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Ignacio Moneo - One of the best experts on this subject based on the ideXlab platform.
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Proteomic profiling and characterization of differential allergens in the nematodes Anisakis Simplex sensu stricto and A. pegreffii
Proteomics, 2014Co-Authors: Susana C. Arcos, Ignacio Moneo, Ana-i. Rodriguez-mahillo, Miguel González-muñoz, Noelia Carballeda-sangiao, Sergio Ciordia, Lee Roberston, Inés Zapico, Yolanda Jiménez-ruiz, Juan Pablo AlbarAbstract:The parasite species complex Anisakis Simplex sensu lato (Anisakis Simplex sensu stricto; (A. Simplex s.s.), A. pegreffii, A. Simplex C) is the main cause of severe anisakiasis (allergy) worldwide and is now an important health matter. In this study, the relationship of this Anisakis species complex and their allergenic capacities is assessed by studying the differences between the two most frequent species (A. Simplex s.s., A. pegreffii) and their hybrid haplotype by studying active L3 larvae parasiting Merluccius merluccius. They were compared by 2D gel electrophoresis and parallel Western blot (2DE gels were hybridized with pools of sera from Anisakis allergenic patients). Unambiguous spot differences were detected and protein assignation was made by MALDI-TOF/TOF analysis or de novo sequencing. Seventy-five gel spots were detected and the corresponding proteins were identified. Differentially expressed proteins for A. Simplex s.s., A. pegreffii, and their hybrid are described and results are statistically supported. Twenty-eight different allergenic proteins are classified according to different families belonging to different biological functions. These proteins are described for the first time as antigenic and potentially new allergens in Anisakis. Comparative proteomic analyses of allergenic capacities are useful for diagnosis, epidemiological surveys, and clinical research. All MS data have been deposited in the ProteomeXchange with identifier PXD000662 (http://proteomecentral.proteomexchange.org/dataset/PXD000662).
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Identification of autoclave-resistant Anisakis Simplex allergens.
Journal of food protection, 2014Co-Authors: Noelia Carballeda-sangiao, Ignacio Moneo, Ana-i. Rodriguez-mahillo, Fabiola Olivares, Mercedes Careche, Margarita Tejada, Miguel González-muñozAbstract:Anisakis Simplex is a fish parasite able to induce allergic reactions in humans infected when eating raw or undercooked fish parasitized with viable third-stage larvae. Some authors claim that exposure to nonviable Anisakis material can result in allergic symptoms in previously sensitized patients, indicating that parasite allergens are resistant to the thermal treatments of usual cooking procedures. Furthermore, some patients report symptoms after eating canned fish. The aim of this work was the analysis of parasite allergen stability in heating to 121°C in an autoclave to simulate the thermal process applied to canned fish. Third-stage larvae were subjected to autoclaving for 20, 40, and 80 min, and parasite crude extracts were analyzed by electrophoresis, immunoblotting, and a flow-cytometric basophil activation test. Allergens resistant to autoclaving were separated by reversed-phase high-performance liquid chromatography and identified by ion trap mass spectrometry. Protein analysis by sodium dodecyl...
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Anisakis Simplex recombinant allergens increase diagnosis specificity preserving high sensitivity.
International archives of allergy and immunology, 2012Co-Authors: María Luisa Caballero, Ignacio Moneo, Ana Umpierrez, Teresa Pérez-piñar, Carmen De Burgos, Juan A. Asturias, Rosa Rodriguez-perezAbstract:Background: So far, the frequency of Anisakis Simplex- specific IgE antibodies has been determined by skin prick tests (SPTs) and the ImmunoCAP system. These comme
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Sensitization to the fish parasite Anisakis Simplex: clinical and laboratory aspects.
Parasitology research, 2007Co-Authors: Ignacio Moneo, María Luisa Caballero, Rosa Rodriguez-perez, Ana-i. Rodriguez-mahillo, Miguel González-muñozAbstract:Hypersensitivity to Anisakis Simplex is a worldwide medical problem. The parasite larvae die after freezing or cooking, but the tolerance of sensitized subjects to eating frozen fish remains a matter of controversy with contradictory findings. The aim of this study was to test if intolerance to properly cooked/frozen fish was due to the recognition of a particular allergen. Sixty-four patients with Anisakis Simplex sensitization were studied by an IgE multiblot using simultaneously five different antigenic extracts. The antigens tested were a crude extract, excretory/secretory allergens, a heated extract, and two gradient ethanol precipitates of the crude extract. Intolerance was reported by 20% of the patients and was not related to the detection of any special allergen, nor to total or specific IgE levels. Intolerant patients only reported a higher frequency of digestive symptoms than the patients who tolerated fish ingestion. The most sensitive immunoblot antigen source was the 50-66% ethanol fraction of a crude extract (10x concentrated) that was found to be positive in 100% of the samples. Interestingly, 95% sensitivity in the IgE-immunoblot assay could be achieved using only two allergens, Ani s 1 and Ani s 4. Allergens from the dead larvae remain a problem for 20% of the sensitized subjects. The use of a fractionated and concentrated crude extract improved the sensitivity of the immunoblot assay.
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Isolation of a heat-resistant allergen from the fish parasite Anisakis Simplex
Parasitology Research, 2005Co-Authors: Ignacio Moneo, María Luisa Caballero, Rosa Rodriguez-perez, Ana-i. Rodriguez-mahillo, Miguel González-muñoz, Augusto SilvaAbstract:The thermal stability of allergenic peptides from the fish parasite Anisakis Simplex has not been fully elucidated. This is of special relevance for physicians who should clearly indicate if sensitized patients should avoid ingestion of raw fish only or whether well-cooked fish should also be avoided, if allergenic peptides derived from the parasite remain immunologically detectable. An allergen was purified after heating a crude parasite extract for 30 min. The allergen was further purified by an ethanol fractionation procedure followed by a reversed-phase HPLC. The N-terminal amino acid sequence was obtained. This allergen was detected by 27% of sensitized subjects. The N-terminal amino acid sequence of the 9 kDa allergen showed no similarities to other known proteins. A minor low molecular weight allergen from A. Simplex is highly resistant to heating and it could therefore have significant clinical relevance.
Carmen Cuéllar - One of the best experts on this subject based on the ideXlab platform.
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Cross-reactivity induced by Anisakis Simplex and Toxocara canis in mice.
Journal of helminthology, 2003Co-Authors: M.j. Perteguer, J L Guillen, S Fenoy, Carmen Del Aguila, Carmen CuéllarAbstract:The aim of this study was to verify whether cross-reactivity appeared between Toxocara canis and Anisakis Simplex in an experimental rodent model. No cross-reactions were detected using sera from mice infected with T. canis eggs. When responses obtained against T. canis ES antigen using sera from BALB/c and C57BL/10 mice infected with T. canis eggs were compared with those obtained by testing sera from mice infected with one A. Simplex L3, an increase in cross-reactions was observed using the C57BL/10 strain.
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Purification of Anisakis Simplex antigen by affinity chromatography
Parasitology Research, 2001Co-Authors: M. Rodero, Tomas Chivato, R Laguna, Antonio Aguilera Jiménez, Carmen CuéllarAbstract:In order to improve the specificity and sensitivity of the techniques for the diagnosis of human anisakidosis, a method of affinity chromatography for the purification of species-specific antigens from Anisakis Simplex third-stage larvae (L3) has been developed. New Zealand rabbits were immunized with A. Simplex or Ascaris suum antigens or inoculated with Toxocara canis embryonated eggs. The IgG-specific antibodies were isolated by means of protein A-Sepharose CL-4B bead columns. IgG anti-Anisakis Simplex, anti-Ascaris suum and anti-T. canis were coupled to CNBr-activated Sepharose 4B. For the purification of the larval Anisakis Simplex antigens, it was loaded into the anti-A. Simplex column and bound antigens were eluted. For the elimination of the epitopes responsible for the cross-reactions, the A. Simplex-specific proteins were loaded into the anti-Ascaris suum and anti-T. canis columns. To prove the specificity of the isolated proteins, immunochemical analyses by polyacrylamide gel electrophoresis and immunoblotting were carried out. Likewise, immunoaffinity columns were prepared using specific IgG from patients with Anisakis Simplex sensitization, previously diagnosed by fluoro-enzymo-immunoassay. The protein patterns of antigen after purification by the human columns were similar to those obtained using the rabbit columns.
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Genomic identification of Anisakis Simplex isolates
Journal of helminthology, 1997Co-Authors: M. Siles, Carmen Cuéllar, M.j. PerteguerAbstract:RAPD technique was used to differentiate individuals of Anisakis Simplex obtained from Merluccius merluccius, Phycis blennoides, Conger conger and Lepidorhombus boscii , from the North Atlantic Ocean. The amplification patterns of the host DNA controls were markedly different from those obtained for the parasitic material. No variation within the same host was detected. The amplification patterns for larvae obtained from fish of the same genus were somewhat different. The amplification patterns of A. Simplex isolates from M. merluccius, P. blennoides, C. conger and L. boscii , were different. These results suggest the possible existence of two populations with a considerable high genetic variability and a different adaptation to different host species.
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Recidivous acute urticaria caused by Anisakis Simplex
Allergy, 1997Co-Authors: A. Montoro, Tomas Chivato, R Laguna, M.j. Perteguer, Carmen CuéllarAbstract:This study aimed to determine the cause of acute recidivous urticaria in patients who usually eat fish or other seafood. Twenty-five patients were studied. The skin prick test with larval Anisakis Simplex extract was performed; total and specific IgE against A. Simplex was measured with the CAP System; specific antibodies to A. Simplex were determined by ELISA; and immunorecognition patterns of the sera were studied by Western blot. Nineteen patients showed specific IgE to A. Simplex, but specific IgE to Ascaris was demonstrated in only two patients. No patients reacted to Toxocara canis or Echinoccocus granulosus antigens with the same test. The skin prick test was positive in 16 patients, in two of them persisting for 48 h. Five patients showed neither skin reaction nor specific IgE to A. Simplex. Sera showed specific immunoglobulin levels against A. Simplex larval crude extract, by both ELISA and Western blot. Likewise, specific immunoglobulin levels against excretory-secretory antigen were also measured by ELISA. Only one patient showed sensitization to fish. A. Simplex was found to be the main cause of acute recidivous urticaria in patients who usually eat fish and are not sensitized to it.
Miguel González-muñoz - One of the best experts on this subject based on the ideXlab platform.
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Proteomic profiling and characterization of differential allergens in the nematodes Anisakis Simplex sensu stricto and A. pegreffii
Proteomics, 2014Co-Authors: Susana C. Arcos, Ignacio Moneo, Ana-i. Rodriguez-mahillo, Miguel González-muñoz, Noelia Carballeda-sangiao, Sergio Ciordia, Lee Roberston, Inés Zapico, Yolanda Jiménez-ruiz, Juan Pablo AlbarAbstract:The parasite species complex Anisakis Simplex sensu lato (Anisakis Simplex sensu stricto; (A. Simplex s.s.), A. pegreffii, A. Simplex C) is the main cause of severe anisakiasis (allergy) worldwide and is now an important health matter. In this study, the relationship of this Anisakis species complex and their allergenic capacities is assessed by studying the differences between the two most frequent species (A. Simplex s.s., A. pegreffii) and their hybrid haplotype by studying active L3 larvae parasiting Merluccius merluccius. They were compared by 2D gel electrophoresis and parallel Western blot (2DE gels were hybridized with pools of sera from Anisakis allergenic patients). Unambiguous spot differences were detected and protein assignation was made by MALDI-TOF/TOF analysis or de novo sequencing. Seventy-five gel spots were detected and the corresponding proteins were identified. Differentially expressed proteins for A. Simplex s.s., A. pegreffii, and their hybrid are described and results are statistically supported. Twenty-eight different allergenic proteins are classified according to different families belonging to different biological functions. These proteins are described for the first time as antigenic and potentially new allergens in Anisakis. Comparative proteomic analyses of allergenic capacities are useful for diagnosis, epidemiological surveys, and clinical research. All MS data have been deposited in the ProteomeXchange with identifier PXD000662 (http://proteomecentral.proteomexchange.org/dataset/PXD000662).
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Identification of autoclave-resistant Anisakis Simplex allergens.
Journal of food protection, 2014Co-Authors: Noelia Carballeda-sangiao, Ignacio Moneo, Ana-i. Rodriguez-mahillo, Fabiola Olivares, Mercedes Careche, Margarita Tejada, Miguel González-muñozAbstract:Anisakis Simplex is a fish parasite able to induce allergic reactions in humans infected when eating raw or undercooked fish parasitized with viable third-stage larvae. Some authors claim that exposure to nonviable Anisakis material can result in allergic symptoms in previously sensitized patients, indicating that parasite allergens are resistant to the thermal treatments of usual cooking procedures. Furthermore, some patients report symptoms after eating canned fish. The aim of this work was the analysis of parasite allergen stability in heating to 121°C in an autoclave to simulate the thermal process applied to canned fish. Third-stage larvae were subjected to autoclaving for 20, 40, and 80 min, and parasite crude extracts were analyzed by electrophoresis, immunoblotting, and a flow-cytometric basophil activation test. Allergens resistant to autoclaving were separated by reversed-phase high-performance liquid chromatography and identified by ion trap mass spectrometry. Protein analysis by sodium dodecyl...
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Sensitization to the fish parasite Anisakis Simplex: clinical and laboratory aspects.
Parasitology research, 2007Co-Authors: Ignacio Moneo, María Luisa Caballero, Rosa Rodriguez-perez, Ana-i. Rodriguez-mahillo, Miguel González-muñozAbstract:Hypersensitivity to Anisakis Simplex is a worldwide medical problem. The parasite larvae die after freezing or cooking, but the tolerance of sensitized subjects to eating frozen fish remains a matter of controversy with contradictory findings. The aim of this study was to test if intolerance to properly cooked/frozen fish was due to the recognition of a particular allergen. Sixty-four patients with Anisakis Simplex sensitization were studied by an IgE multiblot using simultaneously five different antigenic extracts. The antigens tested were a crude extract, excretory/secretory allergens, a heated extract, and two gradient ethanol precipitates of the crude extract. Intolerance was reported by 20% of the patients and was not related to the detection of any special allergen, nor to total or specific IgE levels. Intolerant patients only reported a higher frequency of digestive symptoms than the patients who tolerated fish ingestion. The most sensitive immunoblot antigen source was the 50-66% ethanol fraction of a crude extract (10x concentrated) that was found to be positive in 100% of the samples. Interestingly, 95% sensitivity in the IgE-immunoblot assay could be achieved using only two allergens, Ani s 1 and Ani s 4. Allergens from the dead larvae remain a problem for 20% of the sensitized subjects. The use of a fractionated and concentrated crude extract improved the sensitivity of the immunoblot assay.
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Isolation of a heat-resistant allergen from the fish parasite Anisakis Simplex
Parasitology Research, 2005Co-Authors: Ignacio Moneo, María Luisa Caballero, Rosa Rodriguez-perez, Ana-i. Rodriguez-mahillo, Miguel González-muñoz, Augusto SilvaAbstract:The thermal stability of allergenic peptides from the fish parasite Anisakis Simplex has not been fully elucidated. This is of special relevance for physicians who should clearly indicate if sensitized patients should avoid ingestion of raw fish only or whether well-cooked fish should also be avoided, if allergenic peptides derived from the parasite remain immunologically detectable. An allergen was purified after heating a crude parasite extract for 30 min. The allergen was further purified by an ethanol fractionation procedure followed by a reversed-phase HPLC. The N-terminal amino acid sequence was obtained. This allergen was detected by 27% of sensitized subjects. The N-terminal amino acid sequence of the 9 kDa allergen showed no similarities to other known proteins. A minor low molecular weight allergen from A. Simplex is highly resistant to heating and it could therefore have significant clinical relevance.
Li-yu Chung - One of the best experts on this subject based on the ideXlab platform.
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Larvicidal Constituents of Zingiber officinale (Ginger) against Anisakis Simplex
Planta Medica, 2010Co-Authors: Chin-mei Lu, Li-yu ChungAbstract:In this study, we investigated the anthelmintic ac- tivity of (10)-shogaol, (6)-shogaol, (10)-gingerol and (6)-gingerol, compounds isolated from the roots of Zingiber officinale L., Zingiberaceae (gin- ger), against Anisakis Simplex. The above com- pounds kill or reduce spontaneous movement in A. Simplex larvae. The maximum lethal efficacy of (10)-shogaol and (10)-gingerol was approxi- mately 80% and 100%, respectively. We further examined the time course of compound-induced loss of mobility in A. Simplex. The results showed that various concentrations of (10)-shogaol, (6)- shogaol, (10)-gingerol and (6)-gingerol have max- imum effects on loss of spontaneous movement from 24 to 72h. In addition, the time course of mortality and the percentage of loss of spontane- ous movements were ascertained to determine the minimum effective doses of (10)-gingerol and (10)-shogaol. (10)-Gingerol exhibited a larger maximum larvicidal effect and greater loss of spontaneous movement than (10)-shogaol and albendazole. In addition, these constituents of Zingiber officinale showed effects against 2,2-di- phenyl-1-picrylhydrazyl (DPPH) and peroxyl rad- icals. These constituents of Zingiber officinale are responsible for its larvicidal activity against A. Simplex. " Zingiber officinale L. l " Zingiberaceae (ginger) l " Anisakis Simplex l " larvicidal activity l " anthelmintic activity
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larvicidal constituents of zingiber officinale ginger against Anisakis Simplex
Planta Medica, 2010Co-Authors: Rongjyh Lin, Li-yu Chung, Chungyi Chen, Juneder Lee, Chuanmin YenAbstract:In this study, we investigated the anthelmintic activity of [10]-shogaol, [6]-shogaol, [10]-gingerol and [6]-gingerol, compounds isolated from the roots of Zingiber officinale L., Zingiberaceae (ginger), against Anisakis Simplex. The above compounds kill or reduce spontaneous movement in A. Simplex larvae. The maximum lethal efficacy of [10]-shogaol and [10]-gingerol was approximately 80% and 100%, respectively. We further examined the time course of compound-induced loss of mobility in A. Simplex. The results showed that various concentrations of [10]-shogaol, [6]-shogaol, [10]-gingerol and [6]-gingerol have maximum effects on loss of spontaneous movement from 24 to 72 h. In addition, the time course of mortality and the percentage of loss of spontaneous movements were ascertained to determine the minimum effective doses of [10]-gingerol and [10]-shogaol. [10]-Gingerol exhibited a larger maximum larvicidal effect and greater loss of spontaneous movement than [10]-shogaol and albendazole. In addition, these constituents of Zingiber officinale showed effects against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and peroxyl radicals. These constituents of Zingiber officinale are responsible for its larvicidal activity against A. Simplex.
R Laguna - One of the best experts on this subject based on the ideXlab platform.
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cross reactivity between Anisakis Simplex sensitization and visceral larva migrans by toxocara canis
Acta Tropica, 2003Co-Authors: Maria J Perteguer, Carlos Cuellar, J L Guillen, Carmen Del Aguila, S Fenoy, Tomas Chivato, R LagunaAbstract:Abstract The aim of this work was to study cross-reactivity in the diagnosis of two related ascaridosis. Nineteen patients diagnosed with recidivous acute urticaria (RAU) caused by Anisakis Simplex and 26 patients diagnosed with visceral larva migrans (VLM) caused by Toxocara canis were studied employing commercial diagnostic kits and “in house” assay kits. Cross-reactivity observed was greater when using “in house” assay kits, suggesting that T. canis excretory–secretory antigens were not only recognized by antibodies from patients with RAU but with greater intensity compared to the A. Simplex excretory–secretory antigens.
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Purification of Anisakis Simplex antigen by affinity chromatography
Parasitology Research, 2001Co-Authors: M. Rodero, Tomas Chivato, R Laguna, Antonio Aguilera Jiménez, Carmen CuéllarAbstract:In order to improve the specificity and sensitivity of the techniques for the diagnosis of human anisakidosis, a method of affinity chromatography for the purification of species-specific antigens from Anisakis Simplex third-stage larvae (L3) has been developed. New Zealand rabbits were immunized with A. Simplex or Ascaris suum antigens or inoculated with Toxocara canis embryonated eggs. The IgG-specific antibodies were isolated by means of protein A-Sepharose CL-4B bead columns. IgG anti-Anisakis Simplex, anti-Ascaris suum and anti-T. canis were coupled to CNBr-activated Sepharose 4B. For the purification of the larval Anisakis Simplex antigens, it was loaded into the anti-A. Simplex column and bound antigens were eluted. For the elimination of the epitopes responsible for the cross-reactions, the A. Simplex-specific proteins were loaded into the anti-Ascaris suum and anti-T. canis columns. To prove the specificity of the isolated proteins, immunochemical analyses by polyacrylamide gel electrophoresis and immunoblotting were carried out. Likewise, immunoaffinity columns were prepared using specific IgG from patients with Anisakis Simplex sensitization, previously diagnosed by fluoro-enzymo-immunoassay. The protein patterns of antigen after purification by the human columns were similar to those obtained using the rabbit columns.
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Recidivous acute urticaria caused by Anisakis Simplex
Allergy, 1997Co-Authors: A. Montoro, Tomas Chivato, R Laguna, M.j. Perteguer, Carmen CuéllarAbstract:This study aimed to determine the cause of acute recidivous urticaria in patients who usually eat fish or other seafood. Twenty-five patients were studied. The skin prick test with larval Anisakis Simplex extract was performed; total and specific IgE against A. Simplex was measured with the CAP System; specific antibodies to A. Simplex were determined by ELISA; and immunorecognition patterns of the sera were studied by Western blot. Nineteen patients showed specific IgE to A. Simplex, but specific IgE to Ascaris was demonstrated in only two patients. No patients reacted to Toxocara canis or Echinoccocus granulosus antigens with the same test. The skin prick test was positive in 16 patients, in two of them persisting for 48 h. Five patients showed neither skin reaction nor specific IgE to A. Simplex. Sera showed specific immunoglobulin levels against A. Simplex larval crude extract, by both ELISA and Western blot. Likewise, specific immunoglobulin levels against excretory-secretory antigen were also measured by ELISA. Only one patient showed sensitization to fish. A. Simplex was found to be the main cause of acute recidivous urticaria in patients who usually eat fish and are not sensitized to it.