Abstract: A method for the rapid estimation of hyperplastic and hypertrophic changes in animal airways is an assay which specifically measures acidic and neutral mucoproteins in a linear fashion from 0.5 to at least 10 .mu.g. The assay comprises exposure of a test animal to a suspected metaplastic inducer, removal of the lungs, homogenization in an appropriately buffered solution containing reducing agents and protease inhibitors; removal of particulate matter; and size-fractionation of the SDS treated soluble extract. The high molecular weight material is immobilized and stained for either acidic or neutral mucosubstances and the specific staining is quantitated. The changes observed are consistent with those seen in histological sections of the exposed tissues.

Abstract: A method for determining the complexed forms of immunologically determinable prostate specific antigen (cPSA) in a blood sample, e.g., by two-site immunometric assays, in which the blood sample is treated to render free PSA (fPSA) immunologically nondetectable. A particularly preferred immunometric assay method employs three anti-PSA antibodies: an antibody that binds to both cPSA and fPSA (anti-tPSA), a second anti-tPSA antibody which is characterized by the unique property that binding to fPSA is blocked by binding of fPSA-specific antibodies, and a third antibody which is a fPSA-specific antibody. Thus, binding of the fPSA-specific antibody to PSA in the sample allows only cPSA to be measured in the immunometric assay. Measurement of cPSA blood levels has been found to provide a method for aiding in the diagnosis and monitoring of prostate cancer that is highly sensitive and specific, and eliminates the need for a significant number of patients to undergo unnecessary prostate biopsy.

Abstract: The present invention provides a method of using a reflectance-reading device to determine an initiation time point for measuring the chemical reaction of an analyte from a biological liquid sample on a test surface. The initiation time point is determined by using a device to read a reflectance of a test surface at a plurality of time points, and calculating the K/S ratio of the test surface at each time point according to the Kubelka-Munk equation. As the device continues to calculate a K/S ratio for each time point, the device monitors the rate of change of the K/S ratio with respect to time. The device then determines the initiation time point to be when the rate of change of the K/S ratio is maximal. The present invention also provides a method of measuring the concentration of an analyte on a test surface. The concentration is measured by determining an initiation time point according to the method described above and then measuring the concentration of the analyte at a variable end point.

Abstract: A deepithelialized skin cell diffusion system which can be used to select topical gel and cream formulations containing human other wound healing promoters such as plasma fibronectin. The formulations provide slow release and increased contact time of fibronectin or other wound healing promoters to the wound site leading to its effective absorption.

Abstract: A microscale fluid handling system that permits the efficient transfer of nanoliter to picoliter quantities of a fluid sample from the spatially concentrated environment of a microfabricated chip to "off-chip" analytical or collection devices for further off-chip sample manipulation and analysis is disclosed. The fluid handling system is fabricated in the form of one or more channels, in any suitable format, provided in a microchip body or substrate of silica, polymer or other suitable non-conductive material, or of stainless steel, noble metal, silicon or other suitable conductive or semi-conductive material. The microchip fluid handling system includes one or more exit ports integral with the end of one or more of the channels for consecutive or simultaneous off-chip analysis or collection of the sample. The exit port or ports may be configured, for example, as an electrospray interface for transfer of a fluid sample to a mass spectrometer.

Abstract: A method and apparatus for the quantitative analysis of sample liquids. A sample is dried and irradiated with visible and/or infrared light. Light that is diffusely or specularly reflected from the sample and sample carrier is detected. and analysed.

Abstract: The present invention provides a method for treating the inflammatory response of an established colitis in a subject with inflammatory bowel disease (IBD), comprising administering to a subject diagnosed with an established colitis from an IBD an amount of an antibody to interleukin-12 effective in reducing the colitis-inducing effect of interleukin-12. Also provided is a method for screening a substance for its effectiveness in reducing the inflammatory response of an established colitis comprising obtaining an animal having an established colitis; administering the substance to an animal; and assaying the animal for an effect on interleukin-12 which results in the reduction of the inflammatory response of the colitis, an amount of reduction of the inflammatory response greater than the amount of reduction produced by the administration of antibodies against interferon-gamma or tumor necrosis factor-alpha indicating an effective substance.

Abstract: Methods are described for the identification and preparation of nucleic acid ligands to CG-related glycoprotein hormones. Included in the invention are specific RNA ligands to hCG and hTSH identified by the SELEX method.

Abstract: A glycoprotein having a molecular weight of 28,000 to 32,000 and an isoelectric point of 7.0 to 9.0, both as determined by two-dimensional SDS polyacrylamide gel electrophoresis, is synthesized and secreted specifically by stromal cells of endometriotic origin. Amino acid residues in the region of the N-terminus of the glycoprotein share amino acid sequence identity with a region of tissue inhibitor of metalloproteinases-1 (TIMP-1). A method of screening for endometriosis is disclosed by detection of lower levels of the protein or TIMP-1 in peritoneal fluid or serum samples of women.

Abstract: A method for continuous separation and analysis of glycated and non-glycated proteins in a blood sample by HPLC using a phenylboronic acid resin and a buffered polyol as eluent.

Abstract: A method for antenatal screening for chromosomal and other abnormalities in an unborn child is determined by measuring the gestational age discrepancy of the pregnant mother. This data is determined (a) by reference to the last menstrual period, and (b) a biometric measurement of the fetus. The difference between the ages as determined using (a) and (b) is calculated. This calculated difference is then examined using reference data to determine fetal abnormalities. These data can also be used with assay of maternal fluids for various pregnancy markers.

Abstract: The present invention relates to a novel clinical examination method which comprises analyzing the alteration in the amount of a specific component in oligosaccharides of immunogloblin G. Human diseases such as liver diseases, malignant hypertension, immunogloblin A-nephropathy, pediatric disorders, etc., are examined in terms of the alteration in bisecting N-acetylglucosamine-containing oligosaccharides in immunogloblin G from collected humor. The present invention provides highly accurate information in a manner applicable to practical operation for examination of liver diseases, allergic diseases, malignant hypertension, immunogloblin A nephropathy, pediatric disorders, as well as aging-dependent variations and the therapeutic effect of interferon.

Abstract: A method of detecting nucleic acids, proteins, or protein nucleic acid complexes. The method includes binding an enzyme, such as phosphatase, to a specimen of the nucleic acid, protein, or protein nucleic acid complex. The enzyme is then reacted with a fluorescein derivative phosphate ester to obtain a fluorescein derivative phosphate ester hydrolysate. The hydrolysate is then irradiated with excitation light, and the emitted fluorescein is detected.

Abstract: Methods for detecting abnormalities in prolactin daily rhythms of a subject are provided. Prolactin levels of a subject are compared to levels of healthy subjects and based on the comparison a determination is made of the adjustments necessary to normalize the subject's daily prolactin rhythm. Also provided are methods for normalizing a subject's daily prolactin rhythm.

Abstract: The present invention relates to advanced glycosylation endproducts, and particularly to the use of novel cyclopentenone aminoreductones, 3-alkylamino-2 -hydroxy-4-hydroxymethyl-2-cyclopenten-1-ones. Such AGEs can be used in various diagnostic and therapeutic methods.

Abstract: Methods for detecting heat-stressed wheat, that is, wheat that has experienced elevated temperatures during the grain filling period, and methods to assess end-use properties of wheat grain are disclosed. In the method to detect heat-stressed wheat, wheat heat stress peptide in a sample of wheat grain or flour is measured. Wheat grain or flour that has a level of wheat heat stress peptide two or more times greater that the constitutive level is determined to have experienced elevated temperatures during the grain filling period. In the method to assess an end-use property of wheat, wheat heat stress peptide in a sample of wheat grain or flour is measured, and the level is compared to a calibration curve that correlates the level of wheat heat stress peptide and the end-use property.

Abstract: A novel method for monitoring the success or the yield of chemical reactions involving a reactant bound to a solid phase support, or to forecast the success of such reactions, or to quantify the number of deuterium containing groups present in a solid-phase bound sample, using infrared spectroscopy and deuterium-carbon absorbances, along with novel compounds and intermediates useful for carring out the method.

Abstract: Method for detecting casts in urine by measuring Tamm-Horsfall protein by solid and liquid phase reagents including a method for manufacturing a enzyme specific for Tamm-Horsfall protein which produces a detectable response in the presence of casts in urine.

Abstract: The present invention provides a thrombin-activated platelet protein (TAPP). The protein is selectively expressed on the surface of thrombin-activated platelets. Antibodies which selectively bind to the thrombin-activated platelet protein are also provided. These compositions find use in the detection and treatment of blood clots.

Abstract: A pyridinoline composition in which pyridinoline is derivatized specifically at its aliphatic hydroxyl group by a selected chemical group is disclosed. In various embodiments, the composition may be used as a standard for HPLC or immunoassay of pyridinoline, a pyridinoline immunogen for producing anti-pyridinoline antibodies, and a solid-phase reagent for use in an immunoassay kit. Also disclosed are methods for making and using the composition.

Abstract: The present invention provides a method and apparatus for rapid measurement of a fluid bulk analyte, requiring only microscale volumes. Several fluid bulk analytes can be measured simultaneously and, for biological samples, the cell content can also be measured simultaneously. The invention comprises reporter beads for chemical analysis of fluid bulk properties such as pH, oxygen saturation and ion content. Each reporter bead comprises a substrate bead having a plurality of at least one type of fluorescent reporter molecules immobilized thereon. The fluorescent properties of the reporter bead are sensitive to a corresponding analyte. Reporter beads are added to a fluid sample and the analyte concentration is determined by measuring fluorescence of individual beads, for example in a flow cytometer. Alternatively, reporter molecules which change absorbance as a function of analyte concentration can be employed.

Abstract: In order to quantitatively measure an Amadori compound by a simple optical method utilizing light scattering, a sample containing an Amadori compound which is stored in a cell is irradiated with excitation light from an He--Ne laser unit so that scattered light from the sample is received and separated into its spectral components for obtaining a light scattering spectrum, and a light scattering peak existing at 820 to 840 cm.sup.-1, 1655 to 1660 cm.sup.-1, 2000 to 2020 cm.sup.-1, 2080 to 2100 cm.sup.-1, 2460 to 2470 cm.sup.-1 or 2530 to 2600 cm.sup.-1 in shift wavenumber with respect to the excitation wavelength in the light scattering spectrum is detected by a detector. The saccharide concentration or the saccharification ratio of the Amadori compound is measured by a calibration curve through the peak intensity or the peak integral value of the light scattering peak.

Abstract: The invention is directed to methods to assess connective tissue, especially bone, metabolism in disease or to monitor therapy, which method comprises assessing the levels of native free collagen-derived crosslinks in biological fluids, especially urine. The method can be enhanced by concomitantly determining the levels of an indicator of bone formation in biological fluids of the same individual and assessing the differences between the degradation marker and the formation indicator. Antibodies which are specifically immunoreactive with forms of crosslinks which occur free in biological fluids are also disclosed.

Abstract: The invention is directed to methods to assess connective tissue, especially bone, metabolism in disease or to monitor therapy, which method comprises assessing the levels of native free collagen-derived crosslinks in biological fluids, especially urine. The method can be enhanced by concomitantly determining the levels of an indicator of bone formation in biological fluids of the same individual and assessing the differences between the degradation marker and the formation indicator. Antibodies which are specifically immunoreactive with forms of crosslinks which occur free in biological fluids are also disclosed.

Abstract: Methods of diagnosis of gestational diabetes mellitus are disclosed. In preferred embodiments, a blood sample is obtained from a pregnant female in the 24th to 28th week of pregnancy after an overnight fast, after a 1-hour 50-gram glucose challenge test, or at the 1-hour time point during a 3-hour 100-gram oral glucose tolerance test. The concentrations of fasting plasma glucose and glycosylated plasma proteins in this blood sample are then determined. A fasting plasma glucose concentration equal to or exceeding 90 mg/dL is 100% sensitive and 64% specific in predicting glucose-related macrosomia (i.e., birth weight above 4000 grams). A glycosylated plasma protein concentration equal to or exceeding 23% is 100% sensitive and 52% specific in predicting glucose-related macrosomia. A fasting plasma protein value equal to or exceeding 90 mg/dL and a glycosylated plasma protein value equal to or exceeding 23% is 100% sensitive and 93% specific in predicting glucose-related macrosomia.

Abstract: The invention relates to measuring the decrease in phosphorylation of the fatty acid binding protein (FABP) found in the milk fat globule membrane (MFGM) of bovine or human milk to detect growth hormone treatment in animals. MFGM of cows treated with recombinan+bovine somatotropin (bST) displays weaker FABP autophosphorylation activity than non treated cows. MFGM isolated from cows treated with bST have significantly reduced levels of phosphorylated FABP. The bST test of the present invention can be used not only to determine whether the animal producing the milk has been treated with bST, it can also be used to determine the efficacy of bST on milk production. Dairy managers could thus base their decision on whether to continue bST treatment on such a test.

Abstract: The present invention describes the use of a variety of merocyanine dyes and substituted merocyanine dyes for detecting and quantifying poly(amino acids), including peptides, polypeptides and proteins. The labeled proteins or peptides are highly colored, but are also detected by their strong fluorescence enhancement. Poly(amino acids) are detected in solution, in electrophoretic gels, and on solid supports, including blots and dipsticks. The present method of staining is highly sensitive, extremely facile, and relatively non-selective.

Abstract: An improved method for the mass spectrometric determination of the molecular weight of a highly polyionic analyte is provided. The method employs reagents which are highly polyionic but which are of opposite charge to the analytes. The reagents and analytes form a non-covalent complex which is more easily ionized during mass spectrometry and decreases fragmentation of the analyte. Highly polyionic reagents are also provided. The reagents include a multiplicity of highly ionic groups covalently attached along a flexible molecular backbone.

Abstract: The invention concerns a method for the quantitative analysis of sample liquids. A sample is dried and irradiated with visible and/or infrared light. Light that is diffusely or specularly reflected from the sample and sample carrier is detected and analysed. Furthermore the invention concerns a system for carrying out the method according to the invention and a sample carrier having a diffusely or specularly reflecting surface.

Abstract: A Dot-Blot assay ("spot test") with Bis-N,N,dioctadecylamide (BDA.TDA) as antigen was developed to detect anti-BDA.TDA antibodies in tuberculosis patients. To develop the antigen-antibody reaction, as a first step and in order to enhance the reaction, an anti-human rabbit serum was used followed by incubation with a protein A-colloidal gold conjugate. This assay showed almost equal sensitivity and specificity as the .beta.-galactosidase ELISA test which was conducted in parallel. This simple and fast assay could be used in places where ELISA equipment is not easily available.

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

Abstract: Disclosed are methods for detecting abnormalities in prolactin daily rhythms. The methods involve comparing a prolactin profile of a vertebrate (including a human) subject being tested that has been compiled over a predetermined period to a predetermined standard prolactin profile for healthy subjects. The method also involves determining whether the vertebrate subject has an abnormal daily prolactin rhythm by ascertaining whether (i) at any point during waking hours the prolactin level of the subject being tested is greater than 1 SEM above the normal prolactin profile of said healthy subjects, and/or (ii) at any point during sleeptime the prolactin level of the subject being tested is at least 1 SEM lower than the normal prolactin profile of said healthy subjects. Also provided are methods for determining adjustments needed to an abnormal prolactin profile (or daily rhythm) to cause it to conform to the prolactin profile (or rhythm) of a healthy subject, and methods for effecting such adjustments.

Abstract: An improved method of testing individuals for diabetes, even if they have levels of interfering substances (e.g., uric acid, bilirubin, and glutathione) that would otherwise interfere with such testing, is disclosed. The individual's protein-bound glucose level and glucose level are compared to the analogous values for a reference population to enable the risk of that individual's having diabetes to be assessed. The substances that would otherwise tend to interfere with the assay for protein-bound glucose are removed before the assay, desirably by precipitating the protein-bound glucose using uranyl acetate, which desirably leaves substantially all of the interfering substances in the supernatant, then separating the precipitate from the supernatant, redissolving the precipitate, and conducting the colorimetric assay on the resulting solution. An improved colorimetric test for protein-bound glucose using viologens as the colorimetric electron acceptors is also disclosed.

Abstract: Described are methods for determining the potency of somatotropins. The level of biologically active bovine somatotropin protein in a bovine somatotropin sample is measured by size exclusion HPLC employing as the stationary phase a hydrophilic porous gel having an average particle diameter of about 5 .mu.m to about 15 .mu.m and the a mobile phase a buffered aqueous solution which is non-denaturing to the bovine somatotropin sample. The potency of the bovine somatotropin sample is determined based upon the level of biologically active bovine somatotropin protein so measured. In other embodiments, somatotropin is provided dissolved in a first buffer solution, and then chromatographed in a mobile phase comprised of a second buffer solution having a pH lower than the first buffer solution, to achieve an effective separation of biologically active protein from biologically-inactive large non-covalent soluble aggregates.

Abstract: A multilayer analytical element for assaying fructosamine having a liquid-impermeable support, a dried buffer-containing layer which contains a buffer having pH of 8 to 12 formed on the support, and a tetrazolium salt-containing layer which is laminated on the buffer-containing layer, which element assays fructosamine in a short time with good accuracy.

Abstract: A system for determining the concentration of fructosamine in sera which consists of a first reagent in which a tetrazolium salt which reduces all reactive substances in sera including fructosamine and a second reagent which is responsive to all reactive substance in sera other than fructosamine. The difference in color change as between the two allows the determination of concentration of fructosamine.

Abstract: A rapid diagnostic method for detecting the rupture of fetal membranes is disclosed. The presence of insulin-like growth factor binding protein 1 (IGFBP-1) in a vaginal secretion sample, resulting from the rupture of fetal membranes, is detected with the aid of at least one specific binding substance for IGFBP-1 .

Abstract: The present invention relates to a gelatinase A inhibitor comprising as an active ingredient a peptide analogue consisting of an active minimum unit of gelatinase A inhibition obtained from APP (.beta.-amyloid precursor) or a peptide analogue comprising it. Gelatinase A can be qualified and quantified using any of the gelatinase A inhibitors according to the present invention.

Abstract: Antibodies against highly conserved amino acid sequences of immunogenic substances, a process for the preparation of these antibodies and the use thereof in immunoassays.The invention relates to antibodies which are obtained by immunization with a peptide fragment which represents a highly conserved amino acid sequence of a native protein. The antibodies according to the invention can be used for the preparation of immunoassays, in particular for the preparation of assays for the determination of genetically engineered products such as insulin which arise as sparingly soluble inclusion bodies in microorganisms. The invention particularly relates to a multispecies insulin assay in the form of an RIA.

Abstract: A method for preparing a solventless protein standard is disclosed in which a solventless dye indicator and a predetermined amount of solventless protein are placed in an appropriate receptacle such as the well of a multiwell plate. This results in a solventless protein standard in which the solventless protein and the solventless dye are contained within the same receptacle. When an appropriate solvent is added to the receptacle, the protein and dye react together to produce a color change which is detectable, and which can be used as a standard for a protein assay using that dye. Also claimed is the solventless protein assay standard produced by the process.

Abstract: Processes are provided for pretreating body fluid compositions and subsequently analyzing the pretreated body fluid compositions for analytes of interest. Processes for pretreating the compositions include providing size exclusion gel having a molecular weight fractionation range or a molecular weight exclusion such that the size exclusion gel is capable of excluding or fractionating the analytes of interest, and then causing the composition to contact the size exclusion gel in order to separate the analytes from low molecular weight composition components which interfere with the separation and analysis of the analytes of interest. Processes for analyzing pretreated compositions include electrophoretic methods such as capillary zone electrophoresis which involve the separation and detection of analytes of interest.

Abstract: The present invention is directed to use of an imidazo [4,5b] pyridinium molecule composed of a lysine and an arginine residue crosslinked with a pentose sugar for assessing the biological age of a tissue.

Abstract: A modification of the standard BCA protocol is provided that allows sensitive and reliable protein determinations in a matter of seconds utilizing a microwave oven to irradiate the samples. Methods of determining protein concentrations in a sample are disclosed comprising the steps of combining the sample with a BCA assay reagent in a sample container, placing the sample container into a microwave oven, irradiating the sample and measuring an absorbance value at 562 nm for the sample. Protein concentrations are then determined by comparing the absorbance value with a known value based on a calibration curve. In preferred embodiments, the calibration curve is generated by placing one or more standards in the microwave oven and irradiating the standards. Preferably, the step of irradiating the sample is carried out for a duration of less than about 60 seconds and most preferably, for a duration of about 20 seconds.

Abstract: The present invention is directed to a multi-layer test device and method for analyzing the concentration of fructosamine in a liquid sample. The multi-layer test device has a buffer layer containing a buffer having a pH value of at least 9 which is either superposed above or juxtaposed to an indicator layer containing an indicator capable of being reduced by fructosamine. Supporting the buffer layer, the indicator layer, and any additional layers in the multi-layer device is at least one support member which optionally has a detection aperture for analyzing the concentration of fructosamine on the indicator layer. An additional support member having a sample aperture can be used. Where two support members are used the multi-layers are sandwiched between the first support member having a sample aperture and the second support member optionally having a detection aperture.

Abstract: Character of the envelope transmembrane protein of human immunodeficiency virus type 2 (HIV-2) was carried out using murine polyclonal and monoclonal antibodies or patient sera specific for HIV-2 proteins. A 80-Mr glycoprotein (gp80) was produced in HIV-2 infected cells along with three other glycoproteins that were recently reported: the extracellular glycoprotein (gp125), the envelope glycoprotein precursor (gp140), and the transient dimeric form of gp140 (gp300). The gp125 and gp80 were detectable after the synthesis of gp140 and the formation of gp300. Among these four glycoproteins, only gp80 and gp125 were associated with HIV-2 virions. As the other glycoproteins, gp80 was recognized by all HIV-2 positive sera. A murine polyclonal antibody raised against the purified gp300 recognized all four glycoproteins.

Abstract: The present invention provides an early, biochemical indication of increased risk of impending delivery. The method is particularly useful to identify those pregnant women who are at increased risk for preterm delivery and can also be used to identify those women at risk for a post-date delivery. The method comprises obtaining a cervicovaginal secretion sample from a pregnant patient after week 12 of gestation and determining the level of total fibronectin in the sample. The presence of an elevated fibronectin level in the sample indicates an increased risk of delivery. The test detects greater than 80% of women who deliver prematurely, as early as two to three weeks prior to delivery.

Abstract: The invention provides a method for extending the plasma half-life of heparin-binding proteins by coadministering such proteins with a therapeutically acceptable compound capable of inhibiting their binding to a low affinity heparin-like binding site on the surface of cells. In one embodiment of the invention, the-heparin-binding protein is a selectin. The binding inhibitory compound can, for example, be a purified native heparin preparation, a heparin fragment, or another polyanionic compound, such as dextran sulfate, heparan sulfate, pentosan sulfate, or hyaluronate.

Abstract: An improved method for the N-terminal sequential degradation of proteins and peptides is disclosed. The protein or peptide to be sequenced is reacted with a compound effective to impart a tertiary amine functionality to the thiazolinone derivative of a cleaved terminal amino acid of the peptide.

Abstract: A method is provided for detecting carbohydrate-deficient glycoproteins in samples taken from subjects with metabolic disorders, such as alcohol abuse and subjects who display a syndrome of carrying abnormal levels of carbohydrate deficient glycoproteins. The method involves steps of reglycosylating with a fluorescent-conjugate deglycosylated glycoproteins in a sample of body fluid from a subject. A further step involves fluorometric detection of fluoresceinylated carbohydrates incorporated into truncated serum glycoproteins.

Abstract: The detection of protein is accomplished using novel composition and method involving phenolsulfonephthalein protein error indicator, buffer and an aliphatic ether-polycarbonate present in an amount equal to or less than ten percent by weight.