Abstract: The present invention makes available, inter alia, methods and reagents for modulating smoothened-dependent pathway activation. In certain embodiments, the subject methods can be used to counteract the phenotypic effects of unwanted activation of a hedgehog pathway, such as resulting from hedgehog gain-of-function, ptc loss-of-function or smoothened gain-of-function mutations.

Abstract: Compositions and methods using antibodies which are immunoreactive with specific apolipoproteins to determine the concentrations of lipoproteins such as HDL and LDL, and/or apolipoproteins in human blood, serum or plasma sample, are described. Monoclonal antibodies (MAbs) are described that specifically bind to epitopes present in apolipoproteins and lipoproteins, enabling rapid and reliable determinations of levels of specific blood lipoprotein and/or apolipoprotein levels, including Apo B-100, Apo A-I, Apo A-II, Apo C-III, and Apo E, and thereby determination of relative ratios of HDL and LDL and LpaI and LpaII. In a preferred embodiment, the compositions are strips of a solid phase material coated with one or more of the antibodies and are referred to herein as “dipsticks”. The dipsticks specifically bind a lipoprotein or apolipoprotein when dipped into a protein sample.

Abstract: The present invention herein provides the design, synthesis and characterization of compositions comprising asymmetric bolaamphiphilic lipids that form extended polymeric ribbons and wide sheets. These compositions may be doped, or interspersed, with various compounds to fine-tune the fluidity and rigidity of the bolaamphiphilic lipid composition, and promote other morphologies of the composition, including fluid vesicles and truncated flat sheets. Upon an increase in pH these compositions undergo a calorimetric and morphological transformation.

Abstract: An improved multi-layered diagnostic sanitary test strip for receiving a heterogenous fluid, such as whole blood, to test for presence and/or amount of a suspected analyte in the fluid by facilitating a color change in the strip corresponding to the amount of the analyte in the fluid, wherein the test strip includes fluid volume control dams to prevent spillage of the fluid from the strip and a chemical reagent solution that facilitates end-point testing.

Abstract: The present invention relates to a cell culture system to provide a tool for assessing the potential of a patient's serum for preventing the accumulation of cholesterol in arteries (i.e. serum efflux potential) that leads to atherosclerosis and to screen new drug compositions being developed to reduce the accumulation of cholesterol or enhance the clearance of cholesterol from the vessel wall. The present invention combines two individual assays, which are two different cholesterol assays: an assay measuring scavenger receptor class B type I (SR-BI)-mediated cholesterol efflux and an assay measuring ATP binding cassette protein 1 (ABCA1)-mediated cholesterol efflux, and uses them in parallel to test human and animal sera for their potential to stimulate efflux, as mediated by either of the two receptors described above.

Abstract: A composition useful for separating lipoproteins, such as high density lipoprotein (HDL), low density lipoprotein (LDL), very low density lipoprotein (VLDL) and modified lipoproteins with a high degree of accuracy by ion-exchange based procedures. The composition comprises an organic particle having a hydrophilic surface onto which ion-exchange groups are introduced and which is formulated so as to provide a high enough degree of hydrophilicity to prevent irreversible absorption of lipoproteins, but which is not so high as to block the pores of the ion exchange particles and prevent adequate retention and separation of lipoproteins. A method for making a composition for separating lipoproteins comprising seed polymerization of organic particles, hydrophilic treatment of the organic particles by introducing 0.

Abstract: A method of simultaneously determining the concentrations of cardiovascular risk markers selected from the group consisting of High Density Lipoprotein cholesterol (HDL-C), Low Density Lipoprotein cholesterol (LDL-C), total cholesterol, triglycerides and oxidized LDL using infra-red and/or near infrared light is described.

Abstract: A method for diagnosing Alzheimer's disease even at early stages by assessing the levels of sulfatides or its metabolites in biological fluids is disclosed.

Abstract: An assay to assess thrombotic risk in which oxidized lipids comprising phospholipids are utilized as a membrane source in a clotting assay and the results compared to an assay in which unoxidized phospholipid is used as a membrane source in the presence and absence of activated protein C (“APC”). The assay can monitor for the presence of antibodies in the patient which interfere specifically with the anticoagulant function of APC in an oxidation dependent or independent manner. This can indicate the propensity of the patient to experience episodes of vein thrombosis or arterial thrombosis.

Abstract: The present invention relates to determining the prethrombotic state, in particular determining an amount or presence of circulating microparticles and/or stimulated procoagulant cells.

Abstract: An improved multi-layered diagnostic sanitary test strip for receiving a heterogenous fluid, such as whole blood, to test for presence and/or amount of a suspected analyte in the fluid by facilitating a color change in the strip corresponding to the amount of the analyte in the fluid, wherein the test strip includes fluid volume control dams to prevent spillage of the fluid from the strip and a chemical reagent solution that facilitates end-point testing.

Abstract: The present invention provides a method of diagnosing the presence or severity of liver fibrosis in an individual by detecting ?2-macroglobulin (?2-MG) in a sample from the individual; detecting hyaluronic acid (HA) in a sample from the individual; detecting tissue inhibitor of metalloproteinases-1 (TIMP-1) in a sample from the individual; and diagnosing the presence or severity of liver fibrosis in the individual based on the presence or level of ?2-MG, HA and TIMP-1.

Abstract: Methods of detecting novel therapeutically active compositions based on their ability to modulate the glycolipid metabolism and overcome multidrug resistance are described. These methods are particularly useful in screening for novel chemotherapeutic agents for the treatment of cancer, as well as chemosensitizers that are capable of enhancing the cytotoxicity of such chemotherapeutic agents. A combination of one or more of these compositions can be used in the treatment of a various cancers.

Abstract: A method of lipid assay characterized by assaying the lipids contained in a blood component in the presence of an organic silicon compound. The method can cause specific conditions for direct methods while satisfying requirements such as no influence on precision of assay, no burden on assay apparatus, and easy availability.

Abstract: An improved reagent system for preparation of cells for flow cytometry having a physiologically compatible salt solution composition including a lysing agent, an agent for minimizing white blood cell lysis, and optionally a preservative.

Abstract: The present invention relates to a therapeutic composition and uses thereof for treatment of damaged tissue, wherein the composition comprises at least one extracellular matrix compound, at least one polar surface active lipid, and a plurality of amino acids having a molar ratio which is characteristic of human breast milk protein; and further comprises a fatty acid, gamma-amino butyric acid or L-carnitine.

Abstract: The present invention relates to an assay for determining the levels of sterols, stanols, steryl esters, fatty acid derivatives and combinations thereof in a starch-containing food product. The assay is particularly useful in supporting product health and/or nutritional claims in manufacturing products intended for human or animal consumption. The present invention describes a method for extracting sterols related compounds and uses as an internal standard a steryl ester, preferably cholesterol oleate. By using the present extraction technique the process enables the recovery of substantially all of the sterol related compound in the sample.

Abstract: An electronic system for selectively detecting and identifying a plurality of chemical species, which comprises an array of nanostructure sensing devices, is disclosed. Within the array, there are at least two different selectivities for sensing among the nanostructure sensing devices. Methods for fabricating the electronic system are also disclosed. The methods involve modifiying nanostructures within the devices to have different selectivity for sensing chemical species. Modification can involve chemical, electrochemical, and self-limiting point defect reactions. Reactants for these reactions can be supplied using a bath method or a chemical jet method. Methods for using the arrays of nanostructure sensing devices to detect and identify a plurality of chemical species are also provided.

Abstract: Hydrophobically-modified proteins and methods of making them are described. A hydrophobic moiety is attached to a surface amino acid residue of the protein. The hydrophobic moiety can be a lipid or a peptide. Alternatively, the protein can be derivatized by a wide variety of chemical reactions that append a hydrophobic structure to the protein. The preferred protein is of mammalian origin and is selected from the group consisting of Sonic, Indian, and Desert hedgehog. The hydrophobic moiety is used as a convenient tether to which may be attached a vesicle such as a cell membrane, liposome, or micelle.

Abstract: Disclosed is a method for quantifying cholesterol in HDL, which does not require complex fragmentation and separation operations, and by which the HDL cholesterol in test samples containing HDL and other lipoproteins such as low density lipoprotein (LDL), very low density lipoprotein (VLDL) and chylomicron (CM) may be quantified selectively, simply and accurately. The method for quantifying cholesterol in high density lipoprotein comprises a first step of erasing cholesterol in lipoproteins other than high density lipoprotein in a test sample, and a second step of adding a surfactant which specifically acts on high density lipoprotein to the product of the first step and enzymatically quantifying cholesterol in high density lipoprotein.

Abstract: The present invention relates to a method for inhibiting the etiology of disease in patients having a disease state caused by an excess of 12-lipoxygenase or its products. In particular, the invention provides for administration of a human leukocyte 12-lipoxygenase pathway inhibitor to inhibit disease etiology, to inhibit the proliferation of breast cancer and to increase insulin receptor phosphorylation in a patient having Type II diabetics.

Abstract: A method is disclosed for determining whether a compound binds to a lipoprotein such as LDL or VLDL in a manner which will lower plasma cholesterol. The method provided includes assessing the ability of the compound to form a complex with the lipoprotein, and then determining whether the newly formed complex causes a change in the structure of apoB-100 that results in increased binding affinity to an LDL receptor.

Abstract: An assay device and method for measuring the concentration of HDL-associated cholesterol in a blood-fluid sample are described. The assay design is such that removal of non-HDL lipoproteins from a sample and assay of HDL cholesterol in the sample occur without interruption of the assay. The device also prevents interference by reagents used for the HDL assay with other assays carried out on the same sample.

Abstract: This invention provides novel assays that are prognostic and/or diagnostic for atherosclerosis or risk of atherosclerosis. It was discovered that high density lipoprotein (HDL) or components thereof can prevent the oxidation of lipids (e.g. lipids present in LDLs) and can also repair (reduce) already oxidized lipids and thereby reduce the inflammatory response associated with and characteristic of atherosclerotic plaque formation. Moreover it was a discovery of this invention that individuals vary in the ability of their HDL to afford such protection. Thus an assay of HDL protective and/or repair activity provides a highly effective assay for risk of atherosclerosis and its associated pathologies and such assays are provided herein.

Abstract: A new method for predicting the risk of coronary artery disease (CAD) is disclosed. The method uses a ratio made up of the levels of an individuals LDL-cholesterol (LDL-C), HDL-cholesterol (HDL-C) and serum total bilirubin (bilirubin). The ratio, using a weighted value for bilirubin, is LDL-C/(HDL-C+bilirubin).

Abstract: The invention provides microfabricated silicon substrates and devices having extremely small apertures (termed “nanoapertures”) and methods for producing such nanoapertures. The devices have a nanoaperture (which may have a diameter ranging from about a few millimeters to as small as a few nm) across a substrate effective to connect two regions separated by the substrate. The devices are suitable for the formation of lipid bilayer membranes across the apertures, and for use in devices such as biosensors. Substrates and devices may include multiple nanoapertures, which may each support a lipid bilayer membrane, allowing fault tolerant devices such as fault-tolerant biosensors, and allowing devices able to sense more than one target molecule.

Abstract: This invention provides a method of measuring oxidative response of cells without recourse to preparation of cell culture. The process involves: 1) preparing suspensions of cells from a living host in isotonic solutions, 2) preparing samples of test materials in isotonic solution containing tagged choline, 3) adding the cells suspension prepared in step 1 to the samples prepared in step 2, 4) incubating the product of step 3 with shaking for 2-90 minutes, 5) extracting and drying the lipid phase from the product of step 4, and 6) subjecting the product of step 5 to a scintillation counter to measure choline which has been incorporated into phosphatidylcholine (PC). An increase in incorporation of choline into PC in the short term indicates oxidative stress or free radical induced damage. Because the method of the invention using the cell isolates does not require the expense of cell culture with concomitant expense and possibility of cell change, it is particularly useful for clinical evaluation.

Abstract: The present invention provides an isolated mammalian receptor which specifically binds a high density lipoprotein holoparticle, comprising a subunit of approximately 45-600 kDa molecular weight and one or more subunits selected from the group consisting of a subunit of approximately 40-50 kDa molecular weight, a subunit of approximately 120 kDa molecular weight and a subunit of approximately 400 kDa molecular weight.

Abstract: The present invention relates to a rapid, sensitive, simple, and cost-effective spectrophotometric method of detecting and quantifying mycolic acid in a mycolic acid-fuschin dye complex with absorbance maxima ranging between 490-500 nm in the presence of various test compounds, for screening mycolic acid biosynthesis inhibitors useful as anti-microbial agents and a diagnostic kit thereof comprising basic fuschin dye in the concentration ranging between 0.1-1.0 gm/100 ml, phenol and 95% ethanol in the ratio ranging between 1:4 to 2:1 (v/v), and phenol and distilled water in the ratio ranging between 1:14 to 1:25.

Abstract: A method of simultaneously determining the concentrations of cardiovascular risk markers selected from the group consisting of High Density Lipoprotein cholesterol (HDL-C), Low Density Lipoprotein cholesterol (LDL-C), total cholesterol, triglycerides and oxidized LDL using infra-red and/or near infrared light is described

Abstract: The present invention provides an evaluation method and preparation process of injection containing a lipid A analog. Specifically, it provides, in an injection preparation containing a lipid A analog or a pharmacologically acceptable salt thereof, forecasting and evaluating methods of the pharmacokinetics of the lipid A analog, and quality evaluating method and preparation process of the injection, which each comprises measuring membrane fluidity and/or circular dichroism.

Abstract: An assay pad for measuring the concentration of HDL-associated cholesterol in a sample, a method for using the pad, and a diagnostic assay device for carrying out the method are described. The assay pad includes a polymer adhesive or heat laminate bond. The assay design prevents interference by reagents used for such removal with the HDL quantification reaction or with other assays carried out on the same sample. If desired, removal of non-HDL lipoproteins and assay of HDL cholesterol can be carried out without interruption of the assay.

Abstract: Methods, systems and kits for the simultaneous or sequential analysis of one or more hormones by mass spectrometry are disclosed. The methods require minimal sample size and minimal preparation time. The methods comprise ionizing the hormones and analyzing the hormones by mass spectrometry. In addition, methods, systems and kits for the simultaneous or sequential analysis of steroid hormones are disclosed comprising ionization of the steroid hormones by photoionization.

Abstract: The invention provides a method for identifying patients with normal NCEP lipid levels who are in need of treatment for cardiovascular disease comprising measuring one or more LDL or HDL particle subclass levels and identifying abnormal LDL or HDL subclass levels.

Abstract: The invention provides a non-invasive assay for the detection of a SLOS affected individual by determining the ratio of at least one of the specific SLOS steroid analytes, dehydro-estriol (8-DHE3) and a dehydro-pregnanetriol (7-DHPT) to their normal steroid counterparts estriol (E3) and pregnanetriol (PT), respectively. 8-DHE3 and 7-DHPT represent metabolites which accumulate in the blood and urine of an individual with SLOS or an individual carrying a SLOS affected fetus. The invention provides for a reliable and reproducible method of screening women for SLOS affected fetuses early on in pregnancy.

Abstract: The present invention relates to a method and apparatus for determining blood oxygen transport, and to measure lipid levels by correlating these levels with the rate at which oxygen diffuses through the red blood cell membrane.

Abstract: A method for determination of at least one of the lipid species in a biological sample comprising subjecting the sample to lipid extraction to obtain a lipid extract and subjecting the resulting lipid extract to multidimensional electrospray ionization mass spectrometry using either precursor ion or neutral loss scanning (or both) of all naturally occurring aliphatic chains, lipid fragments and precursor ions leading to observed fragments to generate a multidimensional matrix whose contour densities provides structural and quantitative information directly without chromatography. A method for determination of lipid content and/or lipid molecular species composition and quantity directly from lipid extracts of a biological sample comprising subjecting said lipid extract to electrospray ionization multidimensional mass spectrometry by comparisons to standards and algorithms described herein.

Abstract: An improved reagent system for preparation of cells for flow cytometry having a physiologically compatible salt solution composition including a lysing agent, an agent for minimizing white blood cell lysis, and optionally a preservative.

Abstract: The present invention relates, first, to methods for the modulation of acid sphingomyelinase (ASM)-related processes, including apoptosis. Such apoptosis can include, but is not limited to, environmental stress-induced apoptosis such as, for example, ionizing radiation and/or chemotherapeutic agent-induced apoptosis. Apoptosis can be characterized by a cellular morphology comprising cellular condensation, nuclear condensation or zeiosis. The present invention further relates to methods for the identification of compounds which modulate (i.e., either increase or decrease) sensitivity to ASM-related processes, including apoptosis.

Abstract: A chemical analysis method for determining chemically related differences between subject biological material such as genetically modified plant material and control biological material such as genetically unmodified plant material, which method includes at least the following six steps. The first step is to contact the subject biological material with a fluid extractant, such as a mixture of water, isopropanol and potassium hydroxide, to produce a fluid extract of the subject biological material. The second step is to contact the control biological material with the fluid extractant to produce a fluid extract of the control biological material. The third step is to chromatograph the fluid extract of the subject biological material, for example, gas or fluid chromatography, to produce a chromatogram of the fluid extract of the subject biological material.

Abstract: Methods are disclosed which separate and identify lipoproteins in biological samples. An ultracentrifuge density gradient is used to separate lipoprotein fractions. The fractions are visualized, resulting in a lipoprotein profile. The fractions can be further analyzed by a wide array of laboratory and clinical methods. The lipoprotein profile can be used in clinical diagnoses and other medical applications.

Abstract: A highly sensitive and specific method for the detection and quantification of lipids is provided. Specifically, methods for the simultaneous detection and quantification of phospholipids extracted from mammalian tissues is described. The analytical methods provided disclose a modified one-dimensional thin-layer chromatography technique specifically developed to rapidly and accurantely detect and quantify phospholipids from mammalian cardiac tissues.

Abstract: The present invention relates to the identification of the NSDHL gene product as a 3&bgr;-HSD participating in the conversion of 4,4-dimethylcholest-8(9)-en-3&bgr;-ol to cholest-8(9)-en-3&bgr;-ol in the cholesterol and vitamin D biosynthetic pathway. Based upon this function the present invention contemplates methods for manipulating the biosynthetic pathway at the step of involvement of NSDHL to increase or decrease the levels of cholesterol and/or vitamin D (or downstream products such as steroids) produced by a cell. Also contemplated are methods for manipulating the accumulation of intermediate compounds upstream of the step of NSDHL in the pathway.

Abstract: The invention provides a class of samples that model the human body. This family of samples is based upon emulsions of oil in water with lecithin acting as the emulsifier. These solutions that have varying particle sizes may be spiked with basis set components (albumin, urea and glucose) to simulate skin tissues further. The family of samples is such that other organic compounds such as collagen, elastin, globulin and bilirubin may be added, as can salts such as Na+, K+and Cl−. Layers of varying thickness with known index of refraction and particle size distributions may be generated using simple crosslinking reagents, such as collagen (gelatin). The resulting samples are flexible in each analyte's concentration and match the skin layers of the body in terms of the samples reduced scattering and absorption coefficients, &mgr;'s and &mgr;a. This family of samples is provided for use in the medical field where lasers and spectroscopy based analyzers are used in treatment of the body.

Abstract: The present invention provides a fluorescent HPLC assay for detecting the presence and/or measuring the level of 20-hydroxyeicosatetraenoic acid (20-HETE) and other P-450 metabolites of arachidonic acid in a sample. P-450 metabolites of arachidonic acid are first extracted from the sample and then labeled with 2-(2,3-naphthalimino)ethyl trifluoromethanesulfonate. The labeling reaction is catalyzed by N,N-diisopropylethylamine. Next, the labeled P-450 metabolites are separated on a 4.5×250-mm, 5 &mgr;M particle size C18 reverse-phase HPLC column using a mobile phase of methanol:water:acetic acid (82:18:0.1, v/v/v) and an isocratic elution at a rate of about 1.3 ml per minute. Fluorescence intensities of the column eluent are monitored by a fluorescence detector. Quantitation of P-450 metabolites in a sample can be made by using an internal standard.

Abstract: The present disclosure relates to a method for determining cholesterol in low density lipoprotein comprising the steps of (a) measuring total cholesterol level in a sample containing at least high density lipoprotein, low density lipoprotein, very low density lipoprotein and chylomicron, and (b) measuring cholesterol levels in the high density lipoprotein, very low density lipoprotein and chylomicron in the sample, wherein the cholesterol level in the low density lipoprotein is determined by subtracting a value obtained in the step (b) from a value obtained in the step (a). The present invention enables concurrent determination of cholesterol level in low density lipoprotein and total cholesterol level, facilitating acquisition of two types of biological information at a time.

Abstract: Amounts of components in a specimen can be analyzed with excellent quantitativity. The analysis includes: measuring an amount of a component to be analyzed in a specimen; measuring an amount of a standard component present originally and homeostatically in the specimen other than the component to be analyzed; determining the amount of the specimen from the amount of the standard component thus measured and a known concentration of the standard component in the specimen; and determining a concentration of the component to be analyzed in the specimen from the amount of the specimen thus determined and the amount of the component to be analyzed thus measured. The quantitative analysis of the present invention allows a component to be analyzed to be measured with high quantitativity as shown in FIG. 1.

Abstract: Methods are disclosed which separate and identify lipoproteins in biological samples. An ultracentrifuge density gradient is used to separate lipoprotein fractions. The fractions are visualized, resulting in a lipoprotein profile. The fractions can be further analyzed by a wide array of laboratory and clinical methods. The lipoprotein profile can be used in clinical diagnoses and other medical applications.

Abstract: The invention presents a method designed to simultaneously measure certain unsaturated lipids and certain vitamins present either as single substances or in complex mixtures such as exist in serum and natural oils. Target lipids are free cholesterol, unsaturated cholesteryl esters; free polyunsaturated fatty acids, and their esters as triglycerides, and phospholipids. Distributions of these analytes over the broad range of serum lipoproteins from chylomicrons to high density fractions are determined using a procedure that involves a single step reaction in which the molecular unsaturations are subjected to non-enzymatic color inducing reagents. For natural oils and vitamins, the same method serves as a quality control procedure. Analytical detection is achieved using broad spectrum absorbance and/or fluorescence measurements. Measured spectra are aggregates of the absorbance contributions from each of the analytes. Data analyses follow two paths. One uses raw spectral data.

Abstract: One aspect of the present invention is a multi-well platform for fluorescence measurements, comprising a plurality of wells within a frame, wherein the multi-well platform has low fluorescence background. Another aspect of the present invention is a system for spectroscopic measurements, comprising reagents for an assay and a multi-well platform for fluorescence measurements. A further aspect of the present invention is a method for detecting the presence of an analyte in a sample contained in a multi-well platform by detecting light emitted from the sample. Another aspect of the present invention is a method from identifying a modulator of a biological process or target in a sample contained in a multi-well platform by detecting light emitted from the sample. Another aspect of the present invention is a composition identified by this method. A further aspect of the present invention is a method to identify a therapeutic.