Abstract: The invention discloses a Lac shuttle vector, comprising at least (a) a region which regulates a plasmid copy number; (b) an eukaryotic gene expression cassette, which comprises at least an eukaryotic gene transcriptional promoter sequence, a multiple cloning site and a transcriptional terminator sequence; (c) a lactic acid bacteria plasmid sequence, which comprises a plus origin of replication, and a nucleic acid sequence encoding for a protein which relates to the lactic acid bacteria plasmid replication; and (d) a non-antibiotic resistance selection gene and the promoter sequence thereof. The Lac shuttle vector features a non-antibiotic resistance gene as a selection marker, which is useful in pharmaceuticals and foods.

Abstract: The present invention provides novel in vivo assay systems and methods of using these assays systems to identify compounds that affect microbial cell division. The present invention further provides pharmaceutical compositions that have anti-microbial activity and methods of treating microbial infections.

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: Novel methods for the treatment and/or prophylaxis of diseases caused by tissue-adhering bacteria are disclosed. By interacting with periplasmic molecular chaperones it is achieved that the assembly of pili is prevented or inhibited and thereby the infectivity of the bacteria is diminished. Also disclosed are methods for screening for drugs as well as methods for the de novo design of such drugs, methods which rely on novel computer drug modelling methods involving an approximative calculation of binding free energy between macromolecules. Finally, novel pyranosides which are believed to be capable of interacting with periplasmic molecular chaperones are also disclosed.

Abstract: The present invention relates to an immunogenic conjugate comprising a carrier molecule coupled to an autoinducer of a Gram negative bacteria. The immunogenic conjugate, when combined with a pharmaceutically acceptable carrier, forms a suitable vaccine for mammals to prevent infection by the Gram negative bacteria. The immunogenic conjugate is also used to raise and subsequently isolate antibodies or binding portions thereof which are capable of recognizing and binding to the autoinducer. The antibodies or binding portions thereof are utilized in a method of treating infections, a method of inhibiting autoinducer activity, and in diagnostic assays which detect the presence of autoinducers or autoinducer antagonists in fluid or tissue samples.

Abstract: The invention provides a scintillation proximity assay for detecting peptidoglycan synthesis. The assay is especially suitable for high throughput screening of compounds affecting peptidoglycan synthesis.

Abstract: The invention relates to a vaccine comprising a bacterium attenuated by a non-reverting mutation in a gene encoding a protein which promotes folding of extracytoplasmic proteins. Such mutations were initially identified as being useful in vaccines from a bank of randomly inserted, transposon mutants in which attenuation was determined as a reduction in virulence of the organism in the mouse model of infection. Site directed mutation of the gene results in a strain which shows at least 4 logs of attenuation when delivered both orally and intravenously. Animals vaccinated with such a strain are protected against subsequent challenge with the parent wild type strain. Finally, heterologous antigens such as the non-toxic and protective, binding domain from tetanus toxin, fragment C, can be delivered via the mucosal immune system using such strains of bacteria. This results in the induction of a fully protective immune response to subsequent challenge with native tetanus toxin.

Abstract: Disclosed is a pO157 plasmid-specified polypeptide found in E. coli EDL933and other enterohemorrhagic E. coli that binds to and cleaves C1-esterase inhibitor. Also disclosed are methods employing the polypeptide for diagnosing and treating colitis or hemolytic uremic syndrome, and methods of detecting potential therapeutics.

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 system for the detection of bacteria based on bacteria-antibody complexes. Bacteria attached to antibody are detected with resonance Raman spectroscopy. The bacteria are detected directly in a great numerical excess, e.g. 100 to 10,000 of antibody molecules. A sample to be tested is placed in a medium, the medium containing antibodies attached to a surface for binding to a specific bacteria to form an antigen to antibody complex. The medium is contacted with a beam of light energy. The bacteria, as a lower resonance enhanced Raman backscattered energy, is analyzed for the presence or absence of the bacteria.

Abstract: The present inventors have found that certain preparations containing LPS and/or lipid A variants, derivatives, and/or analogs demonstrate non-pyrogenic properties and exhibit anti-viral activities. In particular, non-pyrogenic preparations of LPS, lipid A, LPS antagonists and lipid A antagonists, and derivatives thereof induce ? chemokine secretion, such as MIP-1?, but not proinflammatory cytokines, such as TNF?, IL-1? and IL-6. Non-pyrogenic preparations of the invention have been demonstrated by the Applicant to suppress HIV replication in human peripheral blood monocytes, as described by way of example herein. The present invention provides preparations of LPS or lipid A variants, analogs and derivatives of decreased or absent pyrogenicity which can be used as therapeutics for the treatment or prevention of immunodeficiency virus infection and its consequences.

Abstract: The present invention relates to a method of screening for compounds that inhibit the virulence of Pseudomonas bacteria and comprises the steps of: providing a culture medium comprising Pseudomonas bacteria, administering a test compound to said bacteria, and then detecting the presence or absence of inhibition of the catabolite repression control (Crc) protein in the bacteria. The inhibition of the Crc protein indicates that the compound has antivirulence activity against Pseudomonas bacteria. Antisense oligonucleotides that inhibit expression of the Crc protein in a Pseudomonas bacteria and is nuclease resistant are also disclosed. Antivirulence compounds and the uses thereof in treating Pseudomonas infections are also disclosed.

Abstract: Methods and compositions comprising immunoassays for the detection of functional antibodies and the analysis of vaccine efficacy are described. In particular, the present invention provides opsonophagocytic assays. The assays are useful for the rapid and simultaneous detection of multiple different functional antibodies. In preferred embodiments, the assays include fluorescent labels of multiple colors and/or intensities.

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 the finding and detection of Helicobacter pylori (H. pylori) antigens in blood of infected individuals. The H. pylori antigens are components of H. pylori cells which include, but not limited to DNA, RNA, and fragments of nucleotides, proteins or peptides. H. pylori DNA, RNA, and fragments of nucleotides can be detected by polymerase chain reaction (PCR), ligase chain reaction (LCR), or DNA hybridization methods or other amplification methods. H. pylori proteins or peptides or other antigenic components thereof can be detected by immunoassays or immunoblot using an antibody against H. pylori, preferably an antibody purified by an affinity column. The present invention further provides immunoassay methods, diagnostic kits, and an immunochromatographic assay device for detection of Helicobacter pylori antigens in serum samples.

Abstract: The invention provides methods for screening for the presence of a clinically relevant amount of bacteria in donor blood or a blood product from a donor mammal, particularly blood or a blood product that will be transferred from the donor mammal to a recipient mammal. The method comprises contacting a sample of the donor blood or a blood product with a set of binding agents that comprises binding agents that specifically bind to Gram-negative bacterial antigen and/or binding agents that specifically bind to Gram-positive bacterial antigen, and determining binding of the set of binding agents to the sample, wherein binding indicates the presence of a clinically relevant amount of Gram-positive bacteria and/or Gram-negative bacteria in the donor blood or blood product and no binding indicates the absence of a clinically relevant amount of Gram-positive bacteria and/or Gram-negative bacteria in the donor blood or blood product.

Abstract: Provided are novel complexes containing a crosslinked avidin, an analyzing method and analyzing reagents and kits whereby a compound to be analyzed can be quickly, conveniently and accurately analyzed while taking advantage of the avidin-biotin reaction. The complexes contain at least two homogeneous or heterogeneous biotin-introduced products and one crosslinked avidin sandwiched therebetween. In the analyzing method, the homogeneous or heterogeneous biotin-introduced products and the crosslinked avidin are used. The analyzing reagent contains the crosslinked avidin. The analyzing kit contains the crosslinked avidin and a biotinylating agent.

Abstract: A method for determining and diagnosing inflammatory enteric disease using an immunochromatographic test device having a multiplicity of test zones. The method tests for the presence of at least one enteric pathogen and at least one of certain inflammatory enteric disease markers. The enteric pathogens tested for can be any number of enteric pathogens such as the pathogens E. coli O157, Campylobacter, Salmonella, Listeria, Shigella, and Yersinia. The inflammatory enteric disease markers tested for are fecal lactoferrin, a bacteria marker, a virus marker, and a protozoa marker. Positive results for any one of the pathogens indicates that pathogen as the cause of the inflammatory enteric disease. Positive results for fecal lactoferrin indicate an inflammatory condition of the intestines. Positive results for the bacteria, virus, protozoa markers indicate respectively a bacterial, viral, or protozoan cause of infection as the cause of the disease.

Abstract: The invention relates to a prokaryotic cell system for monitoring protease activity. The invention also includes assays for identifying protease inhibitors and protease modulators, determining the amino acid sequence of a protease cleavage site for a known protease, identifying and cloning a protease whose cleavage site is known, and rapidly identifying a form of a protease exhibiting increased activity relative to a control protease.

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.

Abstract: A method for detecting Escherichia coli in a sample by providing a sample suspected of containing Escherichia coli; contacting the sample with beads coated with a mixture of antibodies or anti-serum specific to one or more surface antigens of Escherichia coli, each of the one or more surface antigens having a molecular weight of 21±2 KDa, 26±2 KDa, 31±2 KDa, 36±2 KDa, 38±2 KDa, 67±2 KDa, or 77.8±2 KDa; and observing agglutination of the beads, where the presence of the agglutination indicates the presence of Escherichia coli in the sample.

Abstract: The gene of human acidic fibroblast growth factor 155 (haFGF 155) has been obtained by chemical synthesis. The nucleotide sequence of haFGF 155 gene has been deduced on the basis of haFGF 155 amino acid sequence as described in the literature. The amino acid sequence of the synthesized haFGF 155 does not differ from those described in the literature. The nucleotide sequence of haFGF gene differs from those described previously. For chemical synthesis of haFGF 155 gene, codons were used which are the ones most often used by E. coli in highly expressed E. coli proteins. A plasmid with haPGF 155 (phaFGF 155) gene was obtained and was used to transform E. coli. Production of haFGF 154 protein was achieved by cultivation of the producer strain under conditions which slow down the lytic development of lambda phage. The haFGF 154 protein accumulated in culture medium in a soluble condition as a result of the producer strain cells lysis by the lambda phage.

Abstract: Several EHEC proteins which are secreted into the culture supernatant have been discovered. These proteins are not produced by non-pathogenic E. coli, and produce a strong serum antibody response in patients with HUS and bloody diarrhea.

Abstract: The invention is related to antibodies, particularly monoclonal antibodies, which recognize particularly epitopes of the intimin protein of enteropathogenic E. coli and enterohemorrhagic E. coli, methods of detecting such E. coli by use of these antibodies, and kits containing these antibodies for diagnosis.

Abstract: A novel rapid mutational analysis method for mapping protein epitopes is disclosed. This method has been used to identify the binding sites for 16 anti-CD2 and anti-CD4 monoclonal antibodies. The powerful, rapid, and simple method of the present invention allows isolation of a very large number of mutants, and is applicable to any intracellular or surface protein for which a cDNA and monoclonal antibodies are available. The present method is especially useful in ligand binding site studies for the design of new ligands and drugs.

Abstract: A novel method for examining bacterial growth state is carried out by measuring the levels of conserved cytosolic proteins specific for alternative growth states, using bacterial specific antibody fluorochrome-coupled probe. Utilizing the method of the invention, the cellular growth state of individual bacteria can be determined by measuring the abundance of growth state-specific protein homologs. For example, through use of the protein profiling method of the invention, bacterial VNC state can be distinguished by differentiating growing (exponential phase) from nongrowing or dormant (stationary phase) cells.

Abstract: In qualitative or quantitative assays which employ a test device comprising a test strip pretreated for detection of a specified target analyte dissolved in a liquid medium, the presence of solid, semisolid and/or colloidal materials in the liquid medium may interfere with the assay results. The present invention involves collecting the sample from a flowing or quiescent pool of liquid also containing solid, semisolid or colloidal material with a swab comprised of a handle and a mass of fibrous material or foamed, open cell material which, when immersed in and thoroughly wetted by the liquid, entraps solid, semisolid and/or colloidal material and delivers only the liquid to the sample receiving zone of the test strip.

Abstract: Novel methods for the treatment and/or prophylaxis of diseases caused by tissue-adhering bacteria are disclosed. By interacting with periplasmic molecular chaperones it is achieved that the assembly of pili is prevented or inhibited and thereby the infectivity of the bacteria is diminished. Also disclosed are methods for screening for drugs as well as methods for the de novo design of such drugs, methods which rely on novel computer drug modelling methods involving an approximative calculation of binding free energy between macromolecules. Finally, novel pyranosides which are believed to be capable of interacting with periplasmic molecular chaperones are also disclosed.

Abstract: A rapid, simple, and inexpensive sandwich enzyme-linked receptor based immunodot assay detects pathogens or pathogenic products in test samples using receptors for a characteristic component of the pathogen. This assay is widely applicable because it is highly specific, it does not require special equipment, and the results can be obtained within a few hours with the naked eye. Since the lipid-based receptors have a long-shelf life, they can be easily stored and used for a long time.

Abstract: Disclosed is a pO157 plasmid-specified polypeptide found in E. coli EDL933 and other enterohemorrhagic E. coli that binds to and cleaves C1-esterase inhibitor. Also disclosed are methods employing the polypeptide for diagnosing and treating colitis or hemolytic uremic syndrome, and methods of detecting potential therapeutics.

Abstract: A method of determining whether a test microorganism is a known microorganism, involving use of an agent that specifically affects the growth of the known microorganism. The invention also features a method of identifying E. coli O157:H7 that are based on following criteria: a test microorganism is E. coli O157:H7 if the microorganism is (i) E. coli, (ii) incapable of fermenting sorbitol, and (iii) susceptible to infection by AR1 phage.

Abstract: Novel methods for the treatment and/or prophylaxis of diseases caused by tissue-adhering bacteria are disclosed. By interacting with periplasmic molecular chaperones it is achieved that the assembly of pili is prevented or inhibited and thereby the infectivity of the bacteria is diminished. Also disclosed are methods for screening for drugs as well as methods for the de novo design of such drugs, methods which rely on novel computer drug modelling methods involving an approximative calculation of binding free energy between macromolecules. Finally, novel pyranosides which are believed to be capable of interacting with periplasmic molecular chaperones are also disclosed.

Abstract: An immunoassay method comprising bringing an immobilized phase comprising, at different positions on a water-absorbable base material, at least two first immunity substances capable of specifically binding with at least two kinds of assay target substances selected from the group consisting of verotoxin-producing Escherichia coli, verotoxin and human hemoglobin contained in a test sample, into contact with a test sample and a liquid containing labeled immunity substances each comprising a second immunity substance that is labeled with colored particles and capable of binding with said assay target substance, thereby to form an assay target substance-labeled immunity substance complex and to bind said complex with respective first immunity substances at the immobilized phase.

Abstract: A method of measuring the amount of cyclosporin in a sample suspected of containing cyclosporin is disclosed. A method of inactivating interfering cross-reactive material in an assay for measuring the amount of cyclosporin in a sample suspected of containing cyclosporin is also disclosed. Compositions wherein cyclosporin is conjugated to an immunogenic carrier or a label, optionally through a linking group, at an alanine nitrogen atom of the cyclic backbone of cyclosporin are also disclosed. Compositions wherein atiocyclosporin is conjugated, optionally through a linking group, to an immunogenic carrier or a label are also disclosed. Where cyclosporin is conjugated to an immunogenic carrier, the conjugates may be used as immunogens for the preparation of antibodies which are capable of recognizing cyclosporin.

Abstract: The present invention provides an inexpensive and sensitive system and method for detecting analytes present in a medium. The system comprises a diffraction enhancing element, such as functionalized microspheres, which are modified such that they are capable of binding with a target analyte. Additionally, the system comprises a polymer film, which may include a metal coating, upon which is printed a specific, predetermined pattern of a analyte-specific receptors. Finally, the system includes a wicking agent which permits the system to be a single step system which avoids the necessity of any additional rinsing steps. Upon attachment of a target analyte to select areas of the polymer film, either directly or with the diffraction enhancing element, diffraction of transmitted and/or reflected light occurs via the physical dimensions and defined, precise placement of the analyte. A diffraction image, such as a hologram, is produced which can be easily seen with the eye or, optionally, with a sensing device.

Abstract: The present invention provides methods and compositions for detecting the presence of biologically-important analytes by using redox liposome biosensors. In particular, the present invention provides liposome/sol-gel electrodes suitable for the detection of a wide variety of organic molecules, including but not limited to bacterial toxins.

Abstract: The present invention concerns methods of testing water for microbe contamination. The methods of the invention comprise supplementing existing methods with assays using specific reagents such as monoclonal antibodies. The invention also concerns a device for use in the methods of the invention.

Abstract: This invention provides methods, compositions and kits for concentrating target ligands, including microorganisms, from samples, including biological samples. The methods involve the use of magnetic particles to concentrate the target analytes. Also provided are methods, compositions and kits for detecting the presence of target ligands in samples.

Abstract: A family of fatty acid transport proteins (FATPS) mediate transport of long chain fatty acids (LCFAs) across cell membranes into cells. These proteins exhibit different expression patterns among the organs of mammals. Nucleic acids encoding FATPs of this family, vectors comprising these nucleic acids, as well as the production of FATP proteins in host cells are described. Also described are methods to test FATPs for fatty acid transport function, and methods to identify inhibitors or enhancers of transport function. The altering of LCFA uptake by administering to the mammal an inhibitor or enhancer of FATP transport function of a FATP in the small intestine can decrease or increase calories available as fats, and can decrease or increase circulating fatty acids. The organ specificity of FATP distribution can be exploited in methods to direct drugs, diagnostic indicators and so forth to an organ such as the heart.

Abstract: A genetic construct for use in a biosensor comprising: (a) a first nucleic acid molecule including a sequence encoding a reporter molecule having a detectable activity; and (b) a second nucleic acid molecule including a sequence encoding an enzyme which produces a substrate for the reporter molecule, the first sequence being under the control of a first inducible promoter and the second sequence being under the control of a second inducible promoter. A biosensor for measuring an environmental signal comprising a cell including the genetic construct and a means for measuring the activity of the reporter molecule in the cell when the cell has been exposed to the environmental signal.

Abstract: The invention is related to antibodies, particularly monoclonal antibodies, which recognize particularly epitopes of the intimin protein of enteropathogenic E. coli and enterohemorrhagic E. coli, methods of detecting such E. coli by use of these antibodies, and kits containing these antibodies for diagnosis.

Abstract: The present invention provides a method for quantifying the presence of extracellular LBP in body fluids including blood in a subject comprising conducting an LBP immunoassay on plasma obtained from said subject.

Abstract: A method for screening compounds for antimicrobial activity is described that utilizes bacterial protein-protein binding in vitro. The method may be performed using immobilized elements and the immobilization may be carried out using a variety of immobilization means (e.g., columns, beads, adsorbents, nitrocellulose paper, etc.) in order to screen large libraries of compounds.

Abstract: Disclosed is an assay for an analyte in a fluid test sample such as urine which involves combining the fluid test sample with a reagent system comprising an apo-peroxidase, a redox dye, a peroxide and a metal porphyrin which is bound to an analyte/analyte specific binding partner which complex has a combined molecular weight of at least about 180 K Daltons. When this conjugate interacts with analyte in the fluid test sample, a portion of the specific binding partner is dissociated from the complex thereby enabling the metal porphyrin to reconstitute with the apo-peroxidase. The reconstituted peroxidase can interact with the peroxide and redox dye to provide a colored response to analyte in the fluid test sample.

Abstract: A method of determining whether a test microorganism is a known microorganism, involving use of an agent that specifically affects the growth of the known microorganism. The invention also features a method of identifying E. coli O157:H7 that are based on the following criteria: a test microorganism is E. coli O157:H7 if the microorganism is (i) E. coli, (ii) incapable of fermenting sorbitol, and (iii) susceptible to infection by AR1 phage.

Abstract: Several EHEC proteins which are secreted into the culture supernatant have been discovered. These proteins are not produced by non-pathogenic E. coli, and produce a strong serum antibody response in patients with HUS and bloody diarrhea.

Abstract: The present invention relates to methods and compositions for the modulation of bacterial topoisomerase enzymes within bacterial cells. More specifically, the present invention relates to bacterial assays wherein the levels of bacterial topoisomerase enzymes or the levels of target sites within the enzymes are varied within bacterial test strains in order to screen for compounds that target, i.e., interact with the topoisomerase enzymes, causing DNA damage and hence, bacterial growth inhibition and/or cell death. The present methods and compositions are useful for the identification and characterization of novel therapeutic antibacterial compounds.