Abstract: The present invention provides a method and a composition for detecting the levels of a plurality of biomolecular probes in a sample. In particular, the invention relates to a hybridization composition for detecting the presence or levels of different polynucleotide sequences in a sample.

Abstract: A method is disclosed for the rapid diagnosis of disorders characterized by an ischemic event, such as stroke, transient ischemic attack, head trauma, myocardial infarction or other insults resulting in interrupted cranial blood flow. The method involves detection of the presence of the gamma isoform of protein kinase C (PKCg) in peripheral blood, which signals damage to central nervous system tissue and at least transient breakdown of the blood brain barrier. The assay may be performed, e.g., by emergency medical personnel, in a time frame that allows treatment of the patient before permanent damage to the central nervous system occurs.

Abstract: Methods relating to the field of biochemistry and more specifically relating to microparticles for use in diagnostics, therapeutics, and research are provided.

Abstract: This invention provides a novel fluorescent article comprising a core or carrier particle having on its surface a plurality of smaller polymeric particles or nanoparticles, which are stained with different fluorescent dyes. When excited by a light source they are capable of giving off multiple fluorescent emissions simultaneously, which is useful for multiplexed analysis of a plurality of analytes in a sample. The coupled complex particles carrying on their surface fluorescent nanoparticles, methods of preparing such polymer articles, and various applications and methods of using such particles are claimed.

Abstract: The present invention relates to a method of carrying out an immunoassay in a multiphase system. A sample containing an analyte is brought into contact with a receptor A and a tracer. The analyte can either form a complex with the tracer, or counteract the formation of a complex of receptor A and tracer by competing with the tracer for binding to receptor A, or counteract the formation of a complex of receptor A and tracer by competing with receptor A. Receptor B is added and the signal is determined. In this method, receptor A and receptor B are suitably immobilized, ensuring that the tracer either cannot enter into any binding involving the simultaneous participation of receptors A and B or can enter into such a binding to only such a slight extent that it is nevertheless possible to detect and differentiate differing analyte concentrations.

Abstract: A combinatorial chemical library comprising a plurality of members of the Formula [S]—C(O)—L′-Z containing hydroxyamides is disclosed, in which [S] represents a solid support and L′-Z is a linker/compound residue. In this library, Z is and Y is —NR3R4 or —OR9. R3 and R4 are independently selected from the group consisting of H, alkyl, aralkyl, heteroarylalkyl, heterocycloalkyl, —CH(R5)CH(R6)OH, —CH(R5)C(O)NHR6 and —CH(R5)C(O)NHCH(R6)C(O)NHR7. R3 and R4 together are wherein X=O or NR8 and R9 is aryl, aralkyl or R7CH═CH(CH2){overscore (n)}. The combinatorial library can be optionally encoded with identifiers T′-L, which are covalently attached to the solid support. The methods of synthesizing such a library are described. The use of such library in assays to discover biologically active compounds is also disclosed.

Abstract: A microparticle light scattering agglutination assay is disclosed. The assay comprises a mixture of particles having strong light scattering properties with particles having weak light scattering properties. The particles having strong light scattering properties carry a binding partner of high reactivity with the analyte. The particles having weak light scattering properties carry a binding partner of low reactivity with the analyte. Reagents useful in the assay are also disclosed.

Abstract: A method for isolating an active catalyst from a library of compounds that are potential catalysts is disclosed. The method involves providing a library which comprises a plurality of discrete solid supports, each solid support having a different organic compound bound thereto; and providing a reaction solution in a reaction vessel, the reaction solution containing the reactant or reactants necessary for a chemical reaction to occur in the presence of a catalyst for that reaction. The library and the reaction solution are then combined in the reaction vessel, and then one of the discrete solid supports is detected that is characterized by a temperature change in said solution greater than the temperature change of a plurality of other of said discrete solid supports in said solution. The detected solid support carries an active catalyst for the chemical reaction. Continuous flow apparatus for carrying out the method is also disclosed.

Abstract: Disclosed are an optical flow particle apparatus and method for conducting a particle light scatter-based immunoassay for simultaneously measuring the presence or amount of one or more analytes in a fluid sample, which involves the steps of:

Abstract: A biological fluid collection container comprising a cup member, a lid assembly removably mounted to the cup member comprising a housing with a downwardly extending cylindrical skirt, a luer lock with a throughgoing bore extending from one side of the lid housing. A hollow tube extends from the other side of the lid housing adjacent the luer lock and is axially aligned with the throughgoing bore of the luer lock. The hollow tube is provided with a plurality of throughgoing holes leading into its lumen along its surface to provide for a sampling along various liquid level layers of the biological fluid specimen collected in the cup member so that when the biological fluid specimen is removed from the cup member a representative sampling is obtained.

Abstract: The present invention provides monoclonal antibodies specific for kringle 5 of apo(a) and hybridomas secreting such antibodies. The invention also relates to assay methods for directly measuring concentrations of lipoprotein(a) [Lp(a)] in a plasma sample. In one embodiment, the method involves the specific capture of Lp(a) from a plasma sample with a monoclonal antibody developed against kringle 5 of apo(a), which is non-cross-reactive with plasminogen and kringle 4 of apo(a). The quantity of the Lp(a) present in the sample is then measured by detecting the amount of Lp(a)-anti-kringle 5 complex that has formed in the reaction. Alternatively, the Lp(a) may be captured non-specifically and then detected with the monoclonal antibody specific for kringle 5 of apo(a). The invention also provides competitive assays using the above-mentioned kringle 5 specific monoclonal antibodies.

Abstract: A method for rapid purification of a blood component from blood is described in which the blood plasma is first separated from the cellular blood elements by any conventional means, such as centrifugation. An affinity cartridge is then activated with a molecule, such as an amino acid, which binds with a blood component such as plasminogen. The separated blood plasma is then passed through the affinity cartridge such that the blood component is retained by the affinity cartridge. Thereafter, the blood component is eluted from the affinity cartridge by passing a buffer solution containing a releasing agent through the affinity cartridge. This releasing agent disengages the blood component from the affinity cartridge. The releasing agent is then separated from the eluted solution by passing the eluted solution through a device, such as an ion exchange or size exclusion device. The separated blood component, e.g.

Abstract: Disclosed are an optical flow particle apparatus and method for conducting a particle light scatter-based immunoassay for simultaneously measuring the presence or amount of one or more analytes in a fluid sample which involves the use of a reagent set for each analyte including first binding molecule-coated monodisperse microspheres and second binding molecule-coated colloidal particles in which at least one of the first or second binding molecules specifically binds a respective one of the analytes. In the case where more than one analytes are detected, each monodisperse microperse microsphere of a particular reagent set has a light scatter signal resolvable from that of microspheres of any other reagent set. Changes determined in the distributions of the measured light scatter signals for individual microspheres of each of the particular reagent sets are indicative of the presence or amount of the respective analyte(s) in the sample.

Abstract: A method for analyzing blood enables one to isolate, detect, enumerate and confirm under magnification the presence or absence of target cancer cells and/or hematologic progenitor cells which are known to circulate in blood. The analysis is performed in a sample of centrifuged anticoagulated whole blood. The analysis involves both morphometric and epitopic examination of the blood sample while the blood sample is disposed in a centrifuged blood sampling tube. The epitopic analysis of the presence or absence of cancer cells relies on the detection of epitopes which are known to present only on cancer cells; and the epitopic analysis of the presence or absence of hematologic progenitor cells relies on the detection of epitopes which are known to present only on hematologic progenitor cells.

Abstract: An immunochromatography-assisted device is disclosed which facilitates accurate and rapid detection of an antigen contained in a fluid sample and from which background coloring and blank coloring possibly causing a malfunction at detection are successfully eliminated. The immunochromatography-assisted device comprises a porous carrier having an additive-impregnated part between a sample introducing part and a determining part, the additive-impregnated part carrying at least one selected from the group consisting of a surfactant, a water-soluble ammonium salt and a pH buffer such that it dissolves in a sample introduced into the carrier in order to move with the movement of the sample through the carrier. The porous carrier also has a labelled part segmented into plural zones.

Abstract: The invention relates in part to the use of independent assay controls (IACs) for the optical communication between an assay device and an instrument in monitoring and performing assays, preferably immunoassays.

Abstract: The invention relates to a method for detecting specific target-cells in a simple and time saving way, using paramagnetic particles, antibodies recognizing the Fc portions of target-cell associating antibodies and target-cell associating antibodies directed to specific antigen determinants in the target-cell membranes. Incubation of the cell suspension with a mild detergent and/or second set of antibodies or antibody fragments, prelabeled or not with fluorescent agents, metallocolloids, radioisotopes, biotincomplexes or certain enzymes allowing visualization, with dramatically increase the specificity of the method. The method can further be used for isolation of the target-cells by magnetic field application and kit for performing the method according to the invention is described.

Abstract: Disclosed is a method for determining the concentration of an analyte in a fluid test medium by use of an immunochromatographic test strip through which the test fluid can flow by capillarity. The test strip has at least two capture bands and optionally one or more collection bands which capture labeled anti-analyte antibody to provide a detectable signal. The signals from the label is quantitatively detected in each of the bands to provide a pattern of signals which is unique to the concentration of analyte in the test fluid. The pattern of signals are mathematically combined to create a monotonous dose-response curve to thereby factor out the high analyte hook effect which can be present in this type of assay.

Abstract: In order to extend the measuring range and to reduce the Hook effect in an immunological method of determination of an antigen based on the principle of a sandwich assay it is preferable to use a method which is characterized in that the labelled detection molecule is an oligomer of a binding molecule selected from antibody or/and antibody fragment.

Abstract: The present invention relates to a simple immunochemical semi-quantitative assay method according to chromatography, which comprises trapping a certain amount of an analyte in a sample with a predetermined amount of a fixed antibody for the analyte before qualitative analysis of the analyte, the certain amount corresponding to the amount of the fixed antibody, and thereby decreasing a concentration of the analyte to be subjected to subsequent immunochemical qualitative determination, and an apparatus therefor.

Abstract: Assay systems and specialized antibodies for the detection and quantitation of troponin I and troponin T in body fluids as an indicator of myocardial infarction. Since troponin I and T exist in various conformations in the blood, the ratios of the monomeric troponin I an T and the binary and ternary complexes, as well as which form of troponin present in the blood, may be related to the metabolic state of the heart. Disclosed is a system to determine the presence of a troponin form or a group of troponin forms in a sample of whole blood, serum or plasma. Disclosed is a stabilized composition of troponin; the stabilized composition can comprise a stabilized composition of troponin I, wherein the troponin I is oxidized, the troponin I can be unbound or the troponin I can be in a complex.

Abstract: Immunoassay methods and apparatus are provided which utilize flow cytometry, coated latex microspheres, and labelled antibodies, to simultaneously detect the presence and amount of several antigens or antibodies in a sample. Microspheres can be sized by forward angle light scatter (FALS) or electronic volume. By combining FALS and fluorescence, it is practical to use beads of several different sizes, colors or shapes, each bead coated with a different protein or antibody, for the simultaneous detection of multiple analytes in a sample. Available auto-sampling systems make it even more appealing in this regard. In accordance with one embodiment, highly purified RNP. Sm, SS-A, SS-B and Scl-70 antigens are bound to 4, 5, 6, 7 and 10 .mu.m latex beads, respectively and stabilized for extended shelflife. Diluted patient serum is placed into test tubes containing a mixture of the five antigen coated beads and incubated. If an antibody is present for a specific antigen, it will bind to that specific bead.

Abstract: Reagents and methods for the detection of Staphylococcus aureus are provided. The reagents contain an antibody that binds to a capsular polysaccharide of type 5 of Staphylococcus aureus, and can be used in methods for detection of oxacillin resistant Staphylococcus aureus that escapes detection by agglutination in the presence of fibrinogen and antibodies directed against protein A of Staphylococcus.

Abstract: The invention relates in part to methods and compositions for identifying the presence, measuring the amount, stabilizing, and facilitating recovery of troponin complexes or individual troponin isoforms in a sample.

Abstract: A system and method are disclosed for separating particles having different sedimentation velocities. The system includes a container containing a binding substance including first particles and ligands attached to the first particles. When the binding substance is mixed with a liquid carrying at least second and third particles, the ligands bind the first and second particles together to form groups of bound particles. The groups of bound particles are separated from the third particles in a fluid chamber configured to be mounted on a centrifuge rotor. One of the disclosed methods includes forming a saturated fluidized bed of particles to retain the particles groups in the fluid chamber. Another of the disclosed methods includes removing at least some of the first particles after the binding substance is mixed with the liquid carrying second and third particles.

Abstract: An immunoassay for Sarcocystis neurona antibodies in equines is described. The immunoassay uses blocking of Sarcocystis antigens by antibodies to Sarcocystis sp. other than Sarcocystis neurona in connection with the immunoassay.

Abstract: A method for combining magnetic and centrifugal extraction techniques in a manner that improves wash efficiency and reduces the relative disadvantages of stand alone magnetic or centrifugal systems is provided. Centrifugation occurs about the rotational axis of a generally circular container with generally vertical sides featuring an inward protruding physical feature designed to retain material that exceeds the density of supernatent liquid and that is bound to magnetic particles during the centrifugation. During centrifugation, an external magnetic field is applied about the rotational axis of the container such that magnetic lines of force penetrate the container in any desired location. The magnetic and centrifugal forces may be independently modulated or modulated in relative unison to suit the hydrodynamic characteristics of a given complex.

Abstract: The assay devices, assay systems and device components of this invention comprise at least two opposing surfaces disposed a capillary distance apart, at least one of which is capable of immobilizing at least one target ligand or a conjugate in an amount related to the presence or amount of target ligand in the sample from a fluid sample in a zone for controlled fluid movement to, through or away the zone. The inventive device components may be incorporated into conventional assay devices with membranes or may be used in the inventive membrane-less devices herein described and claimed. These components include, flow control elements, measurement elements, time gates, elements for the elimination of pipetting steps, and generally, elements for the controlled flow, timing, delivery, incubation, separation, washing and other steps of the assay process.

Abstract: Sandwich immunoassays for detecting analyte indicative of type II collagen resorption in vivo, by contacting a body fluid sample with a first antibody and a second antibody such that analyte present in the sample forms a first antibody-analyte-second antibody complex, and detecting the presence or amount of the first antibody-analyte-second antibody complex, wherein the first and second antibodies are capable of binding to a telopeptide having a sequence identical to that of a carboxy-terminal telopeptide produced in vivo upon degradation of type II collagen.

Abstract: The invention relates to a method for carrying out heterogeneous immunoassays, in particular to a method for separating a coated solid phase from a liquid phase by means of precipitation and subsequent centrifugation, with a detectable activity remaining in the liquid phase.

Abstract: A compact combination blood cell count and immunoassay apparatus for measuring drawn whole blood specimen includes a load station for receiving the whole blood specimen and a nozzle member that can selectively withdraw whole blood from the blood specimen. A blood cell count section and an immunoassay section are mounted along a unitary path of movement of the nozzle member so that a single nozzle member may be used for transporting the whole blood to the respective blood cell count section and immunoassay section. A controller can automatically direct a transport mechanism to use only the whole blood and to perform various steps of adding dilution liquid and reagents, along with appropriate washing of the nozzle and sample cells to insure an automatic operation. A hematozorit value can be determined from the blood cell count signal and used to correct the immunoassay signal when whole blood is used.

Abstract: Disclosed herein is a method for the detection of a target substance by a colloidal gold immunoassay, which comprises dissolving in an immunoreaction system a metal salt selected from the group consisting of sodium, potassium and lithium fluorides, sodium, potassium, lithium and magnesium iodides, sodium, potassium, lithium and magnesium bromides, lithium and magnesium chlorides, sodium, potassium, lithium and magnesium nitrates, sodium, potassium, lithium and magnesium sulfates, sodium, potassium, lithium and magnesium formates, sodium, potassium, lithium and magnesium acetates, and mixtures of at least two of these metal salts, whereby the metal salt is allowed to exist in a reaction mixture.

Abstract: The present invention provides internally calibrated competitive assays for use on a solid support. Additionally, the invention provides a method of using such assays.

Abstract: A dual particle immunoassay method and kit for detecting analyte in a sample in which the sample to be analyzed, a binding molecule specific for the analyte, and a particle coated with the analyte to be detected or coated with a second binding molecule are reacted and applied to a porous membrane. The competitive immunoassay utilizes an analyte-coated particle, whereas the sandwich immunoassay employs a second binding molecule-coated particle. All of the reagents except for the coated particle are able to pass through the porous membrane. Detectable particles coated with a binding substance that binds to the binding molecule, such as protein A protein G, second antibody reactive to the binding molecule, or a small synthetic affinity ligand, are reacted with coated particles retained on the membrane surface. The detectable particles will pass through the membrane if not complexed with the coated particle.

Abstract: A substrate for solid phase synthesis of the formula: ##STR1## is disclosed. Also disclosed are processes for preparing the substrate and intermediates useful therein. Among the novel intermediates are compounds of the formula: ##STR2## wherein t is 0 or 1; n is 3-20; R is OH, an activated ester or the residue of a solid support having a plurality of amino functionalities; A is --O-- or --NH-- and Q is hydrogen or a protecting group for an amine or alcohol.

Abstract: Microparticles formed by mixing a macromolecule with a polymer at a pH near the isoelectric point of the macromolecule and incubating the mixture in the presence of an energy source for a predetermined length of time. The microparticles are composed of homogeneously distributed, intertwined macromolecule and polymer. Each microparticle allows aqueous fluids to enter and allows solubilized macromolecule and polymer to exit the microparticle and may be formulated to provide a sustained release of macromolecule and polymer from the interior of the microparticle when placed in an appropriate aqueous medium, such as under physiological conditions. Methods of production and methods of use for research, diagnostics and therapeutics are provided.

Abstract: A multilayer dry immunoassay element comprising 1) a spreading layer having a sample application area and a signal read area and 2) a separate receptor layer residing on 3) a radiation-transmissive support characterized in that the spreading layer contains a light absorbing material.

Abstract: The invention relates to libraries of synthetic test compound attached to separate phase synthesis supports. In particular, the invention relates to libraries of synthetic test compound attached to separate phase synthesis supports that also contain coding molecules that encode the structure of the synthetic test compound. The molecules may be polymers or multiple nonpolymeric molecules. Each of the solid phase synthesis support beads contains a single type of synthetic test compound. The synthetic test compound can have backbone structures with linkages such as amide, urea, carbamate (i.e., urethane), ester, amino, sulfide, disulfide, or carbon--carbon, such as alkane and alkene, or any combination thereof. Examples of subunits suited for the different linkage chemistries are provided.

Abstract: A method of detecting an antibody in a sample using a labelling compound and comprising the steps of mixing a ligand antigen, antibody or hapten bound to biotin with the sample; an antibody directed against the antibody to be detected bound to paramagnetic particles; and a chemiluminescent acridinium compound bound to avidin or streptavidin to form a solid phase complex; separating the solid phase from the liquid phase; and analyzing the separated solid phase for the presence of chemiluminescent complex.

Abstract: A method is disclosed for determining the presence or amount of analyte in a fecal sample. The method comprises the steps of (A) contacting the fecal sample with an extraction reagent comprised of at least one detergent and at least one buffer to form a mixture, (B) applying the mixture to an absorbent filter that is in proximity to or in contact with an immunoassay such that analyte present in the sample is transferred to the immunoassay, and (C) detecting the presence of analyte by said immunoassay. In a preferred embodiment, the immunoassay comprises a porous strip containing immunoreagents for detecting analyte.

Abstract: An evaluating method of a testing element having an immobilized antigen or antibody on an immobilizing carrier thereof, which immobilized antigen or antibody has yet to react with another antigen or antibody, which involves contacting the testing element with a solution containing an antibody or antigen reactive with the immobilized antigen or antibody, specifically binding a labelling compound to the reactive antibody or antigen to thereby cause a color-developing reaction of the labelling compound, and determining an amount of the antigen or antibody immobilized on the testing element as a function of a level of produced color. Using this evaluation method, a quantitative determination of the immobilized antigen or antibody can be carried out, and thus the performance of the testing element for antigens or antibodies can be evaluated.

Abstract: A non-competitive method for the determination of analytes. Initially the analyte is bound to a specific binding partner, after which the unoccupied binding sites of the binding partner are inactivated. The bound analyte is then dissociated from the binding partner and replaced by a labeled marker, after which the bound labeled marker is determined. The signal from the bound labeled marker is directly proportional to the initial amount of analyte in the sample, which makes the present method more favorable than the competitive assays.

Abstract: An immunoassay method in which blood can be measured even without pretreatment by a centrifuge etc. In the present invention, antibodies or antigens in a sample are subjected to agglutination reaction with insoluble carriers onto which antigens or antibodies specifically reacting with the antibodies or antigens in the sample have been imrobilized and the resulting agglutination mixture is determined for the change in its absorbance or in its scattered light by irradiation with light, wherein said sample is whole blood and the whole blood is forcibly lysed.

Abstract: A method for identifying a toxicant in an environmental water or soil sample comprising (a) adding to the sample an antibody known to form a complex with a toxicant suspected to be present in the sample; (b) incubating the antibody with the sample for a time and under conditions sufficient to allow complexes to form between the antibody and the toxicant; and (c) comparing the toxicity of the sample treated as in the step (b) with the toxicity of the sample prior to the step (a), whereby a reduced amount of toxicity of the treated sample compared with the untreated sample indicates the presence of the toxicant in said sample. The method described herein provides unexpected and important advantages by providing more accurate correlations between predicted and observed toxicity of a sample than previous methods.

Abstract: The present invention relates to a method of carrying out an immunoassay in a multiphase system. Particularly, a sample containing an analyte is brought into contact with a receptor A and a tracer. The analyte can either from a complex with the tracer, or counteract the formation of a complex of receptor A and tracer by competing with the tracer for binding to receptor A, or counteract the formation of a complex of receptor A and tracer by competing with receptor A. Receptor B is added and the signal is determined. In this method, receptor A and receptor B are suitably immobilized, ensuring that the tracer either cannot enter into any binding involving the simultaneous participation of receptors A and B or can enter into such a binding to only such a slight extent that it is nevertheless possible to detect and differentiate differing analyte concentrations.

Abstract: A substrate for solid phase synthesis of the formula: ##STR1## is disclosed. Also disclosed are processes for preparing the substrate and intermediates useful therein. Among the novel intermediates are compounds of the formula: ##STR2## wherein t is 0 or 1; n is 3-20; R is OH, an activated ester or the residue of a solid support having a plurality of amino functionalities; A is --O-- or --NH-- and Q is hydrogen or a protecting group for an amine or alcohol.

Abstract: A cell population or subpopulation enrichment procedure for separating undesired populations or subpopulations from a biological sample utilizing relatively heavy, dense particles and gravity sedimentation. The particles have one or more reactants bound thereto which are specific to and will bind with the selected population or subpopulation to be eliminated from the sample. The particles preferably are mixed with the sample by repeatedly causing the particles to settle through a substantial portion of the sample to bind to the selected population. The particles with the bound selected population to be eliminated then are allowed to preferentially settle in the sample and the supernatant including the enriched population or subpopulation is separated from the particles with the population to be eliminated bound thereto.

Abstract: Chemiluminescent electron-rich aryl-substituted 1,2-dioxetane compounds are disclosed in which the aryl group is poly-substituted with suitable electron-donating groups such that the light-emitting pattern of the molecule results in a very high luminescent count, thus providing for a sensitive and precise assay for haptens, analytes, polynucleotides and the like. These substituted aryl-containing 1,2-dioxetane compounds can be used as direct labels in an immunoassay or when derivatized with an appropriate leaving group, can be used as a substrate for a enzyme immunoassay. The unusual chemiluminescence of the compounds allows the timing of the luminescent reaction to be exactly controlled.

Abstract: Disclosed is a test cell and a method for detection of a preselected ligand in a liquid sample such as a body fluid. The test cell includes an elongate outer casing which houses an interior permeable material capable of transporting an aqueous solution and defining a sample inlet, a test volume, and a reservoir volume. The reservoir volume is disposed in a section of the test cell spaced apart from the inlet and is filled with sorbent material. The reservoir acts to receive liquid transported along a flow path defined by the permeable material and extending from the inlet and through the test volume. In the test volume is a test site which includes a first protein having a binding site specific to a first epitope of the ligand immobilized in fluid communication with the flow path. The test site can be observed through a window of the casing.

Abstract: Microparticles formed by mixing a macromolecule with a polymer at a pH near the isoelectric point of the macromolecule and incubating the mixture in the presence of an energy source for a predetermined length of time. The microparticles are composed of homogeneously distributed, intertwined macromolecule and polymer. Each microparticle allows aqueous fluids to enter and allows solubilized macromolecule and polymer to exit the microparticle and may be formulated to provide a sustained release of macromolecule and polymer from the interior of the microparticle when placed in an appropriate aqueous medium, such as under physiological conditions. Methods of production and methods of use for research, diagnostics and therapeutics are provided.