Maintaining turbidity and current flow in laboratory aquarium studies, a case study using Sabellaria spinulosa

Many aquatic organisms rely on the suspension of particulate matter for food or for building materials, yet these conditions are difficult to replicate in laboratory mesocosms. Consequently, husbandry and experimental conditions may often be sub-optimal. The Vortex Resuspension Tank (VoRT) is a simple and reliable system for the resuspension of food or sediments using an enclosed airlift. The particle rain from the lift is mixed in the tank by two water inputs that provide directional current flow across the study organism(s). The vortex mixing creates a turbulent lateral water flow that allows the distribution of particulate matter outwards from the sediment outflow. By calibrating a VoRT it is possible to control sedimentation rate by manipulating water and air flow rates. As an example application, three VoRTs were maintained under different sediment loadings to assess the sediment fraction utilisation and tube growth rates of the tube-building polychaete worm Sabellaria spinulosa. S. spinulosa consistently utilised a lower mean particle size than that of the background sediment when provided with well sorted medium sands. Under sediment starved conditions, there was net erosion of colonies whereas under intermediate and high sediment rates there was consistent cumulative growth throughout a 15 d experiment. This highlights the importance of suspended sediment for S. spinulosa and also the suitability of the VoRT system for maintaining organisms with suspended matter requirements.     

A general artificial neural network for the modelization of culture kinetics of different CHO strains

Animal cell cultures are characterized by very complex nonlinear behaviors, difficult to simulate by analytical modeling. Artificial Neural Networks, while being black box models, possess learning and generalizing capacities that could lead to better results. We first trained a three-layer perceptron to simulate the kinetics of five important parameters (biomass, lactate, glucose, glutamine and ammonia concentrations) for a series of CHO K1(Chinese Hamster Ovary, type K1) batch cultures. We then tried to use the same trained model to simulate the behavior of recombinant CHO TF70R. This was achieved, but necessitated to synchronize the time-scales of the two cell lines to compensate for their different specific growth rates. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ericoid mycorrhizal root fungi and their multicopper oxidases from a temperate forest shrub

Ericoid mycorrhizal fungi (ERM) may specialize in capturing nutrients from their host's litter as a strategy for regulating nutrient cycles in terrestrial ecosystems. In spite of their potential significance, we know little about the structure of ERM fungal communities and the genetic basis of their saprotrophic traits (e.g., genes encoding extracellular enzymes). Rhododendron maximum is a model ERM understory shrub that influences the nutrient cycles of montane hardwood forests in the southern Appalachians (North Carolina, USA). We sampled ERM roots of R. maximum from organic and mineral soil horizons and identified root fungi by amplifying and sequencing internal transcribed spacer (ITS) ribosomal DNA (rDNA) collected from cultures and clones. We observed 71 fungal taxa on ERM roots, including known symbionts Rhizoscyphus ericae and Oidiodendron maius, putative symbionts from the Helotiales, Chaetothyriales, and Sebacinales, ectomycorrhizal symbionts, and saprotrophs. Supporting the idea that ERM fungi are adept saprotrophs, richness of root-fungi was greater in organic than in mineral soil horizons. To study the genetic diversity of oxidative enzymes that contribute to decomposition, we amplified and sequenced a portion of genes encoding multicopper oxidases (MCOs) from ERM ascomycetes. Most fungi possessed multiple copies of MCO sequences with strong similarities to known ferroxidases and laccases. Our findings indicate that R. maximum associates with a taxonomically and ecologically diverse fungal community. The study of MCO gene diversity and expression may be useful for understanding how ERM root fungi regulate the cycling of nutrients between the host plant and the soil environment.     

The interaction of Ca2+ with sarcomeric proteins: role in function and dysfunction of the heart

The hallmarks of the normal heartbeat are both rapid onset of contraction and rapid relaxation as well as an inotropic response to both increased end-diastolic volume and increased heart rate. At the microscopic level, Ca(2+) plays a crucial role in normal cardiac contraction. This paper reviews the cycle of Ca(2+) fluxes during the normal heartbeat, which underlie the coupling between excitation and contraction and permit a highly synchronized action of cardiac sarcomeres. Length dependence of the response of the regulatory sarcomeric proteins mediates the Frank-Starling Law of the heart. However, Ca(2+) transport may go astray in heart disease such as in congestive heart failure, and both jeopardize systole and diastole and triggering arrhythmias. The interaction between weak and strong segments in nonuniform cardiac muscle allows partial preservation of force of contraction but may further lead to mechanoelectric feedback or reverse excitation-contraction coupling mediating an early diastolic Ca(2+) transient caused by the rapid force decrease during the relaxation phase. These rapid force changes in nonuniform muscle may cause arrhythmogenic Ca(2+) waves to propagate by the activation of neighboring sarcoplasmic reticulum by diffusing Ca(2+) ions.     

Using structural analysis to generate parasite-selective monoclonal antibodies

Diagnosis of eukaryotic parasitic infection using antibody-based tests such as ELISAs (enzyme-linked immunosorbent assays) is often problematic because of the need to differentiate between homologous host and pathogen proteins and to ensure that antibodies raised against a peptide will also bind to the peptide in the context of its three-dimensional protein structure. Filariasis caused by the nematode, Brugia malayi, is an important worldwide tropical disease in which parasites disappear from the bloodstream during daylight hours, thus hampering standard microscopic diagnostic methods. To address this problem, a structural approach was used to develop monoclonal antibodies (mAbs) that detect asparaginyl-tRNA synthetase (AsnRS) secreted from B. malayi. B. malayi and human AsnRS amino acid sequences were aligned to identify regions that are relatively unconserved, and a 1.9 A crystallographic structure of B. malayi AsnRS was used to identify peptidyl regions that are surface accessible and available for antibody binding. Sequery and SSA (Superpositional Structural Analysis) software was used to analyze which of these peptides was most likely to maintain its native conformation as a synthetic peptide, and its predicted helical structure was confirmed by NMR. A 22-residue peptide was synthesized to produce murine mAbs. Four IgG(1) mAbs were identified that recognized the synthetic peptide and the full-length parasite AsnRS, but not human AsnRS. The specificity and affinity of mAbs was confirmed by Western blot, immunohistochemistry, surface plasmon resonance, and enzyme inhibition assays. These results support the success of structural modeling to choose peptides for raising selective antibodies that bind to the native protein.     

Adhesion, growth and detachment of cells on modified polystyrene surface

By adsorbing poly(N-isopropylacrylamide) (PNIPAAm) from an aqueous solution onto oxidised polystyrene without the need for grafting the polymer to the surface, we showed here that cells(CHO-K1) adhere and grow well at 37 °C and are detached by lowering the temperature to 10 °C without any other deleterious treatment. Both bacterial culture grade polystyrene Petri dishes and polystyrene beads (120 to 250μm diameters) commercially available used in static conditions of growth were tested with similar results. The contact angle of modified Petri dishes with a water droplet increases from 36 to 58° when the temperature is raised from 25 to 37 °C indicating change in hydrophilicity of the surface as a function of temperature. This revised version was published online in July 2006 with corrections to the Cover Date.     

Social and economic policy issues relevant to marine aquaculture

This paper presents a critical review of current social, economic and policy issues relevant to marine aquaculture (mariculture) in Europe. Tools for identifying the full range of social, economic and environmental issues that influence the sustainable development of mariculture are examined. Under present sectoral approaches to policy, investment, development planning and natural resources management, these issues continue to be treated in isolation. The four main challenges presented in this paper are: (i) how to create a more objective information base with which to assess the social, economic and environmental factors that condition the sustainability of mariculture; (ii) how to provide information from different disciplines in an easy to obtain and compatible format; (iii) how to better integrate knowledge and skills from different disciplines to create a holistic and robust framework for assessing options for mariculture development that integrates social, economic and environmental parameters; and (iv) the effective integration of these assessments into the formulation of policy, investment strategies, spatial plans and natural resources management for coastal areas. Specific issues that need to be addressed within the framework for the integrated evaluation of the economic, social and environmental parameters governing the sustainable development of mariculture include: ? development of more accurate information on the economic, social and environmental benefits and costs of well‐planned and managed mariculture; ? clearer definition of gaps in existing knowledge on factors critical to the sustainable and equitable development of mariculture; ? development of pro‐active consideration of the coastal land and water resource requirements of mariculture as part of strategic economic planning, spatial planning and natural resources management; ? the need for more equitable treatment of mariculture regarding rights of access to sites for development and use of resources; ? development of awareness among decision‐makers, planners, and managers from different sectoral agencies of the contribution that mariculture may make in promoting the sustainable use of coastal ecosystems; ? promotion of a shift in emphasis away from controlling the end use of resources and toward a more balanced approach to coastal development where emphasis is also given to maintaining the health and productivity of coastal ecosystems and the resources they generate that sustain different forms of activity, including mariculture.     

Gα<Subscript>16</Subscript> interacts with tetratricopeptide repeat 1 (TPR1) through its β3 region to activate Ras independently of phospholipase Cβ signaling

Background  

G protein-coupled receptors constitute the largest family of cell surface receptors in the mammalian genome. As the core of the G protein signal transduction machinery, the Gα subunits are required to interact with multiple partners. The GTP-bound active state of many Gα subunits can bind a multitude of effectors and regulatory proteins. Yet it remains unclear if the different proteins utilize distinct or common structural motifs on the Gα subunit for binding. Using Gα₁₆ as a model, we asked if its recently discovered adaptor protein tetratricopeptide repeat 1 (TPR1) binds to the same region as its canonical effector, phospholipase Cβ (PLCβ).     

Granuloma Due to Oxidized Regenerated Cellulose in an Aged Rhesus Macaque (Macaca mulatta)

Bioabsorbable hemostatic agents such as oxidized regenerated cellulose are widely used to control intraoperative diffuse capillary bleeding. Compared with electrocautery or ligation, oxidized regenerated cellulose has the advantage of controlling bleeding without occluding the vessel lumen or causing thermal injuries to adjacent tissue. Although the manufacturer recommends removal of the material once hemostasis is achieved, oxidized regenerated cellulose is a bioabsorbable hemostatic agent and is often left in the surgical bed to prevent subsequent bleeding after surgical closure. However, noninvasive imaging techniques have revealed granulomatous foreign-body reactions that mimic infection or tumor recurrence. We present a case report of sterile peritonitis and granuloma formation secondary to the presence of oxidized regenerated cellulose after intestinal resection to excise a colonic adenocarcinoma in an aged rhesus macaque.Bioabsorbable hemostatic agents such as oxidized regenerated cellulose (for example, Surgicel) are widely used to control intraoperative diffuse capillary bleeding. Compared with electrocautery or ligation, oxidized regenerated cellulose has the advantage of controlling bleeding without occluding the vessel lumen or causing thermal injuries to adjacent tissue.¹⁶Oxidized regenerated cellulose is formed by dissolving the α-cellulose of decomposed wood pulp in an alkaline solution and subsequently regenerating it as a continuous fiber. This fiber is then woven into a gauze and oxidized.^17,22 Oxidized regenerated cellulose is supplied as a substrate that is flexible, malleable, and trimable.¹⁶The mechanism of hemostasis of oxidized regenerated cellulose is reportedly associated with its caustic activity.² The oxidation of cellulose produces a low-pH organic acid that reacts with blood, thus forming an artificial clot and causing platelet aggregation.¹⁸Although the manufacturer recommends the removal of oxidized regenerated cellulose once hemostasis is achieved,⁸ the product, a bioabsorbable hemostatic agent, is often left in situ within the surgical bed to prevent bleeding after surgical procedures. The biodegradation and elimination of oxidized regenerated cellulose from the tissue occurs in 2 phases.¹⁴ Polyanhydroglucuronic acid, the major functional unit of oxidized regenerated cellulose, is readily soluble. This acid is degraded extracellulary and systematically cleared from the system approximately 18 h after implantation.^13,14 The remaining fibrous residue, however, requires macrophage phagocytosis for clearance and can be observed within macrophages for at least 48 h after implantation.¹³ Unfortunately, these fibrous residues have a prolonged degradation, and their persistence for as long as 7 mo after surgery has been confirmed histologically.⁷Despite the biocompatibility of oxidized regenerated cellulose, granulomatous foreign-body reactions that imitate infection or tumor recurrence have been revealed by using noninvasive imaging techniques.^{1,11,12,15,17,18,22} Here we describe a case of peritonitis and granuloma formation secondary to the presence of oxidized regenerated cellulose after an intestinal resection to excise a colonic adenocarcinoma in an aged rhesus macaque.