Abstract: Methods of bulk cryopreservation of C. parvum oocysts by vitrification using high aspect ratio cryopreservation devices are disclosed. Cryopreserved oocysts exhibit high viability, maintain infectivity in vitro, and are infectious to interferon-? knockout mice. The course of the infection is comparable to that observed with unfrozen oocysts.

Abstract: Methods for immunizing a subject to an antigen of an infectious agent, a tumor, or an allergen are provided, using vegetative cytoplasmic expression of the antigen or spore surface display of the antigen, and contacting the subject with a composition including a spore or a vegetative cell or both with or without an adjuvant. Also included are thermally-stable vaccine compositions using the method described above and kits including the compositions.

Abstract: A method for the simultaneous concentration of multiple toxins from large volumes of water. The method includes the steps of providing a disposable separation centrifuge bowl, the centrifuge bowl including a positively charged material at it's inner core. A large water sample contaminated with toxins from a group consisting of protozoa, bacteria, bacterial spores, and toxins is delivered to the centrifuge bowl. A centrifugal force is applied to the separation bowl. The water sample is concentrated to remove large particles of the toxins in the bowl due to the centrifugal forces. The concentrated water sample is passes through the positively charged inner core to capture any remaining concentrated targets by electrostatic forces and the concentrated targets are eluted.

Abstract: Compositions, methods and kits are provided for preparing an atoxic C. difficile protein that elicits an immune response in the subject specific for TcdA and TcdB. Atoxic Clostridium difficile toxin proteins were expressed in an endotoxin-free Bacillus system to develop a vaccine to reduce or treat incidence and severity of C. difficile infection (CDI).

Abstract: A method for the simultaneous concentration of multiple toxins from large volumes of water. The method includes the steps of providing a disposable separation centrifuge bowl, the centrifuge bowl including a positively charged material at it's inner core. A large water sample contaminated with toxins from a group consisting of protozoa, bacteria, bacterial spores, and toxins is delivered to the centrifuge bowl. A centrifugal force is applied to the separation bowl. The water sample is concentrated to remove large particles of the toxins in the bowl due to the centrifugal forces. The concentrated water sample is passes through the positively charged inner core to capture any remaining concentrated targets by electrostatic forces and the concentrated targets are eluted.

Abstract: A method for the simultaneous concentration of multiple toxins from large volumes of water. The method includes the steps of providing a disposable separation centrifuge bowl, the centrifuge bowl including a positively charged material at it's inner core. A large water sample contaminated with toxins from a group consisting of protozoa, bacteria, bacterial spores, and toxins is delivered to the centrifuge bowl. A centrifugal force is applied to the separation bowl. The water sample is concentrated to remove large particles of the toxins in the bowl due to the centrifugal forces. The concentrated water sample is passes through the positively charged inner core to capture any remaining concentrated targets by electrostatic forces and the concentrated targets are eluted.

Abstract: Atoxic Clostridium difficile toxin proteins were expressed in an endotoxin-free Bacillus system top develop a vaccine to reduce incidence and severity of C. difficile infection (CDI). Immunogens evaluated as potential vaccine candidates are mutated toxin A (encoded by TcdA) and toxin B (TcdB), and a rationally designed chimeric protein containing full-length TcdB protein except that the receptor binding domain is replaced with that of TcdA (designated as cTxAB). A small deletion (97 amino acids) in the transmembrane domain was used to reduce or eliminate toxicity.

Abstract: Cell-based methods for rapid real time assay of a presence of Clostridium difficile toxin and/or cells are provided, using an assay having a toxin-enhancing antibody and a sensitive cell line carrying FcyR receptors, and kits for this assay. An ultrasensitive cell based immunocytotoxicity assay for detecting less then 1 pg/ml of C. difficile toxins in clinical samples. A TcdA-specific monoclonal antibody, AIH3, was found to significantly enhance the cytotoxicity of TcdA to macrophages and monocytes. The AIH3-dependent enhancement of glucosyltransferase activity, cytoskeleton disruption, and TNF-a production induced by TcdA was demonstrated also in RAW 264.7 cells. Methods for high level recombinant expression of C. difficile toxins in Bacillus cells, and vectors for expression, strains of Bacillus carrying the vectors are provided.

Abstract: Methods for immunizing a subject to an antigen of an infectious agent, a tumor, or an allergen are provided, using vegetative cytoplasmic expression of the antigen or spore surface display of the antigen, and contacting the subject with a composition including a spore or a vegetative cell or both with or without an adjuvant. Also included are thermally-stable vaccine compositions using the method described above and kits including the compositions.

Abstract: Novel human monoclonal antibodies derived from a transgenic mouse are disclosed as well as a process for the preparation of the novel monoclonals and a therapeutic method of treating an individual for hemolytic uremic syndrome or of protecting an individual against hemolytic uremic syndrome by administration of the monoclonals to the individual in need of treatment or protection.

Abstract: Novel human monoclonal antibodies derived from a transgenic mouse are disclosed as well as a process for the preparation of the novel monoclonals and a therapeutic method of treating an individual for hemolytic uremic syndrome or of protecting an individual against hemolytic uremic syndrome by administration of the monoclonals to the individual in need of treatment or protection.

Abstract: Human and humanized monoclonal antibodies which binds specifically to subunit A of Shiga like toxin II have been developed which are effective to prevent or ameliorate one or more symptoms of HUS in a human. Effective dosages for treatment or prevention range from approximately 0.1 to 5.0 mg of antibody/kg of patient weight. The examples demonstrate the preferred dosage ranges based on the pig model, and what is being tested in phase I clinical trials. Antibodies are preferably transfused over a period of two hours, although this will depend on the patient and the disease state at the time of treatment. Preferred dosages for treatment of humans are between 0.1 mg/kg-5.0 mg/kg of 5C120, or an equivalent dosage of another antibody to subunit A of STX2. In the most preferred embodiments, dosages of 0.1 mg/kg, 0.5 mg/kg, or 5.0 mg/kg of 5C12 (low dose, anticipated therapeutic dose based on animal data and high dose) are administered.

Abstract: Cell-based methods for rapid real time assay of a presence of Clostridium difficile toxin and/or cells are provided, using an assay having a toxin-enhancing antibody and a sensitive cell line carrying FcyR receptors, and kits for this assay. An ultrasensitive cell based immunocytotoxicity assay for detecting less then 1 pg/ml of C. difficile toxins in clinical samples. A TcdA-specific monoclonal antibody, AIH3, was found to significantly enhance the cytotoxicity of TcdA to macrophages and monocytes. The AIH3-dependent enhancement of glucosyltransferase activity, cytoskeleton disruption, and TNF-a production induced by TcdA was demonstrated also in RAW 264.7 cells. Methods for high level recombinant expression of C. difficile toxins in Bacillus cells, and vectors for expression, strains of Bacillus carrying the vectors are provided.

Abstract: The present invention relates to a designer or recombinant ubiquitin ligase molecule that includes an antibody fragment that is specific for a toxin active fragment, wherein the toxin active fragment is an enzymatically active fragment of one or more toxins or toxin serotypes; and an E3-ligase domain that comprises an E3-ligase or polypeptide that facilitates E2-mediated ubiquitination of the toxin active fragment. In an embodiment, the composition further includes a delivery system that allow the designer ubiquitin ligase to enter the cell. The present invention further includes methods for treating an individual intoxicated with a toxin by administering the designer ubiquitin ligase of the present invention.

Abstract: A method for the simultaneous concentration of multiple toxins from large volumes of water. The method includes the steps of providing a disposable separation centrifuge bowl, the centrifuge bowl including a positively charged material at it's inner core. A large water sample contaminated with toxins from a group consisting of protozoa, bacteria, bacterial spores, and toxins is delivered to the centrifuge bowl. A centrifugal force is applied to the separation bowl. The water sample is concentrated to remove large particles of the toxins in the bowl due to the centrifugal forces. The concentrated water sample is passes through the positively charged inner core to capture any remaining concentrated targets by electrostatic forces and the concentrated targets are eluted.

Abstract: The present invention relates to methods, microarrays and kits for detecting one or more human astrovirus serotypes in a sample (e.g., a fecal sample) from an individual. The method includes amplifying nucleic acid molecules of the sample with one or more primers, to thereby obtain an amplified nucleic acid product; contacting the amplified nucleic acid product with one or more serotype specific probes having a nucleic acid sequence that is specific for only one astrovirus serotype in the group of astroviruses being assessed, wherein the nucleic acid sequence includes between about 9 and 25 nucleic acid bases (e.g., SEQ ID NO: 5-24); and detecting the hybridization complex. The presence of hybridization complexes with a serotype specific probe indicates the presence of one or more specific astrovirus serotypes, and the absence of hybridization complexes with a serotype specific probe indicates the absence of the specific astrovirus serotype.

Abstract: Human and humanized monoclonal antibodies which binds specifically to subunit A of Shiga like toxin II have been developed which are effective to prevent or ameliorate one or more symptoms of HUS in a human. Effective dosages for treatment or prevention range from approximately 0.1 to 5.0 mg of antibody/kg of patient weight. The examples demonstrate the preferred dosage ranges based on the pig model, and what is being tested in phase I clinical trials. Antibodies are preferably transfused over a period of two hours, although this will depend on the patient and the disease state at the time of treatment. Preferred dosages for treatment of humans are between 0.1 mg/kg-5.0 mg/kg of 5C120, or an equivalent dosage of another antibody to subunit A of STX2. In the most preferred embodiments, dosages of 0.1 mg/kg, 0.5 mg/kg, or 5.0 mg/kg of 5C12 (low dose, anticipated therapeutic dose based on animal data and high dose) are administered.

Abstract: Novel human monoclonal antibodies derived from a transgenic mouse are disclosed as well as a process for the preparation of the novel monoclonals and a therapeutic method of treating an individual for hemolytic uremic syndrome or of protecting an individual against hemolytic uremic syndrome by administration of the monoclonals to the individual in need of treatment or protection.

Abstract: Novel human monoclonal antibodies derived from a transgenic mouse are disclosed as well as a process for the preparation of the novel monoclonals and a therapeutic method of treating an individual for hemolytic uremic syndrome or of protecting an individual against hemolytic uremic syndrome by administration of the monoclonals to the individual in need of treatment or protection.

Abstract: Disclosed is a therapeutic method for the treatment of hemolytic uremic syndrome. More specifically, the method includes the administration of monoclonal antibodies, chimeric monoclonal antibodies and monospecific polyclonal antibodies specific for Shiga like toxin.