Targeting of cyclic AMP degradation to beta 2-adrenergic receptors by beta-arrestins

Catecholamines signal through the beta2-adrenergic receptor by promoting production of the second messenger adenosine 3',5'-monophosphate (cAMP). The magnitude of this signal is restricted by desensitization of the receptors through their binding to beta-arrestins and by cAMP degradation by phosphodiesterase (PDE) enzymes. We show that beta-arrestins coordinate both processes by recruiting PDEs to activated beta2-adrenergic receptors in the plasma membrane of mammalian cells. In doing so, the beta-arrestins limit activation of membrane-associated cAMP-activated protein kinase by simultaneously slowing the rate of cAMP production through receptor desensitization and increasing the rate of its degradation at the membrane.     

Soil physical quality of a Brazilian Oxisol under two tillage systems using the least limiting water range approach

Plant growth is directly affected by soil water, soil aeration, and soil resistance to root penetration. The least limiting water range (LLWR) is defined as the range in soil water content within which limitations to plant growth associated with water potential, aeration and soil resistance to root penetration are minimal. The LLWR has not been evaluated in tropical soils. Thus, the objective of the present study was to evaluate the LLWR in a Brazilian clay Oxisol (Typic Hapludox) cropped with maize (Zea mays L. cv. Cargil 701) under no-tillage and conventional tillage. Ninety-six undisturbed soil samples were obtained from maize rows and between rows and used to determine the water retention curve, the soil resistance curve and bulk density. The results demonstrated that LLWR was higher in conventional tillage than in no-tillage and was negatively correlated with bulk density for values above 1.02 g cm⁻³. The range of LLWR variation was 0–0.1184 cm³ cm⁻³ in both systems, with mean values of 0.0785 cm³ cm⁻³ for no-tillage and 0.0964 cm³ cm⁻³ for conventional tillage. Soil resistance to root penetration determined the lower limit of LLWR in 89% of the samples in no-tillage and in 46% of the samples in conventional tillage. Additional evaluations of LLWR are needed under different texture and management conditions in tropical soils.     

Characterization of single‐nucleotide polymorphism markers in the Mediterranean mussel,Mytilus galloprovincialis

The Mediterranean mussel, Mytilus galloprovincialis, is one of the most important aquaculture species in Europe. Appropriate molecular markers are required to evaluate genetic resources and to trace genealogies in breeding programmes for improving mussel culture. Microsatellites have been commonly applied to this purpose in other species. However, Mediterranean mussel microsatellites have demonstrated high frequencies of null alleles that hamper accurate estimates of population parameters and confident parentage inferences. As alternative markers, we have characterized in silico 25 potential single‐nucleotide polymorphism (SNP) markers in the Mediterranean mussel from expressed sequence tag (EST) public databases. The genotyping of SNPs was performed using a single‐base extension approach. Their polymorphism was evaluated in 47 individuals from an Atlantic population. Out of the 25 potential SNPs tested, 12 were technically feasible (producing a single amplicon) and polymorphic. All were biallelic and had an unbiased heterozygosity ranging from 0.160 to 0.504. One SNP was from a mitochondrial gene. The combined potential of nuclear SNPs for parentage assignment gave an exclusion probability of a false couple of parents of 0.9471. These markers will be useful for evaluating resources and tracing genealogies in genetic breeding programmes implemented to solve the main problems of mussel culture.     

Fish feed can show genotoxic damage

Fish Physiology and Biochemistry - The present study was carried out to evaluate the genotoxic potential of nutritional quality of feed, using erythrocytic nuclear abnormalities assay in Nile...     

Benthic microbial indicators of fish farm impact in a coastal area of the Tyrrhenian Sea

We studied the impact of organic loads due to the biodeposition of a fish farm in a non-impacted coastal area of the Tyrrhenian Sea (Western Mediterranean). Sediment chemistry and benthic microbial community were investigated from July 1997 to February 1998 on monthly basis at two stations: one was located under the fish farm, while the second was about 1 km away, and served as a reference site. The presence of the cage induced rapid changes in the benthic conditions: the sediments were rapidly (after 6 weeks) reduced. A significant accumulation of biopolymeric carbon was observed beneath the cage both 2 weeks after the initial cage deployment, on non-impacted sediments, and 5–7 months after, and appeared to be related to the fish farm production cycle. The density of microbial communities beneath the fish farm increased only during the first month of farming activity (July) and partially 7 months after cage disposal (i.e. in January), when there was a significant sediment organic enrichment. Additional evidence of rapid impact of the fish farm on the benthic bacterial communities is provided by the increase in the numbers of autofluorescent microbial cells. The photosynthetic eukaryotic cells displayed a highly reduced contribution to total autofluorescent microbial density, indicating that their decrease is related with biopolymeric carbon accumulation, and probably was due also to “shadow effect” induced by suspended material, coming from the fish farm, on the environment below. We propose here, to apply the ratio of culturable heterotrophic bacteria to microbial direct counts (CFU/MDC) to detect fish farm impact. In cage sediments, organic enrichment and the consequent modification of the characteristics of the benthic environment, determined an increase in aerobic heterotrophic bacteria and vibrio density indicating that they are efficient colonizers of organic-rich sediments. Densities of Escherichia coli and Enterococci were not significantly higher than in the reference site, and are likely of terrestrial origin. Thus they cannot be used as specific indicators of fish farm impact.     

Significant improvement of shrimp growth performance by growth hormone‐releasing peptide‐6 immersion treatments

Growth hormone‐releasing peptide‐6 (GHRP‐6) is one of the earliest developed synthetic peptidyl growth hormone secretagogue receptor agonists. These compounds mimic the effect of the endogenous ligand ghrelin. In vertebrates, ghrelin is a potent circulating orexigenic hormone with functional roles in controlling food intake, energy expenditure, adiposity, growth hormone secretion and immunity. Ghrelin has been studied mainly in vertebrates; thus, little is known about its role in invertebrates, including crustaceans. We first evaluated the effect of GHRP‐6 injection over feed intake in shrimp and its effects on shrimp growth when the peptide was administrated by successive immersion baths. GHRP‐6 increased feed intake, body weight and size, the number of rostral spines and gill branches, protein concentration and haemocyte number in treated shrimps. We also evaluated the peptide uptake and clearance in a pharmacokinetics, using [H³]GHRP‐6 administered to postlarvae. Given a limited exposure and efficient clearance of the peptide‐associated radioactivity from larvae, our findings suggested that GHRP‐6‐treated Litopenaeus vannamei can be consumed safely by humans after aquaculture applications. These results propose that GHRP‐6 could be an additional tool to study growth physiology in crustaceans and also a promising candidate for development into a new biotechnology product for improving shrimp growth and quality.     

Cloning and expression of chicken anemia virus VP3 protein in Escherichia coli

Purification of chicken anemia virus (CAV) VP3 protein, expressed in a prokaryotic expression system as histidine-tagged fusion protein is demonstrated in the present study. CAV particle was obtained from infected liver of chicken and DNA was extracted. The VP3 protein gene was amplified from the extracted DNA by polymerase chain reaction (PCR) and cloned. The recombinant expression construct (pTrc-VP3) was identified by PCR and sequencing analysis. Expression of VP3 protein with a molecular mass of approximately 21kDa was confirmed by Western blotting analysis with CAV-specific antibodies. The in vitro expressed VP3 protein was purified to near homogeneity by elution from the gel, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The purified VP3 protein was recognized by CAV antibodies in a Western blotting assay. This finding indicates that recombinant VP3 expressed in the pTrcHis2 vector system can be used as antigen to detect anti-CAV antibodies.     

INFLUENCE OF NITROGEN FERTILIZATION ON NICKEL ACCUMULATION AND CHEMICAL COMPOSITION OF COFFEE PLANTS DURING FRUIT DEVELOPMENT

Nutritional and physiological significance of micronutrients in coffee plants, especially with regard to nickel (Ni) is still unknown. The dynamics of nitrogen (N), phosphorus (P), potassium (K) and Ni accumulation in coffee fruits, as well as their relationships with total soluble protein, amino acids, reducing sugars, and starch content during coffee fruit development (green, ripe, and dry fruits), were investigated. Coffee trees received three N fertilizer rates (0, 150, and 300 kg of N ha^?1) as ammonium sulfate split into three applications per year. Nitrogen fertilization increased reducing sugars and starch concentrations in ripe fruits. In contrast, green fruits showed the highest amino acid and Ni concentrations. Fruit Ni concentration decreased in both green and ripe fruits as N rates increased; thus, indicating the possibility of either a N-associated dilution effect on Ni concentration or that Ni uptake by roots and/or transport to developing fruit was limiting. Plant nutritional status and fruit development stage influenced the coffee grain chemical composition. Furthermore, the variation in reducing sugars and starch content was more closely linked to the stage of fruit development than to N supply. A supposed relationship among the decreased of caffeine, starch, amino acids, and proteins with Ni content during green fruit development suggests a fundamental role for Ni in coffee fruit ripening. The interaction between N and Ni metabolism during fruit ripening might influence the chemical parameters involved in the coffee grain quality. This is the first report documenting changes in Ni concentrations of coffee fruit as a function of N fertilization rates and the development stage, but further research is needed to better understand the significance of N-Ni interaction in developing coffee fruit.     

The high biodiversity and vulnerability of two Mediterranean bathyal seamounts support the need for creating offshore protected areas

1. Seamounts host some of the most important deep-sea ecosystems. The unique environmental characteristics of seamounts sustain rich biological hot spots, which, in recent times, have suffered the effects of intense fishing pressure.
2. Biodiversity and vulnerability data are extremely scarce for Mediterranean seamounts, and this, in addition to the complex socio-economic and juridical status of offshore sites, results in difficulty in identifying the best management strategies.
3. An extensive remotely operated vehicle (ROV) survey was used to characterize the megabenthic assemblages of the summits of two upper bathyal seamounts, Ulisse and Penelope (Ligurian Sea, north-west Mediterranean Sea). The biogeographic implications of these findings and the possible environmental factors favouring the occurrence of these communities are discussed.
4. High densities of abandoned, lost, or otherwise discarded fishing gear (mainly longlines) and a reduction in the average size of the fragile and slow-growing habitat-forming gorgonian Callogorgia verticillata indicate the occurrence of high levels of anthropogenic impacts on the summit regions, which are fishing grounds for artisanal and recreational fishers.
5. The recovery of fishing data describing the first catches in the 1970s proved to be useful in inferring the short- and long-term effects of fishing practices in these previously unexploited offshore areas. In particular, the local extinction of demersal top predators, subjected to exceptional catches nearly 50 years ago, highlights the slow recovery rate of such species.
6. Criteria defining vulnerability are discussed for the two study areas, and specific conservation actions, including the creation of regulated protected areas, are proposed.