Structural and chemical defenses of echinoderms from the northern Gulf of Mexico

The feeding deterrent effects of echinoderm body-wall tissues and ethanolic extracts containing mid-polarity compounds were evaluated utilizing generalist fish and crabs as model predators. The body-wall tissues of the echinoderms examined ranged 10-fold from 0.9–9.4 mm in thickness, and four and a half-fold in level of mineralization (17.8–82.7% ash content). Holothuroids had the thickest body-wall tissues and contained the lowest levels of mineralization in their body-walls. Crinoids and ophiuroids had high levels of mineralization in their arms. Asteroid body-wall tissues varied the most in thickness and ash content (0.9–3.9 mm in thickness and 29.2–55.5% in ash content). Body-wall tissues of 19 species of echinoderms were tested for their feeding deterrent properties against the marine fishes Lagodon rhomboides (Linnaeus) and Cyprinodon variegatus (Lacepede), as well as the decapod crustacean Libinia emarginata (Leach). Equivalent sized pieces of fresh body-wall tissue of 16 species of echinoderms caused observable feeding deterrence responses in at least two of the three model predators. There was no significant correlation between body-wall thickness or percent ash and its palatability to any of the three model predators. Agar pellets containing ethanolic body-wall extracts of 12 of 18 echinoderm species caused observable feeding deterrence responses in the fish L. rhomboides. In similar experiments with the arrow crab Stenorhyncus seticornis (Herbst), using carrageenan fish-meal blocks as food models, no differences in consumption of control fish-meal and experimental body-wall extract blocks were detected. Our findings indicate that invertebrate and vertebrate predators may respond quite differently to echinoderm body-wall extracts.     

Use of protein A-coated colloidal gold particles for immunoelectronmicroscopic localization of ACTH on ultrathin sections

Summary ACTH was localized in dissociated porcine adenohypophysial cells using a novel indirect EM immunocytochemical technique. Incubation of ultrathin resin sections in anti-ACTH was followed by incubation with protein A-coated colloidal gold particles. Protein A binds specifically to the Fc part of the IgG molecule, and thus the ACTH-containing secretory granules became labelled with electron-dense gold particles. With this method, the dissociated porcine ACTH cell was identified as containing numerous round or ovoid 170–300 nm, secretory granules.     

Chemical-modification rescue assessed by mass spectrometry demonstrates that gamma-thia-lysine yields the same activity as lysine in aldolase

The role of active site residues in fructose 1,6-bisphosphate aldolase is investigated by chemical-modification rescue. An active-site mutation, K107C, is constructed in a background where the four solvent-accessible cysteine residues are converted to alanine. The resulting mutant, tetK107C, when reacted with bromoethylamine (BrEA), shows a 40-fold increase in activity (to 80% that of wild type). Determination of the sites and their degree of modification using electrospray ionization Fourier transform mass spectrometry (ESI-FTMS) is developed, allowing correlation of activity after chemical modification rescue to the degree of modification. The stoichiometry of the reaction is 2.5 aminoethylations per subunit, as measured by ESI-FTMS. Protein modification with a double-labeled mix (1:1) of natural abundance isotope (d(0)-BrEA) and 2-bromoethyl-1,1,2,2-d4-amine hydrobromide (d(4)-BrEA), followed by dialysis and trypsin digestion, shows aminoethylated peptides as "twin peptides" separated by four mass units in ESI-FTMS analysis. Using this detection procedure under nondenaturing (native) conditions, C107 is aminoethylated, whereas the four buried thiols remain unlabeled. Aminoethylation of other residues is observed, and correlates with those peptides containing histidine, methionine, and/or the amino terminus. Quantification of the aminoethylation reaction is achieved by labeling with nondeuterated d(0)-BrEA under denaturing conditions following double labeling under native conditions. In addition to complete labeling all five thiols, the intensity of the d(0)-BrEA peak for C107 containing peptides increases, and the change in the d(0)/d(4) ratio between native and denaturing conditions shows 82 +/- 4.5% aminoethylation at C107. This correlation of modification with the recovered activity, indicates that gamma-thia-lysine replaces lysine in the catalytic mechanism. Kinetic constants measured for the rescued K107C mutant enzyme with the substrates fructose 1-phosphate and fructose 1,6-bisphosphate are consistent with the role of the positively charged lysine binding to the C6-phosphate. ESI-FTMS, combined with this double-labeling procedure, allows precise identification of sites and measurement of degree of protein modification.     

Osteopathology in Rhinocerotidae from 50 Million Years to the Present

Individual elements of many extinct and extant North American rhinocerotids display osteopathologies, particularly exostoses, abnormal textures, and joint margin porosity, that are commonly associated with localized bone trauma. When we evaluated six extinct rhinocerotid species spanning 50 million years (Ma), we found the incidence of osteopathology increases from 28% of all elements of Eocene Hyrachyus eximius to 65–80% of all elements in more derived species. The only extant species in this study, Diceros bicornis, displayed less osteopathologies (50%) than the more derived extinct taxa. To get a finer-grained picture, we scored each fossil for seven pathological indicators on a scale of 1–4. We estimated the average mass of each taxon using M1-3 length and compared mass to average pathological score for each category. We found that with increasing mass, osteopathology also significantly increases. We then ran a phylogenetically-controlled regression analysis using a time-calibrated phylogeny of our study taxa. Mass estimates were found to significantly covary with abnormal foramen shape and abnormal bone textures. This pattern in osteopathological expression may reflect a part of the complex system of adaptations in the Rhinocerotidae over millions of years, where increased mass, cursoriality, and/or increased life span are selected for, to the detriment of long-term bone health. This work has important implications for the future health of hoofed animals and humans alike.     

Hexagonal-String Phases in Suspension Flow

Abstract

Investigations of the structure of flowing colloidal suspensions have seen structures of predominantly hexagonal strings moving in layered arrangements in the direction of the velocity gradient. A combination of computer simulation trajectories and computer graphics show that the predominant “hexagonal string phase” is the BCC lattice sheared along its (111) slip plane and, moreover, that all the common cubic lattice types contain “hexagonal strings”.     

Adverse Effects of Ball-Chain Radio-Collars on Female Mantled Howlers (<Emphasis Type="Italic">Alouatta palliata</Emphasis>) in Panama

The placement of radio- and GPS-collars on primates has advanced the fields of primate behavior and ecology and yielded considerable conservation gains. However, to ensure that the placement of tracking collars remains a humane practice, the reporting of both positive and negative outcomes is required. Here, we evaluate the impacts of radio-collars on the reproduction, behavior, and health of female mantled howlers (Alouatta palliata) on Barro Colorado Island, Panama. In June 2005, as part of a behavioral study, we fitted 16 female howlers with ball-chain radio-collars weighing 1.2% of the mean adult female weight. We monitored these 16 collared females and 64 noncollared females across an annual cycle, and recaptured the collared females in July 2006 for collar removal. The mortality rate for collared females across the study year was 6.3% (N=1), while the net loss of noncollared females from study groups was 15.6% (N=10). All collared females reproduced. The behaviors of collared and noncollared females were not significantly different. However, on recapture to remove all remaining collars (N = 13), 38% of recaptured collared females presented with damage to the dermal layer on the back of the neck and 31% presented with more severe damage extending into the subcutaneous tissue and muscle. Given this physical damage, which was not apparent through binoculars, we cannot recommend using ball-chain radio-collars on female mantled howlers without careful welfare monitoring.     

A STUDY OF THE SEED BANK OF A SALT MARSH IN NORTHERN SAN FRANCISCO BAY

Abundance, species composition, and distribution of buried seeds in a San Francisco Bay salt marsh were studied by collecting soil samples in October and February and observing seedling emergence in the greenhouse. Results were compared with existing vegetation patterns and field germination. Average numbers of buried viable seed down to a 5-cm depth were 380/ m² in October and 700/m² in February, with field germination averaging 118/m². Salicornia virginica dominated the seed bank in the greenhouse and in field germination. Most other marsh species were present in the seed bank but numbers of seeds were low. A significant correlation was found between highest species diversity and proximity to channels. The nature of this low-diversity, low-density seed bank reflects dominance of long-lived perennial species, seed dispersal patterns and selective environmental pressures.