Thesis (Ph. D.)--University of Wisconsin--Madison, 1968. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M.Sc. (Tropical Medicine))--Mahidol University, 1978.

Thesis. 1977. Ph.D.--Massachusetts Institute of Technology. Dept. of Biology. / Microfiche copy available in Archives and Science. / Bibliography : leaves 272-276. / Ph.D.

Cross-over electrophoresis, an immunological method for analyzing blood residues on archaeological artifacts, is tested. Artifacts from three sites were utilized in the testing of this methodology. The sites are the Dietz site in south-central Oregon (282 artifacts), Konemehu in northern California (48 artifacts tested for Winthrop Associates), and Chimney Shelter in southwestern Oregon (3 artifacts from the Umpqua National Forest).

<p>ABSTRACT</p><p>Allergy to mustard has been reported for many years, in some cases as severe anaphylactic reactions. Recent studies imply that this allergy is increasing. Three major allergens have been isolated and characterised; Sin a 1 and Sin a 2 in yellow mustard (Sinapis alba), and Bra j 1 in oriental mustard (Brassica juncea). Yellow mustard and black mustard (Brassica nigra) are the most common species in Europe, whereas oriental mustard is more frequent outside Europe. Mustard plants belong to the Brassicaceae/Cruciferae family. Mustard is present as an ingredient in different foods, sauces and spices, often in small amounts. According to the European labelling directives, mustard and products thereof must always be declared. To monitor this regulation, methods need to be developed to detect mustard. Polyclonal antibodies, produced in rabbits, against yellow and black mustard were characterised with immunodiffusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and immunoblotting. Rocket-immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA) were developed for the detection and quantification of mustard protein. With indirect competitive ELISA a concentration of 156ng mustard protein per ml food extract was detected, which is more than enough to cover the lowest reported reactive doses.</p>

ABSTRACT Allergy to mustard has been reported for many years, in some cases as severe anaphylactic reactions. Recent studies imply that this allergy is increasing. Three major allergens have been isolated and characterised; Sin a 1 and Sin a 2 in yellow mustard (Sinapis alba), and Bra j 1 in oriental mustard (Brassica juncea). Yellow mustard and black mustard (Brassica nigra) are the most common species in Europe, whereas oriental mustard is more frequent outside Europe. Mustard plants belong to the Brassicaceae/Cruciferae family. Mustard is present as an ingredient in different foods, sauces and spices, often in small amounts. According to the European labelling directives, mustard and products thereof must always be declared. To monitor this regulation, methods need to be developed to detect mustard. Polyclonal antibodies, produced in rabbits, against yellow and black mustard were characterised with immunodiffusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and immunoblotting. Rocket-immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA) were developed for the detection and quantification of mustard protein. With indirect competitive ELISA a concentration of 156ng mustard protein per ml food extract was detected, which is more than enough to cover the lowest reported reactive doses.

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Immunoelectrophoresis revealed an antigen-antibody response between 4 day metabolic products and 8, 12 and 16 day sera and between 4 day trypanosomal extract and 16 day serum. Metabolic products from trypanosomes incubated at room temperature do not appear to be antigenic. The limitations of immunodiffusion are discussed in reference to the results. It is suggested that some of the antibodies to metabolic products may be of the precipitating type while others are not. Since a faint reaction also occurred between 4 day trypanosomal extract and 16 day serum, it may be concluded that metabolic products contribute to only a portion of the antibody response of the rat and are by no means the exclusive agents. They possibly work in conjunction with other metabolics within or on the surface of the trypanosome.

<p>Food allergy is an IgE-mediated immunological disease, which affects almost 4% of the adult population and up to 6% of children. Proteins from milk, egg, peanuts, soybean, wheat, fish and nuts are the main cause of food allergies. A less common allergen is pea protein. The National Food Administration analyses undeclared pea protein and contaminations of pea protein in foods using rocket immunoelectrophoresis and immunodiffusion. For both methods an antiserum against pea protein is needed. The aim of this study has been to characterize a newly developed rabbit-antiserum against pea protein. It is important to know if the antiserum is specific against peas, the detection as well as the quantification limits before it can be taken into use. The results of the study show that the antiserum was not absolutely specific, since it cross-reacted with chickpeas, fenugreek and lenses. However there is an "in-house" established PCR-method that can distinguish between chickpeas, fenugreek and peas and that method can be used as a complement to the rocket immunoelectrophoresis. The PCR-method cannot be used alone because it is not quantitative. Rocket immunoelectro¬phoresis detects 0,003% pea protein with purified IgG-antibodies from the antiserum.</p>

<p>A new European labeling directive (2003/89/EC) states that certain foods and products derived thereof must always be declared. Among the tree nuts specified is Macadamia nut (Macadamia spp.). During the last few years, cases of IgE-allergic reactions, even severe anaphylaxes, have been reported. Reliable methods for the detection of this nut are needed.</p><p>Protein from Macadamia nuts was isolated. Polyacrylamide gel electrophoresis in SDS revealed two main protein bands of about 20 and 50kDa. These protein bands were cut and extracted from the gel and rabbits were immunized with each protein.</p><p>Immunoblotting showed dominant reactivity with the respective antigens. The antisera were further tested for specificity in immunodiffusion and in rocket immunoelectrophoresis.</p><p>In addition, a specific DNA-method was developed, based on Real-Time PCR using Macadamia vicilin as target sequence. Two different primer pairs were tested. Specificity was tested against potentially related nuts. Optimisation of primer and probe concentrations was performed. The limit of detection was 2-4 pg DNA, corresponding to a macadamia nut concentration of 50 to 100 μg per g. In a background of soybean DNA, down to 0,01 % macadamia DNA could be detected.</p>