Fouling-release and chemical activity effects of a siloxane-based material on tunicates.

The antifouling performance of a siloxane-based elastomeric impression material (EIM) was compared to that of two silicone fouling-release coatings, Intersleek 757 and RTV-11. In field immersion trials, the EIM caused the greatest reduction in fouling by the solitary tunicate Ciona intestinalis and caused the longest delay in the progression of fouling by two species of colonial tunicate. However, in pseudobarnacle adhesion tests, the EIM had higher attachment strengths. Further laboratory analyses showed that the EIM leached alkylphenol ethoxylates (APEs) that were toxic to C. intestinalis larvae. The EIM thus showed the longest duration of chemical activity measured to date for a siloxane-based coating (4 months), supporting investigations of fouling-release coatings that release targeted biocides. However, due to potential widespread effects of APEs, the current EIM formulation should not be considered as an environmentally-safe antifoulant. Thus, the data also emphasize consideration of both immediate and long-term effects of potentially toxic constituents released from fouling-release coatings.

In Situ Confocal Raman Microscopy of Hydrated Early Stages of Bacterial Biofilm Formation on Various Surfaces in a Flow Cell.

Bacterial biofilms are precursors to biofouling by other microorganisms. Understanding their initiation may allow us to design better ways to inhibit them, and thus to inhibit subsequent biofouling. In this study, the ability of confocal Raman microscopy to follow the initiation of biofouling by a marine bacterium, Pseudoalteromonas sp. NCIMB 2021 (NCIMB 2021), in a flow cell, using optical and confocal Raman microscopy, was investigated. The base of the flow cell comprised a cover glass. The cell was inoculated and the bacteria attached to, and grew on, the cover glass. Bright field images and Raman spectra were collected directly from the hydrated biofilms over several days. Although macroscopically the laser had no effect on the biofilm, within the first 24 h cells migrated away from the position of the laser beam. In the absence of flow, a buildup of extracellular substances occurred at the base of the biofilm. When different coatings were applied to cover glasses before they were assembled into the flow cells, the growth rate, structure, and composition of the resulting biofilm was affected. In particular, the ratio of Resonance Raman peaks from cytochrome c (CC) in the extracellular polymeric substances, to the Raman phenylalanine (Phe) peak from protein in the bacteria, depended on both the nature of the surface and the age of the biofilm. The ratios were highest for 24 h colonies on a hydrophobic surface. Absorption of a surfactant with an ethyleneoxy chain into the hydrophobic coating created a surface similar to that given with a simple PEG coating, where bacteria grew in colonies away from the surface rather than along the surface, and CC:Phe ratios were initially low but increased at least fivefold in the first 48 h.

Interactions of protamine with the marine bacterium, Pseudoalteromonas sp. NCIMB 2021.

UNLABELLED: Pseudoalteromonas sp. NCIMB 2021 (NCIMB 2021) was grown in synthetic seawater (SSW) containing pyruvate, in the presence (SSW(++) ) and absence (SSW(-) ) of divalent cations. Cultures contained single cells. Addition of the cationic antibacterial peptide (CAP), protamine, did not inhibit, but rather increased, the growth of NCIMB 2021 in SSW(++) and caused the bacteria to grow in chains. Bacterial growth was assessed using turbidity, cell counts and the sodium salt of resazurin. In SSW(-) , NCIMB 2021 was no longer resistant to protamine. The minimum inhibitory concentration (MIC) was 5 mg ml(-1) . SIGNIFICANCE AND IMPACT OF THE STUDY: Protamine is a cationic antimicrobial peptide (CAP), which is active against a variety of bacteria. This is the first in-depth study of the interaction of protamine with a marine bacterium, Pseudoalteromonas sp. NCIMB 2021. Our results show that protamine is only active in seawater in the absence of divalent cations. In the presence of the divalent cations, Mg(2+) and Ca(2+) , protamine enhances the growth of Pseudoalteromonas sp. NCIMB 2021 and produces chains rather than individual cells. These are important considerations when deciding on applications for protamine and in terms of understanding its mechanism of action.

Migration of 18 trace elements from ceramic food contact material: influence of pigment, pH, nature of acid and temperature.

The effect of pH, nature of acid and temperature on trace element migration from ceramic ware treated with 18 commercially available glazes was studied. Besides of the well-studied lead and cadmium, migration of other toxic and non toxic elements such as aluminum, boron, barium, cobalt, chrome, copper, iron, lithium, magnesium, manganese, nickel, antimony, tin, strontium, titanium, vanadium, zinc and zirconium was investigated in order to evaluate their potential health hazards. Trace element concentrations were determined with Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). This study suggests that there is indeed a health risk concerning the possible migration of other elements than lead and cadmium. At low pH (2

The assessment of insemination success in yellow dung flies using competitive PCR.

In spite of considerable interest in postcopulatory sexual selection, separating the effects of sperm competition from cryptic female choice remains difficult because mechanisms underlying postcopulatory processes are poorly understood. One methodological challenge is to quantify insemination success for individual males within the sperm stores of multiply mated females to discover how insemination translates into eventual paternity. Any proposed method must be applicable in organisms without extensive DNA sequence information (which include the majority of model species for sexual selection). Here, we describe the development and application of microsatellite competitive-multiplex-PCR for quantifying relative contributions to a small number of sperm in storage. We studied how DNA template characteristics affect PCR amplification of known concentrations of mixed DNA and generated regressions for correcting observations of allelic signal strength based on such characteristics. We used these methods to examine patterns of sperm storage in twice-mated female yellow dung flies, Scathophaga stercoraria. We confirm previous findings supporting sperm displacement and demonstrate that average paternity for the last mate accords with the mean proportion of sperm stored. We further find consistent skew in storage across spermathecae, with more last male sperm stored in the singlet spermatheca on one side of the body than in the doublet on the opposite side. We also show that the time between copulations may be important for effectively sorting sperm. Finally, we demonstrate that male size may influence the opportunity for sperm choice, suggesting future work to disentangle the roles of male competition and cryptic female choice.

Kinematics, hydrodynamics and force production of pleopods suggest jet-assisted walking in the American lobster (Homarus americanus).

The American lobster (Homarus americanus) displays a diverse set of locomotory behaviours that includes tail flips, walking and paddling. Paddling is carried out by the four pairs of paddle-shaped pleopods on the ventral abdomen. Although it is recognized that pleopod-generated fluid flows have some locomotory role in adults, reports on their relative importance in locomotion are inconsistent. This paper integrates experimental kinematics and hydrodynamics of lobster pleopod beating to determine the mechanism and magnitude of pleopod force production. A kinematic analysis of pleopod beating in live lobsters showed that the pleopods execute an adlocomotory metachronal beating pattern. We modelled in vivo pleopod kinematics with a set of simple trigonometric functions, and used these functions to program a mechanical lobster model consisting of motor-driven pleopods on a lobster abdomen exoskeleton. Based on flow visualizations obtained from applying particle image velocimetry to the lobster model, we propose that the unsteady metachronal kinematics of the pleopods can maximize thrust by exploiting forces arising from individual pleopod activity and interactions among adjacent pairs. The pleopods continuously entrain fluid surrounding the lobster and create a caudally directed fluid jet oriented parallel to the substratum. Inputting wake morphology and velocity data into a simplified model for steady jet thrust showed that the pleopods of the lobster model produced 27-54 mN of thrust, which is comparable to the propulsive forces generated by other proficient swimmers. These results suggest that lobster pleopods are capable of producing forces of a magnitude that could assist the walking legs in forward propulsion.

Molecular and quantitative genetic differentiation across Europe in yellow dung flies.

Relating geographic variation in quantitative traits to underlying population structure is crucial for understanding processes driving population differentiation, isolation and ultimately speciation. Our study represents a comprehensive population genetic survey of the yellow dung fly Scathophaga stercoraria, an important model organism for evolutionary and ecological studies, over a broad geographic scale across Europe (10 populations from the Swiss Alps to Iceland). We simultaneously assessed differentiation in five quantitative traits (body size, development time, growth rate, proportion of diapausing individuals and duration of diapause), to compare differentiation in neutral marker loci (F(ST)) to that of quantitative traits (Q(ST)). Despite long distances and uninhabitable areas between sampled populations, population structuring was very low but significant (F(ST) = 0.007, 13 microsatellite markers; F(ST) = 0.012, three allozyme markers; F(ST) = 0.007, markers combined). However, only two populations (Iceland and Sweden) showed significant allelic differentiation to all other populations. We estimated high levels of gene flow [effective number of migrants (Nm) = 6.2], there was no isolation by distance, and no indication of past genetic bottlenecks (i.e. founder events) and associated loss of genetic diversity in any northern or island population. In contrast to the low population structure, quantitative traits were strongly genetically differentiated among populations, following latitudinal clines, suggesting that selection is responsible for life history differentiation in yellow dung flies across Europe.

The mechanical function and structure of aortic microfibrils in the lobster Homarus americanus.

Marfan syndrome, a connective tissue disorder affecting the cardiovascular system, is caused by mutations of fibrillin-based microfibrils. These mutations often affect the calcium-binding domains, resulting in structural changes to the proteins. It is hypothesized that these Ca+2 binding sites regulate the structure and mechanical properties of the microfibrils. The mechanical properties of fresh and extracted lobster aortic rings in calcium solutions (1, 13 and 30 mM Ca+2) were measured. Samples underwent amino acid compositional analysis. Antibodies were produced against the material comprising extracted aortic rings. The ultrastructure of strained and unstrained samples was examined using transmission electron microscopy. Calcium level altered the tangent modulus of fresh vessels. These rings were significantly stiffer when tested at 30 mM Ca+2 compared to rings tested at 1 mM Ca+2. Amino acid comparisons between extracted samples, porcine and human fibrillin showed compositional similarity. Immunohistochemical analysis showed that antibodies produced against the material in extracted samples localized to the known microfibrillar elements in the lobster aorta and cross-reacted with fibrillin microfibrils of mammalian ciliary zonules. Ultrastructurally, vessels incubated in low calcium solutions showed diffuse interbead regions while those incubated in physiological or high calcium solutions showed interbead regions with more defined lateral edges.

Bergmann and converse bergmann latitudinal clines in arthropods: two ends of a continuum?

Two seemingly opposite evolutionary patterns of clinal variation in body size and associated life history traits exist in nature. According to Bergmann's rule, body size increases with latitude, a temperature effect. According to the converse Bergmann rule, body size decreases with latitude, a season length effect. A third pattern causally related to the latter is countergradient variation, whereby populations of a given species compensate seasonal limitations at higher latitudes by evolving faster growth and larger body sizes compared to their low latitude conspecifics. We discuss these patterns and argue that they are not mutually exclusive because they are driven by different environmental causes and proximate mechanisms; they therefore can act in conjunction, resulting in any intermediate pattern. Alternatively, Bergmann and converse Bergmann clines can be interpreted as over- and undercompensating countergradient variation, respectively. We illustrate this with data for the wide-spread yellow dung fly, Scathophaga stercoraria (Diptera: Scathophagidae), which in Europe shows a Bergmann cline for size and a converse Bergmann cline (i.e., countergradient variation) for development time. A literature review of the available evidence on arthropod latitudinal clines further shows a patterned continuum of responses. Converse Bergmann clines due to end-of-season time limitations are more common in larger species with longer development times. Our study thus provides a synthesis to the controversy about the importance of Bergmann's rule and the converse Bergmann rule in nature.

Comparative equilibrium mechanical properties of bovine and lamprey cartilaginous tissues.

In contrast to all other vertebrate cartilages, the major extracellular matrix protein of lamprey cartilages is not collagen. Instead, there exists a unique family of noncollagenous structural proteins, the significance of which is not completely understood. A custom-built uniaxial testing apparatus was used to quantify and compare equilibrium stress-relaxation behavior (equilibrium moduli, stress decay behavior, recovery times and relaxation times) of (1) lamprey pericardial cartilages with perichondria tested in tension (young adult and aged), (2) annular cartilages without perichondria tested in compression (young adult and aged) and (3) bovine auricular cartilage samples without perichondria tested in both tension and compression. Results of this study demonstrated that all cartilages were highly viscoelastic but with varying relaxation times; approximately 120 min for annular and pericardial cartilages and 30 min for bovine auricular cartilages. For mean equilibrium moduli, young adult lamprey annular cartilages (0.71 MPa) and pericardial cartilages (2.87 MPa) were found to be statistically different. The mean moduli of all bovine auricular cartilages were statistically identical to lamprey cartilages except in the case of aged adult pericardial cartilages, which were statistically larger than all other cartilages at 4.85 MPa. Taken together, the results of this study suggest that lamprey cartilages are able to exhibit mechanical properties largely similar to those of mammalian cartilages despite unique structural proteins and differences in extracellular matrix organization.

The mechanics of locomotion in the squid Loligo pealei: locomotory function and unsteady hydrodynamics of the jet and intramantle pressure.

High-speed, high-resolution digital video recordings of swimming squid (Loligo pealei) were acquired. These recordings were used to determine very accurate swimming kinematics, body deformations and mantle cavity volume. The time-varying squid profile was digitized automatically from the acquired swimming sequences. Mantle cavity volume flow rates were determined under the assumption of axisymmetry and the condition of incompressibility. The data were then used to calculate jet velocity, jet thrust and intramantle pressure, including unsteady effects. Because of the accurate measurements of volume flow rate, the standard use of estimated discharge coefficients was avoided. Equations for jet and whole-cycle propulsive efficiency were developed, including a general equation incorporating unsteady effects. Squid were observed to eject up to 94 % of their intramantle working fluid at relatively high swimming speeds. As a result, the standard use of the so-called large-reservoir approximation in the determination of intramantle pressure by the Bernoulli equation leads to significant errors in calculating intramantle pressure from jet velocity and vice versa. The failure of this approximation in squid locomotion also implies that pressure variation throughout the mantle cannot be ignored. In addition, the unsteady terms of the Bernoulli equation and the momentum equation proved to be significant to the determination of intramantle pressure and jet thrust. Equations of propulsive efficiency derived for squid did not resemble Froude efficiency. Instead, they resembled the equation of rocket motor propulsive efficiency. The Froude equation was found to underestimate the propulsive efficiency of the jet period of the squid locomotory cycle and to overestimate whole-cycle propulsive efficiency when compared with efficiencies calculated from equations derived with the squid locomotory apparatus in mind. The equations for squid propulsive efficiency reveal that the refill period of squid plays a greater role, and the jet period a lesser role, in the low whole-cycle efficiencies predicted in squid and similar jet-propelled organisms. These findings offer new perspectives on locomotory hydrodynamics, intramantle pressure measurements and functional morphology with regard to squid and other jet-propelled organisms.

Microfibrils provide non-linear elastic behaviour in the abdominal artery of the lobster Homarus americanus.

1. Microfibrils are becoming increasingly recognized as an important component of the extra-cellular matrix. However, almost nothing is known about their mechanical role in the diversity of tissues in which they are found. 2. Microfibrils form the principal structural component in the wall of the abdominal artery of the lobster Homarus americanus. We have used previous estimates of the mechanical properties of these microfibrils, estimates of the fraction of the aorta wall volume occupied by the microfibrils, and their angular distribution as a function of strain in a numerical model that predicts the macroscopic mechanical properties of the whole tissue. 3. Microfibrils alone, when their reorientation and deformation are accounted for, characterize the stress-strain behaviour of the vessel. Evidence of the evolutionary conservation of fibrillin between medusans, echinoderms and vertebrates implies that the mechanical properties of lobster microfibrils may apply to microfibrillar function in other taxa. This will have profound implications on the perceived roles of microfibrils in development, physiology and disease.

Scallop Shells Exhibit Optimization of Riblet Dimensions for Drag Reduction.

Drag reduction by streamwise surface grooves, or riblets, has been observed by engineers and has been suggested to apply to certain biological systems. Drag reductions as high as 8% have been observed (1), leading to practical nautical and aeronautical applications (2, 3, 4). The shells of several species of scallop, including Placopecten magellanicus, display riblets arranged radially, and therefore roughly parallel to the flow during swimming (Fig. 1a). The dimensions of these riblets on particular scallops fall within the region necessary for drag reduction at experimentally measured swimming speeds. Moreover, the actual spacing of the riblets gradually migrates into the theoretically optimal spacing region as shell length increases beyond 40 mm (Figs. 2, 3). Specimens of P. magellanicus 40 to 80 mm in length demonstrate the greatest swimming ability (5); our data strongly suggest that streamwise riblets may be a contributing factor to the swimming success in scallops of this size range.

The modulus of elasticity of lobster aorta microfibrils.

The presence of elastic fibres in the extracellular matrix (ECM) provides physiologically important elastic properties for many tissues. Until recently, microfibrils, one component of the ECM, were thought primarily to serve as a scaffolding on which elastin is deposited during development to form elaunin fibres [1]. The most prominent protein that forms mammalian microfibrils is fibrillin. It is known that mutations in the fibrillin gene cause a heterogenous connective tissue disease called Marfan syndrome [2], so information on mechanical properties of microfibrils or their role in tissue function would be useful. Microfibrils are also found in the ECM of some invertebrate tissues, and there is growing evidence that the protein forming the structure is homologous to mammalian fibrillin [3,4]. It has been shown that the microfibril-based arterial wall of the lobster has viscoelastic properties [5], and we have now utilized this primitive artery to measure the modulus of elasticity of microfibrils. It is similar to that of the rubber-like protein elastin.

Dynamics and energetics of scallop locomotion

A dynamic model for a swimming scallop was developed which integrates the mechanical properties of the hinge ligaments, valve inertia, the external fluid-flow reaction, the fluid pressure in the mantle cavity and the muscle contraction. Kinematic data were recorded for a swimming Placopecten magellanicus from high-speed film analysis. Dynamic loading experiments were performed to provide the required mechanical properties of the hinge for the same species. The swimming dynamics and energetics based on data from a 0.065 m long Placopecten magellanicus at 10 °C were analyzed. The main conclusions are as follows. 1. The mean period of a clapping cycle during swimming is about 0.28 s, which can be roughly divided into three equal intervals: closing, gliding and opening. The maximum angular velocity and acceleration of the valve movements are about 182 degrees s-1 and 1370 degrees s-2, respectively. 2. The hysteresis loop of the hinge was found to be close to an ellipse. This may be represented as a simple Voigt body consisting of a spring and dashpot in parallel, with a rotational stiffness of 0.0497 N m and viscosity coefficient of 0.00109 kg m2 s-1 for the 0.065 m long Placopecten magellanicus. 3. The external fluid reaction has three components, of which the added mass is about 10 times higher than the mass of a single valve, and the flow-induced pseudo-viscosity compensates for nearly half of the hinge viscosity for the 0.065 m long Placopecten magellanicus. 4. The locomotor system powered by the muscle can be divided into two subsystems: a pressure pump for jet production and a shell-hinge/outer-fluid oscillator which drives the pumping cycle. The dynamics of the oscillator is determined predominantly by the interaction of the external fluid reaction and the hinge properties, and its resonant frequency was found to be close to the swimming frequencies. 5. The momentum and energy required to run the oscillator are negligibly small (about 1 % for the 0.065 m long Placopecten magellanicus) compared with that for the jet. Almost all the mechanical energy from muscle contraction is used to perform hydrodynamic work for jet production. Thus, the Froude efficiency of propulsion in scallops is nearly the same as the entire mechanical efficiency of the locomotor system. This could be a fundamental advantage of jet propulsion, at least for a scallop. 6. The estimated maximum muscle stress is about 1.06x10(5) N m-2, the cyclic work is 0.065 J and power output is 1.3 W. Using an estimate of the mass of an adductor muscle, the work done by the muscle per unit mass is 9.0 J kg-1 and the peak power per unit mass is 185 W kg-1. 7. The time course of the force generation of the contracting adductor muscle is basically the same as that of the hydrodynamic propulsive force.

The structure and physical properties of invertebrate and primitive vertebrate arteries.

Light and electron microscopy and in vitro inflation experiments were conducted on the aortae of three different invertebrate species: the lobster Homarus americanus, the horseshoe crab Limulus polyphemus and the whelk Busycon contrarium. Inflation experiments were also performed on the aortae of two species of primitive vertebrates, the sea lamprey Petromyzon marinus and the Atlantic hagfish Myxine glutinosa. The inflation experiments demonstrated similar overall biomechanical properties in each case, despite the existence of differences in tissue structure. The vessels were compliant at low strains, but demonstrated nonlinear elasticity, increasing in stiffness as strains increased; this property could act as protection against artery wall rupture. The vessels of the lamprey, hagfish and lobster are capable of acting as fairly efficient elastic reservoirs and of smoothing blood flow during circulation as they had low hysteresis values (13-18%). The aortae of the horseshoe crab and whelk, if performing this function, have much higher energy losses, up to more than 30% per cycle. The microscopy studies of the aortae of the lobster, horseshoe crab and whelk revealed tissue structures which differ widely from each other as well as from the structures of the lamprey and hagfish. None of these arteries contained elastin, but all contained fibrillar material which differed in appearance, size and arrangement between species. These materials were conjectured to contribute to the elastic properties of the tissue.

Interpersonal skills education in entry-level physical therapy programs.

The purpose of this study was to determine the current status of interpersonal skills (IPS) education in entry-level physical therapy programs with respect to content, course work requirements, and evaluation strategies. The disciplines of faculty members who teach IPS and their materials and methods also were explored. Using a 32-item questionnaire, we surveyed in 1984 all accredited baccalaureate (BS), certificate (CERT), and entry-level master's degree (MS) programs (N = 107) in physical therapy. Seventy-two responses (67.2%) were received, reflecting returns from 57 BS (64.7%), 5 CERT (83.3%), and 10 MS (76.9%) program respondents. Nearly all (97.2%, n = 70) of the respondents explicitly taught IPS in the classroom, but fewer of them evaluated learning (74.3%, n = 52). Overall, respondents teaching in MS programs consistently reported including a greater concentration of IPS content than the other two types of programs. All of the MS programs required specific IPS course work, compared with 50 (87.7%) of the BS programs and 3 (60%) of the CERT programs. Predominantly, faculty members who teach IPS are academically based physical therapists who use an extensive variety of methods and materials.

Sociopolitical adjustment among Afghan refugees in Pakistan.

Although international organizations and Pakistanis expect Afghans to act like true refugees--dependent, obedient, and grateful--Afghans consider themselves as temporary exiles who, in protest against an anti-Islamic government, found temporary refuge in Pakistan; or as soldiers in the holy wars who temporarily use their Islamic neighbor as a base before returning to fight in Afghanistan. Conforming to this concept and to these objectives, the refugees seek to preserve a certain autonomy and to lean towards forms of organization which are derived either from their traditional social structure, or as is more common now, from the ideology of the Islamic movements. One can understand that this situation may cause many misunderstandings, especially with international organizations which finance and supervise aid to the Afghan refugees in Pakistan. As for anthropologists, it is necessary to go beyond known concepts, to relativize familiar models and to act on changes which have come about in the structures and ideology of the Afghan people.