Abstract: A process for producing cholesterol oxidase from a recombinant microorganism, the microorganism itself and its recombinant plasmid are disclosed. The cholesterol oxidase has a particular amino acid sequence, a high substrate affinity and a working pH in the acidic range. The gene for this cholesterol oxidase was derived from Brevibacterium sterolicum. The cholesterol oxidase is used for the quantitative determination of cholesterol.

Abstract: In a method for the determination of the fat content of samples, preferably organic samples, fat portions are dissolved out of the sample by means of extraction. A high boiling point solvent is used for extraction. Through the use of a high boiling point solvent, all possible fat portions will be dissolved out and the extraction time will be significantly reduced. Concurrent with extraction, the extracted fat portions are saponified through the addition of a base, as a result of which salts of the fatty acids contained in the sample will be formed. In further procedural steps, these salts of the fatty acids are subjected to further treatment, separated from one another and analysed. The method enables the exact determination of the fat content of organic samples on the basis of the fatty acids contained within the sample.

Abstract: Lipid bilayers which incorporate poly-3-hydroxybutyrate/polyphosphate ion channels. The ion channels are preferably purified polyhydroxybutyrate polyphosphate complexes extracted from organisms or prepared from the individual constituents which have been size selected. The bilayers are useful for testing the affect of various molecules and ions on the channel which correlates to the in vivo result. The bilayers are useful for transporting the channels into cells or other bilayers.

Abstract: Disclosed herein are methods and a diagnostic agent for identifying HIV-infected individuals. The diagnostic agent, termed C-8.2, whose concentration is altered within about 1-3 days of HIV infection is used in assays not dependent on HIV antigens or antibodies. C-8.2 is present in the serum of all mammals and is a phospholipid or a mixture of related phospholipids.

Abstract: Methods and compositions for producing a mammal capable of expressing an exogenously supplied gene in cells of the airway are disclosed. Lipid carrier-nucleic acid complexes are prepared then delivered via aerosol to the lung airway. The invention provides a direct method for transforming pulmonary cells as a means for treating disorders of the lung as for providing a means for delivering substances systematically following expression in the lung.

Abstract: The present invention relates to novel compounds used in an electrode membrane combination. These novel compounds include a linker lipid for use in attaching a membrane including a plurality of ionophores to an electrode and providing a space between the membrane and the electrode, the electrode being either in part or totally made up of the linker lipid. The linker lipid comprises within the same molecule a hydrophobic region capable of spanning the membrane, an attachment group used to attach the molecule to an electrode surface, a hydrophilic region intermediate said hydrophobic region and the attachment group, and a polar head group region attached to the hydrophobic region at a site remote from the hydrophilic region.

Abstract: Disclosed is a method for the quantitative determination of D-3-hydroxybutyric acid and acetoacetic acid, which comprises reacting a biological sample containing D-3-hydroxybutyric acid and acetoacetic acid, with a reagent comprising: (1) a D-3-hydroxybutyrate dehydrogenase, (2) A.sub.1 and (3) B.sub.1, the components (1), (2) and (3) participating in the following cycling reaction: ##STR1## thereby effecting the enzymatic cycling reaction, and measuring a change in the amount of A.sub.2 formed or the amount of B.sub.1 consumed. Also disclosed is an analytical reagent comprising the components (1), (2) and (3) for use in the above method. The method and the analytical reagent ensure rapidness and accuracy in the determination of D-3-hydroxybutyric acid and acetoacetic acid, even with the use of a small quantity of a biological sample, so that they are very useful in application fields, such as clinical diagnosis and food testing.

Abstract: continuous flow analysis system measures the cholesterol distribution among different lipoprotein classes in a blood sample. A blood plasma sample is separated into different lipoprotein classes by single vertical spin density gradient ultracentrifugation. The sample is then introduced in a continuous succession into a reagent stream which flows continuously through a detector. A sensor detects the end of the sample in a sample stream before it is introduced into said reagent stream. A wash solution is introduced into the sample stream in response to detection of the end of said sample. The wash solution back flows through the sample stream into the sample container to wash any residue and/or air bubbles from the sample stream. After a predetermined time period, the sample stream is closed and the process is repeated for the next sample.

Abstract: An antemortem diagnostic test for neurodegenerative disease, particularly Alzheimer's disease, is disclosed. The method of the invention involves the collection of a sample of bodily fluid; the analysis of the concentration of neuronal membrane phospholipids and phospholipid metabolites, or the ratio of a phospholipid to its metabolite, in the sample; and the correlation of the concentration or the ratio to the corresponding value from control samples. The method can be used to determine the presence or the possibility that an individual can develop a neurodegenerative disease, such as Alzheimer's disease; to follow the course of neurodegenerative disease in an individual; and to validate the efficacy of a drug used to treat a neurodegenerative disease.

Abstract: A rapid screening test was used to distinguish between blood of normal infants and that of individuals having the Smith-Lemli-Opitz Syndrome (SLO). Blood was spotted on filter paper and analyzed with no extractions or separations. The method comprises the steps of vaporizing a cholesterol containing organic sample, ionizing the vaporized sample, detecting fragment ions of said vaporized ionized sample, and determining the intensity ratio of said fragment ions for cholesterol/7-dehydrocholesterol in the sample. The method includes performing the analysis by time-of-flight secondary ion mass spectrometry without derivation or chemical steps.

Abstract: Apparatus and method for determining the percentage of free fatty acids in cooking fats and oils. A kit for this analysis includes a test cylinder calibrated for (a) volume of test sample, (b) volume of acid-base indicator solution, and (c) a set of lines marked to be read directly as the percentage of free fatty acids in the sample.

Abstract: An accurate, stable, liquid lipoprotein control composition is provided. The control composition of the present invention preferably comprises lipoprotein, suffactant and a divalent cation. The control may further comprise a chelating agent. It has been surprisingly found that when suffactant and a divalent cation are added to lipoprotein, it performs like native lipoprotein in human serum, i.e. not aggregated or subject to matrix effects. In addition, the control composition of the present invention is stable for up to about 5 months at room temperature (about 22.degree. C.) and up to about 12 months when refrigerated (about 2.degree. C. to about 10.degree. C.). Methods of making and using the control composition are also provided.

Abstract: Low calorie food compositions containing fat-type organoleptic ingredients and non-fat ingredients, wherein natural fats and synthetic fat mimetics are blended in proportion to provide any predetermined amount of fat caloric value, said synthetic fat-type mimetic ingredients comprising esterified epoxide-extended polyols (EEEPs) of the formula P(OH).sub.a+c (EPO).sub.n (FE).sub.b where P is a polyol having a=2-8 primary hydroxyls, c=0-8 secondary and tertiary hydroxyls, a+c is in the range of 3-8, EPO is a C.sub.3 -C.sub.6 epoxide, FE is a fatty acid acyl moiety, n is the minimum epoxylation index average number having a value generally equal to or greater than a and is a number sufficient that greater than 95% of the primary hydroxyls of the polyol are converted to secondary or tertiary hydroxyls, and 2<b.ltoreq.a+c.

Abstract: A lipid antigen found in colon, breast, lung, ovarian and pancreatic human adenocarcinomas identified by reactivity with human monoclonal antibody 81AV78, and the use of the antigen in vaccines.

Abstract: A method of separating a first class of lipoprotein in a sample from a second class of lipoprotein in the sample including: precipitating the second class of lipoprotein; contacting the sample with a magnetically responsive particle; and placing the sample in a magnetic field until the magnetically responsive particle has sedimented, thereby causing the precipitated second class of lipoproteins to sediment, leaving the first class of lipoproteins in the supernatant of the sample.

Abstract: The present invention relates to the direct quantitative determination of cholesterol and involves the formation of a spectrophotometrically active product of cholesterol obtained by contacting cholesterol with an acyl compound and a perchlorate effective to form the spectrophotometrically active product.

Abstract: A multilayer analysis element for determination of total cholesterol is prepared having a light-transmissive water-impermeable support, at least one hydrophilic polymer layer on said support and a spreading layer on said hydrophilic polymer layer(s), and containing in one or more of the layers:(a) at least one enzyme having cholesterol esterase activity,(b) cholesterol oxidase,(c) peroxidase,(d) a color reagent composition, which in the presence of hydrogen peroxide and the peroxidase, produces a color change(e) at least one bile acid compound selected from the group consisting of bile acids, bile acid derivatives and salts of said acids and derivatives, and(f) an alkyl phenoxy polyethoxy ethanol containing in the alkyl group 1 to 20, preferably 7 to 10 carbon atoms, and a polyoxyethylene chain composed of at least 16, preferably 30 to 60 oxyethylene units.

Abstract: A method for stabilizing the functional activity of annexins in a biological fluid sample is provided.

Abstract: The invention is a non-invasive diagnostic test which is performed on the surface of the skin. This test indicates skin cholesterol levels which can provide information about the extent of aortic atherosclerosis. The invention also relates to reagents in the form of affino-enzymatic test compounds for use in the diagnostic test.

Abstract: The present invention relates to polymeric luminophores which contain ruthenium, osmium, and rhenium complexes covalently attached to polymer matrices. The polymeric luminophores are capable of having their luminescence quenched by molecular oxygen and may be coated on optical fibers to form sensing elements to detect oxygen in gases and fluids.

Abstract: There are disclosed a process for detecting and determining an oxidized lipid in a specimen, which is capable of readily and accurately determining a specimen as containing an oxidized lipid, and a process for forming a water-soluble oxidized lipid having a hydroperoxide group which has specific influence on a living body. A specimen is detected and determined to contain an oxidized lipid by adding a lanthanide shift reagent to a specimen, followed by spectroscopic analysis thereof. An oxidized lipid is formed by adding superoxide dismutase (SOD) and CuSO.sub.4 to (1) an emulsion prepared by dissolving linoleic acid or arachidonic acid in deuterated methyl alcohol and adding the solution to a deuterated phosphate buffer under stirring, or to (2) a low density lipoprotein solution sufficiently dialyzed against an undeuterated phosphate buffer; followed by irradiation with long-wave ultraviolet light.

Abstract: The method is directed to assaying for biological components in a sample comprising step (A) generating an oxidase substrate in the presence of an amphoteric surfactant and in the absence of ferrocyanide; (B) initially generating hydrogen peroxide through said oxidase reaction on said substrate of oxidase with subsequent detection of the generated hydrogen peroxide using peroxidase and a color developer capable of being oxidized in the presence of amphoteric surfactant and ferrocyanide; and (C) correlating the amount of color developed to the amount of biological components in the biological sample. Even when the biological component to be detected is present in a very small amount, the interference of bilirubin can be eliminated in assays of biological components in which peroxidase generated from an enzymatic reaction is detected using peroxidase and a color developer capable of being oxidized.

Abstract: A method and apparatus are provided for magnetohydrodynamic acoustic-resonance, near-IR spectroscopy. The method includes a step of applying to a subject under study a magnetic field having a strength between 2,00-10,000 gauss, near-IR radiation having a wavelength between 800-3,000 nm and an acoustic wave having a frequency between 10 khz -1 Mhz. The method also includes the steps of inducing vibration of ions in the magnetic field and detecting an electric wave generated magnetohydrodynamically by the acoustic wave induced vibration of the ions. Next is the collecting of the electrical, acoustical and near-IR spectra and the analyzing of the collected spectra. The spectra is analyzed in a hyphenated, multidimensional fashion.

Abstract: Enzyme electrodes having the enzyme enclosed within liposomes supported on, or in a membrane and immobilized therein are provided. The liposomes may be made using lipids and conveniently by "reverse phase evaporation". The enzyme electrodes incorporate conventional electrode elements, e.g., THE Clark electrode pair (Platinum/Silver) and additional membranes or screens, and used particularly in amperometric measurements. The enzyme can be an oxidase and is useful for measurement, directly or indirectly, of low molecular weight species generated by enzymic action, and effective for study of biological fluids.

Abstract: An antemortem diagnostic test for neurodegenerative disease, particularly Alzheimer's disease, involves the collection of a sample of bodily fluid; the analysis of the concentration of neuronal membrane phospholipids and phospholipid metabolites, or the ratio of a phospholipid to its metabolite, in the sample; and the correlation of the concentration or the ratio to the corresponding value from control samples. The method can be used to determine the presence or the possibility that an individual can develop a neurodegenerative disease, such as Alzheimer's disease; to follow the course of neurodegenerative disease in an individual; and to validate the efficacy of a drug used to treat a neurodegenerative disease.

Abstract: The present invention provides ligand/anti-ligand assays for detecting and/or measuring adherent proteins, including lipophilic serum or plasma proteins, such as serum amyloid A (SAA) and apolipoprotein Al (apoAl); cytokines such as IL-1 beta, IL-6, and TNF alpha; pentraxins, such as CRP; and globular serum or plasma proteins such as albumin.

Abstract: The present invention provides a process for the determination of low density lipoproteins (LDL) in body fluids, wherein high density lipoprotein (HDL) antibodies are added to a sample to be investigated, insolubles formed are separated off and the LDL or one of its components is determined in the supernatant. The present invention also provides a reagent for the determination of the LDL fraction in body fluids, wherein it contains HDL antibodies.

Abstract: This invention provides a method of altering the metabolism of sphingolipids in a cell comprising contacting the cell with a fumonisin, or an analog thereof. The invention also provides a method of detecting the consumption of a fumonisin or a fumonisin analog in a subject comprising (A) detecting, in a sample from the subject, the state of the metabolic pathway of sphingolipids and (B) comparing the state of the metabolic pathway to that of a normal subject, the presence of a change in the state of the metabolic pathway indicating the consumption of a fumonisin or a fumonisin analog. Also provided is a method of detecting the presence of a fumonisin or fumonisin analog contamination in a sample from a food or feed comprising detecting a reaction of the metabolic pathway of sphingolipids, the presence of the reaction indicating the presence of a fumonisin or fumonisin analog contamination. Furthermore, novel fumonisin analogs and compositions comprising fumonisins and fumonisin analogs are provided.

Abstract: A method of assay for an enzyme catalysing the release of long chain fatty acids or, conversely, for an ester substrate for such enzymes is applicable to clinical samples and comprises removing substantially all the albumin from the clinical sample, incubating the sample after albumin removal with, for enzyme assay, an ester substrate or, for substrate assay, the enzyme, under conditions effective to release a fatty acid, causing the fatty acid to bind to a fatty acid binding protein and assaying the binding of the fatty acid to the binding protein.

Abstract: A new method for determining and reporting lipid fatty acids compositions and its use in diagnosing diabetes is disclosed. Adding internal standards, which are true analogs of the fatty acids to be analyzed, to blood serum or plasma samples before performing a gas chromatographic analysis of the fatty acids allows the results of the analysis to be reported in absolute terms as, for example, mg/L, instead of in relative percentages. Reporting the results as absolute concentrations reveals statistically significant differences in levels of various lipid fatty acids between persons having and not having diabetes.

Abstract: An antibody specifically immunoreactive with glycated LDL and methods for using the antibody are provided. The antibody can be used for quantitating amounts of glycated LDL in a sample, for monitoring glycemic control in diabetic patients, for diagnosing disease, for monitoring and diagnosing atherosclerotic cardiovascular disease, and for inhibiting accumulation of LDL cholesterol by cells in tissues subject to atherosclerotic disease.

Abstract: Lipid is determined quantitatively by treating a sample with a surfactant in an aqueous medium to obtain a liquid sample dispersion, then reacting the dispersion with a reagent containing cupric ion and bicinchoninic acid, and measuring a colored state developed by complex formation of bicinchoninic acid with cuprous ion formed from cupric ion in the presence of lipid. This method can be effectively applied to determination of amphiphilic lipid having a tendency of gathering in an aqueous medium. Further, two different species interfering with each other in quantitative determination thereof are determined respectively by conducting two different determination methods on the same sample followed by calculation using the values obtained.

Abstract: The invention is a non-invasive diagnostic test which is performed on the surface of the skin. This test indicates skin cholesterol levels which can provide information about the extent of aortic atherosclerosis. The invention also relates to reagents in the form of affino-enzymatic test compounds for use in the diagnostic test.

Abstract: An apparatus for and a method of on-line detecting and assessing, in situ, he amount and characterization of microbial trace contamination in an associated ultra-pure water system is disclosed. The apparatus comprises, inter alia, a monitor/detector portion and an assessment/identification portion. The monitor/detector portion includes a contamination mass detector which nondestructively senses the presence of microbial biofilms in real time when grown upon the surface of a quartz crystal wafer portion of the contamination mass detector. The assessment/identification portion is configured to provide quantitative analysis of the microbial biofilms formations based on signature biomarker cellular components. A programmable controller is configured to integrate these and other portions into an automated apparatus, which is capable of providing real time chemical countermeasures to prevent the potential deleterious effect of the microbial trace contamination uncovered.

Abstract: Disclosed herein are methods and a diagnostic agent for identifying HIV-infected individuals. The diagnostic agent, termed C-8.2, whose concentration is altered within about 1-3 days of HIV infection is used in assays not dependent on HIV antigens or antibodies. C-8.2 is present in the serum of all mammals and is a phospholipid or a mixture of related phospholipids.

Abstract: The amount of lipid bound sialic acid in a blood plasma or serum sample may be determined by an improved method, which may be automated, involving the following steps to be performed simultaneously on the sample and a standard consisting of commercially available n-acetyl nueraminic acid (NANA); diluting with a buffer; mixing the diluted sample; adding a mixture of a chlorinated lower aklyl hydrocarbon and a lower alkyl alcohol; treating by mixing and centrifuging to yield a substantially clear upper phase; treating the upper phase with a color development reagent; mixing; boiling the admixture; cooling the admixture; and determining the amount of lipid bound sialic acid present in the sample by comparing the optical density of the sample to the optical density of the NANA.

Abstract: An improved device and method of a) separating the cellular components of whole blood from plasma or serum, b) separating the low density lipoprotein (LDL) fraction and the very low density lipoprotein (VLDL) fraction from the plasma or serum, then c) assaying the plasma or serum for cholesterol present in the high density lipoprotein (HDL) fraction are disclosed. The device includes a separation area that separates the cellular components of whole blood from the serum or plasma and separates the LDL and VLDL fractions from the serum or plasma, and a test area that assays the serum or plasma for the concentration of the HDL cholesterol. The method includes introducing a whole blood sample to a test device including a separation area comprising a first zone that separates the cellular components of the whole blood from the plasma or serum; and a second zone that separates the LDL and the VLDL fractions from the plasma or serum.

Abstract: An assay device for measuring the concentration of HDL-associated cholesterol in a blood-fluid sample is disclosed. The device includes a glass-fiber sieving matrix capable of separating soluble and precipitated material migrating through the matrix, and a reagent reservoir in the matrix, for releasing a reagent effective to selectively precipitate LDL and VLDL particles, while permitting soluble HDL to migrate through the matrix. The glass fibers are coated with a polymer coating to prevent binding of HDL to the fibers in the presence of the precipitating reagent.

Abstract: A process is described for generating conjugates of lipids and biologically active agents to produce compositions having therapeutic utility, such as drug delivery vehicles. The process involves mixing the reactive lipid with an appropriate amount of diketene to form an acetoacetylated lipid which is then isolated, dissolved in a suitable medium, and mixed with a nucleophilic-containing biologically active agent to form a biologically active agent-lipid conjugate. Alternatively, the acetoacetylated lipid can be mixed with a polyamine to form a cationic lipid.

Abstract: A method of assaying analytes using a rate procedure is described where the change in density over time has a variable rate. The method features the steps of depositing the sample onto a dried slide-like test element, making an initial rate reading during an early time window, using the initial rate readings in a comparison study with rates from known low and high concentration results to predict whether the sample rate will be sufficiently low as to be ascertainable during a later time window or not, and then calculating a rate of reaction and concentration during either the early time window or the early time window with a portion of the later time window, respectively.

Abstract: A method of quantitative assay for a substrate such as triglyceride capable of undergoing enzymatic hydrolysis to release long chain fatty acids in an albumin-containing clinical sample comprises incubating the albumin-containing clinical sample with an enzyme such as lipase which acts upon the substrate to be assayed under conditions effective to release fatty acid therefrom, causing the fatty acid thus released to bind to a fatty acid binding protein (FABP) and assaying the binding of the fatty acid to the FABP.

Abstract: A general purpose technique called extractive hydrogenation analysis, useful for both trace component and imaging analysis of samples containing organic matter. In processes according to the invention, organic matter is thoroughly decomposed in the presence of a hydrogenation catalyst under substantially aqueous conditions into products, and one or more of the products is then detected.

Abstract: The present invention provides an accurate, rapid and precise methodology for assessing maturity of the lungs in a fetus prior to birth, especially for premature fetuses having a gestation period of 37 weeks or less. The methodology quantitatively measures a specific phosphoglyceride, dipalmitoyl phosphatidyl choline, in samples of amniotic fluid for the assessment of fetal lung maturity. The process enzymatically cleaves such phophoglycerides and preferably detects the resulting diacylglycerols by HPTLC-reflectance spectrodensitometry.

Abstract: The invention relates to an apparatus and a method useful in separating non-high density lipoproteins, referred to as "non-HDLs", from biological fluids containing them. A porous carrier is provided which contains a non-HDL precipitating agent. One need only contact the sample of interest to the carrier for one minute or less, after which the precipitated non-HDLs are no longer present in the sample being tested. The applications of the method include the ability to determine high density lipoproteins in the sample without interference from non-HDLs.

Abstract: A method of separating a first class of lipoprotein in a sample from a second class of lipoprotein in the sample including: precipitating the second class of lipoprotein; contacting the sample with a magnetically responsive particle; and placing the sample in a magnetic field until the magnetically responsive particle has sedimented, thereby causing the precipitated second class of lipoproteins to sediment, leaving the first class of lipoproteins in the supernatant of the sample.

Abstract: Disclosed is a method for the rapid quantitative determination of low density lipoprotein (LDL) in a biological fluid sample containing LDL, very low density lipoprotein (VLDL) and high density lipoprotein (HDL). The sample is first passed through a matrix to bind LDL to the matrix. The LDL is then eluted from the matrix differentially from VLDL and HDL. A parameter representative of the concentration of a solute exiting the matrix during the interval when LDL is eluted is then determined, wherein the parameter is proportional to the amount of the LDL in the sample. The parameter may be determined, e.g., by electronically integrating under a spectrophotometrically detected peak of eluted LDL.

Abstract: Low density lipoproteins are directly determined in fluid samples by selectively precipitating low density lipoproteins from the sample by forming clusters, selectively consuming the high density and very low density lipoproteins, and resolubilizing the low density lipoproteins for direct determination thereof. The clusters are formed by treating the fluid sample with a mixture of a polyanionic compound, a divalent metal, and a nucleating agent. The clusters are redissolved and assayed for low density lipoproteins.

Abstract: Disclosed is an improved process for the recovery of cholesterol rich fractions from mammalian serum or plasma. The process involves adsorbing the fraction on precipitated silica gel agglomerates which are then separated from the serum or plasma whereupon the adsorbed cholesterol rich fraction is eluted from the silica and recovered.

Abstract: A laminated assay device for use in determining the presence or amount of an analyte in a sample involving a bibulous assay strip having a sample application zone, a reagent zone and a pair of liquid impervious barriers defining a measurement zone extending from and in fluid communication with the application zone. The measurement zone has a volume measuring the amount of sample required for the assay. A substantially nonabsorbent first supporting film extends across and is laminated to the front surface of the assay strip, while a substantially nonabsorbent second supporting film extends across and is laminated to the back surface of the assay strip. The assay strip is impregnated with the members of a signal producing system which, upon reaction with a component in the sample, produce a detectable signal.

Abstract: The present invention provides a process for the determination of low density lipoproteins (LDL) in body fluids, wherein high density lipoprotein (HDL) antibodies are added to a sample to be investigated, insolubles formed are separated off and the LDL or one of its components is determined in the supernatant.The present invention also provides a reagent for the determination of the LDL fraction in body fluids, wherein it contains HDL antibodies.