RESUMEN
Crustaceans comprise an ecologically and morphologically diverse taxonomic group. They are typically considered resilient to many environmental perturbations found in marine and coastal environments, due to effective physiological regulation of ions and hemolymph pH, and a robust exoskeleton. Ocean acidification can affect the ability of marine calcifying organisms to build and maintain mineralized tissue and poses a threat for all marine calcifying taxa. Currently, there is no consensus on how ocean acidification will alter the ecologically relevant exoskeletal properties of crustaceans. Here, we present a systematic review and meta-analysis on the effects of ocean acidification on the crustacean exoskeleton, assessing both exoskeletal ion content (calcium and magnesium) and functional properties (biomechanical resistance and cuticle thickness). Our results suggest that the effect of ocean acidification on crustacean exoskeletal properties varies based upon seawater pCO2 and species identity, with significant levels of heterogeneity for all analyses. Calcium and magnesium content was significantly lower in animals held at pCO2 levels of 1500-1999 µatm as compared with those under ambient pCO2. At lower pCO2 levels, however, statistically significant relationships between changes in calcium and magnesium content within the same experiment were observed as follows: a negative relationship between calcium and magnesium content at pCO2 of 500-999 µatm and a positive relationship at 1000-1499 µatm. Exoskeleton biomechanics, such as resistance to deformation (microhardness) and shell strength, also significantly decreased under pCO2 regimes of 500-999 µatm and 1500-1999 µatm, indicating functional exoskeletal change coincident with decreases in calcification. Overall, these results suggest that the crustacean exoskeleton can be susceptible to ocean acidification at the biomechanical level, potentially predicated by changes in ion content, when exposed to high influxes of CO2. Future studies need to accommodate the high variability of crustacean responses to ocean acidification, and ecologically relevant ranges of pCO2 conditions, when designing experiments with conservation-level endpoints.
RESUMEN
Ionizing radiation, from both space and radiation therapy, is known to affect bone health. While there have been studies investigating changes in bone density and microstructure from radiation exposure, the effects of radiation on material properties are unknown. The current study addresses this gap by assessing bone material property changes in rats exposed to helium-4 radiation through spherical micro-indentation. Rats were exposed to a single dose of 0, 5, and 25 cGy whole body helium-4 radiation. Animals were euthanized at 7, 30, 90, or 180-days after exposure. Spherical micro-indentation was performed on axial cross sections of the femur cortical bone to determine instantaneous and relaxed shear moduli. At 90-days after exposure, the 25 cGy exposure caused a significant decline in shear modulus compared to control and 5 cGy groups. The instantaneous modulus decreased 33% and the relaxed modulus decreased 32% as compared to the sham group. This decline was followed by a recovery of both moduli, which was observed by 180-days after exposure; at 180 days, the moduli were no longer statistically different from those at 7 or 30 days. The observed decrease at 90 days, followed by recovery to baseline levels, can be attributed to the biological mechanisms involved in bone formation that were affected by radiation, bone turnover, and systemic changes in hormones due to radiation exposure. Continued assessment of the mechanisms that drive such a response in material properties may enable identification of pathways for therapeutic countermeasures against radiation exposure.
Asunto(s)
Huesos , Helio , Animales , Densidad Ósea , Hueso Cortical , Fémur , RatasRESUMEN
Ocean acidification can affect the ability of calcifying organisms to build and maintain mineralized tissue. In decapod crustaceans, the exoskeleton is a multilayered structure composed of chitin, protein and mineral, predominately magnesian calcite or amorphous calcium carbonate (ACC). We investigated the effects of acidification on the exoskeleton of mature (post-terminal-molt) female southern Tanner crabs, Chionoecetes bairdi Crabs were exposed to one of three pH levels - 8.1, 7.8 or 7.5 - for 2 years. Reduced pH led to a suite of body region-specific effects on the exoskeleton. Microhardness of the claw was 38% lower in crabs at pH 7.5 compared with those at pH 8.1, but carapace microhardness was unaffected by pH. In contrast, reduced pH altered elemental content in the carapace (reduced calcium, increased magnesium), but not the claw. Diminished structural integrity and thinning of the exoskeleton were observed at reduced pH in both body regions; internal erosion of the carapace was present in most crabs at pH 7.5, and the claws of these crabs showed substantial external erosion, with tooth-like denticles nearly or completely worn away. Using infrared spectroscopy, we observed a shift in the phase of calcium carbonate present in the carapace of pH 7.5 crabs: a mix of ACC and calcite was found in the carapace of crabs at pH 8.1, whereas the bulk of calcium carbonate had transformed to calcite in pH 7.5 crabs. With limited capacity for repair, the exoskeleton of long-lived crabs that undergo a terminal molt, such as C. bairdi, may be especially susceptible to ocean acidification.
Asunto(s)
Braquiuros , Dispositivo Exoesqueleto , Animales , Carbonato de Calcio , Femenino , Concentración de Iones de Hidrógeno , Océanos y Mares , Agua de MarRESUMEN
Barnacles are ancient arthropods that, as adults, are surrounded by a hard, mineralized, outer shell that the organism produces for protection. While extensive research has been conducted on the glue-like cement that barnacles use to adhere to surfaces, less is known about the barnacle exoskeleton, especially the process by which the barnacle exoskeleton is formed. Here, we present data exploring the changes that occur as the barnacle cyprid undergoes metamorphosis to become a sessile juvenile with a mineralized exoskeleton. Scanning electron microscope data show dramatic morphological changes in the barnacle exoskeleton following metamorphosis. Energy-dispersive X-ray spectroscopy indicates a small amount of calcium (8%) 1 h post-metamorphosis that steadily increases to 28% by 2 days following metamorphosis. Raman spectroscopy indicates calcite in the exoskeleton of a barnacle 2 days following metamorphosis and no detectable calcium carbonate in exoskeletons up to 3 h post-metamorphosis. Confocal microscopy indicates during this 2 day period, barnacle base plate area and height increases rapidly (0.001 mm2 h-1 and 0.30 µm h-1, respectively). These results provide critical information into the early life stages of the barnacle, which will be important for developing an understanding of how ocean acidification might impact the calcification process of the barnacle exoskeleton.
RESUMEN
The decapod crustacean exoskeleton is a multi-layered structure composed of chitin-protein fibers embedded with calcium salts. Decapod claws display tooth-like denticles, which come into direct contact with predators and prey. They are subjected to more regular and intense mechanical stress than other parts of the exoskeleton and therefore must be especially resistant to wear and abrasion. Here, we characterized denticle properties in five decapod species. Dactyls from three brachyuran crabs (Cancer borealis, Callinectes sapidus, and Chionoecetes opilio) and two anomuran crabs (Paralomis birsteini and Paralithodes camtschaticus) were sectioned normal to the contact surface of the denticle, revealing the interior of the denticle and the bulk endocuticle in which it is embedded. Microhardness, micro- and ultrastructure, and elemental composition were assessed along a transect running the width of the cuticle using microindentation hardness testing, optical and scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS), respectively. In all species tested, hardness was dramatically higher-up to ten times-in the denticle than in the bulk endocuticle. Likewise, in all species there was an increase in packing density of mineralized chitin-protein fibers, a decrease in width of the pore canals that run through the cuticle, and a decrease in phosphorous content from endocuticle to denticle. The changes in hardness across the cuticle, and the relationship between hardness, calcium, and magnesium content, however, varied among species. Although mechanical resistance of the denticles was exceptionally high in all species, the basis for resistance appears to differ among species. STATEMENT OF SIGNIFICANCE: Understanding the diverse mechanisms by which animals attain exceptionally high mechanical resistance may enable development of novel, biologically inspired materials. Decapod crustacean claws, and particularly the tooth-like denticles that these claws display, are of interest in this regard, as they must be especially resistant to wear. We assessed mechanical, elemental, and structural properties of the claw cuticle in five decapod species. Without exception, microhardness was dramatically higher in the denticle than in the bulk endocuticle. Multivariant statistical analyses, however, showed that the relationships among microhardness, elemental content, and structural variables differed among species. Such patterns likely result from strong evolutionary pressure on feeding and defensive structures and a trade-off between mechanical properties and energetic cost of exoskeleton formation.
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Braquiuros , Calcificaciones de la Pulpa Dental , Animales , Dureza , Microscopía Electrónica de Rastreo , Espectrometría por Rayos XRESUMEN
A series of eight novel siloxane-polyurethane fouling-release (FR) coatings were assessed for their FR performance in both the laboratory and in the field. Laboratory analysis included adhesion assessments of bacteria, microalgae, macroalgal spores, adult barnacles and pseudobarnacles using high-throughput screening techniques, while field evaluations were conducted in accordance with standardized testing methods at three different ocean testing sites over the course of six-months exposure. The data collected were subjected to statistical analysis in order to identify potential correlations. In general, there was good agreement between the laboratory screening assays and the field assessments, with both regimes clearly distinguishing the siloxane-polyurethane compositions comprising monofunctional poly(dimethyl siloxane) (PDMS) (m-PDMS) as possessing superior, broad-spectrum FR properties compared to those prepared with difunctional PDMS (d-PDMS). Of the seven laboratory screening techniques, the Cellulophaga lytica biofilm retraction and reattached barnacle (Amphibalanus amphitrite) adhesion assays were shown to be the most predictive of broad-spectrum field performance.
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Biopelículas/crecimiento & desarrollo , Incrustaciones Biológicas/prevención & control , Poliuretanos/química , Siloxanos/química , Animales , Adhesión Celular/fisiología , Dimetilpolisiloxanos/química , Flavobacteriaceae/fisiología , Ensayos Analíticos de Alto Rendimiento , Microalgas/fisiología , Modelos Teóricos , Propiedades de Superficie , Thoracica/fisiologíaRESUMEN
Barnacles permanently adhere to nearly any inert substrate using proteinaceous glue. The glue consists of at least ten major proteins, some of which have been isolated and sequenced. Questions still remain about the chemical mechanisms involved in adhesion and the potential of the glue to serve as a platform for mineralization of the calcified base plate. We tested the hypothesis that barnacle glue contains phosphoproteins, which have the potential to play a role in both adhesion and mineralization. Using a combination of phosphoprotein-specific gel staining and Western blotting with anti-phosphoserine antibody, we identified multiple phosphorylated proteins in uncured glue secretions from the barnacle Amphibalanus amphitrite The protein composition of the glue and the quantity and abundance of phosphoproteins varied distinctly among individual barnacles, possibly due to cyclical changes in the glue secretion over time. We assessed the location of the phosphoproteins within the barnacle glue layer using decalcified barnacle base plates and residual glue deposited by reattached barnacles. Phosphoproteins were found throughout the organic matrix of the base plate and within the residual glue. Staining within the residual glue appeared most intensely in regions where capillary glue ducts, which are involved in cyclical release of glue, had been laid down. Lastly, mineralization studies of glue proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that proteins identified as phosphorylated possibly induce mineralization of calcium carbonate (CaCO3). These results contribute to our understanding of the protein composition of barnacle glue, and provide new insights into the potential roles of phosphoproteins in underwater bioadhesives.
Asunto(s)
Fosfoproteínas/metabolismo , Thoracica/metabolismo , Adhesivos/química , Animales , Western Blotting , Carbonato de Calcio/metabolismo , Fosfoproteínas/aislamiento & purificaciónRESUMEN
BACKGROUND: Ocean acidification as a result of increased anthropogenic CO2 emissions is occurring in marine and estuarine environments worldwide. The coastal ocean experiences additional daily and seasonal fluctuations in pH that can be lower than projected end-of-century open ocean pH reductions. In order to assess the impact of ocean acidification on marine invertebrates, Pacific oysters (Crassostrea gigas) were exposed to one of four different p CO2 levels for four weeks: 400 µatm (pH 8.0), 800 µatm (pH 7.7), 1000 µatm (pH 7.6), or 2800 µatm (pH 7.3). RESULTS: At the end of the four week exposure period, oysters in all four p CO2 environments deposited new shell, but growth rate was not different among the treatments. However, micromechanical properties of the new shell were compromised by elevated p CO2. Elevated p CO2 affected neither whole body fatty acid composition, nor glycogen content, nor mortality rate associated with acute heat shock. Shotgun proteomics revealed that several physiological pathways were significantly affected by ocean acidification, including antioxidant response, carbohydrate metabolism, and transcription and translation. Additionally, the proteomic response to a second stress differed with p CO2, with numerous processes significantly affected by mechanical stimulation at high versus low p CO2 (all proteomics data are available in the ProteomeXchange under the identifier PXD000835). CONCLUSIONS: Oyster physiology is significantly altered by exposure to elevated p CO2, indicating changes in energy resource use. This is especially apparent in the assessment of the effects of p CO2 on the proteomic response to a second stress. The altered stress response illustrates that ocean acidification may impact how oysters respond to other changes in their environment. These data contribute to an integrative view of the effects of ocean acidification on oysters as well as physiological trade-offs during environmental stress.
Asunto(s)
Ácidos/metabolismo , Crassostrea/fisiología , Océanos y Mares , Proteómica , Animales , Crassostrea/metabolismo , Ácidos Grasos/metabolismo , Glucógeno/metabolismoRESUMEN
The continuing increase of carbon dioxide (CO2) levels in the atmosphere leads to increases in global temperatures and partial pressure of CO2 (PCO2) in surface waters, causing ocean acidification. These changes are especially pronounced in shallow coastal and estuarine waters and are expected to significantly affect marine calcifiers including bivalves that are ecosystem engineers in estuarine and coastal communities. To elucidate potential effects of higher temperatures and PCO2 on physiology and biomineralization of marine bivalves, we exposed two bivalve species, the eastern oysters Crassostrea virginica and the hard clams Mercenaria mercenaria to different combinations of PCO2 (~400 and 800µatm) and temperatures (22 and 27°C) for 15weeks. Survival, bioenergetic traits (tissue levels of lipids, glycogen, glucose and high energy phosphates) and biomineralization parameters (mechanical properties of the shells and activity of carbonic anhydrase, CA) were determined in clams and oysters under different temperature and PCO2 regimes. Our analysis showed major inter-species differences in shell mechanical traits and bioenergetics parameters. Elevated temperature led to the depletion of tissue energy reserves indicating energy deficiency in both species and resulted in higher mortality in oysters. Interestingly, while elevated PCO2 had a small effect on the physiology and metabolism of both species, it improved survival in oysters. At the same time, a combination of high temperature and elevated PCO2 lead to a significant decrease in shell hardness in both species, suggesting major changes in their biomineralization processes. Overall, these studies show that global climate change and ocean acidification might have complex interactive effects on physiology, metabolism and biomineralization in coastal and estuarine marine bivalves.
Asunto(s)
Dióxido de Carbono/farmacología , Crassostrea/metabolismo , Metabolismo Energético/efectos de los fármacos , Mercenaria/metabolismo , Minerales/metabolismo , Temperatura , Exoesqueleto/anatomía & histología , Exoesqueleto/efectos de los fármacos , Exoesqueleto/fisiología , Animales , Fenómenos Biomecánicos/efectos de los fármacos , Anhidrasas Carbónicas/metabolismo , Crassostrea/efectos de los fármacos , Crassostrea/enzimología , Activación Enzimática/efectos de los fármacos , Mercenaria/efectos de los fármacos , Mercenaria/enzimología , Especificidad de Órganos/efectos de los fármacos , Análisis de Componente Principal , Análisis de Supervivencia , Agua/químicaRESUMEN
Ocean acidification due to increasing atmospheric CO2 concentrations results in a decrease in seawater pH and shifts in the carbonate chemistry that can negatively affect marine organisms. Marine bivalves such as the hard-shell clam, Mercenaria mercenaria, serve as ecosystem engineers in estuaries and coastal zones of the western Atlantic and, as for many marine calcifiers, are sensitive to the impacts of ocean acidification. In estuaries, the effects of ocean acidification can be exacerbated by low buffering capacity of brackish waters, acidic inputs from freshwaters and land, and/or the negative effects of salinity on the physiology of organisms. We determined the interactive effects of 21 weeks of exposure to different levels of CO2 (~395, 800 and 1500 µatm corresponding to pH of 8.2, 8.1 and 7.7, respectively) and salinity (32 versus 16) on biomineralization, shell properties and energy metabolism of juvenile hard-shell clams. Low salinity had profound effects on survival, energy metabolism and biomineralization of hard-shell clams and modulated their responses to elevated PCO2. Negative effects of low salinity in juvenile clams were mostly due to the strongly elevated basal energy demand, indicating energy deficiency, that led to reduced growth, elevated mortality and impaired shell maintenance (evidenced by the extensive damage to the periostracum). The effects of elevated PCO2 on physiology and biomineralization of hard-shell clams were more complex. Elevated PCO2 (~800-1500 µatm) had no significant effects on standard metabolic rates (indicative of the basal energy demand), but affected growth and shell mechanical properties in juvenile clams. Moderate hypercapnia (~800 µatm PCO2) increased shell and tissue growth and reduced mortality of juvenile clams in high salinity exposures; however, these effects were abolished under the low salinity conditions or at high PCO2 (~1500 µatm). Mechanical properties of the shell (measured as microhardness and fracture toughness of the shells) were negatively affected by elevated CO2 alone or in combination with low salinity, which may have important implications for protection against predators or environmental stressors. Our data indicate that environmental salinity can strongly modulate responses to ocean acidification in hard-shell clams and thus should be taken into account when predicting the effects of ocean acidification on estuarine bivalves.
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Exoesqueleto/química , Bivalvos/metabolismo , Dióxido de Carbono/análisis , Metabolismo Energético/fisiología , Salinidad , Agua de Mar/química , Animales , Fenómenos Biomecánicos , Dióxido de Carbono/metabolismo , Concentración de Iones de Hidrógeno , North CarolinaRESUMEN
Rising levels of atmospheric CO(2) lead to acidification of the ocean and alter seawater carbonate chemistry, which can negatively impact calcifying organisms, including mollusks. In estuaries, exposure to elevated CO(2) levels often co-occurs with other stressors, such as reduced salinity, which enhances the acidification trend, affects ion and acid-base regulation of estuarine calcifiers and modifies their response to ocean acidification. We studied the interactive effects of salinity and partial pressure of CO(2) (P(CO2)) on biomineralization and energy homeostasis in juveniles of the eastern oyster, Crassostrea virginica, a common estuarine bivalve. Juveniles were exposed for 11 weeks to one of two environmentally relevant salinities (30 or 15 PSU) either at current atmospheric P(CO2) (â¼400 µatm, normocapnia) or P(CO2) projected by moderate IPCC scenarios for the year 2100 (â¼700-800 µatm, hypercapnia). Exposure of the juvenile oysters to elevated P(CO2) and/or low salinity led to a significant increase in mortality, reduction of tissue energy stores (glycogen and lipid) and negative soft tissue growth, indicating energy deficiency. Interestingly, tissue ATP levels were not affected by exposure to changing salinity and P(CO2), suggesting that juvenile oysters maintain their cellular energy status at the expense of lipid and glycogen stores. At the same time, no compensatory upregulation of carbonic anhydrase activity was found under the conditions of low salinity and high P(CO2). Metabolic profiling using magnetic resonance spectroscopy revealed altered metabolite status following low salinity exposure; specifically, acetate levels were lower in hypercapnic than in normocapnic individuals at low salinity. Combined exposure to hypercapnia and low salinity negatively affected mechanical properties of shells of the juveniles, resulting in reduced hardness and fracture resistance. Thus, our data suggest that the combined effects of elevated P(CO2) and fluctuating salinity may jeopardize the survival of eastern oysters because of weakening of their shells and increased energy consumption.
Asunto(s)
Dióxido de Carbono/metabolismo , Crassostrea/crecimiento & desarrollo , Animales , Fenómenos Biomecánicos , Calcificación Fisiológica , Crassostrea/anatomía & histología , Crassostrea/metabolismo , Metabolismo Energético , Resonancia Magnética Nuclear Biomolecular , SalinidadRESUMEN
Barnacle cement (BC) was beneficially applied on stainless steel (SS) to serve as the initiator anchor for surface-initiated polymerization. The amine and hydroxyl moieties of barnacle cement reacted with 2-bromoisobutyryl bromide to provide the alkyl halide initiator for the surface-initiated atom transfer radical polymerization (ATRP) of 2-hydroxyethyl methacrylate (HEMA). The hydroxyl groups of HEMA polymer (PHEMA) were then converted to carboxyl groups for coupling of chitosan (CS) to impart the SS surface with both antifouling and antibacterial properties. The surface-functionalized SS reduced bovine serum albumin adsorption, bacterial adhesion, and exhibited antibacterial efficacy against Escherichia coli (E. coli). The effectiveness of barnacle cement as an initiator anchor was compared to that of dopamine, a marine mussel inspired biomimetic anchor previously used in surface-initiated polymerization. The results indicate that the barnacle cement is a stable and effective anchor for functional surface coatings and polymer brushes.
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Antibacterianos/química , Antibacterianos/farmacología , Incrustaciones Biológicas/prevención & control , Materiales Biomiméticos/química , Materiales Biomiméticos/farmacología , Acero Inoxidable/química , Adsorción , Animales , Adhesión Bacteriana/efectos de los fármacos , Bovinos , Quitosano/química , Dopamina/química , Escherichia coli/efectos de los fármacos , Polihidroxietil Metacrilato/química , Polimerizacion , Albúmina Sérica Bovina/química , Propiedades de Superficie , Thoracica/químicaRESUMEN
Microtopography is one of several strategies used by marine organisms to inhibit colonization by fouling organisms. While replicates of natural microtextures discourage settlement, details of larval interactions with the structured surfaces remain scarce. Close-range microscopy was used to quantify the exploration of cyprids of Amphibalanus amphitrite on cylindrical micropillars with heights of 5 and 30 µm and diameters ranging from 5 to 100 µm. While 5 µm-high structures had little impact, 30 µm-high pillars significantly influenced cyprid exploration. An observed step length decrease and step duration increase on 5 µm diameter pillars is attributed to the small dimensions of the voids excluding the cyprid's attachment disc and consequently reducing the area of adhesive contact. When exploring larger diameter pillars, cyprids preferred using the voids to form temporary attachment points. This may enhance their resistance to flow. No-choice assay settlement patterns mirrored this exploration behaviour, albeit in a pattern counter to what was predicted.
Asunto(s)
Conducta Animal/fisiología , Thoracica/fisiología , Animales , Incrustaciones Biológicas/prevención & control , Bioensayo , Larva/fisiología , Biología Marina , Propiedades de Superficie , Thoracica/crecimiento & desarrolloRESUMEN
Cryogenic transmission electron microscopy (cryo-EM) was used to explore the self-assembly of recombinant murine amelogenin (rM179) in vitro. Our cryo-EM data showed that amelogenin self-assembly is a strongly pH-dependent process. At pH 4.4 the main fraction of the protein exists in a monomeric form, although some peculiar structures consisting of chains of monomers were also observed. At pH 5.8 large nanospheres comprising ring-like structures ~50 nm in diameter were the most abundant particle class. Similarly, at pH 8.0 amelogenins self-assembled into ring-like oligomers of different sizes, which subsequently assembled into nanospheres 15-20 nm in diameter. Furthermore, at pH 7.2, which is close to a physiological pH, branched chains of nanospheres were observed. Our results show that amelogenin assembly is a multistep hierarchical process and provides new insight into the control of enamel mineralization.
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Amelogenina/ultraestructura , Microscopía por Crioelectrón , Microscopía Electrónica de Transmisión/métodos , Amelogenina/química , Animales , Concentración de Iones de Hidrógeno , Ratones , Nanosferas/ultraestructura , Estructura Cuaternaria de Proteína , Factores de TiempoRESUMEN
BACKGROUND: Attachment strength of fouling organisms on silicone coatings is low. We hypothesized that low attachment strength on silicones is, in part, due to the interaction of surface available components with natural glues. Components could alter curing of glues through bulk changes or specifically through altered enzyme activity. METHODOLOGY/PRINCIPAL FINDINGS: GC-MS analysis of silicone coatings showed surface-available siloxanes when the coatings were gently rubbed with a cotton swab for 15 seconds or given a 30 second rinse with methanol. Mixtures of compounds were found on 2 commercial and 8 model silicone coatings. The hypothesis that silicone components alter glue curing enzymes was tested with curing barnacle glue and with commercial enzymes. In our model, barnacle glue curing involves trypsin-like serine protease(s), which activate enzymes and structural proteins, and a transglutaminase which cross-links glue proteins. Transglutaminase activity was significantly altered upon exposure of curing glue from individual barnacles to silicone eluates. Activity of purified trypsin and, to a greater extent, transglutaminase was significantly altered by relevant concentrations of silicone polymer constituents. CONCLUSIONS/SIGNIFICANCE: Surface-associated silicone compounds can disrupt glue curing and alter enzyme properties. Altered curing of natural glues has potential in fouling management.
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Adhesivos/metabolismo , Enzimas/metabolismo , Compuestos de Silicona/efectos adversos , Thoracica/enzimología , Thoracica/metabolismo , Adhesividad/efectos de los fármacos , Adhesivos/química , Animales , Cementación , Activación Enzimática/efectos de los fármacos , Pruebas de Enzimas , Enzimas/efectos de los fármacos , Cromatografía de Gases y Espectrometría de Masas , Fenómenos Mecánicos , Metanol/farmacología , Modelos Biológicos , Modelos Teóricos , Compuestos de Silicona/química , Compuestos de Silicona/farmacología , Propiedades de Superficie/efectos de los fármacos , Thoracica/efectos de los fármacos , Thoracica/fisiologíaRESUMEN
Settlement inhibition of barnacle (Amphibalanus amphitrite) cypris larvae resulting from exposure to ultrasound was measured at three frequencies (23, 63, and 102 kHz), applied at three acoustic pressure levels (9, 15, and 22 kPa) for exposure times of 30, 150, and 300 s. The lowest settlement was observed for 23 kHz, which also induced the highest cyprid mortality. Cyprid settlement following exposure to 23 kHz at 22 kPa for 30 s was reduced by a factor of two. Observing surface exploration by the cyprids revealed an altered behaviour following exposure to ultrasound: step length was increased, while step duration, walking pace, and the fraction of cyprids exploring the surface were significantly reduced with respect to control cyprids. The basal area of juvenile barnacles, metamorphosed from ultrasound-treated cyprids was initially smaller than unexposed individuals, but normalised over two weeks' growth. Thus, ultrasound exposure effectively reduced cyprid settlement, yet metamorphosed barnacles grew normally.
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Incrustaciones Biológicas/prevención & control , Ondas de Choque de Alta Energía , Thoracica/efectos de la radiación , Animales , Conducta Animal/efectos de la radiación , Larva/crecimiento & desarrollo , Larva/fisiología , Larva/efectos de la radiación , Movimiento , Thoracica/crecimiento & desarrollo , Thoracica/fisiologíaRESUMEN
Filamentous benthic marine cyanobacteria are a prolific source of structurally unique bioactive secondary metabolites. A total of 12 secondary metabolites, belonging to the mixed polyketide-polypeptide structural class, were isolated from the marine cyanobacterium, Lyngbya majuscula, and were tested to determine if they showed activity against barnacle larval settlement. The assays revealed four compounds, dolastatin 16, hantupeptin C, majusculamide A, and isomalyngamide A, that showed moderate to potent anti-larval settlement activities, with EC(50) values ranging from 0.003 to 10.6 microg ml(-1). In addition, field testing conducted over a period of 28 days (using the modified Phytagel method) based on the cyanobacterial compound, dolastatin 16, showed significantly reduced barnacle settlement as compared to controls at all the concentrations tested. The results of this study highlight the importance of marine cyanobacteria as an underexplored source of potential environmentally friendly antifoulants.
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Amidas/farmacología , Toxinas Bacterianas/farmacología , Incrustaciones Biológicas/prevención & control , Cianobacterias , Depsipéptidos/farmacología , Toxinas Marinas/farmacología , Thoracica/efectos de los fármacos , Amidas/química , Amidas/aislamiento & purificación , Animales , Toxinas Bacterianas/química , Toxinas Bacterianas/aislamiento & purificación , Depsipéptidos/química , Depsipéptidos/aislamiento & purificación , Larva/efectos de los fármacos , Larva/fisiología , Toxinas Marinas/química , Toxinas Marinas/aislamiento & purificación , Thoracica/crecimiento & desarrollo , Thoracica/fisiologíaRESUMEN
The nanoscale morphology and protein secondary structure of barnacle adhesive plaques were characterized using atomic force microscopy (AFM), far-UV circular dichroism (CD) spectroscopy, transmission Fourier transform infrared (FTIR) spectroscopy, and Thioflavin T (ThT) staining. Both primary cement (original cement laid down by the barnacle) and secondary cement (cement used for reattachment) from the barnacle Balanus amphitrite (= Amphibalanus amphitrite) were analyzed. Results showed that both cements consisted largely of nanofibrillar matrices having similar composition. Of particular significance, the combined results indicate that the nanofibrillar structures are consistent with amyloid, with globular protein components also identified in the cement. Potential properties, functions, and formation mechanisms of the amyloid-like nanofibrils within the adhesive interface are discussed. Our results highlight an emerging trend in structural biology showing that amyloid, historically associated with disease, also has functional roles.
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Amiloide/química , Nanoestructuras/química , Thoracica/química , Adhesivos/química , Adhesivos/metabolismo , Amiloide/metabolismo , Animales , Dicroismo Circular , Microscopía de Fuerza Atómica , Estructura Secundaria de Proteína , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Enzymes and biochemical mechanisms essential to survival are under extreme selective pressure and are highly conserved through evolutionary time. We applied this evolutionary concept to barnacle cement polymerization, a process critical to barnacle fitness that involves aggregation and cross-linking of proteins. The biochemical mechanisms of cement polymerization remain largely unknown. We hypothesized that this process is biochemically similar to blood clotting, a critical physiological response that is also based on aggregation and cross-linking of proteins. Like key elements of vertebrate and invertebrate blood clotting, barnacle cement polymerization was shown to involve proteolytic activation of enzymes and structural precursors, transglutaminase cross-linking and assembly of fibrous proteins. Proteolytic activation of structural proteins maximizes the potential for bonding interactions with other proteins and with the surface. Transglutaminase cross-linking reinforces cement integrity. Remarkably, epitopes and sequences homologous to bovine trypsin and human transglutaminase were identified in barnacle cement with tandem mass spectrometry and/or western blotting. Akin to blood clotting, the peptides generated during proteolytic activation functioned as signal molecules, linking a molecular level event (protein aggregation) to a behavioral response (barnacle larval settlement). Our results draw attention to a highly conserved protein polymerization mechanism and shed light on a long-standing biochemical puzzle. We suggest that barnacle cement polymerization is a specialized form of wound healing. The polymerization mechanism common between barnacle cement and blood may be a theme for many marine animal glues.
Asunto(s)
Evolución Biológica , Modelos Biológicos , Polímeros/química , Proteínas/química , Thoracica/química , Secuencia de Aminoácidos , Animales , Calcio/metabolismo , Bovinos , Humanos , Microscopía de Fuerza Atómica , Datos de Secuencia Molecular , Espectrometría de Masas en Tándem , Transglutaminasas/metabolismo , Tripsina/metabolismoRESUMEN
Polymerized barnacle glue was studied by atomic force microscopy (AFM), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy and chemical staining. Nanoscale structures exhibiting rod-shaped, globular and irregularly-shaped morphologies were observed in the bulk cement of the barnacle Amphibalanus amphitrite (=Balanus amphitrite) by AFM. SEM coupled with energy dispersive X-ray (EDX) provided chemical composition information, making evident the organic nature of the rod-shaped nanoscale structures. FTIR spectroscopy gave signatures of beta-sheet and random coil conformations. The mechanical properties of these nanoscale structures were also probed using force spectroscopy and indentation with AFM. Indentation data yielded higher elastic moduli for the rod-shaped structures when compared with the other structures in the bulk cement. Single molecule AFM force-extension curves on the matrix of the bulk cement often exhibited a periodic sawtooth-like profile, observed in both the extend and retract portions of the force curve. Rod-shaped structures stained with amyloid protein-selective dyes (Congo red and thioflavin-T) revealed that about 5% of the bulk cement were amyloids. A dominant 100 kDa cement protein was found to be mechanically agile, using repeating hydrophobic structures that apparently associate within the same protein or with neighbors, creating toughness on the 1-100 nm length scale.