Influence of Gamma Irradiation on Porcine Serum Albumin Structural Properties and Allergenicity.

Pork provides an ideal source of food energy; however, pork can elicit an allergic reaction, and porcine serum albumin (PSA) has been identified as a major allergen. This study examined the impact of gamma irradiation on the allergenicity and structural qualities of PSA; the PSA solution was gamma-irradiated at 1, 2, 4, 6, and 8 kGy. Allergenicity was investigated by immunoblotting and competitive indirect ELISA using serum from patients who are allergic to pork, and conformational changes in irradiated PSA were measured by circular dichroism, sulfhydryl group detection, and fluorescence emission spectra. The secondary and tertiary structures of gamma-irradiated PSA were altered, and the allergenicity of PSA was lowered by boosting the amount of irradiation. In addition, there is high correlation between depletion in the α-helix and immunoglobulin E-binding capability of PSA. The results show a new possibility in using gamma irradiation to reduce the allergenicity of pork products.

Decreased viability and proliferation of CHANG conjunctival epithelial cells after contact with ultraviolet light-irradiated pollen.

CONTEXT: Contact with pollen is the major reason for the development of allergic symptoms on the ocular surface leading to a significant increase of allergic diseases worldwide. Environmental changes such as increased ultraviolet (UV) radiation and air pollution are discussed as contributory causes for this increase. OBJECTIVE: We investigated the effect of UV light on the histamine content of pollen and examined if an irradiation of pollen affects the viability and proliferation of conjunctival cells. MATERIALS AND METHODS: Alder (Alnus glutinosa) and hazel (Corylus avellana) pollen were irradiated for different time periods with sunlight, UV-A or UV-B light and the histamine content was analysed and compared with non-irradiated pollen. Conjunctival epithelial cells (CHANG cells) were exposed to irradiated and non-irradiated pollen followed by an assessment of cell viability with the colorimetric MTS test and the impedance-based measurement of cell proliferation using the xCELLigence real-time analysis system. RESULTS: UV light irradiation increased the histamine level of alder and hazel pollen in a dose-dependent manner. CHANG cells treated with irradiated pollen induced a statistically significant higher decrease of cell viability than treatment with non-irradiated pollen. DISCUSSION AND CONCLUSIONS: Our results indicate that UV light is able to alter pollen thus making them more harmful for conjunctival cells.

Potential allergenicity response to structural modification of irradiated bovine α-lactalbumin.

Bovine α-lactalbumin (α-La) is a major food allergen found in milk and is characterized by high conformational stability because of its four disulfide bridges and being calcium bound. This study aimed to describe the influence of gamma irradiation on the structure and potential allergenicity of α-La. The prepared α-La was irradiated at doses of 1-10 kGy. The changes in structure were characterized through SDS-PAGE, circular dichroism spectroscopy, ultraviolet absorption spectroscopy, and fluorescence spectroscopy. The potential allergenicity of the irradiated α-La was evaluated in vitro through IgG/IgE inhibition ELISA and the human basophil KU812 degranulation assay. The results showed that the secondary and tertiary structures of α-La significantly changed and caused extensive protein denaturation and aggregation. IgG and IgE binding properties remarkably decreased, and the degranulation capacity of basophils weakened. The results suggested that structural damage of α-La induced by irradiation significantly reduces the potential allergenicity of α-La.

Soft-X-ray-enhanced electrostatic precipitation for protection against inhalable allergens, ultrafine particles, and microbial infections.

Protection of the human lung from infectious agents, allergens, and ultrafine particles is difficult with current technologies. High-efficiency particulate air (HEPA) filters remove airborne particles of >0.3 µm with 99.97% efficiency, but they are expensive to maintain. Electrostatic precipitation has been used as an inexpensive approach to remove large particles from airflows, but it has a collection efficiency minimum in the submicrometer size range, allowing for a penetration window for some allergens and ultrafine particles. Incorporating soft X-ray irradiation as an in situ component of the electrostatic precipitation process greatly improves capture efficiency of ultrafine particles. Here we demonstrate the removal and inactivation capabilities of soft-X-ray-enhanced electrostatic precipitation technology targeting infectious agents (Bacillus anthracis, Mycobacterium bovis BCG, and poxviruses), allergens, and ultrafine particles. Incorporation of in situ soft X-ray irradiation at low-intensity corona conditions resulted in (i) 2-fold to 9-fold increase in capture efficiency of 200- to 600-nm particles and (ii) a considerable delay in the mean day of death as well as lower overall mortality rates in ectromelia virus (ECTV) cohorts. At the high-intensity corona conditions, nearly complete protection from viral and bacterial respiratory infection was afforded to the murine models for all biological agents tested. When optimized for combined efficient particle removal with limited ozone production, this technology could be incorporated into stand-alone indoor air cleaners or scaled for installation in aircraft cabin, office, and residential heating, ventilating, and air-conditioning (HVAC) systems.

Pulsed ultraviolet light reduces immunoglobulin E binding to Atlantic white shrimp (Litopenaeus setiferus) extract.

Pulsed ultraviolet light (PUV), a novel food processing and preservation technology, has been shown to reduce allergen levels in peanut and soybean samples. In this study, the efficacy of using PUV to reduce the reactivity of the major shrimp allergen, tropomyosin (36-kDa), and to attenuate immunoglobulin E (IgE) binding to shrimp extract was examined. Atlantic white shrimp (Litopenaeus setiferus) extract was treated with PUV (3 pulses/s, 10 cm from light source) for 4 min. Tropomyosin was compared in the untreated, boiled, PUV-treated and [boiled+PUV]-treated samples, and changes in the tropomyosin levels were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). IgE binding of the treated extract was analyzed via immunoblot and enzyme-linked immunosorbent assay (ELISA) using pooled human plasma containing IgE antibodies against shrimp allergens. Results showed that levels of tropomyosin and IgE binding were reduced following PUV treatment. However, boiling increased IgE binding, while PUV treatment could offset the increased allergen reactivity caused by boiling. In conclusion, PUV treatment reduced the reactivity of the major shrimp allergen, tropomyosin, and decreased the IgE binding capacity of the shrimp extract.

Effects of pulsed UV-light on peanut allergens in extracts and liquid peanut butter.

Pulsed ultraviolet (PUV) light, a nonthermal technology, was used to treat both the peanut extracts and liquid peanut butter. The objective was to determine if such treatment would lead to a reduction in the allergenic properties of the peanut extract and butter. Peanut samples were PUV treated using a Xenon RS-3000C under the following conditions: 3 pulses/s, 14.6 cm from the central axis of the lamp, 4 min (extract) or 3 min (liquid peanut butter). After the treatment, the peanut samples were centrifuged and the supernatants analyzed by SDS-PAGE and competitive inhibition enzyme-linked immunosorbent assay (ciELISA). For comparison, boiling treatments were also performed. SDS-PAGE showed that while boiling treatment had little effect on the peanut allergens, PUV-light-treated samples displayed a reduced solubility or level of peanut allergens (63 kDa). Solubility of another allergen (18 to 20 kDa) was unaffected. Insoluble aggregates formed were responsible for the reduced level of allergens in PUV-light-treated samples. ciELISA showed that untreated samples exhibited an IgE binding 7-fold higher than the PUV-treated samples. It was concluded that PUV light was effective in reducing IgE binding of peanut extracts and liquid peanut butter. The current study provides an approach to the development of a possibly less allergenic peanut product. However, the reduction in actual allergenicity needs to be confirmed by clinical studies.

Microbial population, physicochemical quality, and allergenicity of molluscs and shrimp treated with cobalt-60 gamma radiation.

Frozen molluscs (squid, octopuses, and cuttlefish) and crustaceans (shrimp) were irradiated using a cobalt-60 gamma source, at different doses, in order to investigate the effects of gamma radiation on their microbial population, organoleptic characteristics, lipid profile, and tropomyosin content. Irradiation of shrimp and squid with either 2.5 or 4.7 kGy reduced mesophilic bacteria contamination to low or nondetectable levels, respectively, whereas irradiation of octopus and cuttlefish with the same doses reduced the bacterial population. Irradiation treatment had no significant (P > 0.05) effect on the total lipid content and the major detected classes of polar and neutral lipids, whereas it significantly (P < 0.05) increased the contents of neutral lipids in octopus mantle and in shrimp muscle and cephalothorax samples. The total fatty acid content and the omega-3: omega-6 fatty acid ration was not affected. A dose-dependent significant (P < 0.05) decrease in the ratio of polyunsaturated fatty acids:saturated fatty acids was observed. With the increase in radiation dose, redness (a) and yellowness (b) values showed a variation, whereas the lightness (L) value was significantly (P < 0.05) decreased in mollusc mantles and shrimp muscle and increased in shrimp cephalothorax. The total of color changes ( delta E) increased (P < 0.05) as the dose increased. Significant (P < 0.05) changes in textural properties were observed with radiation treatment in octopus tentacles and in squid and cuttlefish mantle. The amount of tropomyosin, which is the major mollusc and crustacean allergen in the irradiated organisms, was reduced by gamma radiation, depending on the dose.

Deleterious effects of electron beam radiation on allergen extracts.

BACKGROUND: The recent threat to the public posed by the dissemination of Bacillus anthracis through the US postal system has resulted in increased security measures, including electron beam irradiation for the sterilization of some mail. The deleterious effects of electron beam radiation on biological products are not fully understood. OBJECTIVE: The purpose of this investigation was to assess the effect of electron beam radiation, as currently used to sterilize packages and mail in the United States, on several standardized or characterized allergen extracts. METHODS: Selected irradiated extracts were analyzed for allergen content and potency by SDS-PAGE, immunoblot, and ELISA (including inhibition) and compared with untreated extracts. RESULTS: The compositions and immunochemical potencies of these products were altered significantly by irradiation treatment. Physical changes to native protein structures observed after electrophoretic separations coincided with near-complete loss of allergenic and antigenic epitopes present on major and minor allergens, according to ELISA and immunoblot comparisons with untreated extracts. CONCLUSIONS: These results indicate that extracts subjected to electron beam sterilization conditions are likely to contain modified component structures and properties that might compromise the clinical effectiveness of these products.

Effects of gamma radiation on the allergenic and antigenic properties of milk proteins.

This study was carried out to evaluate the application of food irradiation technology as a method for reducing milk allergies. Bovine alpha-casein (ACA) and beta-lactoglobulin (BLG) were used as milk proteins. Using milk-hypersensitive patients' immunoglobulin E (IgE) and rabbit IgGs individually produced to ACA and BLG, the changes of allergenicity and antigenicity of irradiated proteins were observed by competitive indirect enzyme-linked immunosorbent assay. Allergenicity and antigenicity of the irradiated proteins were changed with different slopes of the inhibition curves. The disappearance of the band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and increase of the turbidity showed that solubility of the proteins decreased by radiation, and this decrease might be caused by agglomeration of the proteins. These results indicated that epitopes on milk allergens were structurally altered by gamma irradiation.

Effects of gamma radiation on the conformational and antigenic properties of a heat-stable major allergen in brown shrimp.

This study was performed to evaluate the application of food irradiation technology as a method for reducing shrimp allergy without adverse effects. Shrimp heat-stable protein (HSP) was isolated and gamma irradiated at 0, 1, 3, 5, 7, or 10 kGy in the condition of solution (1 mg/ml), and fresh shrimp was also irradiated. Conformational change of irradiated HSP was monitored by means of spectrometric measures, enzyme-linked immunosorbent assay with mouse monoclonal antibody, or human patients' sera and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The ability of the immunoglobulin E of patients allergic to shrimp to bind to irradiated HSP was dose dependently reduced. The amount of intact HSP in an irradiated solution was reduced by gamma irradiation, depending on the dose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the main band disappeared and the traces induced from coagulation appeared at a higher molecular weight zone. The binding ability of immunoglobulin E to allergens in the extracts from irradiated shrimp decreased, depending on the dose. The results provide a new method so that food irradiation technology can be applied to reduce allergenicity of shrimp.

Immediate hypersensitivity to cockroach. Isolation and purification of the major antigens.

Crude cockroach extract elicited positive skin tests in 50% of patients with positive and in 4% with negative environmental history for cockroach exposure, suggesting a possible role of cockroach in perennial atopic disease. Three major allergens in crude American and German cockroach extracts have been identified using sequential purification steps on Sephadex G-75, diethylaminoethyl (DEAE) cellulose, and agarose gel electrophoresis. Cr-I elicits positive skin tests in 70% of patients sensitive to the crude extracts. It has a molecular weight of approximately 25,500 daltons, is highly acidic, and resists boiling for four hours. Boiling in 4 N acetic acid completely abolishes its allergenicity. The purified allergen elicits positive skin tests at a concentration of 3 mug/ml and is capable of inducing greater than 50% histamine release from sensitive leukocytes at 0.05 ng/ml. A second antigen, Cr-II, elicits positive skin tests also in approximately 70% of cockroach-sensitive individuals, has a molecular weight of approximately 63,000 to 65,000 daltons, and has similar heat stability and acid hydrolysis characteristics to Cr-I. A third, less well-characterized antigen, Cr-III, has a molecular weight less than 10,000 daltons and elicits positive skin tests in 30% of individuals sensitive to the crude extract.