Direct on-filter assay of subtilisin-type proteolytic enzymes for rapid analysis of analyte captured from the workplace atmosphere

A simple assay for some proteolytic enzymes has been developed which can be performed directly on the surface of a cellulose nitrate filter used to capture the analyte during workplace monitoring for health and safety purposes. Following air sampling the analysis is performed on the filter which is retained within the air sampler. This involves two steps: first, a 15 min incubation in which the captured enzyme is dissolved and then digests an alkaline-phosphatase-labelled antibody immobilised as a small dot on the surface of the filter; and second, is a 10 min incubation with substrate solution, which follows an in situ wash under a vacuum. During the incubation colour develops on the spot at the location of the immobilised enzyme antibody conjugate. The intensity of the spot can be assessed visually within the sampler to ascertain the presence or absence of captured enzyme, or alternatively quantitative results can be obtained using an optical scanner. The limit of detection is 5 ng per filter for subtilisin (20 ng for visual discrimination between this standard and the zero). The assay is stable to the effects of ambient air sampling at 31 min(-1) for 18 h.     

Rheology of Lyocell Solutions from Different Cellulose Sources

Rheological measurements were used to characterize the behavior of lyocell solutions, i.e., cellulose dissolved in N-methymorpholine-N-oxide. Cellulose sources included dissolving pulp, kraft pulp, sugar cane fibers, and kenaf fibers. The dominance of viscous behavior, G values, over elastic behavior, G values, is affected by cellulose concentration and molecular weight. At lower concentrations and degrees of polymerization (DP), dissolving pulp solutions show viscous, inelastic behavior at low frequencies. At higher concentration and DP, dissolving pulp solutions are more elastic at higher frequencies. Solutions prepared with kenaf and sugar cane fibers show similar properties to those using pure dissolving pulp, and comparisons suggest the molecular weight and/or the presence of other substances such as lignin in the cellulose from these alternative sources affect the rheology.     

An investigation of adsorption at the air-water and soil-water interfaces for non-micellizing ethylene oxide-propylene oxide surfactants

Adsorption at the air–water interface and soil sorption from aqueous solution have been investigated for a group of ethylene oxide (EO)–propylene oxide (PO) block copolymeric surfactants. The group which have a common structural formula of EO_m PO_n EO_m is distinguished by the fact that they have large critical micelle concentration (CMC) values and therefore do not readily form micelles at common environmental concentrations and temperatures. Adsorption at the air–water interface is readily shown to be driven by the size of the hydrophobic PO block. The size of the reduction in surface tension produced by a common concentration of 10⁻⁵ mol dm⁻³ linearly increases with the size of the PO block as does the efficiency of adsorption at the air–water interface as measured by pC₂₀ – the negative logarithm of the surfactant concentration that produces a reduction in surface tension of 20 mN m⁻¹. Soil sorption data have also been captured for these compounds and the data are readily fitted to the Freundlich adsorption isotherm. However soil sorption is shown to be inversely related to the molecular mass of the molecules and appears to be related to the size of the hydrophilic EO blocks in the molecule.     

Genetic divergence between two morphologically similar varieties of the kuruma shrimp<Emphasis Type="Italic"> Penaeus japonicus</Emphasis>

The kuruma shrimp Penaeus japonicus is widely distributed throughout the Indo-West Pacific. Two morphologically similar varieties, I and II, are recognized from the South China Sea. The two varieties are characterized by different color banding patterns on the carapace, but there are no distinct differences in morphometric traits between them based on measurement of 13 characters. Sequence data and restriction profiles of the mitochondrial genes reveal that these two varieties represent distinct clades, with sequence divergences of about 1% (473 bp) in 16S rRNA, 6–7% (504 bp) in cytochrome oxidase I, and 16–19% (470 bp) in the control region. Analysis of amplified fragment length polymorphism confirms that the two varieties are genetically distinct. We also investigated the geographical distribution of the two varieties in the western Pacific by analyzing specimens collected from Japan and Singapore. Shrimps from Japan and Singapore have been found to belong to varieties I and II, respectively, suggesting that the two varieties have different geographical distribution. Phylogenetic study reveals that the two varieties are more closely related to each other than to the other phylogenetically related Penaeus species. Results from this study suggest the occurrence of two cryptic species in the kuruma shrimp P. japonicus.Communicated by M.S. Johnson, Crawley     

Selecting peptide ligands of microcystin-LR from phage displayed random libraries

In the present study, we investigated to find novel ligands for low molecular weight environmental toxin, microcystin-LR (MC-LR) by using phage display technology. Two random libraries, displaying linear 12-mer peptides and cyclic 7-mer peptides, were screened against the immobilized target respectively. After three rounds of panning, phage clones that recognized microcystin-LR specifically were obtained from both the linear and the constrained libraries, proved by enzyme-linked immumosorbent assays and immunoprecipitation assays. DNA sequencing indicated that peptides displayed on some of the selected clones shared consensus sequences. Compared with traditional methods, this approach provided a cheaper and more rapid alternative to screen specific ligands for microcystin-LR. Moreover, since it is rather difficult to take small molecules as targets of phage display libraries, the success of this experiment expanded the applications of phage display technology, and provided a new avenue to study environmental small molecular toxins.     

Model testing using data on 137Cs from Chernobyl fallout in the Iput River catchment area of Russia

Data collected for 10 years following the Chernobyl accident in 1986 have provided a unique opportunity to test the reliability of computer models for contamination of terrestrial and aquatic environments. The Iput River scenario was used by the Dose Reconstruction Working Group of the BIOMASS (Biosphere Modelling and Assessment Methods) programme. The test area was one of the most highly contaminated areas in Russia following the accident, with an average contamination density of 137Cs of 800,000 Bq m-2 and localized contamination up to 1,500,000 Bq m-2, and a variety of countermeasures that were implemented in the test area had to be considered in the modelling exercise. Difficulties encountered during the exercise included averaging of data to account for uneven contamination of the test area, simulating the downward migration and changes in bioavailability of 137Cs in soil, and modelling the effectiveness of countermeasures. The accuracy of model predictions is dependent at least in part on the experience and judgment of the participant in interpretation of input information, selection of parameter values, and treatment of uncertainties.     

The acetochlor registration partnership: prospective ground water monitoring program

The Acetochlor Registration Partnership conducted a prospective ground water (PGW) monitoring program to investigate acetochlor [2-chloro-N-(ethoxymethyl)-N-(2-ethyl-6-methylphenyl)-acetamide] transport to ground water at eight sites. The distribution of soil textures among these sites was weighted toward coarser soil types, while also including finer-textured soils that dominate most corn (Zea mays L.)-growing areas of the United States. Each site consisted of a 1.2-ha test plot adjacent to a 0.2-ha control plot. Suction lysimeters and monitoring wells were installed at multiple depths within each test and control plot to sample soil-pore water and near-surface ground water. Irrigation was applied to each site during the growing season to ensure water input of 110 to 200% of average historical rainfall. Acetochlor dissipated rapidly from surface soils at all sites with a DT(50) (time for 50% of the initial residues to dissipate) of only 3 to 9 d, but leaching was not an important loss mechanism, with only 0.25% of the 15,312 soil-pore water and ground water samples analyzed containing parent acetochlor at or above 0.05 microg L(-1). However, quantifiable residues of a soil degradation product, acetochlor ethanesulfonic acid, were more common, with approximately 16% of water samples containing concentrations at or above 1.0 microg L(-1). A second soil degradation product, acetochlor oxanilic acid, was present at concentrations at or above 1.0 microg L(-1) in only 0.15% of water samples analyzed. The acetochlor PGW program demonstrated that acetochlor lacks the potential to leach to ground water at detectable concentrations, and when applied in accordance with label restrictions, is unlikely to move to ground water at concentrations hazardous to human health.     

Predictions of binary sorption isotherms for the sorption of heavy metals by pine bark using single isotherm data

The adsorption of three heavy metal ions by pine bark was studied. The study was divided into two parts; single component adsorption of the metals Cu2+, Cd2+ and Ni2+ and bisolute adsorption of the three binary systems Cu2+-Cd2+, Cu2+-Ni2+ and Cd2+-Ni2+. Extended Langmuir model, extended Freundlich model. Sips model and ideal adsorption solution theory (IAST) models were used to predict the equilibrium uptake for Cu2+, Cd2+ and Ni2+ in the binary diluted solutions using the single adsorption constants. The experimental data of single isotherm adsorption process were found to follow Langmuir isotherm model with less accuracy than Freundlich and Sips models. Whereas, the predictions of bisolute adsorption isotherms of the mentioned three systems, Cu2+-Cd2+, Cu2+-Ni2+ and Cd2+-Ni2+, showed good agreement with experimental data when using Extended-Langmuir, Extended-Freundlich and IAST. However, the only good fit of the Sips model was with the Cu2+-Cd2+ system.     

Microcalorimetric studies of the toxic action of La3+ in mitochondria isolated from Star-cross 288 chicken heart tissue cells

The thermogenic curves of the metabolism of mitochondria isolated from the heart of chicken Star-cross 288 and the effect of La3+ on it were studied by using an LKB-2277 BioActivity Monitor, ampoule method, at 37 degrees C. After isolation from the chicken heart tissue, mitochondria still have metabolic activity and can live for a long time by using the stored nutrients. From the thermogenic curves, we obtained the thermokinetic equations under different conditions. The kinetics show that La3+ has changed the metabolism completely.     

Performance of Trickle-Bed Air Biofilter: A Comparative Study of a Hydrophilic and a Hydrophobic Voc

Two lab-scale trickle-bed air biofilters were operated for investigating the difference in performance between a hydrophilic and a hydrophobic volatile organic compound (VOC). Methyl isobutyl ketone (MIBK) and styrene were selected as a model hydrophilic and hydrophobic VOCs, respectively. Effects of loading rates, biofilter re-acclimation, removal profile along biofilter depth, nitrogen consumption, and CO₂ production were compared under three operating conditions, namely, backwashing and two non-use periods (starvation and stagnant). Consistent over 99% removal efficiency up to loading rates of 3.26 kg COD/m³-day was obtained for the MIBK biofilter at 0.76 min empty bed retention time (EBRT) and 1.5 L/d nutrient flow. A similar performance for the styrene biofilter was obtained for loading rates up to 1.9kg COD/m³-day at 2.02 min EBRT and 2.4 L/d nutrient flow. The MIBK biofilter required only an initial acclimation period of 16 days while styrene biofilter required 46 days. Non-use periods can be used as another means of biomass control for both biofilters when the employed loading rate did not exceed 1.27 and 2.17 kg COD/m³-day for styrene and MIBK biofilters, respectively. The re-acclimation of both biofilter was delayed with increase of loading rate. MIBK biofilter re-acclimated in 90 min, while styrene biofilter re-acclimated in more than 600 min. Under similar loading rates, MIBK biofilter utilized less biofilter depth than styrene biofilter. Nitrogen consumption behaviors were apparently different between the two biofilters. Styrene biofilter had higher CO₂ production than MIBK biofilter and its CO₂ production was closely related to the theoretical complete chemical oxidation.