Characterization of partially purified glutathione reductase from cold-hardened and nonhardened spinach leaf tissue

Glutathione reductase (GR) (EC 1.6.4.2) was studied in crude and partially purified extracts from nonhardened (25/20 °C D/N) and hardened (5/5 °C D/N) spinach-leaf tissue. Crude extracts of hardened tissue showed a 66% increase in glutathione reductase activity over that of nonhardened tissue. The enzyme was purified by ammonium sulfate precipitation, Sephadex G-150 chromatography, 2′, 5′ ADP-Sepharose affinity chromatography, and DEAE-Sephadex A-50 ion-exchange chromatography. The partially purified enzyme from the two sources showed different kinetic characteristics, heat inactivation, freezing inactivation, and electrophoretic mobilities. Hardened leaves contain different forms of glutathione reductase than do nonhardened leaves. GR from hardened spinach has greater stability against freezing and a higher affinity for substrates at low temperature than does GR from nonhardened spinach.     

Biomarker of food intake for assessing the consumption of dairy and egg products

Dairy and egg products constitute an important part of Western diets as they represent an excellent source of high-quality proteins, vitamins, minerals and fats. Dairy and egg products are highly diverse and their associations with a range of nutritional and health outcomes are therefore heterogeneous. Such associations are also often weak or debated due to the difficulty in establishing correct assessments of dietary intake. Therefore, in order to better characterize associations between the consumption of these foods and health outcomes, it is important to identify reliable biomarkers of their intake. Biomarkers of food intake (BFIs) provide an accurate measure of intake, which is independent of the memory and sincerity of the subjects as well as of their knowledge about the consumed foods. We have, therefore, conducted a systematic search of the scientific literature to evaluate the current status of potential BFIs for dairy products and BFIs for egg products commonly consumed in Europe. Strikingly, only a limited number of compounds have been reported as markers for the intake of these products and none of them have been sufficiently validated. A series of challenges hinders the identification and validation of BFI for dairy and egg products, in particular, the heterogeneous composition of these foods and the lack of specificity of the markers identified so far. Further studies are, therefore, necessary to validate these compounds and to discover new candidate BFIs. Untargeted metabolomic strategies may allow the identification of novel biomarkers, which, when taken separately or in combination, could be used to assess the intake of dairy and egg products.     

Different pathways for iNOS-mediated toxicity in vitro dependent on neuronal maturation and NMDA receptor expression

Co-localization of activated microglia and damaged neurones seen in brain injury suggests microglia-induced neurodegeneration. Activated microglia release two potential neurotoxins, excitatory amino acids and nitric oxide (NO), but their contribution to mechanisms of injury is poorly understood. Using co-cultures of rat microglia and embryonic cortical neurones, we show that inducible NO synthase (iNOS)-derived NO aloneis responsible for neuronal death from interferon gamma (IFNgamma) +lipopolysaccharide (LPS)-activated microglia. Neurones remain sensitive to NO irrespective of maturation state but, whereas blocking NMDA receptor activation with MK801 has no effect on NO-mediated toxicity to immature neurones, MK801 rescues 60-70% of neurones matured in culture for 12 days. Neuronal expression of NMDA receptors increases with maturation in culture, accounting for increased susceptibility to excitotoxins seen in more mature cultures. We show that MK801 delays the death of more mature neurones caused by the NO-donor DETA/NO indicating that NO elicits an excitotoxic mechanism, most likely through neuronal glutamate release. Thus, similar concentrations of nitric oxide cause neuronal death by two distinct mechanisms: NO acts directly upon immature neurones but indirectly, via NMDA receptors, on more mature neurones. Our results therefore extend existing evidence for NO-mediated toxicity and show a complex interaction between inflammatory and excitotoxic mechanisms of injury in mature neurones.     

Estrella lausannensis, a new star in the Chlamydiales order

Originally, the Chlamydiales order was represented by a single family, the Chlamydiaceae, composed of several pathogens, such as Chlamydia trachomatis, Chlamydia pneumoniae, Chlamydia psittaci and Chlamydia abortus. Recently, 6 new families of Chlamydia-related bacteria have been added to the Chlamydiales order. Most of these obligate intracellular bacteria are able to replicate in free-living amoebae. Amoebal co-culture may be used to selectively isolate amoeba-resisting bacteria. This method allowed in a previous work to discover strain CRIB 30, from an environmental water sample. Based on its 16S rRNA gene sequence similarity with Criblamydia sequanensis, strain CRIB 30 was considered as a new member of the Criblamydiaceae family. In the present work, phylogenetic analyses of the genes gyrA, gyrB, rpoA, rpoB, secY, topA and 23S rRNA as well as MALDI-TOF MS confirmed the taxonomic classification of strain CRIB 30. Morphological examination revealed peculiar star-shaped elementary bodies (EBs) similar to those of C. sequanensis. Therefore, this new strain was called "Estrella lausannensis". Finally, E. lausannensis showed a large amoebal host range and a very efficient replication rate in Acanthamoeba species. Furthermore, E. lausannensis is the first member of the Chlamydiales order to grow successfully in the genetically tractable Dictyostelium discoideum, which opens new perspectives in the study of chlamydial biology.     

Chemotaxonomic evidence suggests that Eriocephalus tenuifolius is the source of Cape chamomile oil and not Eriocephalus punctulatus

The distinct blue essential oil obtained from Eriocephalus sp. (Asteraceae) is known in commerce as Cape chamomile and has become an important ingredient in flavour and fragrance formulations. To date, it is commonly accepted that Cape chamomile oil is obtained from Eriocephalus punctulatus. Samples of E. tenuifolius DC (n = 35) and E. punctulatus DC (n = 17) were collected from different localities in South Africa and Lesotho. The hydrodistilled essential oil was analysed by gas chromatography coupled to mass selective detector and flame ionisation detector (GC–MS-FID). The GC–MS data was exported to MetAlign? for spectral alignment and differentiation. The mid-infrared red (MIR) spectral data was processed using principal component analysis (PCA), partial least squares (PLS) and orthogonal projections to latent structures discriminant analysis (OPLS-DA) algorithms. The oil from E. punctulatus showed a different profile from the commercial sample which mostly contained 1,8-cineole, piperitone, yomogi alcohol and pogostol. MetAlign? successfully discriminated between the oil obtained from the two species and this data together with multivariate analysis showed that the commercial oil was similar to E. tenuifolius oil. Using the MIR data, a three component OPLS-DA model was constructed which convincingly discriminated between E. tenuifolius and E. punctulatus oil. The developed model further predicted the botanical origin of the commercial oils to be E. tenuifolius. The statistical performance of the model was excellent with an R²X (cum) = 0.897; R²Y (cum) = 0.991 and Q²Y (cum) = 0.987.     

Light intensity is a limiting factor to the inland expansion of Cape ivy (<Emphasis Type=Italic>Delairea odorata</Emphasis>)

Cape ivy (Delairea odorata) is a highly invasive climbing perennial vine that is primarily distributed in coastal communities of California and Oregon, with patchy infestations in some inland riparian areas. In this study, we evaluated light as a potential environmental limitation to the spread of Cape ivy into inland regions of the western United States. Cape ivy was collected from four locations representing the north to south range. Plants were grown for 9 to 11 weeks in full sunlight and under two shade regimes (20 and 6% of full sunlight). The experiment was conducted twice at two temperature regimes. Results show some within- and among-population variability, with the southernmost San Diego County population having the highest biomass under the warmer growing conditions and the three northern populations responding most favorably in the cooler growing conditions. Despite the minor differences within and between populations, Cape ivy grew very poorly in full sunlight in both experiments. Although plants growing under 6% light grew better than those in full sunlight, they were far less robust compared to plants growing at 20% light. Our results indicate that while Cape ivy will not persist in areas with prolonged high intensity sunlight, characterized by much of the interior regions of California and Oregon, it is expected to invade and spread in areas with reduced light, including coastal regions frequently exposed to fog or cloudy conditions, or sub-canopy layers of riparian forests or woodlands. These communities should be the target areas for early detection and rapid response programs to prevent further Cape ivy invasion.     

Species of Borrelia distinguished by restriction site polymorphisms in 16S rRNA genes

Abstract Three phyletic groups of Borrelia associated with Lyme disease, B. burgdorferi, B. garinii and group VS461 can be distinguished from each other and other species of Borrelia by Bfa I restriction site polymorphisms in PCR amplified 16S rRNA genes. One strain isolated from an Ixodes pacificus tick in California that was previously unclassifiable was distinguishable from B. burgdorferi by an Mnl I restriction site polymorphism.     

Ecological indicators based on trophic spectrum as a tool to assess ecosystems fishing impacts

Trophic indicators were used to compare two Malian freshwater reservoirs whose main differences are based on their different fishing pressures. Data were collected from a scientific survey of small-scale fishery landings conducted in 2002/2003. The trophic levels of fish species caught by artisanal fisheries are estimated from observations of scientific fishing or from the metabase Fishbase. Important differences exist in the trophic structure of both reservoirs. In Selingue (with high fishing pressure), very few top predators are found in the catches while the low trophic level fishes increase in total catches. In Manantali (with low fishing pressure), the top predators contribute twice as much to catches compared to Selingue. Hence, the mean trophic level of catches in Selingue (2.80) is lower than in Manantali (2.97). When comparing these results with those of study made in 1994/1995, it clearly appears that the effects of the fishing pressure in Selingue are obvious through a decrease of 0.12 in the mean trophic level while in Manantali this mean level has increased by 0.33 due to a recent strategic targeting of top predators. Trophic spectra seem to be relevant tools to characterize exploited fish communities from multi-specific and multi-gear small-scale fisheries catch data.