Abstract: Provided is a method comprising adding anions having a high charge density to a liquid medium; the liquid medium comprising molecules that hydrogen bond with one another; the anions interacting with the molecules of the liquid medium with a force that is stronger than the forces that produce hydrogen bonding between the molecules of the medium; and heating the liquid medium by irradiating it with microwaves.

Abstract: The instant invention provides an economical flow-through method for determining amount of target proteins in a sample. An antibody preparation (whether polyclonal or monoclonal, or any equivalent specific binding agent) is used to capture and thus enrich a specific monitor peptide (a specific peptide fragment of a protein to be quantitated in a proteolytic digest of a complex protein sample) and an internal standard peptide (the same chemical structure but including stable isotope labels). Upon elution into a suitable mass spectrometer, the natural (sample derived) and internal standard (isotope labeled) peptides are quantitated, and their measured abundance ratio used to calculate the abundance of the monitor peptide, and its parent protein, in the initial sample.

Abstract: The invention relates to devices for performing single step assays for the determination of the presence or absence of an analyte in a liquid sample, and methods of determining the presence or absence of such analytes using such devices. Devices disclosed comprise a labeled analyte-binding reagent reversibly-immobilized on a non-porous solid material, which solid material is in physical contact with a dry porous carrier bearing an immobilized analyte-binding reagent. Also provided are quantitative assay devices.

Abstract: Described herein are substrates, methods, articles, and kits that are useful for removing interferents from samples for diagnostic purposes. The interferents are removed with phosphocellulose and cation exchange materials. These materials could also be used in vitro to improve the performance of a diagnostic assay or in vivo to remove the interferents.

Abstract: A device for the separation of small particles or cells from a fluid suspension of the same is described. The device includes a coaxial tubular design in which the inner tube is a micro porous tube that allows the passage of liquids and certain particulates up to a certain size cut-off, and the outer tube allows for the collection of passed fluids. Inlet and outlet ports allow the introduction and flushing of components of interest. Embodiments of the device can be used for the separation of blood components, the sequestering of micro spheres used in micro-sphere-based immuno assay, and sample filtration. Other applications are not precluded. Another field of application for this device is in the separation of plasma from red blood cells. The red blood cells will not pass through the membrane due to their size, but plasma will.

Abstract: A method useful for the enumeration of cell populations in a biological sample includes the steps of reacting in a single reaction mixture a sample, a first antibody labeled with a fluorochrome having a first emission spectrum and an additional antibody. The first antibody binds to an antigenic determinant differentially expressed on leukocytes and non-leukocytes. The additional antibody binds to an antigenic determinant differentially expressed on mature and immature granulocytes or myeloid cells, and is labeled either with the first fluorochrome or an additional fluorochrome having an emission spectrum distinguishable from the first emission spectrum. The reaction mixture can be mixed with a nucleic acid dye having an emission spectrum that overlaps with one of the first or additional emission spectra. The reaction mixture may be treated with a lytic system that differentially lyses non-nucleated red blood cells and conserves leukocytes.

Abstract: A reagent for analyzing immature leukocyte capable of analyzing accurately immature leukocyte such as myeloblast and immature granulocyte, and mature leukocyte, which reagent has broader allowable range of sample processing conditions than conventional ones, and a reagent kit are provided. The present invention relates to a reagent for analyzing immature leukocyte containing a surfactant for giving damage to cell membrane of red blood cell and mature leukocyte in the sample, a solubilizing agent for causing contraction to damaged blood cell, sugar, a dye for staining nucleic acid.

Abstract: Malarial parasite Plasmodium falciparum is responsible for the most severe form of malaria in humans, causing about 2 million deaths every year. Lack of effective vaccines and emergence of drug-resistant strains necessitate the need of novel drug targets to treat the disease. The present invention describes a novel assay method of identifying candidate compounds as anti-malarials based on the property of binding to plasmodial parasite 90 kDa heat shock protein.

Abstract: The present invention provides methods of detecting asymmetric dimethylarginine (ADMA) in a sample, particularly a sample that may contain symmetrical dimethylarginine (SDMA) and/or arginine. The methods generally involve modifying any SDMA and arginine in the sample such that SDMA and arginine are readily distinguishable from ADMA; and detecting ADMA. The invention further provides antibodies specific for ADMA; antibodies specific for modified SDMA; and antibodies specific for modified arginine. The invention further provides kits for practicing the subject methods.

Abstract: A method for spectrometric investigation of a plurality of samples dissolved in a solvent comprises: (a) guiding a first solvent with a sample through an SPE cartridge for concentrating the sample in the cartridge; (b) positioning the cartridge in a carrier for a plurality of cartridges; (c) repeating steps (a) and (b) for a desired number of samples; (d) drying the concentrated samples through removal of the residual first solvent, in particular, dehydration or evaporation; (e) dissolving each sample in a second solvent, transferring these dissolved samples from the cartridges to a spectrometer and acquiring a spectrum of each sample. All samples are dried together in step (d) subsequent to steps (b) and (c) while the cartridges with samples are positioned in the carrier. The drying process is thereby considerably accelerated in a straightforward technical manner.

Abstract: Compositions and methods for the therapy and diagnosis of cancer, particularly ovarian cancer, are disclosed. Illustrative compositions comprise one or more ovarian tumor polypeptides, immunogenic portions thereof, polynucleotides that encode such polypeptides, antigen presenting cell that expresses such polypeptides, and T cells that are specific for cells expressing such polypeptides. The disclosed compositions are useful, for example, in the diagnosis, prevention and/or treatment of diseases, particularly ovarian cancer.

Abstract: The present invention relates to methods of detecting the presence of detergent- or urea-insoluble amyloid-like fibrils or protein aggregates on filters. Preferably, said fibrils or aggregates are indicative of a disease, preferably of a neurodegenerative disease such as Alzheimer's disease or Huntington's disease. In addition, the present invention relates to inhibitors identified by the method of the invention, to pharmaceutical compositions comprising said inhibitors and to diagnostic compositions useful for the investigation of said amyloid-like fibrils or aggregates.

Abstract: The present invention provides a method for preparing a five-part differential white blood cell control and a white blood cell control prepared thereby. The white blood cell control obtained by the method of the present invention perfectly retains the light scattering properties of every type of white blood cells and has much higher stability than that of real white blood cells, and therefore can be used for the quality control of five-part differential hematology analyzers based on the principle of multi-angle light scattering.

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: Methods for enhancement in accuracy of immunochemical analysis of heterogeneous biological fluids containing exogenous substances that can interfere in immunochemical analysis for endogenous analytes of interest. According to this method a heterogeneous biological fluid sample is pretreated with an interferant suppression effective amount of a molybdenum coordination complex, so as to reduce manifestation of the presence of said exogenous material under immunoassay conditions. This invention is suitable for the suppression of manifestation of exogenous substances, specifically metabolites of drugs of abuse, during the immunoassay of biological fluids of infants for detection of endogenous substances indicative of a wellness or disease state. This invention also has application for similar suppression exogenous substances in the biological fluid of adults that have been inadvertently exposed to such substances (e.g. secondhand smoke).

Abstract: A method of rapidly assaying nucleotide receptor P2X7 pore activity in white blood cells contained within a blood sample. A method according to the invention includes the steps of: (a) labeling the white blood cells with a white blood cell-specific label; (b) depolarizing the labeled white blood cells with an isotonic depolarizing solution; (c) contacting the labeled white blood cells with a dye and a P2X7 agonist in an amount sufficient to activate nucleotide receptor P2X7 pore activity; (d) contacting the labeled white blood cells with a divalent cation in an amount sufficient to deactivate nucleotide receptor P2X7 pore activity; and (e) analyzing dye uptake in labeled white blood cells whereby nucleotide receptor P2X7 pore activity is quantified by the amount of dye taken up in labeled white blood cells treated with P2X7 agonist relative to labeled white blood cells in the absence of P2X7 agonist.

Abstract: A test device and method for determining the presence or absence of one or more analytes in a fluid sample, the test device including a support or member bearing a mark thereon, and a matrix or member containing a capture zone. In operation, an observation area in the test device becomes transparent, thereby allowing the user to view a mark that is present on a support that is disposed beneath the observation area. Typically, the mark on the underlying support is configured as a minus (?) sign. In the absence of analyte in the sample, the test device presents a negative result as a minus (?) signal. In the presence of analyte in the sample, however, the mark operates in concert with a perpendicular test line on the observation area to present a positive result as a plus (+) signal that is visible to the user.

Abstract: A fast and simple method for fractional measurement of a small particle LDL entails separating the small particle LDL and HDL from other lipoproteins, and then measuring cholesterol, triglycerides, or proteins in the separated small particle LDL.

Abstract: Analytical reagents and mass spectrometry-based methods using these reagents for the rapid, and quantitative analysis of proteins or protein function in mixtures of proteins. The methods employ affinity labeled protein reactive reagents having three portions: an affinity label (A) covalently linked to a protein reactive group (PRG) through a linker group (L). The linker may be differentially isotopically labeled, e.g., by substitution of one or more atoms in the linker with a stable isotope thereof. These reagents allow for the selective isolation of peptide fragments or the products of reaction with a given protein (e.g., products of enzymatic reaction) from complex mixtures. The isolated peptide fragments or reaction products are characteristic of the presence of a protein or the presence of a protein function in those mixtures. Isolated peptides or reaction products are characterized by mass spectrometric (MS) techniques.

Abstract: A test device and method for determining the presence or absence of an analyte in a fluid sample, the test device including a support bearing a mark thereon, and a matrix defining an axial flow path. In operation, an observation area in the test device becomes transparent, thereby allowing the user to view a mark that is present on a support that is disposed beneath the observation area. Typically, the mark on the underlying support is configured as a minus (?) sign. In the absence of analyte in the sample, the test device presents a negative result as a minus (?) signal. In the presence of analyte in the sample, however, the mark operates in concert with a perpendicular test line on the observation area to present a positive result as a plus (+) signal that is visible to the user.

Abstract: A selective method of dechlorination treatment of circulating water containing disinfectant chlorine in the form of ClO?,HClO, ClO2 or chloramine, where an ascorbic acid aqueous solution is injected from a storage reservoir into the water flow, the mass flow rate of the injected ascorbic acid solution being regulated with respect to the mass flow rate of the chlorine in the circulating water to be treated, in such a way that the ratio (R) of the ascorbic acid and chlorine mass flow rates (R=D1/ D2) is between 2.5 and 4, preferably less than or equal to 3. The method can be used for treatment of water that feeds into a biological detector containing a disinfectant chlorine content that is incompatible with the survival of the biological species used in the detector. The disinfectant chlorine level in the circulating water after mixing is maintained at a value lower than or equal to 0.6 mg/l, and preferably lower than or equal to the detectable limit.

Abstract: A food testing device for testing for the presence of harmful contaminants in a food sample, includes a vessel adapted for holding a liquefied food sample, a liquefier operatively associated with the vessel for converting an unliquefied food sample into a liquefied food sample, at least one test assay dispensable from the device, wherein the test assay includes at least one assay reagent having an affinity for at least one harmful contaminant, and capable of both detecting the presence of the harmful contaminant in the liquefied food sample, and producing a visual cue upon recognition of the harmful contaminant; and a radiation detector disposed proximately to the vessel for indicating the presence of ionizing radiation in the food sample at amounts exceeding normal background levels to detect the presence of a radioactive agent as the harmful contaminant.

Abstract: Rare cells are separated from a sample fluid by a positive selection or negative selection antibody by centrifuging in a tube containing a harvesting float. The harvesting float has an axial passage and a density to settle in the sample fluid and expand the layer of the target component. The positive selection antibody is preferably coupled to a particulate carrier, such as a microbead, to attach to the target component. The negative selection antibody forms a complex or conjugate with a contaminating component. In the positive separation, the particulate carrier is recovered in the axial passage of the float.

Abstract: The present invention relates to a method for determining the condition of a biological fluid by recording the IR spectrum of a sample of the biological fluid. To this end, the biological fluid can be examined in its native form. The method of the invention is usable, for example, for detecting pathological conditions in organisms.

Abstract: In one aspect, an assay test strip includes a test label that specifically binds a target analyte and a control label that is free of any specific binding affinity for the target analyte and has a different optical characteristic than the test label. In another aspect, an assay test strip includes a test label that specifically binds a target analyte and at least one non-specific-binding label that is free of any specific binding affinity for the target analyte. Systems and methods of reading assay test strips also are described.

Abstract: The present invention relates to the preparation of a sample. Preferably, the sample is a sample to be analyzed, for example for ingredient content, etc. Preferred samples include foods, cosmetics, paints, coatings, adhesives, tanning agents, fabrics, chemical compositions, dyestuffs, samples subject to forensic studies, etc. Samples prepared according to the invention method are digested in sulfuric acid, nitric acid, and one or more fluoride salts selected from LiF, NaF, RbF, CsF and KF and then preferably subjected to analysis for metal content, etc, for example using atomic absorption (“AA”) and inductively coupled plasma (“ICP”).

Abstract: An assay test strip includes a flow path, a sample receiving zone, a label, a detection zone that includes a region of interest, and at least one position marker. The at least one position marker is aligned with respect to the region of interest such that location of the at least one position marker indicates a position of the region of interest. A diagnostic test system includes a reader that obtains light intensity measurement from exposed regions of the test strip, and a data analyzer that performs at least one of (a) identifying ones of the light intensity measurements obtained from the test region based on at least one measurement obtained from the at least one reference feature, and (b) generating a control signal modifying at least one operational parameter of the reader based on at least one measurement obtained from the at least one reference feature.

Abstract: The present invention concerns methods and compositions to prepare biological samples to preserve the macromolecules in the samples. Embodiments of the invention concern the use of a soak solution that contains one or more water-miscible solvents. A sample is incubated with the soak solution to the point of saturation at a temperature above the melting temperature of the water-miscible solvent but below 0° C. The use of methods and compositions of the invention allow for subsequent preparation or analysis of the samples.

Abstract: A dust monitor is disclosed that is suitably deployed in dusty environments and capable of providing near real-time indications of exposure to airborne particulates. The monitor includes a filter and filter assembly made of materials that do not interfere with subsequent instrumental (such as spectrometric) analysis for detecting and/or quantitating an analyte. In some disclosed embodiments, the filter is made of nylon or other material that is readily subjected to thermal destruction prior to spectrometric analysis. The dust monitor also includes a humidity correction feature that permits the filter to be made of ashable organic materials even if those materials are not highly hydrophobic. Transport devices are provided for shipment of the filter and/or filter assembly to an analytical laboratory which prevent loss of particulate matter and which facilitate an accurate analysis procedure.

Abstract: An interactive system is provided with at least one active surface of plastic from monomers containing at least one structural element derived from a carbon dioxide (A), and at least one substance associated to a linker with at least one structural element (B) capable of establishing a hydrogen bond, and involving an interaction between the structural elements (A) and (B). The interactive system is suitable for presenting and eliminating substances in liquids.

Abstract: The present invention provides a test device for determining the concentration of LDL-cholesterol in a sample. The test device has a continuous solid surface with at least one reaction area on the continuous solid surface, and the reaction area has, in dry form, a non-LDL inhibitor and a system for determining the concentration of cholesterol.

Abstract: A system and method for cleanup of biological samples from contaminants prior to spectroscopy analysis. The system includes a support configured to hold a sample including a liquid having at least one group of biological molecules with a surface of the support binding the molecules at a surface tension angle to the liquid of less than 180 degrees. The system includes an evaporator configured to evaporate liquid from the support, a solvent applicator configured to apply a solvent for dissolution of the contaminants in the sample, and a solvent removal device configured to remove applied solvent from the sample and thereby at least partially remove the contaminants.

Abstract: Methods of prevention and correction of white blood cell interferences to the measurements of mean cell volume (MCV), red blood cell concentration (RBC) and hematocrit (Hct) of a blood sample are disclosed. The interference to MCV can be prevented by identifying a valley between the red blood cell and white blood cell modes on the red blood cell distribution histogram, defining a red blood cell region using the valley and calculating MCV within the defined region. Alternatively, a curve fit of the white blood cell population on the red blood cell distribution histogram can be used to exclude the white blood cells. RBC can be corrected by subtracting the white blood cell concentration obtained from the analysis of a second aliquot sample, when the white blood cell concentration (WBC) exceeds a predetermined criterion. Hct of the blood sample can be calculated using the obtained MCV and RBC.

Abstract: An analytical test element for blood analyses is provided having at least a substrate body having a channel structure for the flow transport of a blood sample from an application site to at least one analytical site. The channel structure has a dilution channel that can be loaded with the blood sample and is provided with a separation component for retaining corpuscular blood components and has a sample channel which conveys a blood sample aliquot to be diluted and joins the dilution channel at a mixing site.

Abstract: Optimized enzymatic conditions incorporate a single oxygen atom into digested peptides using a peptidase. The incorporation of a single oxygen atom is especially useful for proteolytic 18O labeling in comparative proteomics. The optimized proteolytic 18O labeling minimizes the generation of a mixture of isotopic isoforms of the peptides resulting from incorporation of either one or two 18O atoms. The outcome is accurate quantification of isotopically labeled peptides.

Abstract: Methods, systems and kits for the simultaneous or sequential analysis of a multitude of steroid hormones by mass spectrometry are disclosed. The methods require minimal sample size and minimal preparation time. The methods include 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 including ionization of the steroid hormones by photoionization.

Abstract: Method and arrangement for taking up a first medium, which is present in a first phase, into a capillary device In the capillary device, a reduced pressure is produced which is less than a critical pressure such that, if it is exerted in the capillary device, a surface tension which is produced by the first medium in the capillary device, when the first medium has been taken up fully by the capillary device, would be overcome so that a second medium which is present in a second phase, different from the first phase, would be taken up into the capillary device.

Abstract: A method for classifying leukocytes in animal blood is described. In the method, a measurement sample is prepared by mixing a canine or feline blood sample with a lysing reagent. Erythrocytes are lysed and leukocytes are shrunk in the measurement sample. The data correlated with the size of leukocytes in the measurement sample are measured. The leukocytes, on the basis of the measured data, are classified into a first group containing lymphocytes, a second group containing neutrophils and monocytes and a third group containing eosinophils.

Abstract: Methods are provided for diagnosing and/or characterizing chronic immune disease activity in a subject. In the subject methods, a sample is obtained from a subject suspected of having or known to have a chronic immune disease. The sample is then assayed for the presence of low molecular actin fragments. The assay results are used to diagnose the presence of chronic immune disease activity and/or characterize chronic immune disease activity in the subject, e.g. to confirm an initial chronic immune disease diagnosis, to determine the stage of the disease, to monitor disease progression, to predict disease attacks, and the like. Also provided by the subject invention are kits for practicing the methods.

Abstract: A lytic reagent composition and methods for differential analysis of leukocytes are disclosed. In embodiments, the analysis may utilize optical measurements in flow cytometry based hematology analyzers. The reagent system includes an anionic surfactant in a hypotonic solution, an inorganic buffer to maintain the pH in a range from about 6 to about 10, and optionally a leukocyte stabilizer. The reagent system is used to lyse red blood cells and stabilize the leukocytes to enable multi-part differentiation of leukocytes in a near physiologic pH environment on a flow cytometry based hematology analyzer using axial light loss, light scatter intensity, high-numerical aperture side scatter, and time-of-flight measurements.

Abstract: Disclosed embodiments concern quantifying a biomolecule conjugated to a nanoparticle. Quantifying typically comprises determining the number of biomolecules per nanoparticle. Any suitable biomolecule can be used, including but not limited to, amino acids, peptides, proteins, haptens, nucleic acids, oligonucleotides, DNA, RNA, and combinations thereof. A single type of biomolecule may be conjugated to the nanoparticle, more than one biomolecule of a particular class may be conjugated to the nanoparticle, or two or more classes of biomolecules may be conjugated to the nanoparticle. Certain disclosed embodiments comprise enzymatically or chemically digesting a biomolecule conjugated to the nanoparticle, or displacing a biomolecule using ligand-exchange chemistry.

Abstract: The present invention provides an antibody which specifically recognizes a CNS tau protein but not a peripheral tau protein. More specifically, the present invention provides an antibody obtainable by using a polypeptide comprising an amino acid sequence of a connective portion between the amino acid sequence encoded by Exon 4 of a gene encoding a tau protein and the amino acid sequence encoded by Exon 5 thereof as an epitope specific to the isoform of tau protein predominantly existing in central nervous tissues. The present invention further provides a method of detecting Alzheimer's disease and a reagent kit using the antibody.

Abstract: A mixed potential sensor device and methods for measuring total ammonia (NH3) concentration in a gas is provided. The gas is first partitioned into two streams directed into two sensing chambers. Each gas stream is conditioned by a specific catalyst system. In one chamber, in some instances at a temperature of at least about 600° C., the gas is treated such that almost all of the ammonia is converted to NOx, and a steady state equilibrium concentration of NO to NO2 is established. In the second chamber, the gas is treated with a catalyst at a lower temperature, preferably less than 450° C. such that most of the ammonia is converted to nitrogen (N2) and steam (H2O). Each gas is passed over a sensing electrode in a mixed potential sensor system that is sensitive to NOx. The difference in the readings of the two gas sensors can provide a measurement of total NH3 concentration in the exhaust gas. The catalyst system also functions to oxidize any unburned hydrocarbons such as CH4, CO, etc.

Abstract: This invention provides microfluidic devices and methods for using the same. Microfluidic devices of the present invention comprises a first elastic layer, a fluid flow channel within the elastic layer; and a means for providing a fluid sample from the fluid flow channel to an analytical device. The present invention also provides an analytical apparatus comprising such a microfluidic device and an analytical device.

Abstract: A bulk sample is fed with a liquid into a mixing tank (1) where it is stirred to form a dispersion. A proportion of the dispersion is recycled from the bottom of the tank through a line to the top of the tank so that at least the dispersion in the recycle loop (3) is substantially homogeneous, and a representative sample of the dispersion is taken from the recycle loop, e.g. using a slurry sampler (5).

Abstract: A method for calibrating an analysis instrument, e.g., a gas chromatograph, for water analysis that includes four steps. The first step is to continuously remove water from a liquid that contains water to produce a liquid containing a reduced amount of water such as by membrane pervaporation. The second step is to analyze the liquid containing a reduced amount of water for water by a reference water analysis method such as by Karl Fischer titration. The third step is to analyze the stream of liquid containing a reduced amount of water using the instrument to be calibrated, e.g., a gas chromatograph having a helium photoionization pulse discharge detector. The fourth step is to calibrate the instrument using the analysis of the second step. In a related embodiment, a system for normal phase chromatography.

Abstract: A method for screening of compounds for binding differentiation at various drug target binding sites is used with a device measuring the enthalpy of reaction for the binding. The method includes merging test ligand with target compound and merging test ligand with target compound in the presence of at least one blocking agent. A first heat of reaction is detected for the merged test ligand and target compound solution and a second heat of reaction is detected for the merged test ligand and target compound solution in the presence of a blocking agent. The two heats of reaction are compared to determine whether a reaction has occurred.

Abstract: A method is disclosed for high-throughput screening assay sample preparation and testing for the identification of binding between drug targets and library compounds, for use with a calorimetric device measuring the enthalpy of reaction for the binding. The method includes mixing a library compound with a specified solvent and mixing a target compound solution with a second specified solvent on a calorimetric device. The library compound/solvent is merged with the target compound/solvent solution and the library compound/solvent solution is also merged with a third solvent solution on the calorimetric device. The heats of reaction are detected for both merged solutions and are compared.

Abstract: The present invention relates to a reagent for classifying leukocytes including (a) at least one surfactant capable of lysing erythrocytes and partly damaging the cell membrane of leukocytes, (b) at least one organic compound bearing an anionic group capable of binding to the cationic component present in the leukocytes to provide morphological differences between the leukocytes, and (c) a buffer for adjusting pH to 2˜8. Also disclosed is a method for classifying leukocytes into four groups.

Abstract: A method for restoring immunoreactivity of a tissue, particularly decalcified tissue, fixed width an aldehyde fixative agent and embedded in an embedding medium, usually comprising celloidin, the method comprising the steps of contacting the tissue with an aldehyde releasing reagent solution comprising a solvent and an aldehyde releasing reagent and removing aldehyde released by the aldehyde releasing reagent from contact with the tissue by reacting the aldehyde in a substantially irreversible manner to form a non-aldehyde derivative, and removing excess base from the tissue. A preferred solution for celloidin-embedded decalcified tissue comprises methanolic sodium hydroxide at about one-third saturation.