Changes in fibrinolytic activity in diving grey seals

In order to test the hypothesis that enhanced fibrinolytic activity is a factor which prevents the blood of diving seals from clotting, we instrumented two female grey seals (Halichoerus grypus) with subcutaneous electrodes for measurements of heart rate (HR) and an extradural intravertebral venous catheter for collection of blood samples before, during and after simulated dives of 10 min duration. Blood samples were used for in vitro determination of clot lysis time (CLT), which is a measure of the level of fibrinolytic activity, and for analyses of plasma levels of cortisol, noradrenaline and adrenaline (A). The seals displayed profound diving bradycardia indicative of a substantial reduction in blood flow rates (pre-dive HR: 78 (63–98) bpm; dive HR: 8 (7–10) bpm; (median (range); n=2)) and elevated catecholamine levels (pre-dive A: 121 (98–184) pg·ml⁻¹; peak dive/post-dive A: 3510 (447–6181) pg·ml⁻¹), both of which are factors which promote blood coagulation. Nevertheless, we found that CLT always increased in connection with diving (pre-dive CLT: 436 (356–568) min; peak CLT during diving: 1380 (640–1800) min), which implies a reduced, rather than enhanced, fibrinolytic activity in this situation. These results show that enhanced fibrinolytic activity is not part of the defence system which prevents fatal clotting from occurring in diving grey seals.     

Kinetics and mechanism of dissociation of cooperatively bound T4 gene 32 protein-single-stranded nucleic acid complexes. 2. Changes in mechanism as a function of sodium chloride concentration and other solution variables

The dissociation kinetics of bacteriophage T4 coded gene 32 protein-single-stranded nucleic acid complexes have been examined as a function of monovalent salt concentration, temperature, and pH in order to investigate the details of the dissociation of cooperatively bound protein. Fluorescence stopped-flow techniques were used, and irreversible dissociation was induced by a combination of [NaCl] jumps and mixing with excess nucleic acid competitor. This made it possible to directly investigate the irreversible dissociation process over a wide range of NaCl concentrations [e.g., from 50 mM to 0.60 M for the gene 32 protein-poly(A) complex], in the absence of reassociation. Over the entire salt range, the only dissociable species observed is the singly contiguously bound gene 32 protein which dissociates from the ends of protein clusters. However, the [NaCl] dependence of the dissociation rate constant suggests that two competing pathways exist for dissociation of cooperatively bound gene 32 protein from the ends of protein clusters. At high monovalent salt concentrations, dissociation is dominated by a single-step process, with log ke/log [NaCl] = 6.5 +/- 0.5; i.e., the dissociation rate constant increases with increasing NaCl concentration due to the uptake of approximately six monovalent ions upon dissociation. This indicates that singly contiguous protein dissociates directly into solution. However, at much lower [NaCl] the data suggest that gene 32 protein, when bound at the end of a protein cluster, dissociates by first sliding off the end to form a noncooperatively bound intermediate which subsequently dissociates. A quantitative model which incorporates the sliding pathway [Berg, O. G., Winter, R. B., & von Hippel, P. H. (1981) Biochemistry 20, 6929-6948] in the dissociation mechanism fits the data reasonably well and suggests that noncooperatively bound monomers of gene 32 protein may be capable of one-dimensional translocation along single-stranded nucleic acids as suggested by independent kinetic data on the association reaction [Lohman, T. M., & Kowalczykowski, S. C. (1981) J. Mol. Biol. 152, 67-109]. It is also observed that both the absolute dissociation rate constant for T4 gene 32 protein and its salt dependence are sensitive to the average molecular weight and polydispersity of the nucleic acid sample used. This is a general phenomenon exhibited by proteins that bind to nucleic acids in a highly cooperative manner.     

Terminal degeneration in the mediodorsal cerebral cortex of the lizard Agama agama: Light and electron microscopy

The degeneration of axon terminals in the small-celled part of the mediodorsal cortex (sMDC) of the lizard Agama agama has been studied after lesions in the dorsal cortex at various survival periods. The Fink-Heimer stain was used to map and demonstrate terminal degeneration with the light and electron microscope. Electron microscopy was used to identify and describe degenerating boutons ultrastructurally. One sham-operated and three unoperated animals served as controls. Between 6 and 21 days postsurgically, degenerating terminals can be seen through 80% of the superficial plexiform layer, the zone adjacent to the cellular layer remaining free of degeneration. Swelling of dendrites in the outer part of the superficial plexiform layer and increased numbers of vacuolar invaginations, both present at short (24 hr–6 days; peak at 48–54 hr) survival periods, can be regarded as reaction to the surgical trauma. Degeneration of axon terminals takes three forms, all of the electron-dense type: gray boutons, degenerating bouton-dendritic spine complexes surrounded or engulfed by glia, and degeneration debris inside glial processes. Several forms of terminal degeneration occur concomitantly at any short (3–12 days) survival time. At longer survival times (15–21 days) only debris is present. From 6 days on, considerable numbers of degenerating structures are present, but the majority of degenerating boutons and debris are associated with reactive glia rather than with dendrites. From these observations it is concluded that in this lizard application of the combined degeneration-Golgi-EM technique would probably lead to little success. Electron microscopy of Fink-Heimer-stained sections suggests that degenerating bouton-dendritic spine complexes and degeneration debris accumulate silver particles, whereas gray boutons do not.     

Afrotropics on the wing: phylogenomics and historical biogeography of awl and policeman skippers

The Old‐World Tropics encompass many unique biomes and associated biotas shaped by drastic climate and geological changes throughout the Cenozoic. Disjunct distributions of clades between the Afrotropics and the Oriental regions are testament to these changes. Awl and policeman skippers (Hesperiidae: Coeliadinae) are disjunctly distributed with some genera endemic to the Afrotropics and others restricted to the Oriental and Australian regions. We reconstruct the phylogeny of these butterflies using target exon capture phylogenomics. We also generate a dated framework for this clade that uses the putatively oldest known butterfly fossil to estimate the historical biogeography of Coeliadinae using a model‐based approach. We infer a stable and robust phylogeny for the subfamily, with all but one Afrotropical lineage forming a derived clade. The African genus Pyrrhiades syn. nov. is placed in synonymy with Coeliades to accommodate the new phylogeny. Our comparative dating exercise casts doubt on the assignment of the fossil Protocoeliades kristenseni as a derived Coeliadinae and suggests, along with our biogeographic estimation, a split of Coeliadinae from the rest of skippers in the Palaeocene ca. 70 million years ago. The origin of crown Coeliadinae skippers is estimated in Indomalaya during the late Eocene ca. 36 million years ago, with subsequent Oligocene colonisation events toward the Australian region and the Afrotropics. Colonisation of the Afrotropics from the Indian region occurred during climatic transition, associated biome shifts, and the closure of the Tethys Ocean, which likely allowed geodispersal through the Arabian Peninsula. The current disjunct distribution of Coeliadinae in the Old World Tropics may result from the emergence of savannahs in the Miocene that progressively replaced woodlands and forests in the Arabian Peninsula and western Asia. Coeliadinae skippers are almost exclusively dicot feeders and were likely extirpated as grasslands became dominant, resulting in the present‐day disjunct distribution of these butterflies.