Detection limits of a biological monitoring system for chemical water pollution based on mussel activity.
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The efficacy of lymphography and computed tomography (CT) in staging nonseminomatous testicular cancer was analyzed in 41 patients. Retroperitoneal lymphadenectomy in 30 patients with early tumor stage revealed 56% sensitivity, 90% specificity, and 80% overall accuracy for lymphography; CT was less accurate (44% sensitivity, 81% specificity, 70% overall accuracy). The presence of advanced disease in 11 patients was depicted equally by lymphography and CT, but CT better demonstrated the anatomic extent of the metastases. CT is valuable for discriminating between advanced and early tumor stages; the similar inaccuracy of lymphography in early disease militates against its routine and complementary use; retroperitoneal lymphadenectomy then remains the only accurate staging modality.
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Simplified procedures for immunoelectron microscopy (IEM) and electron microscopy (EM) are described. The procedures employ the principle of agar filtration and pseudoreplication. The modification consisted of the use of microwells for storage of gels with or without antiserum (for IEM or EM, respectively) and an array of containers in which pseudoreplication and negative staining were performed. The containers were prepared from 5 ml syringes from which the needle holding parts were cut. This device enabled simultaneous and rapid handling of specimens. With Sindbis virus as a model, our microwell pseudoreplica IEM (MW-PR-IEM) was compared to six other IEM techniques and was found to be the most rapid and sensitive technique. With the MW-PR-IEM technique, the specific minimal detection limit (detection of clumps) was 1.5 x 10(7) virus particles per ml, and the non-specific detection limit (detection of single virions) was 1.8 x 10(6) virus particles per ml.
Currie Hypothesis testing is applied to gamma-ray spectral data, where an optimum part of the peak is used and the background is considered well known from nearby channels. With this, the risk of making Type I errors is about 100 times lower than commonly assumed. A programme, PeakMaker, produces random peaks with given characteristics on the screen and calculations are done to facilitate a full use of Poisson statistics in spectrum analyses. SHORT TECHNICAL NOTE SUMMARY: The Currie decision limit concept applied to spectral data is reinterpreted, which gives better consistency between the selected error risk and the observed error rates. A PeakMaker program is described and the few count problem is analyzed.
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Methyl tert-butyl ether (MTBE) is a common groundwater contaminant, introduced to the environment by leaking petroleum storage tanks, urban runoff, and motorized watercraft. In this study. a simplified (static) headspace analysis method was adapted for determination of MTBE in water samples and soil water extracts. The MDL of the headspace method was calculated to be 2.0 microg L(-1) by the EPA single-concentration design method(1) and 1.2 microg L(-1) by a calibration method developed by Hubaux and Vos (Hubaux, A.; Vos, G. Anal. Chem. 1970,42, 849-855). The MDL calculated with the Hubaux and Vos method was favored because it considers both a true positive and a false positive. The static headspace method was applied to analysis of a tap water sample and a monitoring well sample from a gasoline service station, a river sample, and aqueous extracts from soil excavated during removal of a leaking underground storage tank (LUST). The water samples examined in this study had MTTBE concentrations ranging from 6 to 19 microg L(-1). Aqueous extracts of a soil sample taken from the LUST site had 8 microg L(-1) MTBE.
Highly sensitive real-time pyrosequencing seems promising for constructing an inexpensive and small DNA sequencer with a low running cost. A DNA sample of a picomole level is usually used in the conventional pyrosequencing based on a luciferase assay coupled with an APS-ATP surfurylase reaction for producing ATP from pyrophosphate (PPi). Although the luminescence intensity could be increased by increasing the amount of luciferase, it was impossible to reduce the target DNA amount because of a large background luminescence due to the luciferase-APS reaction. In this report, a novel approach using a new conversion reaction of PPi to ATP is proposed. This method has a very low background and can produce high signals in the presence of a large amount of luciferase; thus, the sample amount required for sequencing is significantly reduced. The ATP production from PPi is catalyzed with pyruvate orthophosphate dikinase (PPDK) using AMP and phosphoenolpyruvate as the substrates, which are inactive for the luciferase-catalyzed reaction. All of the components in the AMP-PPDK-based pyrosequencing system are suitable for highly sensitive DNA sequencing in one tube. Real-time DNA sequencing with a readable length up to 70 bases was successfully demonstrated by using this system. By increasing the amount of luciferase, as low as 2.5 fmol of DNA templates was accurately sequenced by the proposed method with a novel simple and inexpensive DNA sequencer having a photodiode array as a sensor instead of a PMT or CCD camera. A sample amount as low as 2 orders of magnitude smaller than that used in the conventional pyrosequencer can be used.
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We review statistical methodology for estimating mean concentrations of potentially toxic pollutants in water, for small samples that are not normally distributed and often contain substantial numbers of nondetects, i.e. samples that are only known to be below some set of fixed thresholds. Maximum likelihood estimation (MLE) and regression on order statistics (ROS) are two main approaches that dominate the literature, with transformation bias under non-normality that increases with the severity of censoring being the main problem. We consider exact maximum likelihood estimators in conjunction with the Box-Cox transformation and propose the Quenouille-Tukey Jackknife as a method for bias reduction and variance estimation. Exact maximum likelihood estimators resulting from the expectation-maximization (EM) algorithm are exhibited in a simple heuristic form that also provides estimated values for the nondetects as subsidiary outputs. We show in simulationsthatthetwo main approaches perform well for the log-normal and gamma distributions as long as the jackknife is employed to reduce bias. Bias corrections to MLE used in the literature are shown to correct in the wrong direction under severe censoring. The jackknife is also used for estimating the variance of the both the MLE and ROS estimators. Robustness is improved by searching a class of power transformations (Box-Cox) for the best approximating normal distribution. We conclude that both the exact MLE and ROS procedures can be useful under varying experimental conditions. Limited simulations indicate that the ROS procedure is unbiased and has a smaller variance than the MLE under the log-normal distribution and is robust. The MLE performed better in simulations involving the gamma as the underlying distribution. We also compare the estimators for the mean and variance that one obtains from typical sets of water quality data, analyzing for copper, alumnium, arsenic, chromium, nickel, and lead.
In this communication, single molecules of beta-galactosidase were captured on a 1 mm femtoliter array using biotin-streptavidin binding. The femtoliter arrays, containing 24 000 individual reaction chambers, permit digital concentration readout as the percentage of reaction vessels that successfully capture a target molecule is correlated to the bulk target concentration. This capture and readout approach should prove useful for DNA and antibody assays that utilize an enzyme label to catalyze the generation of a fluorescent signal.
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