Airborne dust in swine and poultry confinements was analyzed to determine concentrations of total and gram-negative bacteria, total fungi, Aspergillus fumigatus and endotoxin. Airborne concentrations of total and gram-negative bacteria in swine and confinement units have been found to be as high as, or higher, than those found in other environments, such as wastewater treatment plants and cotton card rooms, where microbiologically contaminated organic dusts were present. Airborne endotoxin concentrations in the swine units (average 0.12 micrograms/m3) and poultry units (average 0.31 micrograms/m3) were in the range where clinical effects have occurred in other populations. Therefore, health studies of poultry and swine confinement workers with concurrent estimation of the individual daily exposure dose are warranted.
Results of the analyses of occupational and environmental samples are frequently reported as "less than a specified value," a practice followed by many analytical laboratories. A left-censored distribution occurs when analytical laboratories do not report results that fall below their limits of detection or quantification. Approximately 37% of the household interior dust lead loadings collected in a large-scale, multisite, longitudinal study of lead-based paint hazard controls were reported to be below the "method detection limit." These unreported values are unusable in any statistical analysis of the data and must be replaced by a valid dust lead loading estimate, a process called data imputation. This investigation tested how well data imputed using a newly formulated procedure for estimating the data below the method detection limit were correlated with dust lead loadings reported by the participating laboratories after special request. These results were also compared with those obtained by imputing the minimum detectable level by the square root of 2. Imputation of the low lead loadings was accomplished by substituting the value associated with the median percentile below each laboratory's method detection limit. A correlation of r = 0.50 was calculated between the predicted and reported dust lead loadings, with only slight bias (2.9%) in the predicted values. An alternative imputation procedure that used the predicted value from structural equation models fit to the noncensored dust lead loadings performed about as well, although the predictions had to be "centered" to correspond to the censored data. An estimator that combined both of these imputation procedures only slightly improved the correlation between the predicted and laboratory values (r = 0.51). These results support the use of the new procedure rather than the commonly used imputed values of the method detection limit divided by 2 or by the square root of 2. Imputing values based on either of these common approaches may result in much more biased predictions for the censored data; in the case of these data, the dust lead loadings were overestimated by 348%. The results also suggest that analytical laboratories should provide a numerical result for all analyzed samples, with a "flag" of those values below their detection limit, since these results may be more accurate than any imputed value, particularly those provided by the commonly used method of dividing the minimum detection limit by the square root of 2.
The mechanisms by which many plant growth promoting rhizobacteria (PGPR) affect plants are unknown. We recently isolated a rhizosphere bacterium (Bacillus thuringiensis NEB17), that promotes soybean growth and screened the liquid growth medium in which it grew for plant growth stimulating materials. We have also shown that it produces a bacteriocin (named by us as thuricin-17 and a member of the recently described class IId bacteriocins). Here we show that application of this bacteriocin to leaves (spray) or roots (drench) directly stimulates the growth of both a C(3) dicot (soybean) and a C(4) monocot (corn). This growth stimulation is similar in nature to that previously seen when plants are treated with Nod factors. Strain NEB17 contains three copies of the gene for thuricin 17 that code for identical amino acid sequences. These two lines of evidence suggest that the dual functions of these proteins may have constrained their evolution. This is the first report of direct plant growth enhancement by a bacteriocin.
This protocol describes an efficient method to site-specifically label cell-surface or purified proteins with chemical probes in two steps: PRobe Incorporation Mediated by Enzymes (PRIME) followed by chelation-assisted copper-catalyzed azide-alkyne cycloaddition (CuAAC). In the PRIME step, Escherichia coli lipoic acid ligase site-specifically attaches a picolyl azide derivative to a 13-amino acid recognition sequence that has been genetically fused onto the protein of interest. Proteins bearing picolyl azide are chemoselectively derivatized with an alkyne-probe conjugate by chelation-assisted CuAAC in the second step. We describe herein the optimized protocols to synthesize picolyl azide, perform PRIME labeling, and achieve CuAAC derivatization of picolyl azide on live cells, fixed cells, and purified proteins. Reagent preparations, including synthesis of picolyl azide probes and expression of lipoic acid ligase, take 12 d, while the procedure to perform site-specific picolyl azide ligation and CuAAC on cells or on purified proteins takes 40 min-3 h.
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