Abstract: Whole-cell vaccines and methods for their use in producing protective immune responses in vertebrate hosts subsequently exposed to pathogenic bacteria. The present invention involves a method of enhancing antigen presentation by intracellular bacteria in a manner that improves vaccine efficacy. After identifying an enzyme that has an anti-apoptotic effect upon host cells infected by an intracellular microbe, the activity of the enzyme is reduced, thereby modifying the microbe so that it increases immunogenicity. Also, the present invention provides a method of incrementally modifying enzyme activity to produce incrementally attenuated mutants of the microbe from which an effective vaccine candidate can be selected.

Abstract: Purified cellobiohydrolase I (glycosyl hydrolase family 7 (Cel7A)) enzymes from Penicillium funiculosum demonstrate a high level of specific performance in comparison to other Cel7 family member enzymes when formulated with purified EIcd endoglucanase from A. cellulolyticus and tested on pretreated corn stover. This result is true of the purified native enzyme, as well as recombinantly expressed enzyme, for example, that enzyme expressed in a non-native Aspergillus host. In a specific example, the specific performance of the formulation using purified recombinant Cel7A from Penicillium funiculosum expressed in A. awamori is increased by more than 200% when compared to a formulation using purified Cel7A from Trichoderma reesei.

Abstract: This invention pertains to methods, kits and/or compositions for the determination of analytes by mass analysis using unique labeling reagents or sets of unique labeling reagents. The labeling reagents can be isomeric or isobaric and can be used to produce mixtures suitable for multiplex analysis of the labeled analytes.

Abstract: This invention provides a method for rapidly measuring an premature yeast flocculating factor of malt contained in a brewing material, comprising the following steps of: (1) mixing yeast in the late logarithmic growth phase or thereafter with a water-extracted high molecular fraction prepared from a test material sample in a buffer solution and suspending the mixture in the buffer solution; and (2) irradiating the suspension obtained in the step (1) with visible light, photographing the scattered light with a camera device, image-analyzing the image data thus obtained and determining the degree of whiteness of the suspension to measure the degree of sedimentation of the yeast in the suspension. According to the present invention, the premature yeast flocculating factor of malt contained in the brewing material can be measured in a highly accurate, rapid and simple manner, and, further, even a plurality of analytes can be rapidly measured.

Abstract: According to the invention, there is provided null-LOX-1 barley and plant products produced thereof, such as malt manufactured by using barley kernels defective in synthesis of the fatty acid-converting enzyme lipoxygenase-1. Said enzyme accounts for the principal activity related to conversion of linoleic acid into 9-hydroperoxy octadecadienoic acid, a lipoxygenase pathway metabolite, which—through further enzymatic or spontaneous reactions—may lead to the appearance of trans-2-nonenal. The invention enables brewers to produce a beer devoid of detectable trans-2-nonenal-specific off-flavors, even after prolonged storage of the beverage.

Abstract: An edible product in accordance with the present invention involves an edible inner component made from a first material and an edible outer cover component substantially completely encapsulating the edible inner component. In one form of the invention, the inner component is in the form of a hairball treatment for cats and the outer component is a dough-like material. A system for producing the edible product includes a first material source for supplying a first edible material to form the inner component of the edible product, a second material source for supplying a second edible material to form the outer component of the edible product, and an extruder for continuously coextruding the first edible material and the edible second material to produce an elongated rope of edible product. A crimping apparatus receives the longitudinal rope extruded from the extruder for crimping the longitudinal rope to separate the longitudinal rope into individual edible products.

Abstract: The present invention relates to methods for producing a heterologous polypeptide, comprising: (a) cultivating a mutant of a parent filamentous fungal cell under conditions conducive for the production of the heterologous polypeptide, wherein (i) the mutant cell comprises a nucleic acid sequence encoding the heterologous polypeptide and (ii) the mutant produces less of the cyclohexadepsipeptide than the parent filamentous fungal cell when cultured under the same conditions; and (b) isolating the heterologous polypeptide from the cultivation medium. The present invention also relates to mutants of filamentous fungal cells and methods for obtaining the mutant cells. The present invention also relates to isolated cyclohexadepsipeptide synthetases and isolated nucleic acid sequences encoding the cyclohexadepsipeptide synthetases.

Abstract: The present invention provides a process for economically producing N-acetylneuraminic acid without using expensive materials such as pyruvic acid and phosphoenolpyruvic acid. The process comprises: allowing (i) a culture of a microorganism having N-acetylneuraminic acid aldolase activity or N-acetylneuraminic acid synthetase activity, or a treated matter of the culture, (ii) a culture of a microorganism capable of producing pyruvic acid or a treated matter of the culture, or a culture of a microorganism capable of producing phosphoenolpyruvic acid or a treated matter of the culture, (iii) N-acetylmannosamine, and (iv) an energy source which is necessary for the formation of pyruvic acid or phosphoenolpyruvic acid to be present in an aqueous medium to form and accumulate N-acetylneuraminic acid in the aqueous medium; and recovering N-acetylneuraminic acid from the aqueous medium.

Abstract: Barley plants having reduced lipoxygenase-1 enzyme activity are provided, for example, barley plants expressing mutant LOX-1 protein. The barley plants of the invention are useful in the production of plant products such as malt and brewed beverages, particularly beer, having increased stability and reduced T2N potential.

Abstract: The invention provides isolated nucleic acid compounds encoding a glycosyltransferase enzyme of Amycolatopsis orientalis. Also provided are vectors carrying genes that encode said enzyme, transformed heterologous host cells for expressing said enzyme, and methods for producing glycopeptide compounds using the cloned genes that encode said enzyme.

Abstract: A method for extraction of heterologous protein from plant seed comprises extracting the germ portion of the seed and extracting and purifying the protein from the germ. Enhanced expression in the germ is provided, and allows for improved efficiency in production, and cost savings. Directing expression to the germ portion further increases expression levels of the protein. The ubiquitin promoter preferentially directs expression to the germ portion of plant seed.

Abstract: The T cell activation marker, granulysin, is demonstrated to be an effective antimicrobial agent. It is used in vitro and in vivo to reduce the population of viable cells in a microbial population. Of particular interest is the use of the active fragment of human granulysin, or modified forms thereof, to treat bacterial infections.

Abstract: The present invention is directed to oxygen sensing. The present invention provides methods for correcting (or providing for) oxygen sensing in cells. Preferably, the methods involve supplying the SDHD gene or cybS protein, or any of the other components of the mitochondrial complex II cytochrome b oxygen sensing complex in an amount which restores oxygen sensing to affected cells, and thus facilitates normoxic conditions. More specifically, the present invention relates to mutations in the SDHD gene in human cancers and their use in the diagnosis and prognosis of mammalian cancer.

Abstract: A method of producing enzymes in which plant seeds are subjected to a softening process and a germination process in an aqueous environment and wherein a substance is added to the aqueous environment at least before and during the softening process, the substance selected from the group consisting of liposomes and niosomes and mixtures thereof, the substance containing a germination enhancing factor, the substance relatively easily and rapidly permeating a biomembrane and the germination enhancing factor enhancing the germination process.

Abstract: An enzymatic method for producing N-formyl-.alpha.-L-aspartyl-L-phenylalanine methyl ester by a condensation reaction between N-formyl-L-aspartic acid and L-phenylalanine methyl ester or D,L-phenylalanine methyl ester, which comprises: supplying an organic phase comprising a water-immiscible solvent containing N-formyl-L-aspartic acid and L- or D,L-phenylalanine methyl ester onto an aqueous phase comprising a thermolysin-like metalloprotease; proceeding the condensation reaction in the aqueous phase to produce N-formyl-.alpha.-L-aspartyl-L-phenylalanine methyl ester; and extracting the N-formyl-.alpha.-L-aspartyl-L-phenylalanine methyl ester thus produced from the aqueous phase to the organic phase.

Abstract: A method and apparatus for correcting refractive errors of the eye are disclosed. Accelerated reshaping of the corneal tissue is accomplished by administering one or more enzymes and/or other agents to the eye which temporarily soften the cornea. The cornea is thereafter fitted with a rigid contact lens or a series of lenses which have a concave curvature that will correct a refractive error. The softened cornea then rapidly reshapes its convex curvature to the concave curvature of the contact lens or series of lenses, thereby rendering the eye emmetropic. The enzymes and/or other agents then dissipate from the cornea, and the cornea "hardens" to retain the new emmetropic shape. After "hardening" has occurred, the lens rendering the eye emmetropic is removed.

Abstract: The present invention discloses a method for the enzymatic synthesis of a peptide. A protected peptide having a C-terminal carboxylate group or a protected, N-acyl amino acid having an alpha carboxylate group is reacted with a protected peptide having an N-terminal ammonium group or a protected amino acid having an alpha ammonium group in the presence of a condensation enzyme under conditions in which the carboxylate group and the ammonium group condense to form a protected, uncharged, peptide product. This peptide product is transported across a water-immiscible hydrophobic phase into an aqueous product phase and prevented from back diffusing across the water-immiscible hydrophobic phase. The peptide product can be converted, chemically or enzymatically, to a charged species that cannot back diffuse across the water-immiscible phase into the aqueous reaction phase.

Abstract: The invention provides novel DNA and peptide sequences encoding a family of phospholipase A.sub.2 enzymes, with specific activities of approximately 20 .mu.mol/min/mg in the mixed micelle assay. These enzymes are useful in methods for detecting the anti-inflammatory potential of various chemical agents. The invention also details novel methods for determining such potential using the novel sequences, methods for making the novel peptides, and methods for developing new anti-inflammatory drugs.

Abstract: A chemical product and method for accelerated biodegradation of petroleum on water. The chemical product includes a fermentation product portion and a surfactant containing emulsifier portion which has a monosodium glutamate additive.

Abstract: A derivative of a fibrinolytic enzyme in which the catalytic site on the enzyme which is responsible for fibrinolytic activity is blocked by a human protein attached thereto by way of a reversible linking group.

Abstract: There is described a method of inactivating reproducible pathogens in preparations containing plasmatic enzymes and proenzymes, activated or non-activated coagulation factors, such as Factors II, V, VII, VIII, IX, X, XIII, "FEIBA", prothrombin complex preparations, plasmatic inhibitors, immunoglobulins or other blood products, such as fibronectin and fibrinogen. In order to break and overcome the protective effect of proteins on pathogens by simultaneously preserving the biological activity and the molecular integrity of the proteins, the preparation is adjusted to a salt concentration of more than 0.5 molar by the addition of ammonium sulfate and is thermally treated, whereupon the ammonium sulfate is removed from the preparation.

Abstract: The present invention comprises a method for preparing a commercial .beta.-amylase product from whole or at least partially dehusked barley grain by extracting the grain with water. The water may optionally contain a reducing agent. The invention further comprises the purification, concentration and stabilization of the .beta.-amylase solution and the resultant commerical product.

Abstract: A fibrinolytically active protein conjugate comprising at least one optionally blocked fibrinolytic enzyme linked by way of a site other than the catalytic site responsible for fibrinolytic activity to at least one human protein.Processes from making the conjugates and pharmaceutical compositions containing them are also described.

Abstract: A derivative of a fibrinolytic enzyme in which the catalytic site on the enzyme which is responsible for fibrinolytic activity is blocked by a human protein attached thereto by way of a reversible linking group.

Abstract: During the kilning of germinated barley in the course of a malting operation nitrosamines are usually formed and ultimately these end up in beer obtained from such malt. It has been found that such formation of nitrosamines can be suppressed even in the absence of sulphur dioxide when the germinated barley is treated with a solution or dispersion of saccharides before it is kilned.

Abstract: Germination of cereal grain in malting is carried out by passing steeped grain to and through a series of six closed spaced discrete vessels in succession. The grain is maintained in each vessel for about a day and in each vessel is subjected to an upward flow of humidified attemperated air. The grain is turned either in a vessel or through transference to the next vessel. Transference from one vessel to the other is carried out by discharging the grain from each vessel along a lower conveyor to an elevator which raises the grain to an upper conveyor that discharges the grain down into the next vessel. Grain leaves the last vessel of the series as green malt and then passes to a malt kiln where it is dried to a desired moisture level.

Abstract: Malt having a high content of alpha-1,6-hydrolase activity is prepared by germinating previously steeped barely until the average acrospire reaches a length of at least 11/4 times the average length of the kernals and it contains at least 55 units of alpha-1,6-hydrolase activity per gram of malt. The malt provides more complete hydrolyzing of starch to glucose and maltose, and it is useful in producing low carbohydrate beer, in producing distilled alcoholic beverages and in producing maltose syrup.