Efficient Screening of Marine Extracts for Protease Inhibitors by Combining FRET Based Activity Assays and Surface Plasmon Resonance Spectroscopy Based Binding Assays

The screening of extracts from marine organisms is a widely used strategy to discover new drug leads. A common problem in the screening process is the generation of false positive hits through unspecific effects from the complex chemical composition of the crude extracts. In this study, we explored a combination of a fluorescence resonance energy transfer (FRET) based activity assay and a surface plasmon resonance (SPR) based binding assay to avoid this problem. An aqueous extract was prepared from rest raw material of the Norwegian spring spawning herring, and further fractionated by methanol solubility and solid phase extraction. FRET based activity assays were used to determine the influence of each extract on the activity of different proteases. Several extracts showed more than 50% inhibition. The inhibition mechanisms were elucidated by SPR based competition experiments with known inhibitors. For the secreted aspartic proteases 1, 2, 3 and HIV-1 protease, the results indicated that some extracts contain inhibitors interacting specifically with the active site of the enzymes. The study shows that a combination of an activity assay and an SPR based binding assay is a powerful tool to identify potent inhibitors in marine extracts. Furthermore, the study shows that marine vertebrates offer an interesting source for new bioactive compounds, although they have rarely been explored for this purpose.     

Ultrasonographic findings in 2 cows with duodenal obstruction

Two cases where clinical findings suggestive of proximal ileus resulting in hypokalemic and hypochloremic alkalosis are presented. Ultrasonographs showed distension of the sigmoid flexure along the ventral border of the liver and between the serosal lamellae of the greater omentum in the flank, allowing exact preoperative obstruction localization.     

A non-competitive chemiluminescence enzyme immunoassay for the equine acute phase protein serum amyloid A (SAA) -- a clinically useful inflammatory marker in the horse.

A non-competitive chemiluminescence enzyme immunoassay for measuring serum amyloid A (SAA) in equine serum was developed. A polyclonal anti-equine-amyloid A antiserum specific for equine SAA was utilized, and the assay was standardized using highly purified equine SAA. An acute phase horse serum was calibrated against the purified SAA and was used as standard when running the assay. Serum SAA concentrations in the range of 3-1210 mg/l could be measured. The reference range of SAA in clinically healthy adult horses was <7 mg/l. The clinical validation of the assay comprised the SAA responses after surgery and experimentally induced aseptic arthritis, and those associated with viral and bacterial infections. The SAA response after surgery (castration) was consistent, with peak concentrations on day 2 and a return to normal SAA concentrations within eight days. The aseptic arthritis produced an SAA response with a pattern similar to that seen after surgery, with peak concentrations of SAA 36-48 h after induction. Seven horses showed a biphasic pattern, with a second rise in SAA concentrations on day 4 and 5. All animals had SAA levels <7 mg/l on day 15. All horses with viral and bacterial infections had SAA concentrations above 7 mg/l. The ranges of SAA concentrations following the different types of inflammation overlap, being consistent with the unspecific nature of the SAA response. This study revealed that SAA is a sensitive and unspecific marker for inflammation, and describes the dynamics of the SAA response after standardized and well defined tissue damage.     

Ecosystem‐based management of Amazon fisheries and wetlands

Infrastructure development and overfishing in the Amazon make it imperative to define adequate scales for the ecosystem‐based management of commercial fisheries and the wetlands on which they depend. We mapped fisheries and fish ecology data from Brazil, Peru, Bolivia and Colombia to an explicit GIS framework of river basins and mainstems. Migratory species account for more than 80% of the known maximum catches of commercial fisheries across the Amazon. Of these migratory species, we nominated six long‐distance migratory fish taxa as flagship species to define the two main commercial fishery regions. The migrations of at least one goliath catfish species define a large‐scale longitudinal link joining the Andes, Amazon Lowlands and Amazon River estuary. Migratory Characiforms demonstrate interbasin wetland connectivity between nutrient‐rich and nutrient‐poor rivers over at least 2 million km², or about one‐third of the Amazon Basin. We show that flooded forest area is the most important wetland variable explaining regional variations in migratory characiforme biomass as indicated by maximum annual fishery catches. The sustainable management of Amazon fisheries will require transnational cooperation and a paradigm shift from local community management alone to a more integrated approach that considers both rural and urban consumers and challenges, and the realistic life histories of migratory species.     

Reference bulk density and critical degree-of-compactness for no-till crop production in subtropical highly weathered soils

The concept of degree of compactness (DC), referred to as field bulk density (BD) as a percentage of a reference bulk density (BD_ref), was developed to characterize compactness of soil frequently disturbed, but for undisturbed soil such as under no-tillage critical degree of compactness values have not been tested. The objective of this study was to compare methods to determine BD_ref and limits of DC and BD for plant growth under no-tillage in subtropical soils. Data from the literature and other databases were used to establish relationships between BD and clay or clay plus silt content, and between DC and macroporosity and yield of crops under no-tillage in subtropical Brazil. Data of BD_ref reached by the soil Proctor test on disturbed soil samples, by uniaxial compression with loads of 200 kPa on disturbed and undisturbed soil samples, and 400, 800 and 1600 kPa on undisturbed soil samples, were used. Also, comparisons were made with critical bulk density based on the least limiting water range (BDc LLWR) and on observed root and/or yield restriction in the field (BDc Rest). Using vertical uniaxial compression with a load of 200 kPa on disturbed or undisturbed samples generates low BD_ref and high DC-values. The standard Proctor test generates higher BD_ref-values, which are similar to those in a uniaxial test with a load of 1600 kPa for soils with low clay content but lower for soils with high clay content. The BDc LLWR does not necessarily restrict root growth or crop yield under no-tillage, since field investigations led to higher BDc Rest-values. A uniaxial load greater than 800 kPa is promising to determine BD_ref for no-tillage soils. The BD_ref is highly correlated to the clay content and thus pedotransfer functions may be established to estimate the former based on the latter. Soil ecological properties are affected before compaction restricts plant growth and yield. The DC is an efficient parameter to identify soil compaction affecting crops. The effect of compaction on ecological properties must also be further considered.     

良好的种蛋贮存有助于获得更多的雏鸡

种蛋的孵化率很大程度取决于种鸡群的繁殖力,但是种蛋的贮存也是一个主要决定因素（图1）。防止出现胚胎死亡是种蛋贮存的首要任务,而且这取决于许多影响因素。     

Temperature‐dependent feed requirements in farmed blue mussels (Mytilus edulis L.) estimated from soft tissue growth and oxygen consumption and ammonia‐N excretion

Feed requirements were estimated from specific growth rates in standardized soft tissue dry weight (SGR_DW’) and atomic O:N ratios for mussels fed seven rations of microalgae (5–735 μg C h^?1 ind^?1) at 7 and 14°C respectively. The mean oxygen consumption and ammonia‐N excretion rates were significantly higher at 14°C (0.29 μg O² and 27.3 μg N ind^?1 h¹) compared with those at 7°C (0.16 μg O² and 11.4 μg N ind^?1 h^?1) (P < 0.05), resulting in O:N ratios between 3 and 45 at 7°C and 7 and 28 at 14°C. Low O:N ratios indicate protein catabolism and an unfavourable condition, whereas high ratios indicate that carbohydrate is the primary energy source. The measured SGR_DW’ suggests minimum feed requirements of ~240 and ~570 μg C ind^?1 h^?1 for weight maintenance at 7 and 14°C, with corresponding O:N ratios of 24 and 16, respectively, indicating a more stressed condition at 14°C. A 0.5% SGR_DW’ day^?1 was obtained by ~565 (O:N = 29) and ~680 (O:N = 23) μg C ind^?1 h^?1 at 7 and 14°C respectively. A positive and significantly higher SGR_DW’, with the lowest feed ration at 7°C compared with a negative SGR_DW’ at 14°C (P < 0.05), indicated that storage time can also possibly be prolonged at low temperatures if the mussels are not fed.     

THE VALUE OF INFORMATION IN SALMON FARMING. HARVESTING THE RIGHT FISH AT THE RIGHT TIME

Research on management practices in fish farming has traditionally focused on two topics: production planning and forecasting of prices. This article combines these two areas of research, and illustrates how information on price patterns can change production plans, and hence increase the value of the farm enterprise. It will present a model farm and illustrate, with different levels of price information, how information on future prices alters the original production plan and hence creates extra value for the farmer. Although the specifics and empirical application of the paper are on salmon and salmon farming, the ideas and general results could be applied to all farmed species.