Abstract: The method consists in the use of one compound of formula: X--NH--R in which X represents one 5-bromoindole-3-yle group and R represents the acyl radical of one amino acid selected between leucine and alanine, as tracer for demonstrating, by the formation of a colored product, a peptidase activity in a culture of micro-organisms.

Abstract: The present invention relates to a method for detection of atopic dermatitis, and to a kit for use in the detection of atopic dermatitis. The present invention is useful in cases where it is difficult or almost impossible to distinguish atopic dermatitis from other allergoses when detected by the conventional methods, or where it is difficult to macroscopically distinguish atopic dermatitis from other dermatopathies. Moreover, the detection method of the present invention is also useful as a method of primary screening for atopic dermatitis.

Abstract: D-enzyme compositions are described comprising an amino acid residue sequence that defines an polypeptide able to catalyze an enzymatic reaction. The D-enzyme has an amino acid residue sequence consisting essentially of D-amino acids.

Abstract: The present invention provides novel purified and isolated nucleotide sequences encoding the cGMP-binding, cGMP-specific phosphodiesterase designated cGB-PDE. Also provided by the invention are methods and materials for the recombinant production of cGB-PDE polypeptide products and methods for identifying compounds which modulate the enzymatic activity of cGB-PDE polypeptides.

Abstract: The present invention relates to a method for the detection and for the determination of enzymatic reactions in urine, wherein certain substances which improve the optical measurability of the enzymatic reaction are added to the reaction.

Abstract: Proteins such as enzymes and antibodies are immobilized by crosslinking crystals of the proteins such as microcrystals having a cross-section of 10.sup.-1 mm or less with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. Crystals of an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease may be crosslinked to provide crosslinked enzyme crystals that retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred Pronase.TM.:enzyme ratio is 1:40.

Abstract: This invention relates to a hydrophobic layer formed on a solid support, the hydrophobic layer incorporating by hydrophobic interactions an amphipathic enzyme substrate labeled with a reporter on its hydrophilic region, and the use of the same to conduct various enzyme assays without extraction steps.

Abstract: The instant invention encompasses isolated stable esterase enzymes characterized by the ability to remain stable at certain temperatures, substrate specificities, and activity profile.

Abstract: Methods are disclosed for detecting one or more mutations in an isolated test nucleic acid by forming a heteroduplex with a homologous control DNA and contacting the heteroduplex with a resolvase capable of recognizing at least one single base pair mismatch within the heteroduplex. In preferred embodiments of the invention, the resolvase is bacteriophage T4 endonuclease VII.

Abstract: The presence of neuro-cognitive disorders associated with systemic immunological malfunction are assessed in human patients by determining the level of intracellular RNase L in a sample of the patient's peripheral blood and comparing that level to predetermined levels of RNase L in healthy individuals. Aberrant RNase L levels as compared with those in healthy individuals indicate the presence of neuro-cognitive disorders associated with systemic immunological malfunction. This procedure is useful to distinguish systemic immunological malfunction from primary psychological or neuropsychiatric disorders presenting otherwise similar clinical symptoms.

Abstract: The amount of cholesterol in low density lipoproteins in a sample can be measured by contacting the sample with one or more reagent solutions to carry out the reaction in the presence of a polyanion and an amphoteric surfactant, followed by optical measurement of the reaction product.

Abstract: A CHO-K1 cell line stably expressing a recombinant full-length human PDE IVa (rhPDEIVa) enzyme was established under hygromycin B selection. Inhibition of the expressed PDE IVa activity by selective PDE IV inhibitors was evaluated. The rank order of potencies of the inhibitors in elevating cAMP in the whole-cell assay was quite different from that on the soluble enzyme. The whole-cell rank order of potencies was also maintained when PKA activity ratios were measured in place of cAMP levels. When inhibition of soluble PDE IVa activity was examined in the presence of 100 mM MgCl.sub.2, the rank order of potencies of the inhibitors mirrored that obtained for the whole-cell assays (both cAMP and PKA). The observed "high-affinity" conformation of the enzyme was not dependent upon a specific cation or anion.

Abstract: Disclosed are methods and compositions for identifying, monitoring and treating premalignant and malignant conditions in a human subject. The present invention further discloses methods and compositions for determining cells undergoing apoptosis, and for increasing the efficacy of a cancer therapy. The methods involve the use of apurinic/apyrimidinic endonuclease (APE), independently, as a marker for (pre)malignant conditions and for apoptosis. Also described are polyclonal antibody preparations for use in methods for detecting APE and methods for modulating expression susceptibility of cells to apoptosis.

Abstract: A method for determining adenosine 5' diphosphate (ADP) contained in a liquid sample by means of an enzymatic reaction, comprising reacting the sample at 15 to 45.degree. C. at least in the presence of glucose, ADP-dependent hexokinase, and oxidized NAD(P), a glucose-6-phosphate dehydrogenase, and one or more salts releasing ions selected among magnesium, cobalt, and manganese ions and then determining the ADP contained in the sample together with the AMP resulting from the reaction based on the amount of the reduced NAD(P) yielded. This method has advantages in that the limit of determination is high because ADP is determined based on the amount of the reduced NAD(P) yielded, and since the reduced NAD(P) has a definite molecular extinction coefficient, the value found is highly reliable and uninfluenced by the reducing substances contained in the sample.

Abstract: A MicroColonyImager instrument and solid phase methods to screen cells expressing mutagenized enzymes for enhanced activity. The MicroColonyImager instrument and methods permit high throughput screening of enzyme libraries by time course analyses of single-pixels, using either absorption, fluorescence or FRET. This high throughput assay can detect small differences in enzyme rates within microcolonies grown at a nearly confluent density on an assay disk. Each microcolony is analyzed simultaneously at single-pixel resolution, requiring less than 100 ml substrate/measurement. By simultaneously assaying different substrates tagged with spectrally distinct chromogenic or fluorogenic reporters, the substrate specificity of an enzyme can be changed.

Abstract: This method for non-invasively, rapidly and simply distinguishing between allergies, viral infections and sinusitis involves testing nasal secretions, preferably with commercially available (Ames Division, Miles Laboratories, Inc., Elkhart, Ind. 46515; or from Boehringer Mannheim Corporation, Advanced Diagnostics, 9115 Hague Road, P.O. Box 50457, Indianapolis, Ind. 46250-0457) or novel, modified reagent test strips. The commercially available strips, also referred to as dipsticks, test for pH, protein, nitrite, glucose, ketone, white blood cell esterase, bilirubin and blood. In the method of this invention, a sample of a patient's nasal secretions is tested and, based on the pH, amount of protein, nitrite, esterase and a measure of eosinophil infiltration, it can quickly be determined if the patient is suffering from an allergy, from a viral infection or a bacterial infection. The method has the potential to supplant much more expensive and invasive clinical procedures.

Abstract: A method and apparatus for measuring enzyme activity by supplying a substrate solution and an enzyme solution into a reaction vessel combined with a total reflection absorption prism is disclosed. A measurement solution (40) in which the enzyme solution and the substrate solution are combined together is kept at a constant temperature within the reaction vessel (26) by a temperature control unit (23), and is stirred by a stirrer (21). Infrared light (36) emitted from an infrared light source (3) is made incident on the interface between a total reflection absorption prism (28) disposed in contact with the measurement solution (40) and the measurement solution (40) from the side of the total reflection absorption prism (28) and is totally reflected by the interface. The spectrum of transmitted infrared light (37) thus totally reflected and emitted is detected by an infrared light detector (5).

Abstract: The present invention relates to a universal test systems and methods of use thereof for identifying a microorganism among at least two groups of widely divergent microorganisms. The universal test system comprises a predetermined combination of non-redundant biochemical tests comprising a substrate for at least one enzyme wherein the substrate, if acted on by the enzyme results in formation of a detectable product. Detectable products from the combination of biochemical tests are then used to identify the microorganism.

Abstract: Cholesterol in low-density lipoprotein (LDL) or very low-density lipoprotein (VLVL) in a sample is determined in the presence of a sugar compound and/or a protein-solubilizing agent.

Abstract: Provided is a method of determining the amount of cholesterol in high-density lipoprotein (HDL), which comprises reacting an HDL-containing sample with cholesterol esterase and cholesterol oxidase or cholesterol dehydrogenase in the presence of a reagent for aggregating lipoproteins- except HDL, and determining the amount of hydrogen peroxide or reductive co-enzyme formed therein.

Abstract: The present invention is to provide a method for measuring an amount of cholesterol in low density lipoproteins (LDL-cholesterol) in a sample specifically at high accuracy and a reagent used in this method, and the present invention can attain such effect that direct measuring an amount of LDL-cholesterol by widely used automatic analyzers can be conducted by using the invention, which has not been possible after known methods.

Abstract: The present invention provides a method for measuring an amount of LDL-cholesterol in samples specifically and accurately and also a reagent used for the method, and by using the present invention, such effect can be attained that LDL-cholesterol can be measured directly using so far widely used automatic analyzer, which has been impossible in known measurement methods.

Abstract: A peptide consisting of the following amino acid sequence, or an analogue or a fragment thereof, has an inhibitory activity on cholesteryl ester transfer protein:Glu Asp Thr Ser Pro Glu Asp Lys Met Gln Asp Tyr Val Lys Gln Ala Thr Arg Thr Ala Gln Asp Ala Leu Thr Ser Val Lys.

Abstract: Nonsteroidal anti-inflammatory drugs cause a dramatic increase in intracellular ceramide, which induces apoptosis. The ceramide increase is likely mediated by cyclooxygenase inhibition, which elevates arachidonic acid, which stimulates sphingomyelinase, which produces ceramide. Contacting members of this pathway with test compounds and observing their effects provides a method of screening for potential cancer chemopreventative agents.

Abstract: This invention provides a method to identify compounds potentially useful for the treatment of neoplasia in mammals. The phosphodiesterase inhibitory activity of a compound is determined along with COX inhibitory activity. Growth inhibitory and apoptosis inducing effects on cultured tumor cells are also determined. Compounds that exhibit phosphodiesterase inhibiton, growth inhibition and apoptosis induction, but not substantial prostaglandin inhibitory activity, are desirable for the treatment of neoplasia.

Abstract: Method for the rapid diagnostic of an infection of the urinary tract (UTI), by a specific examination of urine.The method of analyzing a urine specimen, for the rapid and simultaneous counting of both viable bacteria and leucocytes (white blood cells or WBC), consists of:(1) simultaneous labeling of said bacteria and said white blood cells (WBC) present in said urine specimen, with a fluorescent viability marker of bacteria, said marker tagging and accumulating in both bacteria and white blood cells, under the same conditions and leading to different fluorescent elements;(2) automatically counting all the fluorescent elements by photoelectric detection of the fluorescent marker; and(3) simultaneously classifying said fluorescent elements in fluorescent viable bacteria and in fluorescent white blood cells, in view to obtain in one part the total number of bacteria and on the other part, the total number of white blood cells, on the basis of their size.

Abstract: Processes for detecting or quantitating a particular phosphoglyceride in a sample containing the phosphoglyceride and other lipids and clinical uses for the process (e.g. for diagnosing improperly functioning lungs and other conditions) are disclosed.

Abstract: Recombinant human phosphodiesterase type IVC is described, and DNA coding for it. Particular conformers of the enzyme are identified, including a R- and S-rolipram stereoselective conformer which is obtainable by expression of human phosphodiesterase type IVC DNA in mammalian or insect cells. The recombinant enzyme may be used in a screen to select a compound capable of modulating the action of the enzyme, or as an immunogen to generate an antibody.

Abstract: A method of detecting the presence of a substance being monitored in a medium, selected from the group of substances including organophosphorus compounds and the metal ions Zn, Be and Bi, including the steps of: providing a 1,2-dioxetane phenyl phosphate compound; providing a phosphatase that catalytically degrades the 1,2-dioxetane phenyl phosphate compound to produce light, the catalytic activity of the phosphatase toward 1,2-dioxetane phenyl phosphate compound being altered by the substance being monitored; exposing the 1,2-dioxetane phenyl phosphate compound and the phosphatase together to a medium which may contain the substance being monitored; detecting light produced after the exposing step; and determining, from the detected light, the presence and concentration in the medium of the substance being monitored.

Abstract: The present invention involves a composition and method for the determination of enzymatic activity or enzyme substrate concentration in a fluid test sample in which the enzyme catalyzes a reaction in a reagent system to provide a detectable response indicative of the concentration of the enzyme or enzyme substrate in the test sample. A measured amount of a protease capable of eliminating the enzymatic activity is included in the reagent composition, so that when a measured amount of test sample is treated, the reaction producing the detectable response will have a definite end point. The invention is particularly useful in conjunction with the determination of leucocytes in urine wherein the leukocytes are lysed to release their human leucocyte elastase which hydrolyzes a chromogenic ester to provide a phenol which then reacts with a diazonium salt to form a colored azo dye.

Abstract: The invention provides a novel calcium-independent cytosolic phospholipase A.sub.2 /B enzyme, polynucleotides encoding such enzyme and methods for screening unknown compounds for anti-inflammatory activity mediated by the arachidonic acid cascade.

Abstract: This invention provides (1) a reagent for endotoxin assay which comprises aprotinin and a limulus amebocyte lysate reagent, (2) a kit for endotoxin assay which comprises the limulus amebocyte lysate reagent and a reagent containing aprotinin, (3) a method for assaying endotoxin in a sample using the limulus amebocyte lysate reagent in which aprotinin is added to the lysate reagent and/or the sample, (4) a method for assaying endotoxin in a serine protease-containing sample using the limulus amebocyte lysate reagent in which the sample is allowed to contact with an aprotinin-immobilized insoluble carrier in advance of endotoxin assay, (5) a carrier for pretreating a serine protease-containing sample on which aprotinin is immobilized, (6) a method for inhibiting factor G activation in which aprotinin is added to the limulus amebocyte lysate reagent and (7) a factor G activation inhibitor which comprises aprotinin as an active ingredient.

Abstract: A portable apparatus for detecting the presence of at least one methylxanthine chemical species such as caffeine or theophylline in a beverage comprises a first portion comprising an effective concentration of phosphodiesterase enzyme, a second portion comprising cyclic AMP, and means for indicating inhibition of degradation of the cyclic AMP by the phosphodiesterase due to the presence of the methylxanthine species. A method for determining the presence of at least one methylxanthine chemical species in a beverage comprises contacting at least one test portion of the beverage with effective concentrations of at least one phosphodiesterase enzyme and cyclic AMP, and further contacting the test portion with means for indicating the inhibition of the degradation of the cyclic AMP by the phosphodiesterase due to the presence of the methylxanthine species.

Abstract: Cholesterol in low density lipoproteins can be measured specifically and precisely by optically measuring a reaction product of a living sample with cholesterol oxidase or cholesterol dehydrogenase in the presence of an amphoteric surfactant. In addition to the above processes for measurement, the present invention provides reagent compositions and kits for measuring cholesterol in low density lipoprotein.

Abstract: The present invention relates to a method for the determination of cholesterol in low-density lipoprotein (LDL) in a sample containing LDL, which comprises eliminating cholesterol in high-density lipoprotein in the sample, subjecting the sample to a reaction utilizing the action of a cholesterol ester-hydrolyzing enzyme and the action of a cholesterol-oxidizing enzyme or of cholesterol oxidoreductase, and determining the amount of hydrogen peroxide or a reduced type coenzyme generated by the reaction.

Abstract: A method of screening for a genetic predisposition to anticholinesterase exposure. The method includes the steps of obtaining a peripheral blood sample, and then analysing serum from the blood sample for BuChE levels and inhibitor-susceptibilites. The DNA of peripheral white blood cells from the blood sample is also screened for the presence of BuChE alleles thereby identifying patients who have a genetic predisposition to anticholinesterase exposure.

Abstract: A rapid single step assay suitable for the detection or quantification of enzymes, in particular, hydrolases, especially, aminopeptidases and esterases. The enzymatic reaction causes the cleavage of a metal ligand labelled hydrolase substrate. The cleaved ligand alters the electrochemiluminescence of bidentate aromatic heterocyclic nitrogen-containing ligand reagent. The change in electrochemiluminescence correlates to the presence of hydrolase activity present in the sample. The assay can be performed on an IGEN Origen.RTM. Analyzer.

Abstract: A method is provided for determining the presence and/or amount of a microorganism and/or its intracellular material present in a sample performed by: (a) exposing the sample to a specific binding agent that has been immobilized upon a solid substrate, the specific binding agent being capable of binding to the microorganism or its intracellular material such that it becomes associated with the solid substrate, (b) exposing the solid substrate to an agent capable of making adenylate kinase associated with the microorganism and/or its intracellular material accessible to solutions applied to the substrate, (c) applying a solution containing adenosine diphosphate (ADP) to the substrate under conditions whereby adenosine triphosphate (ATP) may be produced by any adenylate kinase present, and (d) measuring the amount of adenosine triphosphate (ATP) and relating that to the presence and/or amount of microorganism or intracellular contents.

Abstract: Disclosed is a method for stably storing a liquid diagnostic reagent, comprising air-hermetically keeping the liquid diagnostic reagent in a closed container in the presence of a disoxidant therein. Preferably, at least one of the liquid diagnostic reagent and the disoxidant is covered with a separating container made of a material pervious to oxygen but not to solutions. The liquid diagnostic reagent may comprise an enzyme or an indicator.

Abstract: The invention relates to novel mannosidase inhibitors, processes for their preparation and their use as therapeutic agents. The invention also relates to the use of compounds of the formula II as described herein as prodrugs.

Abstract: A method for quantitatively determining cholesterol in high density lipoproteins, in which, prior to the determination of cholesterol by an enzymatic method, a surfactant and a substance which forms a complex with lipoproteins other than high density lipoproteins are added to a sample containing lipoproteins.The method does not require any pretreatments such as centrifugal separation. With a simple operation, cholesterol in HDLs can be measured effectively. Also, this method can be adopted in a variety of automated analyzers, and thus is very useful in the field of clinical assays.

Abstract: The subject invention pertains to a combination of enzyme activities comprising an ATP-degrading enzyme and one or more enzymes capable of degrading substances, other than ATP, that are substrates for the light-emitting reaction of firefly luciferase. These activities can be used to treat a growth medium in order to reduce background to negligible amounts, in a subsequent bioluminescence assay.

Abstract: With the recognition that like esterase, the thiazole derivatives show a strong nucleophilicity owing to sufficient electronic density in heterocycle, when its ester is degraded by some enzymes. The composition for detecting leucocyte and proteinase in urine has been developed, which contains a thiazole derivative. According to this invention, the thiazole derivative may be synthesized as an ester form by combining both the well-known thiazolone derivative and amino acid derivatives as starting materials. Now that the method for manufacturing the thiazole derivative is simple and general, the mass-scale production may be available within a short period of time. Further, in case of the measuring device produced according to this invention, the application method is simple and diagnostic performance is superior.

Abstract: Provided are a method of determining the amount of cholesterol in high-density lipoprotein (HDL), which comprises measuring the amount of cholesterol in low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL) and chylomicron (CM) in a sample in the presence of a sugar compound and/or a protein solubilizing agent, and calculating the difference between the amount of cholesterol in LDL, VLDL and CM and the total amount of cholesterol in the sample, and a method of determining the amount of cholesterol in HDL, which comprises measuring the amount of cholesterol in HDL in a sample in the presence of a sugar compound and/or a protein solubilizing agent.

Abstract: A means for cleaving a nucleic acid cleavage structure in a site-specific manner is disclosed. A cleaving enzyme having 5' nuclease activity without interfering nucleic acid synthetic ability is employed as the basis of a novel method of detection of specific nucleic acid sequences.

Abstract: A method and test kit for determining insect resistance based upon an esterase in the body fluid of the insect in the field. The method and kit includes an absorbent material pad (12) containing a dry substrate for the esterase which is preferably mounted on a fluid impermeable sheet (11). A roller (20) is preferably used to express the fluids from the insect onto the pad 12. The fluids are sufficient to enable the esterase to react with the substrate to produce an intermediate compound which is reacted with a chromogen to produce a visually detectable image with an intensity which varies directly as a function of the amount of the esterase and which is easily determinable by the farmer. The method and kit is particularly useful with homopteran, particularly aphids which are a problem in potatoes.

Abstract: Novel light producing 1,2-dioxetanes are described of the formula ##STR1## wherein ArOX is an aryl ring substituted with an X oxy group and A are passive organic groups which allow the 1,2-dioxetane to produce light when triggered by removing X. X is a chemically labile group which is removed by an activating agent. The 1,2-dioxetane compounds can be triggered to produce light at room temperatures.

Abstract: The invention concerns a method for the non-radioactive detection of metabolically labelled DNA, in one aspect detecting labelled DNA indicative of cell lysis or apoptosis.

Abstract: In an assay method in which a member of a specific binding pair is detected by means of an optically detectable reaction, the improvement wherein the optically detectable reaction includes the reaction, with an enzyme, of a dioxetane having the formula ##STR1## where T is a cycloalkyl or polycycloalkyl group bonded to the 4-membered ring portion of the dioxetane by a spiro linkage; Y is a fluorescent chromophore; X is hydrogen, alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl, cycloheteroalkyl, or enzyme-cleavable group; and Z is hydrogen or an enzyme-cleavable group, provided that at least one of X or Z must be an enzyme-cleavable group, so that the enzyme cleaves the enzyme-cleavable group from the dioxetane to form a negatively charged substituent bonded to the dioxetane, the negatively charged substituent causing the dioxetane to decompose to form a luminescent substance that includes group Y of said dioxetane.