Single-molecule kinetics of lambda exonuclease reveal base dependence and dynamic disorder

We used a multiplexed approach based on flow-stretched DNA to monitor the enzymatic digestion of lambda-phage DNA by individual bacteriophage lambda exonuclease molecules. Statistical analyses of multiple single-molecule trajectories observed simultaneously reveal that the catalytic rate is dependent on the local base content of the substrate DNA. By relating single-molecule kinetics to the free energies of hydrogen bonding and base stacking, we establish that the melting of a base from the DNA is the rate-limiting step in the catalytic cycle. The catalytic rate also exhibits large fluctuations independent of the sequence, which we attribute to conformational changes of the enzyme-DNA complex.     

Impacts of climate change on Swiss biodiversity: An indicator taxa approach

We present a new indicator taxa approach to the prediction of climate change effects on biodiversity at the national level in Switzerland. As indicators, we select a set of the most widely distributed species that account for 95% of geographical variation in sampled species richness of birds, butterflies, and vascular plants. Species data come from a national program designed to monitor spatial and temporal trends in species richness. We examine some opportunities and limitations in using these data. We develop ecological niche models for the species as functions of both climate and land cover variables. We project these models to the future using climate predictions that correspond to two IPCC 3rd assessment scenarios for the development of ‘greenhouse’ gas emissions. We find that models that are calibrated with Swiss national monitoring data perform well in 10-fold cross-validation, but can fail to capture the hot-dry end of environmental gradients that constrain some species distributions. Models for indicator species in all three higher taxa predict that climate change will result in turnover in species composition even where there is little net change in predicted species richness. Indicator species from high elevations lose most areas of suitable climate even under the relatively mild B2 scenario. We project some areas to increase in the number of species for which climate conditions are suitable early in the current century, but these areas become less suitable for a majority of species by the end of the century. Selection of indicator species based on rank prevalence results in a set of models that predict observed species richness better than a similar set of species selected based on high rank of model AUC values. An indicator species approach based on selected species that are relatively common may facilitate the use of national monitoring data for predicting climate change effects on the distribution of biodiversity.     

Molecular and serological detection of Babesia spp. in neotropical and exotic carnivores in Brazilian zoos

Large and small piroplasms have been observed in the blood smears of various wild carnivores, but few studies utilizing molecular characterization have been done. The goal of this present study was to investigate the presence of Babesia sp. by molecular and serologic techniques in exotic and neotropical carnivores maintained in captivity at Brazilian zoos. Blood and sera samples were collected from 146 Brazilian wild felids, 21 exotic felids, 1 genet (Genetta tigrina), 3 European wolves (Canis lupus), and 94 Brazilian wild canids in Brazilian zoos in the S?o Paulo and Mato Grosso states and in the Federal District. A total of 53 wild felids (31.74%) and 10 wild canids (10.31%) were seropositive for Babesia canis by Indirect Fluorescent Antibody Test (IFAT). Antibodies were detected in ocelots, little-spotted cats, margays, pampas cats, jaguars, pumas, jaguarundis, crab-eating foxes, and bush dogs. Babesia sp. DNA, with high similarity to B. leo, was detected in one pampas cat and one genet.     

Lamellodiscus (Monogenea: Diplectanidae) parasites of Dentex macrophthalmus (Teleostei: Sparidae) from the North Atlantic coast of Africa, with a redescription of L. dentexi Aljoshkina, 1984, and description of three new species

Lamellodiscus dentexi Aljoshkina, 1984, a gill parasite of Dentex macrophthalmus (Bloch), is redescribed based on new material from the northwest coast of Africa (Senegal and Morocco). Three new species of Lamellodiscus Johnston et Tiegs, 1922 from D. macrophthalmus are described, Lamellodiscus toguebayei sp. n., L. vicinus sp. n., and L. triacies sp. n., all belonging to the "ignoratus" group. They can be distinguished from all other species of this group by the size and shape of male copulatory organ and sclerotised parts of the haptor. Considering the peculiar morphology of the male copulatory organ (long and thin tube) we propose to put together L. dentexi, L. virgula Euzet et Oliver, 1967 and L. obeliae Oliver, 1973 to form the "elongatus" type within the "elegans" group.     

Possible role of plant phenolics in the production of trichothecenes by Fusarium graminearum strains on different fractions of maize kernels

Four trichothecene-producing strains of Fusarium graminearum were grown on three maize grain fractions, whole grain, degermed grain, and the germ, to determine the effect of natural substrates on mycotoxin production. Monitoring the ergosterol content after 25 days of incubation indicated that fungal growth on all grain fractions was comparable. Trichothecene (TCT) production was highest on degermed grain, less on whole grain, and very low or nondetectable on the germ; similar results were found with four different strains. It was concluded that inhibitor(s) of TCT biosynthesis were present in maize germ. The presence of phenolic compounds was investigated in the different fractions. The hydroxamate 4-acetylbenzoxazolin-2-one (4-ABOA), a known inhibitor of mycotoxin production, was found in the degermed and whole grain fractions but not in the germ. Therefore, the TCT inhibition observed on the maize germ fraction used in our study is clearly not linked to 4-ABOA. Other soluble phenolic compounds were found at a much higher concentration in the germ than in the two other fractions. The inhibition property of the soluble ester-bound extracts was tested in liquid culture. A possible role for these compounds is discussed.     

Management of human health risk in the context of kitchen gardens polluted by lead and cadmium near a lead recycling company

Purpose

At the global scale, gardening activities are often performed in urban areas with a historical background of pollution. In this study, a participatory program was developed with citizens concerned by gardening activities near a 50-year-old regulated lead recycling company, with the aim of co-constructing the tools for the assessment and management of potential sanitary risks induced by historic pollution with persistent (eco) toxic metals: lead and cadmium.

Materials and methods

Soils and vegetables (lettuce, leek, celery, carrot, chard, pumpkin, and celeriac) samples were collected from four kitchen gardens neighboring a 50-year-old secondary lead smelter. Both total and in vitro human bioaccessible metal concentrations in the cultivated plants were measured in relation to soil characteristics.

Results and discussion

The results showed that the soils of these gardens were slightly contaminated by metals (Pb, 77 to 236 mg kg^?1; and Cd, 0.5 to 1 mg kg^?1) in comparison with the natural geologic background. However, significant pollution of vegetables can occur especially with lead (Pb up to 9.8 mg kg^?1 in lettuce) and certainly as a result of direct foliar transfer. The washing of plants before consumption is therefore recommended in the context of atmospheric fallout of ultrafine particles enriched with metals.

Conclusions

Metal bioaccessibility measure integrates the influence of metal type, plant type, and soil physico-chemical properties. Based on the results, it is proposed that human bioaccessible fraction of metals may also be currently taken into account as well as total metal quantities and bioaccumulation factors in risk assessment studies performed in gardens. Overall, this study has led to reflections and functional recommendations aimed at reducing human exposure and to finally developing sustainable gardening practices.

     

Effect of pH and temperature on comparative antioxidant activity of nonenzymatically browned proteins produced by reaction with oxidized lipids and carbohydrates

The antioxidative activity of nonenzymatically browned bovine serum albumin (BSA) produced by reaction with ribose (RI), hydroperoxides of methyl linoleate oxidation (HP), and secondary products of methyl linoleate oxidation (SP), at different pHs (4, 7, and 10) and temperatures (25, 37, 50, 80, and 120 degrees C), was studied to compare the antioxidative effects of carbohydrate- and oxidized lipids-modified proteins. The modified proteins (RIBSA, HPBSA, and SPBSA) were tested for antioxidative activity (at 100 ppm) in soybean oil using the thiobarbituric acid-reactive substances (TBARS) assay. All of them decreased significantly (p < 0.05) the TBARS formation in the oil and exhibited different effectiveness as a function of the temperature and the pH of the medium. In addition, there was a good correlation between the antioxidative activity of the protein and the amino acid losses produced during the nonenzymatic browning. These results are in agreement with an analogous and complimentary contribution of both Maillard and oxidized lipid/protein reactions to the antioxidative activity produced in foods during processing and storage.     

Analysis of bitter limonoids in citrus juices by atmospheric pressure chemical ionization and electrospray ionization liquid chromatography-mass spectrometry

Improved analytical techniques for bitter limonoids in citrus and citrus juices can expedite the evaluation of freeze-induced citrus damage for citrus growers and juice quality for citrus juice producers. Microbore normal-phase and reverse-phase chromatography coupled to a mass spectrometer operating in a positive ion atmospheric pressure chemical ionization and electrospray ionization modes were found to be rapid, selective, and sensitive methods for the analysis of the bitter limonoids limonin and nomilin in citrus juices. Analysis was performed on a chloroform extract of citrus juice to which an internal standard was added. The methods are capable of detecting citrus limonoids in citrus juice in the 60-200 picogram range and quantifying citrus juice limonoids in concentrations as low as 120 picograms. An accurate "total limonoid bitterness" in citrus juice, as represented by the combined occurrence of limonin and nomilin, is easily determined by these methods.     

Analytical validation of a portable human Accu-Chek glucometer in honeybee hemolymph

Glucose and trehalose are the main energy sources used by honeybees (Apis mellifera) for daily activities. However, there is no validated point-of-care method to reliably measure both sugars. We performed an analytical validation of a portable human glucometer (Accu-Chek; Roche) for glucose measurement in honeybee hemolymph compared to a reference method (GluCH, UniCel DxC 600; Beckman Coulter). We used 30 pooled hemolymph samples collected from the antennae of anesthetized honeybees and diluted 1:4 in 0.9% saline. We evaluated dilution linearity, spike recovery, and inter- and intra-assay imprecision. Glucose concentration was measured over time (2 h, 4 h, 8 h, 12 h, 1 d, 2 d, 3 d, 7 d, 21 d, 28 d) at various storage temperature (25°C, 4°C, −20°C, −80°C). The trehalose concentration was measured indirectly by trehalase hydrolyzation. Glucose concentrations measured by both instruments had a strong correlation (0.985, p < 0.0001) and a bias of −7.33 mmol/L (±1.96SD: 13.70 to −28.36), with linear agreement at <20 mmol/L (physiologic value: 100 mmol/L). The accuracy of the glucometer decreased at >20 mmol/L. Recovery of 115–130% of diluted spikes indicated good specificity. Inter- and intra-assay imprecision were 2.50% and 2.21%, respectively. Glucose concentrations fluctuated in stored samples dependent on time and temperature; however, glucose concentrations were constant with storage at −80°C for ≥28 d. The Accu-Chek glucometer is an adequate instrument to measure honeybee glucose concentration in hemolymph diluted with 0.9% NaCl, with good accuracy and precision at <20 mmol/L. Hemolymph storage at −80°C is suitable for long-term conservation of glucose.