Abstract: The invention provides methods useful for identifying polynucleotides expressed in vivo by a microbe during infection of a host. Antibodies against antigens that are expressed by the microbe in vivo and in vitro are adsorbed with cells or cellular extracts of the microbe that have been grown in vitro. Unadsorbed antibodies are isolated and are probed against a phage display library of the microbe's DNA or RNA. A polynucleotide of the microbe that is expressed in vivo is isolated and identified.

Abstract: Disclosed are methods for identifying molecular interactions (e.g., protein/protein, protein/DNA, protein/RNA, or RNA/RNA interactions). All of the methods within the invention employ counterselection and at least two hybrid molecules. Molecules which interact reconstitute a transcription factor and direct expression of a reporter gene, the expression of which is then assayed. Also disclosed are genetic constructs which are useful in practicing the methods of the invention.

Abstract: The present invention includes a method for detecting and isolating sugarcane proteins that interact with the HC-Pro and P1 proteins of SrMV and other proteins involved in gene silencing, particularly in sugarcane. The method uses a two hybrid assay with an HC-Pro, P1, or other silencing-related protein-containing bait protein and a prey protein containing a polypeptide encoded by a DNA molecule in a cDNA library. The method also includes identification of false positives through reverse two-hybrid assays and using in vitro techniques such as farwestern blots or pull down assays where plant physiological conditions may be replicated. Finally, interactions may be confirmed in planta. Some novel proteins used in and discovered using the these methods are also identified. Methods of using viral and plant proteins to regulate silencing in plants such as sugarcane are also discussed.

Abstract: The present invention provides a method for identifying a modulator or mediator of a biological activity, which activity includes antigenicity and or immunogenicity, said method comprising the step of: (i) producing a gene fragment expression library derived from defined nucleotide sequence fragments; and (ii) assaying the expression library for at least an amino acid sequence derived from step (i) for a biological activity wherein that activity is different from any activity the amino acid sequence may have in its native environment.

Abstract: The present invention relates, in general, to biosensors and, in particular, to bioelectronic sensors comprising a macromolecule immobilized on an electrode surface so that a redox cofactor that is site-specifically attached to the surface of the macromolecule is between the macromolecule and electrode surface ligand-mediated conformational changes alter the geometry of interaction between the redox cofactor and the electrode surface resulting in a change in electronic coupling between the cofactor and electrode.

Abstract: A kit and method of predicting a refractory response in a subject diagnosed as having periodontal disease by measuring serum concentrations of actinomyces antibodies, streptococcal antibodies and lysine decarboxylase antibodies and using the measurement along with other subject information in a set of derived equations.

Abstract: The present invention relates to isolated polypeptides having oxaloacetate hydrolase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.

Abstract: A correct viable cell number of a microorganism in a biological tissue is measured by cultivating a fragment of the biological tissue such as skin infected with the microorganism and administered with an antimicrobial agent, in a medium containing a phospholipid, a nonionic surfactant, or both of the phospholipid and the nonionic surfactant.

Abstract: The present invention relates to a method and a means of diagnosing Candida infection. In particular, the present invention relates to a method of diagnosing Candida infection comprising the steps of obtaining a biological sample from a subject at risk of, or suspected to be suffering from, Candida infection, contacting the sample with a mannose depleted antigen composition comprising a soluble cytoplasmic antigen preparation comprising antigens of molecular weight 55 kDa, 30 kDa and 20 kDa and detecting the antigen/antibody reaction.

Abstract: The present invention is directed to a method for selecting a prey polypeptide that is able to interact with a bait polypeptide of interest, to a prey polynucleotide encoding the prey polypeptide as well as to the prey polypeptide itself. The invention also concerns plasmids used for performing the method of the invention as well as prokaryotic or eukaryotic recombinant host organisms containing such plasmids and also a collection of said recombinant host organisms consisting in a DNA library, such as a collection of recombinant haploid Saccharomyces cerevisiae. Finally, the invention is also directed to a technical medium containing the whole information concerning the interactions between metabolically related bait and prey polypeptides and/or polynucleotides coding for bait and prey polypeptides.

Abstract: A method and apparatus for the differentiation of Crohn's disease from other gastrointestinal illnesses, such as ulcerative colitis and irritable bowel syndrome, using the presence of fecal anti-Saccharomyces cerevisiae antibodies (ASCA) as a marker for Crohn's disease are provided. The apparatus includes an enzyme-linked immunoassay or other immunoassay that utilizes antibodies specific to human immunoglobins for the measurement of total endogenous ASCA in a human fecal sample. The method and apparatus may be used by healthcare providers to distinguish Crohn's disease from other gastrointestinal illnesses, such as ulcerative colitis and irritable bowel syndrome.

Abstract: The present invention relates to oxidized fungal antigens and methods of making and using thereof. More particularly, the present invention provides a method for producing an oxidized fungal antigen in culture filtrate. The present invention also provides for the produced oxidized fungal antigens. Devices comprising such oxidized fungal antigens, methods for testing for fungal antibodies using the oxidized fungal antigens and methods for producing anti-fungal antibodies using oxidized fungal antigens are further provided. Antigen detection devices comprising anti-fungal antibodies raised against oxidized fungal antigens produced by the present methods are further provided.

Abstract: A method for determining the presence of food allergy or food intolerance and their cross-reactive tissue antigens is disclosed. The method includes determining a level of antibodies against a dietary antigen in a mucosal sample from the patient and comparing the level with normal levels of the antibodies. Dietary antigens that were tested include milk and milk products; eggs and egg products; meat and meat products; fish, mollusks, and crustaceans and their products; oils, fats, and their products; grains and grain products; pulses, seed, kernels, nuts, and their products; vegetable and vegetable products; fruit and fruit products; sugar, sugar product, chocolate products, and confectionary; and spices.

Abstract: A high throughput toxicology screening method is provided. In the subject method, at least 10 different compound compositions are tested simultaneously. Each compound composition is tested by contacting it with a plurality, e.g. from about 10 to 1000, of non-mammalian multi-cellular organisms and determining the effect of the compound composition on the organisms. The multi-cellular organisms employed in the subject methods are small, have differentiated tissues and organs and have a rapid generation time. The subject high throughput screening methods find use in a variety of applications, and are particularly suited for use in the toxicology screening of libraries of compounds, such as libraries of combinatorially produced compounds.

Abstract: Disclosed is a transformed yeast cell containing a first heterologous DNA sequence which codes for a mammalian G protean coupled receptor and a second heterologous DNA sequence which codes for a mammalian G protein ? subunit (mammalian G?). The first and second heterologous DNA sequences are capable of expression in the cell, but the cell is incapable of expressing an endogenous G protein ?-subunit (yeast G?). The cells are useful for screening compounds which affect the rate of dissociation of G? from G?? in a cell. Also disclosed is a novel DNA expression vector useful for making cells as described above. The vector contains a first segment comprising at least a fragment of the extreme amino-terminal coding sequence of a yeast G protein coupled receptor. A second segment is positioned downstream from the first segment (and in correct reading frame therewith), with the second segment comprising a DNA sequence encoding a heterologous G protein coupled receptor.

Abstract: A transposable element is provided that as a 3? and a 5? end. The transposable element includes a 5? recombining site 5? of a nucleic acid sequence encoding a selectable marker, a 3? recombining site 3? of the nucleic acid sequence encoding a selectable marker, a nucleic acid sequence encoding and MHC epitope 5? to the 5? recombining site or 3? to the 3? recombining site, and an insertion end comprising an inverted repeat sequence sufficient for integration of the transposable element at the 5? and the 3? end of the transposable element. In one embodiment, a transposable element is provided that has a 5? and a 3? end. The transposable element includes a 5? loxP sequence 5? of a nucleic acid encoding a selectable marker, a 3? loxP sequences 3? of a nucleic acid encoding the selectable marker, an MHC epitope 5? to the 5? loxP sequences or 3? of the 3? loxP sequence, an insertion end at the 5? end of the transposable element, and an insertion end at the 3? of the transposable element.

Abstract: The invention is directed to isolated DNAs having nucleic acid sequences which encode proteins which regulate aureobasidin sensitivity. Also disclosed are recombinant plasmids containing the DNAs, transformants containing the plasmids, and methods of producing the proteins.

Abstract: This invention is directed to mutant SPE-C toxins or fragments thereof, vaccine and pharmaceutical compositions, and methods of using the vaccine and pharmaceutical compositions. The preferred SPE-C toxin has at least one amino acid change and is substantially non-lethal compared with the wild type SPE-C toxin. The mutant SPE-C toxins can form vaccine compositions useful to protect animals against the biological activities of wild type SPE-C toxin.

Abstract: Methods for the diagnosis of ABPA in a human individual comprise determining if the individual carries antibodies reactive with one or more ABPA-related recombinant allergens, which one or more ABPA-related recombinant allergens discriminate between ABPA and allergic sensitization to A. fumigatus. Suitable allergens include rAsp F4, rAsp F6, rAsp F8, and ABPA-related fragments thereof which bind with IgE or IgG antibody.

Abstract: A method for detecting protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other. The interaction between the two fusion proteins leads to protein trans-splicing, generating an active and detectable reporter.

Abstract: The present invention provides for the production of ASA from yeast capable of producing ASA from KLG. The present invention provides methods for the production of ASA as well as recombinant yeast capable of producing ASA from a carbon source.

Abstract: The invention relates to a process for identifying microorganisms by means of mass spectrometry, especially by means of MALDI-TOF-US. According to the invention, a data base (DB) is used that comprises synthetic reference spectra (REFs) of known microorganisms, which are formed by combination of a number, reduced relative to natural mass spectra (REFN), of signals (S) that are specific to the respective microorganism, as well as difference spectra (DIF), which are formed by offsetting in each case two synthetic reference spectra (REFS) of the known microorganisms. The process also comprises a first analysis step, in which a similarity analysis of a sample mass spectrum (SAM) of a microorganism that is to be identified is performed with synthetic reference spectra (REFS) that are contained in data base (DB), and a second analysis step, in which a similarity analysis of sample spectrum (SAM) is performed with at least a portion of difference spectra (DIF) that are contained in data base (DB).

Abstract: The invention provides isolated nucleic acid and amino acid sequences of TL-&ggr;, antibodies to TL-&ggr;, methods of screening for TL-&ggr; modulators using biologically active TL-&ggr;, and kits for screening for TL-&ggr; modulators.

Abstract: The present invention relates to a method for determining the ability of a compound to modify the interaction between parkin and the p38 protein, and in particular to a method for screening for or detecting compounds intended for the prevention and/or treatment of neurodegenerative pathological conditions.

Abstract: The invention relates to a method for quantitation of recombinant proteins of interest in the absence of specific antibodies, which is particularly suitable for screening cell clones expressing heterologous genes.

Abstract: Yeast cells are engineered to express both a surrogate of a pheromone system protein (e.g., enzymes involved in maturation of &agr;-factor, transporters of a-factor, pheromone receptors, etc.) and a potential peptide modulator of the surrogate, in such a manner that the inhibition or activation of the surrogate affects a screenable or selectable trait of the yeast cells. Various additional features improve the signal-to-noise ratio of the screening/selection system.

Abstract: The invention provides for methods of determining the presence, absence, or amount of microbial contamination in an animal by-product. The invention further provides for methods of monitoring animal by-products before, during, or after processing of the animal by-product into, for example, feed. The invention also provides for articles of manufacture for carrying out the claimed methods.

Abstract: A method for identifying small acid-soluble proteins (SASPs) by generating an increased number of biomarkers upon controllably triggering enzymatic digestion in an intact spore is disclosed. An additional embodiment of the method includes oxidizing an unknown protein in a microorganism by pre-selected oxidation facilitating agent, which causes a predetermined mass gain in Methionine, thus serving as an indicator of a particular family of proteins.

Abstract: Antibodies to various fungal antigens are disclosed, including monoclonal antibody 9B4 that selectively binds an antigen of Stachybotrys chartarum spores not found in Stachybotrys chartarum mycelium. The antibodies may be used in a variety of methods, such as detecting the presence of fungal antigens in the environment or within a sample obtained from an animal or plant, or testing the effectiveness of an agent in binding an antigen.

Abstract: The present invention is directed to methods for extracting markers from biological samples, and to systems, devices, kits and reagents for use in such methods. The invention is also to methods, kits, reagents and compositions for measuring a plurality of different organism types in a sample. One of the specific advantages of the present invention is the ability to simultaneously extract more than one microorganism or viral particle marker in one volume from a single sample containing a complex biological matrix.

Abstract: Methods of isolating mutant yeast cells with increased life span, as well as mutant yeast cells isolated by the methods, are disclosed. Also described are methods of identifying agents which increase life span of yeast cells, and methods of isolating genes which affect senescence in organisms.

Abstract: The present invention relates to a method and a means of diagnosing Candida infection. In particular the present invention relates to a method of diagnosing Candida infection by measuring the levels of antibody to Candida cytoplasmic antigen present in a biological sample taken from a subject at risk of, or suspected to be suffering from a Candida infection.

Abstract: The present invention provides a method and a device for the identification of a pathogenic agent in a sample. The method comprises the steps of contacting the pathogenic agent in the sample with a plurality of ligands selected from the group consisting of an oligosaccharide, a siderophore, a ferrisiderophore, a vitamin, analogs or derivatives thereof, and combinations thereof, wherein the plurality of ligands are non-diffusably bound to a solid support in a spatially defined manner, detecting binding of the pathogenic agent to at least one of the ligands, wherein the pathogenic agent binds specifically to the at least one ligand to generate a spatially defined binding pattern, and identifying the pathogenic agent based on the spatially defined binding pattern.

Abstract: The invention relates to compounds of formula (I) 1

Abstract: Ku is a protein found in a wide range of organisms. It comprises two tightly-associated subunits termed Ku70 and Ku80. The present invention relates to the discovery and characterisation of an interaction between Ku70 and Ku80 and DNA-PKCS. Various applications based on this interaction are provided. These are relevant to numerous cellular processes which are of interest in therapeutic contexts.

Abstract: The present invention involves a method and an immuno-analytical device for the rapid and simultaneous detection of multiple micro-organisms in the biological fluids from milk-producing animals suffering from mastitis. This method is based on a lateral flow immuno-assay technique performed to detect antigens specific for multiple infectious agents which are known to cause and/or be encountered in cases of mastitis. Mastitis is an inflammatory condition affecting the udders of milk-producing animals as a result of microbial infections.

Abstract: The invention provides isolated nucleic acid compounds encoding a multiple drug resistance protein of Aspergillus nidulans. Vectors and transformed host cells comprising the multiple drug resistance-encoding DNA of Aspergillus nidulans atrD are also provided. The invention further provides assays which utilize these transformed host cells.

Abstract: A method of screening a mixture for active entities, which method comprises: providing a plurality of ligands, wherein each ligand is attached to a support to form a plurality of ligand-support complexes, contacting the ligand-support complexes with a mixture comprising a plurality of entities under conditions that allow at least one entity to bind to at least one ligand-support complex, thereby forming at least one entity-ligand-support complex, separating at least one entity-ligand-support complex from the unbound entities, assaying at least one entity of at least one entity-ligand-support complex for an activity, detecting the activity, and selecting at least one entity-ligand-support-complex having the entity, which exhibited the detected activity, whereupon a mixture is screened for active entities; and related methods.

Abstract: The present invention concerns methods for detecting the presence of a micro-organism in a fluid, gaseous and solid samples, together with apparatus for use in same. The invention comprises the use of a plurality of hollow fibres to filter the fluid and the cpture of the micro-organisms on the membrane by means of an specific binding pair.

Abstract: A method for a screening of an agent acting on a cell wall, which comprises the steps of (1) culturing each of microorganisms having a reporter protein fixed on a cell wall or a cell membrane as a GPI-anchored protein in the presence of a test agent; (2) determining the reporter protein released into each culture fluid of the microorganism cultured; and (3) judging that the test agent has a selective inhibitory action on the cell wall when the reporter protein is released from the microorganism having the reporter protein fixed on the cell wall into the culture fluid and the reporter protein is not substantially released from the microorganism having the reporter protein fixed on the cell membrane into the culture fluid.

Abstract: The present invention relates to growing and testing microorganisms in a multitest format which utilizes a gel forming matrix for the rapid screening of clinical and environmental cultures. The present invention is suited for the characterization of commonly encountered microorganisms (e.g., E. coli, S. aureus, etc.), as well as commercially and industrially important organisms from various and diverse environments (e.g., the present invention is particularly suited for the growth and characterization of the actinomycetes and fungi). The present invention is also particularly suited for comparative analysis of phenotypic differences between cell types, including strains of microorganisms that have been designated as the same genus and species, as well as other cell types (e.g., mammalian, insect, and plant cells).

Abstract: A method for identifying a protein antigen to a target fungus is disclosed. The method comprises screening expressed proteins from a cDNA gene expression library with antisera to the target fungus and cross-screening with antisera to a nontarget fungus. Antibodies to the protein antigen are also disclosed. Methods for detecting the presence or absence of the antibodies or of the protein antigen are also disclosed, as well as kits for performing such assays. In preferred embodiments, the target fungus is H. capsulatum.

Abstract: Compositions, methods, and kits are provided for efficiently generating and screening humanized antibody with high affinity against a specific antigen. The library of humanized antibody is generated by mutagenizing a chimeric antibody template that combines human antibody framework and antigen binding sites of a non-human antibody. Alternatively, the library of humanized antibody is generated by grafting essential antigen-recognition segment(s) such as CDRs of the non-human antibody into the corresponding position(s) of each member of a human antibody library. This library of humanized antibody is then screened for high affinity binding toward a specific antigen in vivo in organism such as yeast or in vitro using techniques such as ribosome display or mRNA display. The overall process can be efficiently performed in a high throughput and automated manner, thus mimicking the natural process of antibody affinity maturation.

Abstract: The invention provides a method for screening a test compound for the ability of the test compound to induce a response from human naive T-cells. The method comprises admixing human naive T cells, macrophages/monocytes, immortalized B cells lacking class I and class II major histocompatibility antigens, and a test compound; and determining whether the test compound induces a response from the human naive T cells. The invention further provides a method for primary in vitro sensitization of human naive T-cells. The method comprises admixing human naive T cells, macrophages/monocytes, immortalized B cells lacking class I and class II major histocompatibility antigens, and an antigen.

Abstract: A predictive test for rheumatoid arthritis comprises the detection of antibodies to collagen in a biological sample from a patient by contacting the biological sample with an antigen comprising the CB10 peptide of mammalian type II collagen, or an antibody-binding fragment or variant thereof, for a time and under conditions for an antibody-antigen complex to form, and detecting the antibody-antigen complex, for example by immunoassay. A diagnostic test kit is also disclosed.

Abstract: The present invention is a very significant way to determine exposure to toxic mold, or the disease called mycotoxicosis. Urine samples are extracted for trichothecene mycotoxins to confirm the disease of mycotoxicosis in humans. Diagnosis of mycotoxicosis is confirmable even when toxic mold exposure cannot be ascertained to a reasonable degree of certainty.

Abstract: The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 104 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. In effect, yeast has evolved a platform for protein-protein binding without steric hindrance, from cell wall components.

Abstract: Apparatus comprising G-protein coupled receptor (GPCR) for detecting ligands or substances in liquid or vapor media. The GPCR is based on in a cell or in a synthetic membrane or polymer system, and combined with means for obtaining a sample of a liquid or vapor medium, and with automatic optical detection system and monitoring system for detecting a ligand of interest. Methods are disclosed for detecting a ligand of interest using the GPCR apparatus.

Abstract: A method and test kit for an agglutination assay for antibodies in human or other mammal serum is described. The kit preferably contains latex particles and uses extracellular antigen(s) of Pythiosis insidiosum which bind to the particles and which in turn bind to the serum antibodies to produce the agglutination. The assay is particularly important for detecting early stages of Pythiosis.

Abstract: A method for determining a cause for digestive and immune disorders is disclosed. The method determines the levels of antibodies against normal intestinal microflora and food antigens. It then compares the results to normal levels to determine the cause. The test can be used to diagnose food allergy or intolerance, microflora imbalance, gut barrier dysfunction, bacterial translocation, immunodeficiencies, candidiasis and autoimmunities.