Development of a multianalyte enzyme-linked immunosorbent assay for permethrin and aroclors and its implementation for analysis of soil/sediment and house dust extracts

Development of a multianalyte enzyme-linked immunosorbent assay (ELISA) for detection of permethrin and aroclors 1248 or 1254 and implementation of the assay for analysis of soil/sediment samples are described. The feasibility of using the multianalyte ELISA to monitor aroclors 1254 and permethrin simultaneously was tested with permethrin and aroclor standards and with aroclor- and permethrin-containing soil/sediment and house dust samples. Comparison of the I?? and I?? values of the multianalyte with those of a single-analyte assay revealed similar results, and multianalyte ELISA determination of analyte amounts in soil/sediment dust samples yielded similar results to those of a single-analyte assay. A single-analyte assay of permethrin content in permethrin-containing dust samples showed that the ELISA can determine the analyte accurately in samples with dust matrix contents ranging from 6.25 to 100 mg as indicated by the good correlation between the results of the immunoassay and those of the gas chromatography analysis.     

Optimization of Hydrolysis Conditions for Production of Angiotensin-Converting Enzyme Inhibitory Peptides from Basa Fish Skin Using Response Surface Methodology

ABSTRACT

Basa fish skin free from fat and noncollagenous proteins was hydrolyzed by different proteinases to produce angiotensin-converting enzyme (ACE) inhibitory peptides. Protamex hydrolysates were found to have the highest ACE inhibitory activity. Response surface methodology (RSM) was applied to optimize the hydrolysis conditions of Protamex—including enzyme-to-substrate ratio, hydrolysis time, and solid (fish skin)-to-liquid (water for hydrolysis)—at a temperature of 50°C and a pH of 7.0. The mathematical model demonstrated a good fit with the experimental data. An enzyme-to-substrate ratio of 6,108 U/g, a hydrolysis time of 1.24 h, and a solid-to-liquid ratio of 1:8.05 g/mL were found to be the optimal conditions to achieve the highest value of ACE inhibitory activity. The IC₅₀ of the hydrolysates for ACE inhibitory activity was 1.417 mg/mL under these conditions.     

Characterization and Cytotoxicity Studies of the Rare 21:4 n-7 Acid and Other Polyunsaturated Fatty Acids from the Marine Opisthobranch Scaphander lignarius,Isolated Using Bioassay Guided Fractionation

The marine opisthobranch Scaphander lignarius has been analyzed in the systematic search for novel bioactive compounds in Arctic marine organisms using bioassay guided fractionation. A number of highly cytotoxic fractions were shown to contain mainly polyunsaturated fatty acids (PUFAs). Selected PUFAs were isolated and identified using both liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR). It was shown that the opisthobranch contained unusual PUFAs such as several ω3 fatty acids and the ω7 heneicosa-5,8,11,14-tetraenoic acid (21:4 n-7) not isolated before. The organism was shown to be a very rich source of PUFAs and the activity of the isolated compounds against a range of human cancer cell lines (melanoma, colon carcinoma and breast carcinoma) is further reported. The ω7 PUFA was significantly more cytotoxic in comparison with reference ω6 arachidonic and ω3 eicosapentaenoic acid. A noteworthy non-selective cytotoxicity against normal lung fibroblasts was also established. The paper contains isolation protocols in addition to cytotoxicity data of the isolated compounds. The potential of marine mollusks as a source for rare PUFAs is also discussed.     

Distribution of monoterpenes between organic resources in upper soil horizons under monocultures of Picea abies, Picea sitchensis and Pinus sylvestris

The aim of this study was to compare the monoterpene content and distribution in litters and roots of three conifer species: Picea abies (L.) Karst, Picea sitchensis (Bong.) Carr. and Pinus sylvestris (L.). We analysed the monoterpene content of green needles, needle litter, F (fermentation) layer material and roots collected from monoculture plots. The rate of loss of monoterpenes from freshly fallen litter in the field was also studied at two monthly intervals over 10 months, to assess the length of time that monoterpenes entering the litter layer remain. Monoterpene analysis was carried out by extracting homogenised samples in hexane and identifying and quantifying the resulting monoterpenes using gas chromatography with flame ionisation detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Mean total monoterpene concentrations varied significantly between the three species examined (e.g. in freshly fallen litter 1531 ± 96, 100 ± 5 and 1175 ± 122 μg g⁻¹ d. wt for P. abies, P. sitchensis and P. sylvestris); each species had distinctive and consistent monoterpene profiles associated with each type of tissue, and total monoterpene concentrations in green needles varied between individual trees of the same species, particularly for P. sitchensis. A substantial proportion of the monoterpene content of green needles remained in the needles after litter fall for P. abies (42%), P. sitchensis (11%) and P. sylvestris (30%). Although rates of monoterpene loss from needle litters varied initially (P. sylvestris > P. abies > P. sitchensis), the majority of the monoterpene content was lost after 4-6 months. Maximum monoterpene emission rates from decaying litter were calculated of 39 (P. abies), 1.7 (P. sitchensis) and 39 μg m⁻² h⁻¹ (P. sylvestris). Monoterpene concentrations in F layer material were very low (<10 μg g⁻¹ d. wt). Roots, particularly in P. sylvestris, represented a significant pool of monoterpenes (185 ± 16, P. abies; 258 ± 54, P. sitchensis; 2133 ± 200 μg g⁻¹ d. wt, P. sylvestris). The monoterpene profile was similar between roots and litter of P. sylvestris (α-pinene most abundant), and for P. sitchensis, (limonene and α-pinene most abundant), although a different pattern was observed between needle litter (most abundant β-pinene) and roots (most abundant myrcene) of P. abies. The relatively high concentrations and different profiles of monoterpenes characterised in upper organic soil horizons here emphasise the need for their influence on soil ecological processes to be assessed.     

A qPCR MGB probe based eDNA assay for European freshwater pearl mussel (Margaritifera margaritifera L.)

1. Environmental (e)DNA assays are becoming increasingly used to detect rare or invasive aquatic species. 2. The Critically Endangered freshwater pearl mussel Margaritifera margaritifera is undergoing range‐wide reduction in population numbers and distribution. 3. An eDNA assay to detect the presence of M. margaritifera was developed, based on the mitochondrial cytochrome oxidase I gene, utilizing species‐specific primers, a minor groove binding (MGB) probe and quantitative (q)PCR approaches. 4. The results from this pilot study demonstrated high sensitivity both in laboratory and field trials, and provide a valuable non‐invasive tool for detecting M. margaritifera.     

Infectious prions in the saliva and blood of deer with chronic wasting disease

A critical concern in the transmission of prion diseases, including chronic wasting disease (CWD) of cervids, is the potential presence of prions in body fluids. To address this issue directly, we exposed cohorts of CWD-na?ve deer to saliva, blood, or urine and feces from CWD-positive deer. We found infectious prions capable of transmitting CWD in saliva (by the oral route) and in blood (by transfusion). The results help to explain the facile transmission of CWD among cervids and prompt caution concerning contact with body fluids in prion infections.