Evaluation and comparison of commercially available latex extracts for skin prick tests.

BACKGROUND: Crude latex extracts are commonly used in skin prick tests (SPT) for the diagnosis of natural rubber latex (NRL) allergy. Nevertheless, variations in protein and allergen composition between latex extracts from different manufacturers can hamper a correct diagnosis. OBJECTIVES: To analyze the heterogeneity of proteins and allergens in latex extracts from 7 different manufacturers and to assess its relevance in the diagnosis of latex allergy. METHODS: Seven latex SPT extracts were analyzed for protein content using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The 4 major allergens Hev b 1, Hev b 3, Hev b 5, and Hev b 6.02 were also quantified using enzyme immunoassay. All commercial extracts were tested for their in vitro allergenic capacity using microarray inhibition assays and for their ability to induce biological reactivity in latex-allergic patients undergoing SPT. RESULTS: The protein content of the extracts varied widely from 8.0 microg/mL to 526.5 microg/mL. SDS-PAGE revealed broad differences in protein profiles between the extracts. Marked variability in the contents of all 4 major allergens was observed, and Hev b 3 and Hev b 5 were undetectable in some extracts. Microarray inhibition assays and SPT demonstrated relevant differences in allergenic capacity between the extracts. CONCLUSIONS: The marked heterogeneity in protein and allergen content of latex extracts from different manufacturers could explain the broad spectrum of SPT results recorded. Our findings suggest that the extracts used for the diagnosis of latex allergy should be improved and standardized.

Different in vivo reactivity profile in health care workers and patients with spina bifida to internal and external latex glove surface-derived allergen extracts.

BACKGROUND: Allergy to natural rubber latex is a well-recognized health problem, especially among health care workers and patients with spina bifida. Despite latex sensitization being acquired in health institutions in both health care workers and patients with spina bifida, differences in allergen sensitization profiles have been described between these two risk groups. OBJECTIVE: To investigate the in vivo reactivity of health care workers and patients with spina bifida to extracts of internal and external surfaces of latex gloves and also to specific extracts enriched in major allergens for these risk groups. METHODS: Gloves from different manufacturers were used for protein extraction, and salt precipitation and hydrophobic interaction chromatography (HIC) were applied to obtain the enriched latex extracts. The major latex allergens were quantified by an enzyme immunoassay. The extracts obtained were tested in 14 volunteers using skin prick tests (SPT). RESULTS: Latex glove extracts enriched in the hydrophobic allergens that are most often seen in patients with spina bifida were obtained by selective precipitation, whereas HIC produced extracts enriched in the hydrophilic allergens commonly found in health care workers. The health care workers had positive SPTs to glove extracts from internal surfaces and to the hydrophilic allergen-enriched extracts. By contrast, patients with spina bifida had larger skin reactions both to external glove extracts and to the extracts enriched with the hydrophobic major allergens for this risk group. Despite the protein concentration of these extracts being less than half the concentration of the commercial extract, the weal-and-flare reactions were of similar magnitude. CONCLUSION: Using novel latex extracts, our study showed a different in vivo reactivity pattern in health care workers and in patients with spina bifida to extracts of the internal and external surfaces of gloves, which suggests that sensitization may occur by different routes of exposure, and that this influences the allergen reactivity profiles of these risk groups.

Ultrasound biomicroscopy for biomechanical characterization of healthy and injured triceps surae of rats.

This work describes the use of ultrasound biomicroscopy (UBM) to follow up the degeneration-regeneration process after a laceration injury induced in the lateral gastrocnemius (LG) and soleus (SOL) muscles of rats. UBM (40 MHz) images were acquired and used for biomechanical characterization of muscular tissue, specifically using pennation angle (PA) and muscle thickness (MT). The animals were distributed in three groups: the variability group (VG; N=5), the gastrocnemius injured group (GG; N=6) and the soleus injured group (SG; N=5). VG rats were used to assess data variability and reliability (coefficients of variation of 9.37 and 3.97% for PA and MT, respectively). GG and SG rats were submitted to the injury protocol in the LG and SOL muscles of the right legs, respectively. UBM images of muscles of both legs were acquired at the following time points: before and after injury (immediately, 7, 14, 21 and 28 days). We observed an increase in PA for the non-injured leg 28 days after injury for both GG and SG rats (GG=10.68 to 16.53 deg and SG=9.65 to 14.06 deg; P<0.05). Additionally, MT presented a tendency to increase (GG=2.92 to 3.13 mm and SG=2.12 to 2.35 mm). Injured legs maintained pre-injury PA and MT values. It is suggested that a compensatory hypertrophic response due to the overload condition imposed to healthy leg. The results indicate that UBM allows qualitative and quantitative muscle differentiation among healthy and injured muscle at different stages after lesion.

Deformação relativa e frouxidão do tendão calcanear durante mobilização articular passiva através de ultra-sonografia por imagem / Strain and slackness of achilles tendon during passive joint mobilization via imaging ultrasonography

CONTEXTUALIZAÇÃO: O estudo do comportamento das propriedades mecânicas do tendão in vivo pode trazer avanços na avaliação do impacto de programas de intervenção para flexibilidade e força, nas áreas clínica e desportiva. OBJETIVO: O objetivo deste trabalho foi quantificar a deformação (strain) e a frouxidão (slackness) relativas do tendão calcanear, durante mobilização passiva para quatro ângulos articulares do tornozelo e dois do joelho. MATERIAIS E MÉTODOS: O deslocamento da junção miotendínea foi quantificado através de imagens ultra-sonográficas capturadas durante a mobilização passiva do tornozelo, com o auxílio de um eletrogoniômetro e um eletromiógrafo, para garantir as angulações requeridas e a inatividade muscular, respectivamente. RESULTADOS: Os valores de deformação relativa encontrados variaram de 4,28±2,37 a -0,94±1,58 por cento para o joelho estendido e de 2,38±1,63 a -2,32±2,16 por cento para o joelho fletido. CONCLUSÕES: Os valores encontrados ratificam os da literatura, demonstrando a participação do tendão calcanear na variação do comprimento da unidade músculo-tendão, durante movimentação passiva. Estes resultados sugerem que as propriedades mecânicas dos tecidos tendinosos afetam a relação entre o comprimento das fibras e o ângulo articular, até mesmo nesse tipo de movimento.

BACKGROUND: In vivo study of the mechanical behavior of tendons may bring advances in evaluating the impact of intervention programs for flexibility and strength, in clinical practice and sports. OBJECTIVE: The aim of this study was to quantify the relative strain and slackness of achilles tendons during passive mobilization, for four ankle joint angles and two knee angles. METHODS: The displacement of the muscle-tendon junction was quantified by means of ultrasound images acquired during passive ankle mobilization, with the aid of an electrogoniometer and an electromyograph to ensure the achievement of the required angles and muscle inactivity, respectively. RESULTS: The strain values ranged from 4.28 percent±2.37 to -0.94 percent±1.58 for the fully extended knee, and from 2.38 percent±1.63 to -2.32 percent±2.16 percent for the flexed knee. CONCLUSIONS: The values found in this study confirm those in the literature and demonstrate how the Achilles tendon participates in length changes in the muscle-tendon unit during passive movement. These results suggest that the mechanical properties of tendinous tissues affect the relationship between the length of muscle fibers and the joint angle, even during this type of movement.

Latex allergy: new insights to explain different sensitization profiles in different risk groups.

BACKGROUND: Differences in latex allergen sensitization profiles have been described between children subjected to repetitive surgical interventions and health care workers (HCW). 'Major' allergens for patients with spina bifida are Hev b 1, 3 and 7, while for HCW, 'major' allergens are Hev b 2, 5, 6.01 and 13. The reason for these differential sensitization profiles is currently unknown. OBJECTIVES: To investigate latex allergen profiles on internal and external surfaces of natural rubber latex gloves. METHODS: Eighty-two samples of commonly used surgical gloves (41 glove brands) were used for analysis. Specific allergen levels of Hev b 1, 3, 5 and 6.02 on both surfaces of the gloves were quantified using an enzyme immunometric assay, a FITkit (FIT Biotech, Tampere, Finland). RESULTS: Differences in allergen levels were observed between internal and external surfaces of all glove types. Concentrations of Hev b 1 and Hev b 3 were significantly higher on external surfaces, while internal surfaces had higher allergen levels of Hev b 5 and Hev b 6.02. Analysis of surgical and examination gloves, powdered and nonpowdered gloves also showed that the content of Hev b 5 and Hev b 6.02 was significantly higher on internal surfaces while that of Hev b 1 and Hev b 3 was higher on external surfaces. CONCLUSIONS: Our study showed different allergen profiles on internal and external surfaces of natural rubber latex gloves. These results may suggest a relationship between latex allergen localization and sensitization routes in different risk groups.

Differential expression of allergens on the internal and external surfaces of latex surgical gloves.

BACKGROUND: Differences in latex allergen sensitization profiles have been described between children undergoing repeated surgical interventions and health care workers. The purpose of this study was to determine whether such sensitization profiles are associated with differences in the expression of latex allergen between the internal and external surfaces of surgical gloves. METHODS: Extracts were obtained from whole surgical gloves as well as from their external and internal surfaces. The extracts were centrifuged, filtered, concentrated, dialyzed and lyophilized. The protein profile of the extracts was analyzed using hydrophobic interaction chromatography (HIC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblotting was performed using sera from two patients with confirmed latex allergy. Latex recombinant allergen-specific IgE in these two patients was determined using a fluorescence enzyme immunoassay (FEIA) method. Latex allergen quantification was determined on both glove surfaces using an ELISA method. RESULTS: HIC and SDS-PAGE showed qualitative and quantitative differences in proteins between the internal and external glove surfaces, with the former being much richer in proteins. Immunoblotting of glove extracts using sera from two latex-allergic health workers showed differences between glove surface extracts. ELISA quantification of latex allergens demonstrated that the internal glove surface had high amounts of Hev b 5 and Hev b 6.02 whereas the external surface showed Hev b 1, Hev b 3, and Hev b 6.02. CONCLUSIONS: Our results reveal substantial differences in the composition of latex allergen profiles between the internal and external surfaces of surgical latex gloves, which may suggest a relationship between latex allergen localization and sensitization routes in different risk groups.

Differential expression of allergens on the internal and external surfaces of latex surgical gloves / Diferente expresión de alergenos en la superficie interna o externa de los guantes quirúrgicos de látex

Background: Differences in latex allergen sensitization profiles have been described between children undergoing repeated surgical interventions and health care workers. The purpose of this study was to determine whether such sensitization profiles are associated with differences in the expression of latex allergen between the internal and external surfaces of surgical gloves. Methods: Extracts were obtained from whole surgical gloves as well as from their external and internal surfaces. The extracts were centrifuged, filtered, concentrated, dialyzed and lyophilized. The protein profile of the extracts was analyzed using hydrophobic interaction chromatography (HIC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblotting was performed using sera from two patients with confirmed latex allergy. Latex recombinant allergen-specific IgE in these two patients was determined using a fluorescence enzyme immunoassay (FEIA) method. Latex allergen quantification was determined on both glove surfaces using an ELISA method. Results: HIC and SDS-PAGE showed qualitative and quantitative differences in proteins between the internal and external glove surfaces, with the former being much richer in proteins. Immunoblotting of glove extracts using sera from two latex-allergic health workers showed differences between glove surface extracts. ELISA quantification of latex allergens demonstrated that the internal glove surface had high amounts of Hev b 5 and Hev b 6.02 whereas the external surface showed Hev b 1, Hev b 3, and Hev b 6.02. Conclusions: Our results reveal substantial differences in the composition of latex allergen profiles between the internal and external surfaces of surgical latex gloves, which may suggest a relationship between latex allergen localization and sensitization routes in different risk groups

Antecedentes: se han descrito diferentes perfiles en la sensibilización a los alergenos del látex en niños repetidamente intervenidos quirúrgicamente y en los profesionales sanitarios. El propósito de este estudio ha sido determinar si tales perfiles de sensibilización estaban asociados a una expresión diferente de los alergenos del látex entre la superficies interna y externa de los guantes quirúrgicos. Métodos: se obtuvieron extractos totales de guantes quirúrgicos, así como de las superficies externa e interna, que se centrifugaron, filtraron, concentraron y luego fueron dializados y liofilizados. El perfil protéico de los extractos se analizó por cromografía de interacción hidrofóbica (CIH) y SDS-PAGE. La inmunotransferencia se realizó con el suero de dos pacientes que tenían confirmada la alergia al látex. La IgE específica para el alergeno recombinante del látex de estos dos pacientes se determinó mediante el método FEIA. La cuantificación del alergeno del látex se determinó en ambas superficies del guante por el método ELISA. Resultados: La CIH y SDS-PAGE mostraron diferencias cuantitativas y cualitativas de proteinas entre ambas superficies, siendo mucho más elevada en la interna que en la externa. La inmunotransferencia con los extractos de guante, usando suero de los dos sanitarios alérgicos al látex, demostró diferencias entre los extractos de ambas superficies del guante. La cuantificación por ELISA de los alergenos del látex demostró que la superificie interna del guante tenía un alto contenido de Hev b 5 y Hev b 6.02, mientras que la superficie externa tenía Hev b 1, Hev b 3 y Hev b 6.02. Conclusiones: nuestros resultados demuestran diferencias sustanciales en la composición del perfil de alergenos del látex entre las superficies interna y externa de los guantes quirúrgicos, lo que sugiere una relación entre la localización del alergeno del látex y las rutas de sensibilización en los diferentes grupos de riesgo