Abstract: Compounds termed "vitaletheine modulators" which include beta-alanyl-taurine and carbobenzoxy beta-alanyl-taurine are synthesized and added to culture media for in vitro culture of cells such as mammalian or plant cells. The compounds support cell vitality, and provide increased cellular life span, increased cellular bioproductivity, improved cellular function, and adaption of resistant cells to culturing. Carbobenzoxy beta-alanyl-taurine is produced by coupling .beta.-alanine, which has it's terminal amine protected with a carbobenzoxy (CBZ) group, to N-hydroxysuccinimide to produce an active ester of .beta.-alanine, coupling two of the active esters to cystamine to produce a CBZ-protected .beta.-alethine having an internal disulfide bond, isolating and purifying the CBZ-protected .beta.-alethine, and reacting the CBZ-protected .beta.-alethine with iodine to oxidize the disulfide bond to obtain carbobenzoxy beta-alanyl-taurine.

Abstract: Vaccines for diseases caused by normally encapsulated organisms are produced by genetically modifying those organisms by deleting the genes encoding for capsule synthesis or a portion thereof sufficient to produce non-capsulated mutants of the organisms. As an example, a live, attenuated strain of Actinobacillus pleuropneumoniae genetically modified with a large deletion in a chromosomal regions of the DNA which encodes for capsule synthesis is a safe and effective vaccine against swine pleuropneumonia.

Abstract: This invention relates to flagella-less strains of Borrelia and to novel methods for use of the microorganisms as vaccines and in diagnostic assays. Although a preferred embodiment of the invention is directed to Borrelia burgdorferi, the present invention encompasses flagella-less strains of other microorganisms belonging to the genus Borrelia. Accordingly, with the aid of the disclosure, flagella-less mutants of other Borrelia species, e.g., B. coriacei, which causes epidemic bovine abortion, B. anserina, which causes avian spirochetosis, and B. recurrentis and other Borrelia species causative of relapsing fever, such as Borrelia hermsii, Borrelia turicatae, Borrelia duttoni, Borrelia persica, and Borrelia hispanica, can be prepared and used in accordance with the present invention and are within the scope of the invention.

Abstract: Stable reference nucleic acid for use in all steps of molecular screening and diagnostic assays and method of making. A desired nucleic acid sequence is amplified, ligated into an expression vector, and used to transform a vehicle. A cellular vehicle is subsequently killed without affecting the encapsulated nucleic acid. The vehicle membrane is stabilized under controlled conditions to a stability substantially matching that of a test cell membrane. The recovered nucleic acid is used as a standard or control in molecular diagnostic and genetic testing assays.

Abstract: This invention relates to the isolation of thermophilic phytase-producing microorganisms, method for producing phytase using such microorganisms, phytase obtained therefrom, and usage of the phytase to hydrolyze phytic acid or phytate. In particular, this invention relates to phytase-producing microorganisms, which belong to Streptomyces sp., Pseudonocardia sp. or Microbispora sp., and which produce phytase available for recovery in an efficient and practical manner.

Abstract: Methods are disclosed which employ cold-shock proteins, such as Trigger Factor, to confer cold-tolerance to cells and to enhance viability of cells at temperatures at which the cells would not normally remain viable. Proteins having peptidyl-proline isomerase activity are also disclosed as being useful to confer cold-tolerance to cells. Cells which overexpress cold-shock proteins are useful in cell-based expression systems which are intended to express proteins at low temperatures.

Abstract: Disclosed herein are novel medical devices, particular well-suited for sustained delivery of therapeutically-significant substances. Also disclosed are methods of making and using these delivery devices. Using these devices and methods the present invention teaches sustained, targeted and reversible delivery of immunostimulating agents, as well as therapeutic agents such as enzymes, hormones and neurotransmitters, to name but a few.

Abstract: This invention provides a purified immortalized human endothelial cell infected with Ehrlichia chaffeensis or Ehrlichia canis. Also provided is a method of simultaneously screening a sample from a human subject for the presence of E. chaffeensis and Rickettsia rickettsii comprising contacting the sample with immortalized human endothelial cells under conditions which allow infection of the cells and detecting the presence of infection, the presence of infection indicating the presence of E. chaffeensis and/or R. rickettsii. The invention also provides a method of screening a sample from a human subject for the presence of E. chaffeensis comprising contacting the sample with endothelial cells under conditions which allow infection of the cells by E. chaffeensis and detecting the presence of infection by E. chaffeensis, the presence of infection by E. chaffeensis indicating the presence of E. chaffeensis in the sample. Finally, the invention provides a method of culturing E. chaffeensis or E.

Abstract: The invention relates to a method for making an inactivated vaccine of Mycoplasma hyopneumoniae by inactivating the bacteria with Thimerosal. The resulting bacterin is mixed with an adjuvant of aluminum hydroxide and DEAE dextran and injected into pigs. The resulting bacterin and adjuvant mixture can also be mixed with other bacteria such as Bordetella and Pasteurella, for further adjuvant effect. Protective immunity against mycoplasmal pneumonia is elicited in swine using these vaccines.

Abstract: A method of biochemically transforming macromolecular compounds found in solid carbonaceous materials, such as coal is provided. The preparation of new microorganisms, metabolically weaned through challenge growth processes to biochemically transform solid carbonaceous materials at extreme temperatures, pressures, pH, salt and toxic metal concentrations is also disclosed.

Abstract: A method for large scale cultivation and attenuation of L. intracellularis bacteria by inoculating cells with L. intracellularis bacteria to infect the cells, incubating the infected cells in a reduced oxygen concentration and maintaining the infected cells in suspension. Anti-L. intracellularis vaccines are prepared from cultures grown in suspension. Diagnostic agents are also disclosed.

Abstract: The invention relates to a mutant of Bacillus thuringiensis which produces a larger amount of crystal delta-endotoxin with a greater pesticidal activity as compared to the corresponding parental strain. The mutant may also have a larger crystal size as compared to the corresponding parental strain. The crystal delta-endotoxin produced by the mutant Bacillus thuringiensis will have an activity directed towards the same pest(s) as its parental Bacillus thuringiensis crystal delta-endotoxin. The invention further relates to a method for producing such a mutant, compositions comprising such a mutant as well as methods for controlling a pest(s) using these compositions.

Abstract: A process for selection of oxygen-tolerant, H.sub.2 -producing algal mutant cells comprising:(a) growing algal cells photoautotrophically under fluorescent light to mid log phase;(b) inducing algal cells grown photoautrophically under fluorescent light to mid log phase in step (a) anaerobically by (1) resuspending the cells in a buffer solution and making said suspension anaerobic with an inert gas; (2) incubating the suspension in the absence of light at ambient temperature;(c) treating the cells from step (b) with metronidazole, sodium azide, and added oxygen to controlled concentrations in the presence of white light.(d) washing off metronidazole and sodium azide to obtain final cell suspension;(e) plating said final cell suspension on a minimal medium and incubating in light at a temperature sufficient to enable colonies to appear;(f) counting the number of colonies to determine the percent of mutant survivors; and(g) testing survivors to identify oxygen-tolerant H.sub.2 -producing mutants.

Abstract: A carrier for holding a microorganism in soil releases, from a constituting material thereof, an inducer for production of an enzyme of the microorganism for soil remediation. The carrier for holding a microorganism may comprise a combination of a microorganism holding carrier composed of a hydrophilic polymer for holding the microorganism with an inducer-holder composed of another polymer for holding inducer manifesting a biological action to the microorganism adjacent to each other. A method for remediation of soil comprises application of the microorganism-holding carrier into the soil. A soil-remedying agent comprises the microorganism-holding carrier and a microorganism held thereon which exhibits enzyme activity for decomposition of a polluting substance in the soil under action an inducer released by the carrier.

Abstract: A process for biochemical conversion of heavy crude oils is provided. The process includes contacting heavy crude oils with adapted biocatalysts. The resulting upgraded oil shows, a relative increase in saturated hydrocarbons, emulsions and oxygenates and a decrease in compounds containing in organic sulfur, organic nitrogen and trace metals. Adapted microorganisms which have been modified under challenged growth processes are also disclosed.

Abstract: An organic compound is biodegraded using JM1 (FERM BP-5352) by contacting the JM1 with an organic compound and culturing the JM1 in a medium including source of conjugated carbon which is citric acid, maleic acid or a salt thereof.

Abstract: We describe a bacterial delivery system for the delivery of DNA and antigens into cells. We constructed an attenuated bacterial vector which enters mammalian cells and ruptures delivering functional plasmid DNA, such as a mammalian expression plasmid, and antigens into the cell cytoplasm. This Shigella vector was designed to deliver DNA to colonic surfaces, thus opening the possibility of oral and other mucosal DNA immunization and gene therapy strategies. The attenuated Shigella is also useful as a vaccine for reducing disease symptoms caused by Shigella.

Abstract: The present invention provides a process for the in vitro culture of parasites of the genus Babesia, processes for the in vitro attenuation of the virulence of these parasites, processes for the in vitro cloning of these parasites, attenuated parasites, clonal lines of the genus Babesia, nucleotide probes derived from the clonal lines, the use of these probes for the identification of Babesia parasites and to vaccinces against babesioses.

Abstract: This aerobic waste pretreatment process comprises inoculating a milk industry effluent with a mixture of bacteria and yeasts both classes of microorganisms capable of living and growing in symbiosis in the effluent, the population of the bacteria being, in most cases, several times greater than the population of the yeasts, maintaining the temperature and pH of the inoculated effluent between 0.degree. C. and 50.degree. C. and between 1.7 and 9, aerating the effluent while varying, if necessary, the pH at maximum rate of 1.5 pH units per minute and also, if required, modulating the aeration of the inoculated effluent at a maximum rate of 130 micromoles of oxygen per minute. A biomass is obtained which has a good nutritional value suitable for animal feed.

Abstract: An attenuated microorganism harboring two mutated genes, each of which is located in the organisms aromatic pathway is provided. These organisms can usefully form the basis of a vaccine. They can be genetically engineered so as to express antigens from other pathogens and thus form the basis of a range of multi-valent vaccines.

Abstract: An accelerated micro-organism culture method is for decomposing harmful substances. For this purpose, the cultivation, mutagenic treatment and stabilization of the micro-organisms are carried out simultaneously in a mutation fermenter containing a nutrient-rich basic medium (basic medium 1); selection is carried out in a selection fermenter containing a basic medium which is low in nutrients and is enriched with the harmful substance (basic medium 2); small amounts of biomass released from the nutrient-rich basic medium (basic medium 1) of the mutation fermenter are continuously removed from the mutation fermenter and transferred to the selection fermenter. Small amounts of biomass released from the basic medium which is low in nutrients (basic medium 2) of the selection fermenter are removed from the selection fermenter and transferred to the mutation fermenter. The entire process is carried out continuously in alternating selection and mutation phases, in the manner of a repeated cycle.

Abstract: A live Salmonella vaccine having an increased stability is disclosed. Further, immunization methods of a host such as chickens against Salmonella diseases are carried out by the use of the vaccine. The vaccine may be administered orally or parenterally and includes one or more of the live vaccines disclosed herein. The live vaccine is produced from one or more live vaccine strains with metabolic drift attenuation via an increased generation time. The vaccine is an immunogenic, stable and single or multiple marker live vaccine strain with or without envelope mutation; and the live vaccine is a suppressor mutant of an original live vaccine strain still having the attenuation marker(s) of the original live vaccine strain but a shortened generation time and having macrolide tolerance.

Abstract: The present invention is concerned with Marek's Disease Virus of serotype 1 adapted for growth on an avian cell line. A preferred cell line for adaption of this virus is the quail cell line QT-35. These adapted viruses can be grown on the avian cell line in order to obtain virus material for the preparation of vaccines.

Abstract: This invention involves a method of altering and regulating the gene expression of cells, by means of contacting the cells with a nuclease capable of degrading extra-cellular DNA and/or RNA. The nuclease will degrade the extra-cellular nucleic acids (NA) into nucleotides or oligonucleotides which are too short to have substantial affinity for the chromosomal DNA.By means of this method, cultures of cells have been created with desirable properties, including greater phenotypic uniformity and higher levels of cell reproduction.This invention also comprises a culture of cells which has been treated by this method, and cells descended from cells which have been treated according to this method.

Abstract: An attenuated microorganism harbouring two mutated genes, each of which is located in the organisms aromatic pathway is provided. These organisms can usefully form the basis of a vaccine. They can be genetically engineered so as to express antigens from other pathogens and thus form the basis of a range of multi-valent vaccines.

Abstract: A method for modifying a wild strain of an entero-invasive Shigella to produce a modified strain of Shigella that can be used for making a vaccine against the wild strain of Shigella. The genome of the wild strain of Shigella is transformed so that it cannot substantially invade cells of a human host and cannot spread substantially within infected cells and from infected to uninfected cells of the host and cannot produce toxins which will kill substantial numbers of the host's infected, as well as uninfected, cells. A first gene of the wild strain of Shigella, coding for a protein necessary for the Shigella to invade cells of the host, and a second gene, coding for a protein necessary for the Shigella to spread within infected cells and between the infected and uninfected cells of the host, are mutagenized.

Abstract: Regulatable RNA molecules such as regulatable ribozymes, nucleic acids encoding such regulatable ribozymes, and methods of making and using such regulatable ribozymes are disclosed. Regulatable ribozymes comprise a ligand-binding RNA sequence and a ribozyme sequence capable of cleaving a separate targeted RNA sequence, wherein upon binding of the ligand to the ligand-binding RNA sequence, the activity of the ribozyme sequence against the targeted RNA sequence is altered. The ligand may be either an inorganic or an organic molecule and may be a co-drug which can be administered to specifically regulate the ribozyme activity. Regulatable RNA molecules other than ribozymes are also disclosed, such as regulatable mRNA molecules which comprise a ligand-binding RNA sequence separate from the coding sequence, wherein upon binding of a ligand to the ligand-binding RNA sequence, translation of the regulatable mRNA is altered.

Abstract: A modified Salmonella, wherein the wild type pag gene such as pagC has been replaced by a gene encoding a truncated pag gene such as pagC gene fused to a heterologous DNA segment, is disclosed. This modification significantly attenuates the virulence of the Salmonella. In addition, immune reaction to the portion of the fusion protein encoded by the heterologous gene can be generated. Uses of the vector, including its use in a drug screen are also disclosed.

Abstract: Stabilized bacteria and bacterial formulations which can survive long term storage at high temperature are described. Bacteria are dried until they reach a dormant state. Oxygen is then removed from the environment surrounding the bacteria to prevent oxidative damage to the dormant cells. The bacteria is packaged and stored in material impermeable to gas and water vapor until such time as it is ready for use. Bacteria stored under these conditions will remain stable and efficacious for at least a year.

Abstract: A method for large scale cultivation and attenuation of IS intracellularis bacteria by inoculating cells with IS intracellularis bacteria to infect the cells, incubating the infected cells in a reduced oxygen concentration and maintaining the infected cells in suspension. Anti-IS intracellularis vaccines are prepared from attenuated strains. Diagnostic agents are also disclosed.

Abstract: A bacterial cell the virulence of which is attentuated by a first mutation in a PhoP regulon and a second mutation in an aromatic amino acid synthetic gene and bacterial cells the virulence of which is attenuated by a mutation in one or more PhoP-activated genes or one or more PhoP-repressed genes.

Abstract: A vector capable of integrating into the chromosome of Salmonella including a first DNA sequence encoding a heterologous protein, a second DNA sequence encoding a marker, and a third DNA sequence encoding a phoP regulatory region regulated gene product necessary for virulence, the third DNA sequence being mutationally inactivated

Abstract: A vector capable of integrating into the chromosome of Salmonella including a first DNA sequence encoding a heterologous protein, a second DNA sequence encoding a marker, and a third DNA sequence encoding a phoP regulatory region regulated gene product necessary for virulence, the third DNA sequence being mutationally inactivated.

Abstract: Live vaccines are provided and methods for preparing the live vaccines for protection of a host from a pathogenic microorganism. The vaccines are prepared by introducing at least one modification in a gene involved in at least one, normally at least two, biosynthetic pathways involving the production of products which are unlikely to be found in the disease susceptible host. The modification results in a gene change which cannot be repaired by a single step, e.g. polynucleotide deletions and inversions. Where the aro gene suffers such a change, the resultant auxotrophic mutants require aromatic amino acids, p-aminobenzoic acid and 2,3-dihydroxybenzoic acid or a highly concentrated source of absorbable iron. The auxotrophic mutations have substantially reduced or nonexistent virulence while retaining the desired immunogenicity to initiate the immunogenic response. Various techniques can be employed for providing the desired change.

Abstract: This invention relates to flagella-less strains of Borrelia to novel methods for use of the microorganisms as vaccines and in diagnostic assays. Although a preferred embodiment of the invention is directed to Borrelia burgdorferi, the present invention encompasses flagella-less strains of other microorganisms belonging to the genus Borrelia. Accordingly, with the aid of the disclosure, flagella-less mutants of other Borrelia species, e.g., B. coriacei, which causes epidemic bovine abortion, B. anserina, which causes avian spirochetosis, and B. recurrentis and other Borrelia species causative of relapsing fever, such as Borrelia hermsii, Borrelia turicatae, Borrelia duttoni, Borrelia persica, and Borrelia hispanica, can be prepared and used in accordance with the present invention and are within the scope of the invention. Therefore, a preferred embodiment comprises a composition of matter comprising a substantially pure preparation of a strain of a flagella-less microorganism belonging to the genus Borrelia.

Abstract: Disclosed herein are methods for attenuating virulent gram negative bacteria to produce avirulent bacteria. The methods comprise passaging the wild-type bacteria through phagocytic cells, such as macrophages or polymorphonuclear leukocytes, or through lysosomes derived from such cells, a sufficient number of times until the bacteria become avirulent to the animal host. The bacteria are preferably from the family Enterobacteracea and most preferably from the genus Salmonellae. The invention further comprises the avirulent bacteria produced by the methods, pure cultures of such bacteria, and methods of using the bacteria, preferably in a vaccine for administration to an animal host to induce an immune response to the wild-type gram negative bacteria in the host.

Abstract: This invention provides methods for implanting encapsulated cells in a host comprising exposing cells to restrictive conditions for a sufficient period of time to establish a desired cell property in response to the restrictive conditions and implanting the encapsulated cells in a host, the cell property being substantially maintained following implantation. Also provided are cells produced by exposure to restrictive conditions.

Abstract: The present invention provides a chondroitinase attenuated Edwardsiella ictaluri bacteria. Further, this invention provides a vaccine comprising a protective amount of a chondroitinase attenuated strain of Edwardsiella ictduri bacteria and a method for protecting a fish from Enteric Septicemia comprising administering the vaccine to the fish.

Abstract: Disclosed is a process for growing living cellular material in a reaction zone having a membrane in contact with an aqueous medium. The concentration of nutrient in samples from the process is measured using a spectrometric instrument to obtain spectral data characteristic of nutrient components. This spectral data is analyzed using a chi-squared mathematical technique to determine the unknown concentration of nutrient components in said samples. Then the rate at which nutrient is fed to the reaction zone is altered based on the determination of concentration of nutrient components as required to optimize the process.

Abstract: Novel attenuated strains of Aeromonas salmonicida are disclosed that are effective as live effective vaccines against furunculosis in fish. These vaccines may be administered by the immersion of fish in a solution of the vaccine. Methods of producing these strains and other strains having the identifying characteristics of these strains are also disclosed.

Abstract: This invention relates to the preparation of new, modified organisms, through challenge growth processes, that are viable in the extreme temperature, pressure and pH conditions and salt concentrations of an oil reservoir and that are suitable for use in microbial enhanced oil recovery. The modified microorganisms of the present invention are used to enhance oil recovery and remove sulfur compounds and metals from the crude oil. The processes are comprised of steps which successively limit the carbon sources and increase the temperature, pressure and salinity of the media. This is done until microbial strains are obtained that are capable of growing in essentially crude oil as a carbon source and at a temperature range from about 70.degree. C. to 90.degree. C., at a pressure range from about 2,000 to 2,500 psi and at a salinity range from about 1.3 to 35%.

Abstract: This invention relates to flagella-less strains of Borrelia and to novel methods for use of the microorganisms as vaccines and in diagnostic assays. Although a preferred embodiment of the invention is directed to Borrelia burgdorferi, the present invention encompasses flagella-less strains of other microorganisms belonging to the genus Borrelia. Accordingly, with the aid of the disclosure, flagella-less mutants of other Borrelia species, e.g., B. coriacei, which causes epidemic bovine abortion, B. anserina, which causes avian spirochetosis, and B. recurrentis and other Borrelia species causative of relapsing fever, such as Borrelia hermsii, Borrelia turicatae, Borrelia duttoni, Borrelia persica, and Borrelia hispanica, can be prepared and used in accordance with the present invention and are within the scope of the invention.

Abstract: Disclosed herein are methods for attenuating virulent gram negative bacteria to produce avirulent bacteria. The methods comprise passaging the wild-type bacteria through phagocytic cells, such as macrophages or polymorphonuclear leukocytes, or through lysosomes derived from such cells, a sufficient number of times until the bacteria become avirulent to the animal host. The bacteria are preferably from the family Enterobacteracea and most preferably from the genus Salmonellae. The invention further comprises the avirulent bacteria produced by the methods, pure cultures of such bacteria, and methods of using the bacteria, preferably in a vaccine for administration to an animal host to induce an immune response to the wild-type gram negative bacteria in the host.

Abstract: Noncapsulated mutants of normally encapsulated bacteria in which the capsule is required for virulence are useful as vaccines for disease states caused by the normally encapsulated bacteria. In particular, a non-capsulated mutant of Actinobacillus (Haemophilus) pleuropneumoniae is useful in protecting against swine pleuropneumonia.

Abstract: The invention described herein consists of a process for preparing an antigenic reagent useful for the indirect determination of Salmonella typhi, the organism that is the causal agent of typhoid fever (TF). The invention consists on the following steps: to grow Salmonella typhi in a culture medium, characterized by containing a free-iron chelator, which generates a specific S. typhi outer membrane protein (OMP) pattern, OMPs that are used as a selective antigen for the detection of specific serum antibodies, by an immunoassay technique (ELISA).

Abstract: Disclosed are methods and compositions for treating neuroblastoma cells. The methods include contacting the neuroblastoma cells with a neurotrophic factor and less than a lethal dose of an inhibitor of cell proliferation for about 1 to 15 days, and then maintaining the neuroblastoma cells in contact with the neurotrophic factor for an additional 1 to 15 days. The composition includes a neurotrophic factor such as the neurotropin, nerve growth factor, and an inhibitor of cell proliferation such as aphidicolin, thymidine, or hydroxyurea. Also disclosed are methods for inducing the remission or differentiation of, or eliminating, neuroblastoma cells.

Abstract: An insect cell line has been established and characterized, derived from embryonic tissue (BTI-TN-5B1-4, ATCC CRL 10859) of Trichoplusia ni (cabbage looper). The line is susceptible to various baculoviruses, including TnSNPV and AcMNPV, and may be used to replicate such viruses for use as insecticides or otherwise.

Abstract: This invention relates to the preparation of new, modified organisms, through challenge growth processes, that are viable in the extreme temperature, pressure and pH conditions and salt concentrations of an oil reservoir and that are suitable for use in microbial enhanced oil recovery. The modified microorganisms of the present invention are used to enhance oil recovery and remove sulfur compounds and metals from the crude oil.

Abstract: Method and composition for vaccination of a pig against swine dysentary caused by T. hyodysenteriae infection characterized by parenteral, preferably intramuscular, administration to the pigs of a live strain or of an oxygen-treated, non-viable strain of T.hyodysenteriae.

Abstract: A vaccine against bovine respiratory disease is provided containing an attenuated strain of Pasteurella haemolytica isolated from an asymptomatic calf. The vaccine effectively triggers an immunological system response to whole cell, denuded, cytotoxin and capsular antigens.