Biomedical subjects

R O Harrison

Publications and source records attributed to R O Harrison.

10 recordsLinked to original sources

PubMed2000

Comparison of an enzyme-linked immunosorbent assay (ELISA) to gas chromatography (GC)--measurement of polychlorinated biphenyls (PCBs) in selected US fish extracts.

The analysis of PCBs in fish tissues by immunoassay methods was evaluated using fish collected from a US monitoring program, the National Contaminant Biomonitoring Program of the US Department of Interior, Fish and Wildlife Service. Selected composite whole fish samples, which represented widely varying concentrations and sources of PCBs, were extracted and subjected to congener PCB analysis by gas chromatography (GC) and total PCB analysis using an ELISA (ePCBs) calibrated against technical Aroclor 1248. PCB congener patterns in these fishes were different from the patterns found in commercial Aroclors or their combinations as demonstrated by principal component analysis of normalized GC congener data. The sum of the PCB congeners measured by GC (total-PCBs) ranged from 37 to 4600 ng/g (wet weight). Concentrations of PCBs as determined by the ELISA method were positively correlated with total-PCBs and the ePCBs/total-PCBs ratios for individual samples ranged from 1 to 6. Ratios of ePCBs/total-PCBs for dilutions of Aroclors 1242, 1254, and 1260 and for matrix spikes range from 0.6 for 1242 to 2.5 for 1254 and 1260. These results suggest that higher chlorinated PCB congeners have higher affinity for the anti-PCB antibodies. Partial least squares with latent variable analysis of GC and ELISA data of selected Aroclors and fish samples also support the conclusion that ELISA derived PCB concentrations are dependent on the degree of chlorination.

Animals↗

PubMed1999

Immunochemical analysis for dioxins--progress and prospects.

Recent developments in antibody design and sample preparation have considerably enhanced the use of enzyme immunoassay (EIA) as an alternative to the conventional techniques based on gas chromatography and mass spectroscopy for the analysis of the trace organic pollutants polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in source and environmental samples. The EIA-specific sample preparation technique for solid matrices using a compatible extracting solvent coupled with an assay using a more sensitive antibody permits screening of source and environmental samples to be undertaken with minimal sample preparation. EIA has been validated on an increasing range of environmental and source samples as a cost effective I-TEQ screening tool, exploiting the commonly recognised general strengths of immunoassays such as speed, simplicity, low cost and parallel processing of many samples, greatly reducing the number of samples requiring conventional GCMS analysis. Further research and developmental work is required in the areas of sample extraction and extract cleanup to ensure compatibility with the immunoassay from the outset, and to standardise as far as is possible an assay format relevant to the application of interest, covering sample preparation, sample cleanup, quality control, assay procedure and data analysis.

Antibodies↗

PubMed1991

Comparison of enzyme-linked immunosorbent assay and high-performance liquid chromatography for the analysis of atrazine in water from Czechoslovakia.

Twenty-two water samples from Bohemian Paradise in Czechoslovakia were analyzed for atrazine (2-chloro-4-ethylamino-6- isopropylamino-1,3,5-triazine) by enzyme immunoassay (EIA) and high-performance liquid chromatography (HPLC). Three samples were drinking water, the others were creek water. The amount of atrazine found by EIA ranged from 0.2 to 12,700 ng/ml and 0.10 to 10,300 ng/ml by HPLC. Agreement between the methods was good. Nine samples containing the highest concentrations of atrazine were used to check the day-to-day reproducibility of both methods at high atrazine levels. The percent coefficients of variation (% CVs) ranged from 5.9 to 26.2 for the EIA and 1.5 to 33.2 for the HPLC. Day to day % CV values for each calibrator concentration ranged from 5.7 to 10.9 for EIA and 9.9 to 13.9 for HPLC. Sensitivity of the EIA was 0.2 ng/ml while it was 0.1 ng/ml for solid phase extraction HPLC and slightly less than 60 ng/ml for direct injection HPLC.

Atrazine↗

PubMed1990

Statistical design of ELISA protocols.

This paper shows how to obtain accuracy and efficiency in an ELISA analysis by allocating the wells on a 96-well microplate between calibration and determination of unknowns, and by choosing the known concentrations for calibration. The method also can determine how much is lost in precision by using a convenient but non-optimal protocol.

Enzyme-Linked Immunosorbent Assay↗

PubMed1990

Increased milk production versus reproductive and energy status of high producing dairy cows.

Two groups of 10 Holstein cows were chosen by pairs from a 20-yr genetic selection project that used either breed average or breed high sires chosen only for Predicted Differences in milk production. Milk production (305-d mature equivalent) was 10,814 kg and 6912 kg for the high and average groups of cows. Days to first visual estrus and number of ovulations before first visual estrus were greater for the high versus the average group (66 vs. 43 d and 1.6 vs. .7 ovulations). No differences were significant between groups for the interval from parturition to uterine involution or for days to first ovulation. Energy balance was less for the high group during wk 1, 2, 10, and 11. Plasma glucose concentration was lowest during wk 2 for both groups, and nonesterified fatty acids and beta-hydroxybutyrate were greatest for both groups during wk 1 and 2. Liver glycogen content was lower at d 15 postpartum for the high group, and liver triglyceride content was greater on d 30 for the high group. The data for reproductive functions support the concept that high milk production is antagonistic to expression of estrous behavior but not to reactivation of ovarian function.

3-Hydroxybutyric Acid↗

PubMed1984

Plasma and liver metabolites and glucose kinetics as affected by prolonged ketonemia-glucosuria and fasting in steers.

For 28 days, four steers received 1,3-butanediol, which causes ketonemia, and phlorizin, which causes glucosuria. Steers also were fasted for 9 days. Effects of treatments on concentrations of metabolites in blood and liver and on kinetics of glucose metabolism were determined. Treatments were: control, control with dietary butanediol plus injected phlorizin, and fasting. Fasting caused hypoinsulinemia and decreased liver glycogen by 60%. Butanediol plus phlorizin and fasting caused 18 and 19% decreases of plasma glucose and 2.5- and 6-fold increases of free fatty acid concentrations in blood plasma. Glucose irreversible loss averaged 371, 541, and 182 g/day during control, butanediol plus phlorizin treatment, and fasting. Butanediol plus phlorizin increased liver ketone body concentrations, caused glucosuria, ketonuria, and ketonemia, but did not affect insulin, glucagon, or growth hormone concentrations in plasma or triglyceride and glycogen contents in liver. Steers given butanediol plus phlorizin did not show all the usual signs of lactation ketosis, but the treatment still offers promise for studying causes and effects of ketosis.

Acidosis↗

PubMed1981

Monoclonal antibodies against simian virus 40 tumor antigens: analysis of antigenic binding sites, using adenovirus type 2-simian virus 40 hybrid viruses.

The antigenic binding sites of two monoclonal antibodies are located in the COOH-terminal region (clone 412) and probably in an internal region (clone 7) of simian virus 40 large T antigen. A third monoclonal antibody (clone 122), which has been shown to bind nonviral T antigen, does not react with HeLa cells infected with nondefective adenovirus type 2 (Ad2)-simian virus 40 hybrid viruses Ad2+ND1, Ad2+ND2, or Ad2+ND4.

Adenoviruses, Human↗

PubMed1980

Monoclonal antibodies against simian virus 40 T antigens: evidence for distinct sublcasses of large T antigen and for similarities among nonviral T antigens.

We have isolated three clones of hybrid cells which synthesize antibodies specific for determinants on simian virus 40 (SV40) T antigens. Mouse myeloma NS1 cells were fused with spleen cells from mice that had been immunized with SV40-transformed mouse cells. Hybrid cells were selected in HAT medium and cloned in soft agar. We used an enzyme-linked immunosorbent assay for detection and quantification of mouse antibodies against SV40 T antigens. Monoclonal antibodies from 3 of the 24 clones that scored as positive in the enzyme-linked immunosorbent assay were verified by immunoprecipitation to be specific for SV40 T antigens. Two clones (7 and 412) produced antibodies that recognized denaturation-sensitive antigenic determinants unique to large T antigen. Antibodies from clone 7 appeared to have a low affinity for large T antigen. Antibodies from clone 412 had a higher affinity for large T antigen but did not recognize a subclass of large T antigen that was recognized by tumor serum. Antibodies of the third clone, clone 122, recognized a denaturation-stable antigenic determinant of the 53,000-dalton mouse nonviral T antigen in SV40-transformed cells. Antibodies from clone 122 also recognized similar (51,000- to 56,000-dalton) nonviral T antigens in SV40-transormed or lytically infected cells from five mammalian species and in four uninfected mouse lines. From these observations, we have concluded that (i) the 94,000-dalton SV40 large T antigen may exist as immunologically distinguishable subclasses, and (ii) the nonviral T antigens of five mammalian species share at least one antigenic determinant.

Animals↗

PubMedDate not supplied

An immunoassay for TEQ screening of dioxin/furan samples: current status of assay and applications development.

A simple and easy to use system has been developed for rapid screening of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) in a variety of situations. The system uses a competitive inhibition Enzyme ImmunoAssay (EIA) based on a previously described mouse monoclonal antibody which is specific for 2,3,7,8-TCDD and related congeners. Two formats have been developed, a rapid tube test and a microplate test. Respective sensitivities are 100 pg/tube and 25 pg/well of 2,3,7,8-TCDD. Congener specificity of the tube test roughly correlates with 1-TEF. The concept of TEQ screening is validated by comparing TEQ values for a set of soil samples to the EIA responses predicted for those samples. Actual analysis of crude extracts of some of the same soils indicated some clean-up is required to remove interferences. Rapid extraction and sample clean-up methods for soil and other matrices are being developed. Potential implementation of the EIA in a fixed lab is discussed. The EIA system offers significant improvements in speed, sample throughput, and cost compared to GC-MS. This system should be useful for screening environmental samples in many situations.

Dioxins↗

PubMedDate not supplied

Location dependent biases in automatic 96-well microplate readers.

Procedures performed in 96-well microplates and quantitated by automatic readers assume instruments to be precise, accurate, and free of well location dependent bias. Instrument specifications generally focus on precision and accuracy without specifically addressing biases which are dependent on well location. These biases appear to be meniscus dependent and can be demonstrated in varying degrees in automatic readers of many designs by using a reverse plate wet test, which compares repeated readings of a dye loaded plate in normal and reversed positions. This test analyzes differences between readings and is, therefore, independent of pipetting error or other experimental variables such as protein binding or immunoassay variability. Different plates increased or decreased the magnitude of observed errors but did not themselves cause the errors measured by the reverse plate wet test. Error patterns were consistent for each reader and varied widely among the 16 instruments tested. Only 4 of 16 instruments passed an existing manufacturer's specification for precision, and only one of the 16 readers tested passed a similar specification for accuracy. The severest location dependent bias was found in an instrument which exhibited excellent repeatability and consistently passed its built-in diagnostic tests. One reader with significant bias was returned to the manufacturer for routine service and calibration, but it was not demonstrably improved. The reverse plate wet test is an extremely useful diagnostic tool for quality control at all stages of instrument manufacture and use.

Evaluation Studies as Topic↗