Growth dilution of metals in microalgal biofilms

Despite the key role microalgae play in introducing toxicants into aquatic food webs, little is known about the effects of environmental factors on metal accumulation by these primary producers. Environmental factors such as light and nutrients alter growth rates and may consequently influence metal concentrations in microalgae through growth dilution. Laboratory experiments suggested that metal uptake and elimination by microalgal biofilms were gradual enough to enable dilution of metals within the biofilms by photosynthetically accrued carbon, and a simple kinetic model of metal accumulation predicted significant variation in metal content due to growth dilution over the natural range of microalgal growth rates. The ratio of metal uptake to carbon uptake by microalgal biofilms decreased exponentially with increasing light in short-term laboratory experiments because photosynthesis was much more sensitive to a light gradient than was metal uptake. The effect of light on biofilm metal concentrations was confirmed in situ with a long-term experiment in which experimental shading of biofilms in a metal-contaminated stream decreased biofilm growth rates and caused a 3x increase in biofilm concentrations of twelve metals, including methylmercury. Slow growth at the primary producer level is a likely contributor to higher biotic metal concentrations in shaded, oligotrophic, or cold ecosystems.     

Uncertainty in NAPL volume estimates due to random measurement errors during partitioning tracer tests

The uncertainty in NAPL volume estimates obtained through partitioning tracers can be quantified as a function of random errors in volume and concentration measurements when moments are calculated from experimentally measured breakthrough curves using the trapezoidal rule for numerical integration. The methodology is based upon standard stochastic methods for random error propagation. Monte Carlo simulations using a synthetic data set derived from the one-dimensional solution of the advective-dispersive equation serve to verify the process. It is shown that the uncertainty in NAPL volume predictions nonlinearly increases as the retardation factor decreases. An important result of this observation is that there is a large degree of uncertainty in using partitioning tracers to conclude NAPL is absent from the swept zone. Under the conditions investigated, random errors in concentration measurements are shown to have a greater impact on NAPL volume uncertainty than random errors in volume measurements, and it is also shown that uncertainty in NAPL volume decreases as the resolution of the breakthrough curves increases. The impact of uncertainty in background retardation (i.e., sorption of partitioning tracers in the absence of NAPL) was also investigated, and it likewise indicated that the relative uncertainty in NAPL volume estimates increases as the retardation factor decreases.     

Automation of long-path absorption cell measurements

Recent advances have been made in the operation of long-path absorption cells which make them easier to align and improve the accuracy of measurements made with them. Only one person is required now for routine measurements of low absorption coefficients of atmospheric absorbers. Unique gear designs for the adjustment of the cell mirrors are described which utilize low-torque linear drives and make possible rapid changes in pathlength and precision repositioning of the cell output beam at long pathlengths. Automation of cell operation by the use of remote Selsyn controls is described. Several techniques are discussed for precision optical alignment of long-path absorption cells, including the use of infrared radiation sources. The system accuracy which results from these refinements in operation is included.     

Generalized overlap measures for evaluation and validation in medical image analysis

Measures of overlap of labelled regions of images, such as the Dice and Tanimoto coefficients, have been extensively used to evaluate image registration and segmentation algorithms. Modern studies can include multiple labels defined on multiple images yet most evaluation schemes report one overlap per labelled region, simply averaged over multiple images. In this paper, common overlap measures are generalized to measure the total overlap of ensembles of labels defined on multiple test images and account for fractional labels using fuzzy set theory. This framework allows a single "figure-of-merit" to be reported which summarises the results of a complex experiment by image pair, by label or overall. A complementary measure of error, the overlap distance, is defined which captures the spatial extent of the nonoverlapping part and is related to the Hausdorff distance computed on grey level images. The generalized overlap measures are validated on synthetic images for which the overlap can be computed analytically and used as similarity measures in nonrigid registration of three-dimensional magnetic resonance imaging (MRI) brain images. Finally, a pragmatic segmentation ground truth is constructed by registering a magnetic resonance atlas brain to 20 individual scans, and used with the overlap measures to evaluate publicly available brain segmentation algorithms.     

Effect of surface roughness and stainless steel finish on Listeria monocytogenes attachment and biofilm formation

The purpose of this study was to evaluate the effect of surface roughness (Ra) and finish of mechanically polished stainless steel (Ra = 0.26 +/- 0.05, 0.49 +/- 0.10, and 0.69 +/- 0.05 microm) and electropolished stainless steel (Ra = 0.16 +/- 0.06, 0.40 +/- 0.003, and 0.67 +/- 0.02 microm) on Listeria adhesion and biofilm formation. A four-strain cocktail of Listeria monocytogenes was used. Each strain (0.1%) was added to 200 ml of tryptic soy broth (TSB), and coupons were inserted to the mixture for 5 min. For biofilm formation, coupons with adhesive cells were incubated in 1:20 diluted TSB at 32 degrees C for 48 h. The experiment was performed by a randomized block design. Our results show that the level of Listeria present after 48 h of incubation (mean = 7 log CFU/cm2) was significantly higher than after 5 min (mean = 6.0 log CFU/cm2) (P < 0.01). No differences in initial adhesion were seen in mechanically finished (mean = 6.7 log CFU/cm2) when compared with electropolished stainless steel (mean = 6.7 log CFU/cm2) (P > 0.05). Listeria initial adhesion (values ranged from 5.9 to 6.1 log CFU/cm2) or biofilm formation (values ranged from 6.9 to 7.2 log CFU/cm2) was not significantly correlated with Ra values (P > 0.05). Image analysis with an atomic force microscope showed that bacteria did not colonize the complete surface after 48 h but were individual cells or grouped in microcolonies that ranged from 5 to 10 microm in diameter and one to three cell layers in thickness. Exopolymeric substances were observed to be associated with the colonies. According to our results, electropolishing stainless steel does not pose a significant advantage for food sanitation over mechanically finished stainless steel.     

Kinetic modeling of oxidation of antibacterial agents by manganese oxide

Several groups of popular antibacterial agents (i.e., phenols, fluoroquinolones, aromatic N-oxides, and tetracyclines) were demonstrated in earlier studies to be highly susceptible to oxidation by manganese oxides, a common oxidant in soils. However, because of the high complexity, the reaction kinetics were not fully characterized. A mechanism-based kinetic model has now been developed to successfully describe the entire range of kinetic data for a total of 21 compounds of varying structural characteristics (with R2 > 0.93). The model characterizes the reaction kinetics by two independent parameters, the reaction rate constant (k) and total reactive surface sites (S(rxn)). The model fitting indicates that the reaction kinetics of antibacterials with MnO2 are controlled by either the rate of surface precursor complex formation (for tetracyclines) or by the rate of electron transfer within the precursor complex (for phenols, fluoroquinolones, and aromatic N-oxides). The effect of reactant concentration, pH, and cosolutes on the reaction kinetics was evaluated and correlated to kand S(rxn). All the trends are consistent with the proposed rate-limiting steps. This new model improves the ability to quantitatively evaluate the kinetics of oxidative transformation of organic contaminants by manganese oxides in well-defined systems.     

Enhanced visualization of histological samples with an adjustable RGB contrast system with application for tissue used in photodynamic therapy

The analysis of histological sections has long been a valuable tool in the pathological studies. The interpretation of tissue conditions, however, relies directly on visual evaluation of tissue slides, which may be difficult to interpret because of poor contrast or poor color differentiation. The Chromatic Contrast Visualization System (CCV) combines an optical microscope with electronically controlled light-emitting diodes (LEDs) in order to generate adjustable intensities of RGB channels for sample illumination. While most image enhancement techniques rely on software post-processing of an image acquired under standard illumination conditions, CCV produces real-time variations in the color composition of the light source itself. The possibility of covering the entire RGB chromatic range, combined with the optical properties of the different tissues, allows for a substantial enhancement in image details. Traditional image acquisition methods do not exploit these visual enhancements which results in poorer visual distinction among tissue structures. Photodynamic therapy (PDT) procedures are of increasing interest in the treatment of several forms of cancer. This study uses histological slides of rat liver samples that were induced to necrosis after being exposed to PDT. Results show that visualization of tissue structures could be improved by changing colors and intensities of the microscope light source. PDT-necrosed tissue samples are better differentiated when illuminated with different color wavelengths, leading to an improved differentiation of cells in the necrosis area. Due to the potential benefits it can bring to interpretation and diagnosis, further research in this field could make CCV an attractive technique for medical applications.     

Performance characteristics of a compact position-sensitive LSO detector module

We assembled a compact detector module comprised of an array of small, individual crystals of lutetium oxyorthosilicate:Ce (LSO) coupled directly to a miniature, metal-can, position-sensitive photomultiplier tube (PSPMT). We exposed this module to sources of 511-keV annihilation radiation and beams of 30- and 140-keV photons and measured spatial linearity; spatial variations in module gain, energy resolution, and event positioning; coincidence timing; the accuracy and sensitivity of identifying the crystal-of-first-interaction at 511 keV; and the effects of intercrystal scatter and LSO background radioactivity. The results suggest that this scintillator/phototube combination should be highly effective in the coincidence mode and can be used, with some limitations, to image relatively low-energy single photon emitters. Photons that are completely absorbed on their first interaction at 511 keV are positioned by the module at the center of a crystal. Intercrystal scatter events, even those that lead to total absorption of the incident photon, are placed by the module in a regular "connect-the-dot" pattern that joins crystal centers. As a result, the accuracy of event positioning can be made to exceed 90%, though at significantly reduced sensitivity, by retaining only events that occur within small regions-of-interest around each crystal center and rejecting events that occur outside these regions in the connect-the-dot pattern.