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Biomedical subjects

J A Callow

Publications and source records attributed to J A Callow.

18 recordsLinked to original sources
PubMed2006

Mussel (Mytilus edulis) byssus deposition in response to variations in surface wettability.

Mussels (Mytilus edulis) are economically important in their role as an aquaculture species and also with regard to marine biofouling. They attach tenaciously to a wide variety of submerged surfaces by virtue of collagenous attachment threads termed 'byssi'. The aim of this study was to characterize the spreading of the byssal attachment plaque, which mediates attachment to the surface, on a range of surfaces in response to changes in wettability. To achieve this, well characterized self-assembled monolayers of omega-terminated alkanethiolates on gold were used, allowing correlation of byssal plaque spreading with a single surface characteristic--wettability. The present results were inconsistent with those from previous studies, in that there was a positive correlation between plaque size and surface wettability; a trend which is not explained by conventional wetting theory for a three-phase system. A recent extension to wetting theory with regard to hydrophilic proteins is discussed and the results of settlement assays are used to attempt reconciliation of these results with those of similar previous studies and, also, with recent data presented for the spreading of Ulva linza spore adhesive.

Adhesiveness↗
PubMed2006

Biofilms.

Biofilms of bacteria, frequently in association with algae, protozoa and fungi, are found on all submerged structures in the marine environment. Although it is likely that for the majority of organisms a biofilmed surface is not a pre-requisite for settlement, in practice, colonization by spores and larvae of fouling organisms almost always takes place via a biofilmed surface. Therefore, the properties of the latter may be expected to influence colonization, positively or negatively. Biofilms are responsible for a range of surface-associated and diffusible signals, which may moderate the settling behaviour of cells, spores and larvae. However, there is no consensus view regarding either cause and effect or the mechanism(s) by which biofilms moderate settlement. Studies with mixed biofilms, especially field experiments, are difficult to interpret because of the conflicting signals produced by different members of the biofilm community as well as their spatial organisation. Molecular techniques highlight the deficiencies of culture methods in identifying biofilm bacteria; hence, the strains with the most impact on settlement of spores and larvae may not yet have been isolated and cultured. Furthermore, secondary products isolated from cultured organisms may not reflect the situation that pertains in nature. The evidence that bacterial quorum sensing signal molecules stimulate settlement of spores of the green macroalga, Ulva, is discussed in some detail. New molecular and analytical tools should provide the opportunity to improve our fundamental understanding of the interactions between fouling organisms and biofilms, which in turn may inform novel strategies to control biofouling.

4-Butyrolactone↗
PubMed2005

The influence of surface energy on the wetting behaviour of the spore adhesive of the marine alga Ulva linza (synonym Enteromorpha linza).

The environmental scanning electron microscope has been used to image the adhesive pads secreted by zoospores of the marine alga Ulva linza as they settle on a range of self-assembled and grafted monolayers of different wettability, under natural, hydrated conditions. Results reveal that the diameter of the adhesive pad is strongly influenced by surface wettability, the adhesive spreading more (i.e. wetting the surface better) on the more hydrophilic surfaces. This is in direct contrast to previous observations on the spreading of marine bioadhesives and is in apparent contradiction to the predictions of the Young-Dupre equation for three-phase systems. In this paper, we attempt an explanation based upon thermodynamic analysis of the wetting properties of hydrophilic proteins.

Adhesiveness↗
PubMed2004

Activity of commercial enzymes on settlement and adhesion of cypris larvae of the barnacle Balanus amphitrite, spores of the green alga Ulva linza, and the diatom Navicula perminuta.

Fouling species produce adhesive polymers during the settlement, adhesion and colonization of new surfaces in the marine environment. The present paper tests the hypothesis that enzymes of the appropriate specificity may prevent biofouling by hydrolysing these adhesive polymers. Seventeen commercially available enzyme preparations designed originally for bulk use in a range of end-use applications were tested for their effects on the settlement and/or adhesion of three major fouling species, viz. the green alga Ulva linza, the diatom Navicula perminuta and the barnacle Balanus amphitrite. The serine-proteases were found to have the broadest antifouling potential reducing the adhesion strength of spores and sporelings of U. linza, cells of N. perminuta and inhibiting settlement of cypris larvae of B. amphitrite. Mode-of-action studies on the serine-protease, Alcalase, indicated that this enzyme reduced adhesion of U. linza in a concentration-dependent manner, that spores of the species could recover their adhesive strength if the enzyme was removed and that the adhesive of U. linza and juvenile cement of B. amphitrite became progressively less sensitive to hydrolysis as they cured.

Adhesives↗
PubMed2004

Adhesion and motility of fouling diatoms on a silicone elastomer.

Recent demands for non-toxic antifouling technologies have led to increased interest in coatings based on silicone elastomers that 'release' macrofouling organisms when hydrodynamic conditions are sufficiently robust. However, these types of coatings accumulate diatom slimes, which are not released even from vessels operating at high speeds (>30 knots). In this study, adhesion strength and motility of three common fouling diatoms (Amphora coffeaeformis var. perpusilla (Grunow) Cleve, Craspedostauros australis Cox and Navicula perminuta Grunow) were measured on a poly-dimethylsiloxane elastomer (PDMSE) and acid-washed glass. Adhesion of the three species was stronger to PDMSE than to glass but the adhesion strengths varied. The wall shear stress required to remove 50% of cells from PDMSE was 17 Pa for Craspedostauros, 24 Pa for Amphora and >>53 Pa for Navicula; the corresponding values for glass were 3, 10 and 25 Pa. In contrast, the motility of the three species showed little or no correlation between the two surfaces. Craspedostauros moved equally well on glass and PDMSE, Amphora moved more on glass initially before movement ceased and Navicula moved more on PDMSE before movement ceased. The results show that fouling diatoms adhere more strongly to a hydrophobic PDMSE surface, and this feature may contribute to their successful colonization of low surface energy, foul-release coatings. The results also indicate that diatom motility is not related to adhesion strength, and motility does not appear to be a useful indicator of surface preference by diatoms.

Adhesives↗
PubMed2004

Roughness-dependent removal of settled spores of the green alga Ulva (syn. Enteromorpha) exposed to hydrodynamic forces from a water jet.

Topographic features change the hydrodynamic regime over surfaces subjected to flow. Hydrodynamic microenvironments around topographic structures may have consequences for recruitment and removal of propagules of marine benthic organisms. The settlement and adhesion of zoospores from the green alga Ulva linza (syn. Enteromorpha linza) to defined topographies was investigated. A range of topographic size scales (Rz: 25-100 microm) was manufactured from plankton nets, creating patterns with ridges and depressions. The topographic scales span a roughness similar to that of natural substrata and antifouling coatings. Spores were removed from the surfaces by a calibrated water jet. Fewer spores were removed from the smallest topographic structure tested (Rz: 25 microm) compared to both the smooth (Rz: 1) and the roughest (Rz: 100 microm) structures. Zoospores that settled in depressions were less likely to be removed compared to spores on the ridges. The results in terms of the interaction between surface topography and hydrodynamic forces have implications for both natural substrata exposed to wave action and antifouling surfaces on ships' hulls. The possible effects of topography on increasing zoospore adhesion and offering a refuge from hydrodynamic forces are discussed.

Adhesiveness↗
PubMed2003

Three models to relate detachment of low form fouling at laboratory and ship scale.

Since fouling-release coating systems do not prevent settlement, various methods to quantify the tenacity of adhesion of fouling organisms on these systems have been offered. One such method is the turbulent channel flow apparatus. The question remains how the results from laboratory scale tests relate to the self-cleaning of a ship coated with a fouling-release surface. This paper relates the detachment strength of low form fouling determined in the laboratory using a turbulent channel flow to the conditions necessary for detachment of these organisms in a turbulent boundary layer at ship scale. A power-law formula, the ITTC-57 formula, and a computational fluid dynamics (CFD) model are used to predict the skin-friction at ship scale. The results from all three methods show good agreement and are illustrated using turbulent channel flow data for sporelings of the green macrofouling alga Enteromorpha growing on a fouling-release coating.

Adhesiveness↗
PubMed2001

Brefeldin A affects adhesion of zoospores of the green alga Enteromorpha.

Primary adhesion of zoospores of the green macroalga Enteromorpha to substrata involves a massive release of adhesive glycoproteins from Golgi-derived, membrane-bounded vesicles in the anterior region of the spore, followed by rapid curing. This process is sensitive to low concentrations (5-10 microg x ml(-1)) of the secretion-inhibiting antibiotic, brefeldin A (BFA). The proportion of cells that settled in BFA was reduced by approximately 50%, but the effect was fully reversed by washing in seawater to remove the BFA. Ultrastructural observations showed that BFA caused the breakdown of Golgi stacks in the majority of cells examined. When settled cells were subjected to shear stress, a greater proportion of those settled in the presence of BFA were detached, compared with controls, indicating reduced adhesion strength in the presence of the antibiotic. The most likely reason for this is that strong adhesion to substrata either requires the synthesis of extra adhesive materials beyond those present in the swimming spore, or the secretion of an additional component required for adhesive curing. The novel use of atomic force microscopy in force modulation mode demonstrated that the adhesive secreted by most spores in the presence of BFA did not undergo the rapid curing process typical of control spores. However, some variation between zoospores was observed, with some cells showing no ultrastructural changes and normal adhesive curing. These results are discussed in relation to variations observed in the propensity and competence of spores to settle, which may be reflected in differential requirements for de novo synthesis and secretion of materials needed for full adhesion.

Antifungal Agents↗
PubMed2000

The application of atomic force microscopy to topographical studies and force measurements on the secreted adhesive of the green alga Enteromorpha.

Atomic force microscopy (AFM) enables the topographical structure of cells and biological materials to be resolved under natural (physiological) conditions, without fixation and dehydration artefacts associated with imaging methods in vacuo. It also provides a means of measuring interaction forces and the mechanical properties of biomaterials. In the present study, AFM has been applied for the first time to the study of the mechanical properties of a natural adhesive produced by a green plant cell. Swimming spores of the green alga Enteromorpha linza (L.) J. Ag. (7-10 microm) secrete an adhesive glycoprotein which provides firm anchorage to the substratum. Imaging of the adhesive in its hydrated state revealed a swollen gel-like pad, approximately 1 microm thick, surrounding the spore body. Force measurements revealed that freshly released adhesive has an adhesion strength of 173 +/- 1.7 mN m(-1) (mean +/- SE; n=90) with a maximum value for a single adhesion force curve of 458 mN m(-1). The adhesive had a compressibility (equivalent to Young's modulus) of 0.54 x 10(6) +/- 0.05 x 10(6) N m-2 (mean +/- SE; n=30). Within minutes of release the adhesive underwent a progressive 'curing' process with a 65% reduction in mean adhesive strength within an hour of settlement, which was also reflected in a reduction in the average length of the adhesive polymer strands (polymer extension) and a 10-fold increase in Young's modulus. Measurements on the spore surface itself revealed considerably lower adhesion-strength values but higher polymer-extension values than the adhesive pad, which may reflect the deposition of different polymers on this surface as a new cell wall is formed. The study demonstrates the value of AFM to the imaging of plant cells in the absence of fixation and dehydration artefacts and to the characterisation of the mechanical properties of plant glycoproteins that have potential utility as adhesives.

Cell Adhesion↗
PubMed2000

Use of self-assembled monolayers of different wettabilities to study surface selection and primary adhesion processes of green algal (Enteromorpha) zoospores.

We investigated surface selection and adhesion of motile zoospores of a green, macrofouling alga (Enteromorpha) to self-assembled monolayers (SAMs) having a range of wettabilities. The SAMs were formed from alkyl thiols terminated with methyl (CH(3)) or hydroxyl (OH) groups or mixtures of CH(3)- and OH-terminated alkyl thiols and were characterized by measuring the advancing contact angles and by X-ray photoelectron spectroscopy. There was a positive correlation between the number of spores that attached to the SAMs and increasing contact angle (hydrophobicity). Moreover, the sizes of the spore groups (adjacent spores touching) were larger on the hydrophobic SAMs. Video microscopy of a patterned arrangement of SAMs showed that more zoospores were engaged in swimming and "searching" above the hydrophobic sectors than above the hydrophilic sectors, suggesting that the cells were able to "sense" that the hydrophobic surfaces were more favorable for settlement. The results are discussed in relation to the attachment of microorganisms to substrata having different wettabilities.

Cell Adhesion↗
PubMed1995

Cloning of a pea cDNA encoding a polypeptide of the light-harvesting complex associated with photosystem I using a monoclonal antibody.

A monoclonal antibody (MAb UB42) is described that binds to thylakoids in pea chloroplasts, as shown by EM-immunogold labelling. The antibody recognised proteins of ca. 23-29 kDa in western blots of a pea leaf homogenate. A cDNA library was prepared from pea epidermal cells in the vector lambda ZAP II, and immunoscreening of the library with UB42 led to the isolation of a clone, pUB42. This was sequenced and had an open reading frame of 269 codons encoding a predicted polypeptide of 28.9 kDa. The sequence showed extensive homology with three closely related polypeptides belonging to a family of chlorophyll a/b-binding proteins from the light harvesting complex of photosystem I (LHCI). Collectively, the results suggest that MAb UB42 recognises an epitope on the type II chlorophyll a/b-binding protein from LHCI and that clone pUB42 encodes this protein.

Amino Acid Sequence↗
PubMed1985

Sexual recognition and fertilization in brown algae.

Fertilization in the brown marine algae known as fucoids, is oogamous. The naked egg cell (80 micron diam.) is fertilized by small biflagellate spermatozoids and both monoecious and dioecious species are found. Fertilization is highly species-specific and this appears to be controlled during plasmogamy. Following fusion of egg and sperm, a rapid (less than 1 min) release of polyuronide cell wall material takes place from cytoplasmic vesicles within the egg. This is easily visualized using the fluorescent brightener Calcofluor, which therefore provides the basis of a quantitative fertilization bioassay. It has not proved possible to measure direct sperm binding to eggs. In experiments to investigate the molecular basis of egg-sperm recognition, the effect of exogenous agents on the initial rate of fertilization was examined. Predigestion of eggs with low concentrations of two glycosidases, alpha-fucosidase and alpha-mannosidase, caused inhibition of fertilization. The lectins concanavalin A and RCA120 bound strongly to egg surfaces, as detected using fluorescent labels, but not to sperm. The binding to eggs inhibited fertilization. On the other hand, Fucose Binding Protein bound only weakly to eggs but strongly to sperm, again causing inhibition of fertilization. It has not proved possible to quantitate lectin binding since high levels of lectin-nonspecific binding were detected using iodinated lectins. These inhibition experiments suggest that specific sugar residues may be involved in egg-sperm recognition, but the effects of lectins must be treated with caution since a large amount of variability in the sensitivity of gametes was detected. Attempts to isolate receptor fractions from egg cells have been partially successful. Egg membrane preparations bind sperm and sodium dodecyl sulphate-solubilized fractions, purified by concanavalin A affinity chromatography, have yielded low levels of a soluble receptor-like fraction that has not yet been fully characterized. Antisera raised against surface antigens of Fucus serratus sperm flagella, cause inhibition of fertilization in a species-specific manner, possibly by binding directly to the sperm fertilization receptor. A number of flagellar antigens were detected and future attempts to pinpoint the sperm receptor will make use of monoclonal antibodies.

Concanavalin A↗
PubMed1983

Fertilization in brown algae. V. Further investigations of lectins as surface probes.

The binding of FITC- (fluorescein isothiocyanate), Au- and 125I-labelled lectins (conA, RCA120 (Ricinus communis agglutinin, MW 120 000) and FBP (fucose-binding protein from Lotus tetragonobolus)) to gametes of Fucus serratus and their physiological effects on fertilization have been studied. Results indicate that eggs strongly bind FITC- and Au-labelled conA and RCA120, whilst FITC-FBP binds strongly to sperm. All three iodinated lectins bound to eggs but this was apparently non-specific and similar in magnitude to the binding of iodinated bovine serum albumin. The results suggested the possibility of two distinct types of lectin receptor on egg surfaces: non-specific, highly abundant receptors and less abundant, specific receptors, possibly locally aggregated. All three lectins inhibit fertilization, FBP being the most effective.

Eukaryota↗
PubMed1983

Fertilization in brown algae. IV. Appearance of sperm-specific antigens on fertilized eggs.

Flagella antigens from sperm of Fucus serratus have been used to raise antibodies in rabbits. The immunoglobulin G fraction inhibits fertilization with some degree of species specificity. The antigens detected on sperm are not present on unfertilized egg membranes, but appear after fertilization. The common antigens on the fertilized egg can be distinguished from the cell wall material that is also released on fertilization.

Epitopes↗
PubMed1980

Fertilization in brown algae. III. Preliminary characterization of putative gamete receptors from eggs and sperm of Fucus serratus.

Membrane fractions have been isolated from eggs of Fucus serrratus which inhibit fertilization in a species-specific manner. This activity is destroyed by alpha-fucosidase and alpha-mannosidase. Some 6% of the protein of this membrane fraction binds to Con A-agarose, following SDS solubilization, and when eluted with alpha-methyl mannoside inhibits fertilization when preincubated with sperm, but not eggs. This inhibitory activity is species-specific and destroyed by alpha-fucosidase but not by trypsin. SDS-gel electrophoresis reveals 1 band staining strongly with Coomassie Brilliant Blue G and weakly with the PAS reagent. This major band represents a glycoprotein with an approximate molecular weight of 30 000 Daltons. Membrane fractions from sperm of Fucus serratus solubilized in KC1 yielded a protein-containing fraction, after affinity chromatography on desulphated focoidan-Sepharose. This fraction is 100-fold more effective in inhibiting fertilization after preincubation with eggs than either Con A or fucose-binding protein. It is species-specific and inhibition is reversed when pretreated eggs are washed with fucoidan. Activity is destroyed by heat and trypsin and only one diffuse band is apparent on SDS gels. This stains positively with Coomassie Brilliant Blue G but not with PAS and has a molecular weight of approximately 60 000 Daltons. Tentative calculation of the numbers of putative receptor molecules gives a figure of 2.5 X 10(9) receptors per egg and 1.8 X 10(6) receptors per sperm.

Cell Membrane↗
PubMed1979

Fertilization in brown algae. II. Evidence for lectin-sensitive complementary receptors involved in gamete recognition in Fucus serratus.

Fertilization in Fucus serratus is directly proportional to the number of sperm added, saturating at approximately 250 sperm per egg with an apparent Km of 120 sperm per egg. The effect of a range of lectins on fertilization has been tested. Preincubation of gametes with Con A and fucose-binding protein (FBP) inhibited fertilization. At low concentrations this was by specifically binding to eggs; at high concentrations pretreatment of either gametes inhibited fertilization probably due to cytotoxicity. Fertilization was not inhibited by simple sugar haptens, but polysaccharides containing fucosyl or mannosyl residues (yeast mannan, fucoidan, ascophyllan) inhibited fertilization by binding to sperm. Pretreatment of eggs with alpha-fucosidase or alpha-mannosidase was effective in inhibiting fertilization. All the results indirectly demonstrate that fertilization in Fucus serratus is based on an association between fucosyl- and mannosyl-containing ligands on the egg surface and specific carbohydrate-binding receptors on the sperm surface.

Carbohydrates↗
PubMed1978

Fertilization in brown algae. I. SEM and other observations on Fucus serratus.

The cell wall secreted immediately following sperm entry into an egg can be visualized by the fluorescent dye Calcofluor white. Cell wall secretion precedes nuclear fusion by 10-20 min. SEM observations of the surface of unfertilized and fertilized eggs and sperm attachment to eggs are described. These results are discussed in relation to fertilization in sea urchins and the biochemical phenomena associated with egg-sperm recognition in Fucus.

Cell Wall↗