Coccolithophores: functional biodiversity, enzymes and bioprospecting

Emiliania huxleyi is a single celled, marine phytoplankton with global distribution. As a key species for global biogeochemical cycling, a variety of strains have been amassed in various culture collections. Using a library consisting of 52 strains of E. huxleyi and an 'in house' enzyme screening program, we have assessed the functional biodiversity within this species of fundamental importance to global biogeochemical cycling, whilst at the same time determining their potential for exploitation in biocatalytic applications. Here, we describe the screening of E. huxleyi strains, as well as a coccolithovirus infected strain, for commercially relevant biocatalytic enzymes such as acid/alkali phosphodiesterase, acid/alkali phosphomonoesterase, EC1.1.1-type dehydrogenase, EC1.3.1-type dehydrogenase and carboxylesterase.     

Promoting microbial immobilization of soil nitrogen during restoration of abandoned agricultural fields by organic additions

Application of organic materials to soils to enhance N immobilization into microbial biomass, thereby reducing inorganic N concentrations, was studied as a management option to accelerate the reestablishment of the native vegetation on abandoned arable fields on sandy soils the Kiskunság National Park, Hungary. Sucrose and sawdust were used at three different topographic sites over 4 years. N availability and extractable inorganic N concentrations were significantly reduced in all sites. Soil microbial biomass C and microbial biomass N increased significantly following C additions, but the microbial C to microbial N ratio remained unaffected. It is concluded that the combined application of the rapidly utilized C source (sucrose) promoted N immobilization, whereas the addition of the slowly utilized C source (sawdust) maintained the elevated microbial biomass C and microbial biomass N in the field.     

Integration of β-Glucan Fibre Rich Fractions from Barley and Mushrooms to Form Healthy Extruded Snacks

β-glucan is a commonly researched plant cell wall component that when incorporated into food products has been associated with cholesterol and glycaemic response reductions. This study focusses on β-glucan rich fractions from barley and mushroom used in the production of extruded ready to eat snacks. Inclusion of barley β-glucan rich fractions and mushroom β-glucan fractions at 10 % levels increased the total dietary fibre content of extrudates compared to the control (P?<?0.05). Product expansion increased with the introduction of both barley and mushroom fraction (P?<?0.05) which in turn resulted in a reduction in product hardness (P?<?0.05). In vitro digestion protocol illustrated that inclusion of barley and mushroom β-glucan rich fractions manipulated the starch digestibility profile and hence rate of glucose release during digestion compared to the control sample. This in turn resulted in a significant (P?<?0.05) reduction in potential glycaemic response of the samples of between 20 and 25 % for barley β-glucan rich fractions and between 17 and 25 % for mushroom β-glucan rich fractions. We conclude that the inclusion of these fractions could be utilised by the food industry to manipulate the glycaemic response of extruded snack products.     

Rapid,effective and user-friendly immunophenotyping of canine lymphoma using a personal flow cytometer

Background

Widespread use of flow cytometry for immunophenotyping in clinical veterinary medicine is limited by cost and requirement for considerable laboratory space, staff time, and expertise. The Guava EasyCyte Plus (Guava Technologies, Hayward, CA, US) is the first, personal, bench-top flow cytometer designed to address these limitations.

Objective

The aim of this study was to adapt the immunohistochemical protocol used for immunophenotyping of canine lymphoma to the personal flow cytometer for rapid, effective and user-friendly application to the diagnosis and prognosis of canine lymphoma and to demonstrate its practicality for widespread veterinary application. Performance of the personal flow cytometer for immunophenotyping T and B lymphocytes in blood and lymph nodes from normal dogs and dogs with lymphoproliferative disease, was assessed using only two monoclonal antibodies (against CD3 and CD21), and by comparison with analysis using two conventional flow cytometers.

Methods

26 dogs with lymphoproliferative disease (23 with lymphoma, 3 with lymphocytic leukaemia) were studied along with 15 controls (2 non-lymphoma lymph nodes and 13 non-leukemic bloods. Lymphocytes were immunostained with fluorescent-labeled, monoclonal antibodies against CD3 and CD21. To assess the effectiveness of the personal flow cytometer in discrimination between T and B cell immunophenotypes, T and B cell counts for half the samples (14 blood and 11 lymph node) were also determined using the same method and conventional flow cytometers (FACSCalibur, Cyan Dako). To assess the effectiveness of the personal flow cytometer in discriminating between leukocyte types, lymphocyte differential counts were determined for 21 blood samples and compared with those from automated hematology analyzers (CELL-DYN 3500, n=11 and ADVIA 2120, n=10). Quality and sub-cellular distribution of immunostaining was assessed using fluorescence microscopy.

Results

The protocol for immunophenotyping took 2 to 3 hours to complete from the point of receipt of sample to reporting of immunophenotype. The personal flow cytometer differential lymphocyte counts correlated highly (n=20; r=0.97, p<0.0001) with those of automated haematology analyzers. The personal flow cytometer counts consistently, but mildly, underestimated the percentages of lymphocytes in the samples (mean bias of -5.3%.). The personal flow cytometer immunophenotype counts were indistinguishable from those of conventional flow cytometers for both peripheral blood samples (n=13; r=0.95; p<0.0001; bias of -1.1%) and lymph node aspirates (n=11,r=0.98; p<0.001; bias of 1%). All but one leukemic and one lymphomatous lymph node sample, out of 26 samples of dogs with lymphoproliferative disease analyzed, could be immunophenotyped as either B or T cells.

Conclusions

We conclude that use of only 2 monoclonal antibodies is sufficient for immunophenotyping most cases of canine lymphoma by flow cytometry and enables rapid immunophenotyping. The personal flow cytometer may be as effectively used for immunophenotyping canine lymphoma as conventional flow cytometers. However, the personal flow cytometer is more accessible and user-friendly, and requires lower sample volumes.     

The representation of landscapes in global scale assessments of environmental change

Landscape ecology has provided valuable insights in the relations between spatial structure and the functioning of landscapes. However, in most global scale environmental assessments the representation of landscapes is reduced to the dominant land cover within a 0.5 degree pixel, disregarding the insights about the role of structure, pattern and composition for the functioning of the landscape. This paper discusses the contributions landscape ecology can make to global scale environmental assessments. It proposes new directions for representing landscape characteristics at broad spatial scales. A contribution of landscape ecologists to the representation of landscape characteristics in global scale assessments will foster improved information and assessments for the design of sustainable earth system governance strategies.     

Quantitative analysis of soluble exudates produced by ectomycorrhizal roots as a response to ambient and elevated CO2

Despite its potential impact on soil carbon flow, few studies have attempted to quantify the effects of elevated carbon dioxide (CO₂) on production of exudates by mycorrhizal plants. In this study we quantified low molecular weight (LMW) organic compounds exuded by non-mycorrhizal (NM) and ectomycorrhizal (ECM) plants in relation to exposure to elevated CO₂. Scots pine seedlings, either colonized by one of eight different ECM fungi or non-mycorrhizal (NM), were exposed to either ambient (350 ppm) or elevated (700 ppm) concentrations of CO₂. Exudation of LMW organic acids (LMWOAs), amino acids, dissolved monosaccharides and total dissolved organic carbon (DOC) was determined and exudation rates were calculated per g root and fungal dry mass. CO₂ had a significant impact on exudation. Under elevated CO₂, exudation of total LMWOAs increased by 120-160%, amino acids by 250%, dissolved monosaccharides by 130-270% and DOC by 180-220% compared to ambient CO₂ treatment. Net CO₂ assimilation rates increased significantly by 41-47% for seedlings exposed to elevated CO₂. Exuded C calculated as a percentage of assimilated CO₂ increased by 41-88% in the elevated CO₂ treatment compared to ambient CO₂ treatment.     

Comparison of ozone and UV-C treatments on the postharvest stilbenoid monomer, dimer, and trimer induction in var. 'Superior' white table grapes

Postharvest treatment of seedless white table grapes (var. 'Superior') with different gas ozone concentrations (3.88 and 1.67 g/h) for 1, 3, and 5 h induced an increase in stilbenoid biosynthesis [trans-resveratrol, piceatannol, and viniferinas (resveratrol dehydrodimers and dehydrotrimers)] during storage at 22 degrees C and 95% relative humidity. The maximal resveratrol concentration was reached after 2 days of storage, and this amount was similar to that induced by optimized UV-C treatments (1 min, 510 W, 40 cm). Although similar resveratrol concentrations accumulated in grapes after both UV-C and O3 treatments (maximum ozone production and time), the ozone treatment was more efficient in inducing viniferins accumulation in grape berries. A sequence in the biosynthesis of stilbenoids was observed, starting with the resveratrol monomer, continuing with the resveratrol dehydrodimers epsilon-viniferin and delta-viniferin, and ending with four different resveratrol dehydrotrimers. These trimers were different from alpha-viniferin, a trimer previously reported to be induced in grapes after biotic and abiotic stresses. Two alpha-viniferin isomers were also detected in the ozone-treated grapes, although at very low concentrations that prevented their quantification.     

Extracellular matrix expression by equine oral and limb fibroblasts in in vitro culture

Wounds on the limbs of horses are notoriously difficult to heal, with over production of TGFβ1 thought to be responsible for excessive scarring; in contrast, wounds in the oral cavity heal rapidly with minimal scarring. This experiment aimed to determine the effect of TGFβ1 on the production of mRNA and proteins for various extracellular matrix components by two equine fibroblast cell lines isolated from the oral mucosa and distal limb. Fibronectin mRNA was up-regulated by TGFβ1 in the limb but not the oral cells. TGFβ1 increased the ratio of mRNA for collagen types I-III for the oral cells only. mRNA expression for TGFβ receptors-I and -II was significantly lower in limb fibroblasts, and treatment of either cell line with TGFβ1 down-regulated mRNA expression for both receptors. These differences may account for the improved healing seen in oral wounds compared to the excessive scarring seen in limb wounds.