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At least 1,333 records · Page 74Linked to original sources
PubMed1997

Use of cloned excretory/secretory low-molecular-weight proteins of Cooperia oncophora in a serological assay.

The potential of Cooperia oncophora excretory/secretory (ES) proteins as antigens in a serological assay which aims to establish exposure levels in cattle was assessed. ES proteins were analyzed by one- and two-dimensional polyacrylamide gel electrophoresis and immunoblotting. The N-terminal domains of two ES proteins were sequenced, and the corresponding cDNAs were cloned. Two cDNAs, designated CoES14.0 and CoES14.2, were expressed in Escherichia coli. The recombinant proteins were tested in an indirect enzyme-linked immunosorbent assay (ELISA) in which crude worm antigen (CWA) was used as a reference standard. In total, 67 reference serum samples were used: 27 negative serum samples, 29 C. oncophora-specific serum samples, 7 Dictyocaulus viviparus-specific serum samples, and 4 Ostertagia ostertagi-specific serum samples. This showed respective sensitivities and specificities of 17 and 84%, 0 and 100%, and 100 and 100% by the ELISAs with the three different types of proteins (CWA, CoES14.0, and CoES14.2, respectively). Since the CoES14.2 ELISA had the best sensitivity and specificity with reference sera, its specificity was further validated in an antigen inhibition ELISA. In this assay CoES14.2 and CWA preparations of C. oncophora, Cooperia curticei, O. ostertagi, Nematodirus helvetianus, Fasciola hepatica, D. viviparus, Haemonchus placei, and Trichostrongylus colubriformus were used as competitor antigens. This experiment showed that only the homologous antigens C. oncophora CWA and CoEs14.2 resulted in 100% inhibition. The CWA preparations of all other nematodes did not affect the ELISA, even if concentrations of 250 times the 50% inhibitory concentration of C. oncophora CWA were used. These results indicate that CoES14.2 does not share cross-reactive epitopes with heterologous CWAs. Finally, we tested the CoES14.2 ELISA with sequential serum samples from naturally infected groups of animals. The optical density values that were obtained correlated well with exposure levels based on cumulative egg excretion. Thus, the CoES14.2 ELISA seems to be a very sensitive tool for estimating exposure levels in cattle.

Amino Acid Sequence↗
PubMed2005

Middle cerebral artery spasm after subarachnoid hemorrhage: detection with transcranial color-coded duplex US.

PURPOSE: To prospectively determine the accuracy of transcranial color-coded duplex ultrasonography (US) used alone and in conjunction with carotid artery US for diagnosis of middle cerebral artery (MCA) spasm, with intraarterial digital subtraction angiography (DSA) used as the reference standard. MATERIALS AND METHODS: The institutional ethics committee approved the study. Each patient, or members of the patient's family, gave informed consent. One hundred twenty consecutive patients (64 women, 56 men; mean age, 45.5 years +/- 13.6 [standard deviation]) were routinely referred for DSA after subarachnoid hemorrhage. Vasospasm was graded as mild (< or =25% reduction in vessel diameter), moderate (>25% to 50% reduction), or severe (>50% reduction). US was performed 2 hours or less before angiography. The ratio of flow velocity in the middle cerebral artery (V(MCA)) to flow velocity in the ipsilateral extracranial internal carotid artery (V(ICA)) was calculated. Diagnostic accuracy was evaluated by calculating the area under the receiver operating characteristic curve (Az). The significance of the difference between the two Az values (for US vs DSA) was determined by using the z test with correction for correlated data. RESULTS: Nine of 120 patients were excluded because of inadequacy of acoustic windows in the squama of temporal bones. Spasm was mild in 17, moderate in 16, and severe in only nine of 222 arteries studied. Arteries with moderate or severe vasospasm were combined in one group. The best-performing parameters were peak systolic velocity and V(MCA)/V(ICA) ratio. Az values for these two parameters in diagnosis of moderate-to-severe vasospasm were 0.93 and 0.95, and in diagnosis of mild vasospasm, 0.90 and 0.91. Accuracy of the V(MCA)/V(ICA) ratio calculated on the basis of end-diastolic velocity for diagnosis of mild MCA narrowing was significantly better than that of end-diastolic MCA velocity alone (Az = 0.88 vs 0.84, P < .05). The stepwise approach with use of the V(MCA)/V(ICA) ratio after flow velocity measurements in the MCA resulted in a decreased number of false-negative findings in both vasospasm subgroups. The thresholds of highest efficiency were at a mean velocity of 94 and 108 cm/sec and a peak systolic V(MCA)/V(ICA) ratio of 3.6 and 3.9 for diagnosis of mild and moderate-to-severe vasospasm, respectively. CONCLUSION: Transcranial color-coded duplex US alone or in conjunction with carotid artery US has excellent accuracy for angiographic detection of vasospasm. Use of MCA velocity measurements and V(MCA)/V(ICA) ratio can increase the accuracy of Doppler US.

Adolescent↗
PubMed2000

Assessment of fresh pork color with color machine vision.

Currently, fresh pork color is visually evaluated using either the Japanese Pork Color Standards (JPCS) or the National Pork Producers Council Pork Quality Standards (NPPC) as a reference. Although useful, visual evaluation of meat color can vary with evaluator and may be quite expensive. In this study, three separate studies were used to compare the ability of color machine vision (CMV) and untrained panelists to evaluate pork color. Panels visually evaluated over 200 pork loin chops using either the JPCS or NPPC reference standards. Results from each panel were used to evaluate the ability of the CMV to sort pork loin chops based on the same criteria. Representative samples, typical of each color class, were used to train neural-network-based image processing software. After training, the CMV system was used to evaluate quality classes of pork samples based on color distribution. Classification by CMV was compared with the average panel score, rounded to the nearest integer. Training the CMV system using images of actual meat samples resulted in a stronger correlation to panel scores than training with either set of artificial color standards. Agreement between the CMV system and the panels was as high as 90%. Agreement between individual panelists and the integer panel average (52 to 85%) was less than that observed for CMV classification. Finally, the on-line performance of CMV using a laboratory conveyor system was simulated by repeatedly classifying 37 samples at a speed of 1 sample per second. Collectively, these results demonstrate that CMV is a rapid and repeatable means of evaluating pork color.

Animals↗
PubMed1976

The serology of Pseudomonas aeruginosa analysed by means of quantitative immunoelectrophoretic methods. V. Thermostability, resistance to degradation by plasmin activity and storage conditions of a polyvalent P. aeruginosa reference standard-antigen.

The stability of a polyvalent Pseudomonas aeruginosa antigen (St-Ag) was analysed under different experimental conditions by quantitative immunoelectrophoretic methods. St-Ag did not undergo proteolytic degradation by plasmin which is present in the corresponding polyvalent rabbit antiserum (St-Ab) against St-Ag. Storage at -30 degrees C for up to 4 years caused no qualitative changes in St-Ag and, after 1 year, only minor quantitative changes could be revealed. Repeated thawing and freezing caused no changes in St-Ag whereas storage at 4 degrees C, or at room temperature, caused gradual degradation of most of the antigens of St-Ag, beginning after 1 week. Heating up to 40 degrees C for 2 min did not change St-Ag, but heating for longer periods of time or at higher temperatures gradually destroyed most of the antigens of St-Ag. The most thermostable antigen in St-Ag was number 37 which is related to the group specific O antigens.

Antigens, Bacterial↗
PubMedDate not supplied

A standardized enzyme-linked immunosorbent assay for infectious bronchitis virus: comparison with hemagglutination-inhibition and virus-neutralization assays for measuring protective antibody levels in chickens.

An enzyme-linked immunosorbent assay (ELISA) was developed to measure antibodies to infectious bronchitis virus (IBV) in chickens. The results are reported in IBV standard ELISA values calculated by comparing antibody levels in test sera with antibody levels in a series of standard reference sera. The IBV standard ELISA values were good indicators of responses to vaccination and the immune status of experimentally challenged birds. Although the assay was not serotype-specific, the sensitivity makes it ideally suited for determining the immune status of poultry flocks. The assay results compared favorably with other laboratory results, including virus-neutralization titers, hemagglutination-inhibition levels in sera, virus isolation from vaccinated/challenged birds, and the tracheal ring test results.

Animals↗