Api m 6 and Api m 10 as Major Allergens in Patients With Honeybee Venom Allergy.

BACKGROUND: Component-resolved diagnosis plays a key role in the diagnosis and treatment of honeybee venom allergy (HVA). Our aim was to study whether any of the allergens not included in the usual diagnostic platforms are relevant in our population. MATERIAL AND METHODS: The allergenic sensitization profile of Spanish patients who experienced a systemic reaction after a honeybee sting and were diagnosed with HVA was studied by immunoblotting based on raw autochthonous Apis mellifera venom characterized using SDS-PAGE and mass spectrometry and a commercial assay (ImmunoCAP). RESULTS: Allergens in the International Union of Immunological Societies database were detected in the raw A mellifera venom extract used, except Api m 12. Sera from 51 patients with a median (IQR) age of 46.2 years (35.6-54.6) were analyzed. ImmunoCAP revealed Api m 1 and Api m 10 to be major allergens (88.2% and 74.5%, respectively). Moreover, Api m 6 (85.4%) was detected by immunoblotting. CONCLUSION: Api m 1, Api m 6, and Api m 10 are major A mellifera venom allergens in our population.

Api m 6 and Api m 10 as Major Allergens in Patients With Honeybee Venom Allergy

Background: Component-resolved diagnosis plays a key role in the diagnosis and treatment of honeybee venom allergy (HVA). Our aimwas to study whether any of the allergens not included in the usual diagnostic platforms are relevant in our population.Patients and Methods: The allergenic sensitization profile of Spanish patients who experienced a systemic reaction after a honeybee stingand were diagnosed with HVA was studied by immunoblotting based on raw autochthonous Apis mellifera venom characterized usingSDS-PAGE and mass spectrometry and a commercial assay (ImmunoCAP).Results: Allergens in the International Union of Immunological Societies database were detected in the raw A mellifera venom extract used,except Api m 12. Sera from 51 patients with a median (IQR) age of 46.2 years (35.6-54.6) were analyzed. ImmunoCAP revealed Api m 1and Api m 10 to be major allergens (88.2% and 74.5%, respectively). Moreover, Api m 6 (85.4%) was detected by immunoblotting.Conclusion: Api m 1, Api m 6, and Api m 10 are major A mellifera venom allergens in our population (AU)

Antecedentes: El diagnóstico molecular puede ser una herramienta valiosa en el diagnóstico y el tratamiento de la alergia al veneno deabeja. Este estudio investiga si alguno de los alérgenos no incluidos en las plataformas diagnósticas habituales son relevantes en nuestrapoblación.Pacientes y métodos: Estudiamos mediante immunoblotting el perfil de sensibilización alergénica en pacientes españoles diagnosticadosde alergia al veneno de abeja. Los resultados se compararon con los obtenidos usando un ensayo comercial (ImmunoCAP). El venenocrudo de Apis mellifera autóctona se obtuvo y caracterizó mediante SDS-PAGE y espectrometría de masas.Resultados: Los alérgenos descritos en la base de datos International Union of Immunological Societies (IUIS) fueron detectados enel extracto crudo de veneno de A. mellifera utilizado. Se analizaron sueros de 51 pacientes con una edad media de 46,2 años (rangointercuartil 35,6–54,6). Api m 1 y Api m 10 fueron detectados como alérgenos mayoritarios (88,2% y 74,5%, respectivamente) usandoImmunoCAP. Además, se encontró Api m 6 (85,4%) mediante immunoblotting.Conclusión: Nuestra población reconoce Api m 1, Api m 6 y Api m 10 como alérgenos mayoritarios del veneno de A. mellifera (AU)

Transcriptional and proteomic stress responses of a soil bacterium Bacillus cereus to nanosized zero-valent iron (nZVI) particles.

Nanosized zero valent iron (nZVI) is emerging as an option for treating contaminated soil and groundwater even though the potentially toxic impact exerted by nZVI on soil microorganisms remains uncertain. In this work, we focus on nanotoxicological studies performed in vitro using commercial nZVI and one common soil bacterium (Bacillus cereus). Results showed a negative impact of nZVI on B. cereus growth capability, consistent with the entrance of cells in an early sporulation stage, observed by TEM. Despite no changes at the transcriptional level are detected in genes of particular relevance in cellular activity (narG, nirS, pykA, gyrA and katB), the proteomic approach used highlights differentially expressed proteins in B. cereus under nZVI exposure. We demonstrate that proteins involved in oxidative stress-response and tricarboxilic acid cycle (TCA) modulation are overexpressed; moreover proteins involved in motility and wall biosynthesis are repressed. Our results enable to detect a molecular-level response as early warning signal, providing new insight into first line defense response of a soil bacterium after nZVI exposure.

Proteomic analysis by two-dimensional differential gel electrophoresis (2D DIGE) of a high-pressure effect in Bacillus cereus.

High hydrostatic pressure (HHP) is a new method used to reduce or eliminate microorganisms that are present in food. Proteins are known to be the most important target of high pressure in living organisms. The main goal of this investigation was focused on the changes that occur on the proteins of Bacillus cereus under HHP stress conditions. The two-dimensional differential gel electrophoresis (2D DIGE) technique allows for a simultaneous resolution of thousands of proteins based on fluorescent prelabeling of the samples with spectrally resolvable fluorescent CyDyes. The results of proteomics profiling show an average of 1300 spots being detected. The analysis revealed 75 spot proteins whose abundance is modified after the application of high pressure, of which 66 were decreased after the HHP treatment. Among them, flagellin was the protein that changed the most. The differential expression of some proteins after HHP treatment at 700 MPa may suggest a reduction of virulence and protective response against oxidative stress in flagellated Bacillus .

CL100/MKP-1 modulates JNK activation and apoptosis in response to cisplatin.

Treatment of cells with cisplatin induces a sustained activation of the stress activated protein kinase SAPK/JNK and the mitogen-activated protein kinase p38. Activation of JNK by cisplatin is necessary for the induction of apoptosis. Expression of the MAPK phosphatases CL100/MKP-1 and hVH-5 selectively prevents JNK/SAPK activation by cisplatin in a dose dependent fashion and results in protection against cisplatin-induced apoptosis. In contrast, expression of the ERK-specific phosphatase Pyst1 inhibits JNK/SAPK activity only when expressed at very high levels and does not confer protection against cisplatin. Furthermore, expression of a catalytically inactive mutant of CL100 in 293 cells decreases the IC50 for cisplatin and increases the toxicity of transplatin. This effect seems to be mediated by an increase in JNK activity since p38 activity is unaffected. These results suggest that dual-specificity MAPK phosphatases may be candidate drug targets in order to optimize cisplatin based therapeutic protocols.