic techniques such as the Chi square; verifying significant association between variables. The level of signification was 5 percent (p˂0.05). The third decade of life was the average age of the patients in this study; male sex and, urban residents were predominant. Around half of the patients had rhinitis and family history of asthma. All patients had local reactions; most of the patients were re-exposed to stings. Only 20 percent of patients reported systemic allergic reaction after immunotherapy. Skin reactivity to bee extract was reduced with the immunotherapy(AU)

Epidemiology of envenomation by Africanized honeybees in the state of Rio Grande do Norte, Northeastern Brazil / Epidemiologia dos envenenamentos por abelhas africanizadas no Estado do Rio Grande do Norte, Nordeste do Brasil

ABSTRACT: Introduction: In the American continent, accidents caused by honeybees are a public health problem due to the high incidence and severity of the cases. Despite its medical importance, there are few epidemiological studies on this topic in Brazil, especially referring to the Northeastern states. The present study aims to describe the epidemiological features of honeybee envenomation cases in the state of the Rio Grande do Norte, Northeastern Brazil, from 2007 to 2014. Methods: Data were collected from the Notifiable Diseases Information System database of the Health Department of Rio Grande do Norte. Results: We analyzed a total of 2,168 cases. Cases occurred in all months of the years studied, reaching higher frequencies from June to October. Most incidents happened in urban areas and involved men, with victims aged between 20 and 39 years. Victims were mainly stung on the head and hand, and they received medical care predominantly within 3 hours after the injury. Local manifestations were more frequent than systemic ones. Clinically, most cases were mild and progressed to cure. Conclusion: The high number of honeybee sting cases shows that Rio Grande do Norte may be an important risk area for such injury.

RESUMO: Introdução: No continente americano, os acidentes causados por abelhas são um problema de saúde pública devido à alta incidência e severidade dos casos. Apesar de sua importância médica, há poucos estudos epidemiológicos sobre esse tema no Brasil, especialmente referentes aos estados do Nordeste. O presente estudo tem como objetivo descrever as características epidemiológicas dos casos de envenenamento por abelhas no estado do Rio Grande do Norte, no Nordeste do Brasil, de 2007 a 2014. Metodologia: Os dados foram coletados da base de dados do Sistema de Informação de Agravos de Notificação da Secretaria Estadual de Saúde do Rio Grande do Norte. Resultados: Um total de 2.168 casos foram analisados. Os casos mostraram distribuição em todos os meses dos anos estudados, com maior frequência de junho a outubro. A maioria dos casos ocorreu em áreas urbanas e envolveu homens, com vítimas entre 20 e 39 anos de idade. As vítimas foram principalmente picadas na cabeça e na mão, e receberam assistência médica predominantemente dentro de 3 horas após serem picadas. As manifestações locais eram mais frequentes do que as sistêmicas. Clinicamente, a maioria dos casos foi leve e progrediu para cura. Conclusão: O alto número casos de picadas de abelhas mostra que o Rio Grande do Norte pode ser uma importante área de risco para tal incidente.

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.

Honey bee envenoming in Santa Catarina, Brazil, 2007 through 2017: an observational, retrospective cohort study

Abstract INTRODUCTION: We evaluated the epidemiological and clinical profile of reported cases of bee sting incidents in Santa Catarina, Brazil. METHODS: This retrospective cohort study included all reported cases of bee sting incidents among the population of Santa Catarina from 2007 to 2017. RESULTS: In total, 8,912 cases were reported, corresponding to an overall rate of 12.3/100,000 population. The mean age was 29,8 years with 60.2% men. The lethality rate was 0.2%. CONCLUSIONS: Santa Catarina has a high incidence rate of bee stings, which is higher than the national average. The data presented in this study may be underestimated.

Multiple bee stings, multiple organs involved: a case report

Abstract Accidents related to Africanized honey bees are growing globally and are associated with multiple stings owing to the aggressive behavior of this species. The massive inoculation of venom causes skin necrosis and rhabdomyolysis leading to renal failure. Anaphylactic manifestations are more common and are treated using well-defined treatment protocols. However, bee venom-induced toxic reactions may be serious and require a different approach. We report the case of a 3-year-old child, which would help clinicians to focus on the treatment approach required after an incident involving multiple bee stings.

Peptidomic analysis of the venom of the solitary bee Xylocopa appendiculata circumvolans

Background: Among the hymenopteran insect venoms, those from social wasps and bees - such as honeybee, hornets and paper wasps - have been well documented. Their venoms are composed of a number of peptides and proteins and used for defending their nests and themselves from predators. In contrast, the venoms of solitary wasps and bees have not been the object of further research. In case of solitary bees, only major peptide components in a few venoms have been addressed. Therefore, the aim of the present study was to explore the peptide component profile of the venom from the solitary bee Xylocopa appendiculata circumvolans by peptidomic analysis with using LC-MS. Methods: A reverse-phase HPLC connected to ESI-OrbiTrap MS was used for LC-MS. On-line mass fingerprinting was made from TIC, and data-dependent tandem mass spectrometry gave MSMS spectra. A major peptide component was isolated by reverse-phase HPLC by conventional way, and its sequence was determined by Edman degradation, which was finally corroborated by solid phase synthesis. Using the synthetic specimen, biological activities (antimicrobial activity, mast cell devaluation, hemolysis, leishmanicidal activity) and pore formation in artificial lipid bilayer were evaluated. Results: On-line mass fingerprinting revealed that the crude venom contained 124 components. MS/MS analysis gave 75 full sequences of the peptide components. Most of these are related to the major and novel peptide, xylopin. Its sequence, GFVALLKKLPLILKHLH-NH2, has characteristic features of linear cationic α-helical peptides; rich in hydrophobic and basic amino acids with no disulfide bond, and accordingly, it can be predicted to adopt an amphipathic α-helix secondary structure. In biological evaluation, xylopin exhibited broad-spectrum antimicrobial activity, and moderate mast cell degranulation and leishmanicidal activities, but showed virtually no hemolytic activity. Additionally, the peptide was able to incorporate pores in artificial lipid bilayers of azolectin, confirming the mechanism of the cytolytic activity by pore formation in biological membranes. Conclusions: LC-ESI-MS and MS/MS analysis of the crude venom extract from a solitary bee Xylocopa appendiculata circumvolans revealed that the component profile of this venom mostly consisted of small peptides. The major peptide components, xylopin and xylopinin, were purified and characterized in a conventional manner. Their chemical and biological characteristics, belonging to linear cationic α-helical peptides, are similar to the known solitary bee venom peptides, melectin and osmin. Pore formation in artificial lipid bilayers was demonstrated for the first time with a solitary bee peptide.(AU)

Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata

Background Mass spectrometry-guided venom peptide profiling is a powerful tool to explore novel substances from venomous animals in a highly sensitive manner. In this study, this peptide profiling approach is successfully applied to explore the venom peptides of a Japanese solitary carpenter bee, Xylocopa appendiculata (Hymenoptera: Apoidea: Apidae: Anthophila: Xylocopinae: Xylocopini). Although interesting biological effects of the crude venom of carpenter bees have been reported, the structure and biological function of the venom peptides have not been elucidated yet. Methods The venom peptide profiling of the crude venom of X. appendiculata was performed by matrix-assisted laser desorption/ionization-time of flight mass spectroscopy. The venom was purified by a reverse-phase HPLC. The purified peptides were subjected to the Edman degradation, MS/MS analysis, and/or molecular cloning methods for peptide sequencing. Biological and functional characterization was performed by circular dichroism analysis, liposome leakage assay, and antimicrobial, histamine releasing and hemolytic activity tests. Results Three novel peptides with m/z 16508, 1939.3, and 1900.3 were isolated from the venom of X. appendiculata. The peptide with m/z 16508 was characterized as a secretory phospholipase A2 (PLA2) homolog in which the characteristic cysteine residues as well as the active site residues found in bee PLA2s are highly conserved. Two novel peptides with m/z 1939.3 and m/z 1900.3 were named as Xac-1 and Xac-2, respectively. These peptides are found to be amphiphilic and displayed antimicrobial and hemolytic activities. The potency was almost the same as that of mastoparan isolated from the wasp venom. Conclusion We found three novel biologically active peptides in the venom of X. appendiculata and analyzed their molecular functions, and compared their sequential homology to discuss their molecular diversity. Highly sensitive mass analysis plays an important role in this study.(AU)

Acute kidney injury complicating bee stings - a review

ABSTRACT Bee stings can cause severe reactions and have caused many victims in the last years. Allergic reactions can be triggered by a single sting and the greater the number of stings, the worse the prognosis. The poisoning effects can be systemic and can eventually cause death. The poison components are melitin, apamin, peptide 401, phospholipase A2, hyaluronidase, histamine, dopamine, and norepinephrine, with melitin being the main lethal component. Acute kidney injury (AKI) can be observed in patients suffering from bee stings and this is due to multiple factors, such as intravascular hemolysis, rhabdomyolysis, hypotension and direct toxicity of the venom components to the renal tubules. Arterial hypotension plays an important role in this type of AKI, leading to ischemic renal lesion. The most commonly identified biopsy finding in these cases is acute tubular necrosis, which can occur due to both, ischemic injury and the nephrotoxicity of venom components. Hemolysis and rhabdomyolysis reported in many cases in the literature, were demonstrated by elevated serum levels of indirect bilirubin and creatine kinase. The severity of AKI seems to be associated with the number of stings, since creatinine levels were higher, in most cases, when there were more than 1,000 stings. The aim of this study is to present an updated review of AKI associated with bee stings, including the currently advised clinical approach.

Melittin induces in vitro death of Leishmania (Leishmania) infantum by triggering the cellular innate immune response

Abstract Background Apis mellifera venom, which has already been recommended as an alternative anti-inflammatory treatment, may be also considered an important source of candidate molecules for biotechnological and biomedical uses, such as the treatment of parasitic diseases. Methods Africanized honeybee venom from Apis mellifera was fractionated by RP-C18-HPLC and the obtained melittin was incubated with promastigotes and intracellular amastigotes of Leishmania (L.) infantum. Cytotoxicity to mice peritoneal macrophages was evaluated through mitochondrial oxidative activity. The production of anti- and pro-inflammatory cytokines, NO and H2O2 by macrophages was determined. Results Promastigotes and intracellular amastigotes were susceptible to melittin (IC50 28.3 μg.mL−1 and 1.4 μg.mL−1, respectively), but also showed mammalian cell cytotoxicity with an IC50 value of 5.7 μg.mL−1. Uninfected macrophages treated with melittin increased the production of IL-10, TNF-α, NO and H2O2. Infected melittin-treated macrophages increased IL-12 production, but decreased the levels of IL-10, TNF-α, NO and H2O2. Conclusions The results showed that melittin acts in vitro against promastigotes and intracellular amastigotes of Leishmania (L.) infantum. Furthermore, they can act indirectly on intracellular amastigotes through a macrophage immunomodulatory effect.

Massive attack by honeybees in a German shepherd dog: description of a fatal case and review of the literature

In the present study, a fatal case caused by honeybee (Apis cerana) stings was documented in a female German shepherd dog that was presented at the Veterinary Teaching Hospital, University of Agriculture Faisalabad, Pakistan. Characteristic clinical signs included hematuria, hematemesis, incoordination and convulsions along with evidence of massive honeybee attack supported the diagnosis of envenomation. The dog was treated with dexamethasone and diphenhydramine, but it did not respond to therapy and died. This outcome could be avoided if we had a bee antivenom available for treating envenomated patients.

Effect of bee venom on IL-6, COX-2 and VEGF levels in polycystic ovarian syndrome induced in Wistar rats by estradiol valerate

Background : Polycystic ovarian syndrome (PCOS) is a low-grade inflammatory disease characterized by hyperandrogenemia, hirsutism, chronic anovulation and vascular disorder. Interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) are triggered by inflammatory stimuli and lead to angiogenesis and pathogenesis of the ovary. Honeybee venom (HBV) contains an array of biologically active components possessing various pharmaceutical properties. This study was designed to assess the possibility of HBV application as an anti-inflammatory therapeutic agent to suppress levels of the main inflammatory mediators IL-6, COX-2 and VEGF. To induce PCOS, 1 mg of estradiol valerate (EV) per 100 g of body weight was subcutaneously (SC) injected into eight-week-old rats. After 60 days, 0.5 mg/kg of HBV was administered Intraperitoneal (IP) for 14 consecutive days, and the results of PCOS treatment were investigated. Rats were then anesthetized with CO2, and the ovaries were surgically removed. Serum IL-6 was detected by the ELISA kit. Immunoexpression of COX-2 and VEGF were examined in three groups: EV-induced PCOS, HBV-treated PCOS and control animals. Results : Thickness of theca layer, number and diameter of cysts and levels of IL-6 significantly decreased in HBV group relative to PCOS group. The immunohistochemical analysis showed an increase in COX-2 and VEGF expression in PCOS group whereas HBV-treated rats presented weak and irregular immunostaining. Conclusions : Our results suggest that the beneficial effect of HBV may be mediated through its inhibitory effect on serum IL-6 level and ovarian COX-2 and VEGF expression.(AU)

Potentiation of a novel palladium (II) complex lethality with bee venom on the human T-cell acute lymphoblastic leukemia cell line (MOLT-4)

Background : Although honeybee venom (BV) has been reported to induce apoptosis in different types of cancerous cells, its synergistic effects with customary anti-cancer drugs remain largely unknown. In the present study, we evaluated the cytotoxic effect of BV alone (as a natural product) and the synergistic cytological effects of this component in combination with [Pd (bpy) (Pi-Pydtc)]NO3 - a novel palladium complex on human T-cell lymphoblastic leukemia cells. To investigate the cytotoxic effect of the BV alone and in combination with palladium complex on MOLT-4 cells MTT assay was performed. In order to determine the apoptotic effects of BV separately and in combination with Pd (II) complex on these cells and its ability to induce apoptosis, morphological examination, flowcytometric analysis and caspase-3 colorimetric assay were done. Results : We found that BV induced morphological changes, namely nuclear shrinkage, and inhibited MOLT-4 cell proliferation; both effects were dose- and time-dependent. Flow cytometry by Annexin-V antibody demonstrated that BV induced apoptosis in MOLT-4 cells. Furthermore, BV induced apoptosis independently of caspase-3 in these cells. In addition, we proved a clear synergistic effect of BV on [Pd (bpy) (Pi-Pydtc)]NO3. The apoptotic pathway activated by BV in combination with Pd complex was caspase-3-dependent. Conclusions : These observations provide an explanation for the anti-proliferative properties of BV, and suggest that this agent may be useful for treating lymphoblastic leukemia alone or in combination with chemotherapy drugs pending further investigations on animal models as preclinical tests.(AU)

Lesions caused by Africanized honeybee stings in three cattle in Brazil

We report three cases of stings by Africanized bees in cattle in the state of Rio de Janeiro, Brazil. Erythema, subcutaneous edema, necrosis accompanied by skin detachment, and subsequent skin regeneration were observed, especially on the head and dewlap. Histopathological examinations performed 45 days later revealed complete skin reepithelialization with moderate dermal fibrosis. The clinical picture and differential diagnosis are discussed in the present manuscript, with a focus on photosensitization, which causes cutaneous lesions on the head (sequela) with cicatricial curving of the ears and can be very similar to what is observed in cattle attacked by swarms of bees. The distinction between photosensitization and bee sting lesions can be made with a focus on history and clinical and pathological aspects.