Three-dimensional electron microscopy of myriapod hemocyanin, planorbid snail acetylcholine-binding protein and cyanobacterial Vipp1

Three-dimensional electron microscopy (3-D EM) provides a framework for the analysis of large protein quaternary structures. The advantage over the generally higher resolving meth- od of X-ray crystallography is the embedding of the proteins in their physiological environ- ment. However, results of the two methods can be combined to obtain superior structural information. In this work, three different protein types – (i) Myriapod hemocyanin, (ii) vesi- cle-inducing protein in plastids 1 (Vipp1) and (iii) acetylcholine-binding protein (AChBP) – were structurally analyzed by 2-D and 3-D EM and, where possible, functionally interpreted.rnMyriapod hemocyanins have been previously shown to be 6x6-meric assemblies that, in case of Scutigera coleoptrata hemocyanin (ScoHc), show two 3x6-mer planes whith a stag- gering angle of approximately 60°. Here, previously observed structural differences between oxy- and deoxy-ScoHc could be substantiated. A 4° rotation between hexamers of two dif- ferent 3x6-mer planes was measured, which originates at the most central inter-hexamer in- terface. Further information about allosteric behaviour in myriapod hemocyanin was gained by analyzing Polydesmus angustus hemocyanin (PanHc), which shows a stable 3x6-mer and divergent histidine patterns in the inter-hexamer interfaces when compared to ScoHc. Both findings would conclusively explain the very different oxygen binding properties of chilopod and diplopod hemocyanin.rnVipp1 is a protein found in cyanobacteria and higher plants which is essential for thyla- koid membrane function and forms highly variable ring-shaped structures. In the course of this study, the first 3-D analysis of Vipp1 was conducted and yielded reconstructions of six differently sized Vipp1 rings from negatively stained images at resolutions between 20 to 30 Å. Furthermore, mutational analyses identified specific N-terminal amino acids that are essential for ring formation. On the basis of these analyses and previously published results, a hypothetical model of the Vipp1 tertiary and quaternary structure was generated.rnAChBP is a water-soluble protein in the hemolymph of mollusks. It is a structural and functional homologue of the ligand-binding domain of nicotinic acetylcholine receptors. For the freshwater snail Biomphalaria glabrata, we previously described two types of AChBP (BgAChBP1 and BgAChBP2). In this work, a 6 Å 3-D reconstruction of native BgAChBP is presented, which shows a dodecahedral assembly that is unprecedented for an AChBP. Single particle analysis of recombinantely expressed BgAChBP types led to preliminary results show- ing a dodecahedral assembly of BgAChBP1 and a dipentameric assembly of BgAChBP2. This indicates divergent biological functions of the two types.

Clinical, laboratory and pathological findings in dogs experimentally infected with Angiostrongylus vasorum

The aim of this comparative study was to investigate the development of clinical signs and accompanying haematological, coproscopic and pathological findings as a basis for the monitoring of health condition of Angiostrongylus vasorum infected dogs. Six beagles were orally inoculated with 50 (n=3) or 500 (n=3) A. vasorum third stage larvae (L3) obtained from experimentally infected Biomphalaria glabrata snails. Two dogs were treated with moxidectin/imidacloprid spot-on solution and two further dogs with an oral experimental compound 92 days post infection (dpi), and were necropsied 166 dpi. Two untreated control dogs were necropsied 97 dpi. Prepatency was 47-49 days. Dogs inoculated with 500 L3 exhibited earlier (from 42 dpi) and more severe respiratory signs. Clinical signs resolved 12 days after treatment and larval excretion stopped within 20 days in all four treated dogs. Upon necropsy, 10 and 170 adult worms were recovered from the untreated dogs inoculated with 50 and 500 L3, respectively. Adult worms were also found in two treated dogs, in the absence of L1 or eggs. Despite heavy A. vasorum infection load and severe pulmonary changes including vascular thrombosis, only mild haematological changes were observed. Eosinophilia was absent but the presence of plasma cells was observed. Neutrophilic leucocytes showed a transient increase but only after treatment. Signs for coagulopathies were slight; nevertheless coagulation parameters were inoculation dose dependent. Ten weeks after treatment pulmonary fibrosis was still present. Infections starting from 50 L3 of A. vasorum had a massive impact on lung tissues and therefore on the health of affected dogs, particularly after prepatency, although only mild haematological abnormalities were evident.

Planktonic foraminifera in the latest Maastrichtian of Walvis Ridge, South Atlantic

An abrupt global warming of 3-4°C occurred near the end of the Maastrichtian at 65.45-65.10 Ma. The environmental effects of this warm event are here documented based on stable isotopes and quantitative analysis of planktonic foraminifera at the South Atlantic DSDP Site 525A. Stable isotopes of individual species mark a rapid increase in temperature and a reduction in the vertical water mass stratification that is accompanied by a decrease in niche habitats, reduced species diversity and/or abundance, smaller species morphologies or dwarfing, and reduced photosymbiotic activity. During the warm event, the relative abundance of a large number of species decreased, including tropical-subtropical affiliated species, whereas typical mid-latitude species retained high abundances. This indicates that climate warming did not create favorable conditions for all tropical-subtropical species at mid-latitudes and did not cause a massive retreat in the local mid-latitude population. A noticeable exception is the ecological generalist Heterohelix dentata Stenestad that dominated during the cool intervals, but significantly decreased during the warm event. However, dwarfing is the most striking response to the abrupt warming and occurred in various species of different morphologies and lineages (e.g. biserial, trochospiral, keeled globotruncanids). Dwarfing is a typical reaction to environmental stress conditions and was likely the result of increased reproduction rates. Similarly, photosymbiotic activity appears to have been reduced significantly during the maximum warming, as indicated by decreased delta13C values. The foraminiferal response to climate change is thus multifaceted resulting in decreased species diversity, decreased species populations, increased competition due to reduced niche habitats, dwarfing and reduced photosymbiotic activity.

Chemobiostratigraphy of the Cretaceous/Paleocene boundary at ODP Hole 120-750A

An integrated biostratigraphic and stable isotope investigation was conducted on a high-latitude sequence across the Cretaceous/Paleogene (K/P) boundary recovered in Hole 750A in the southern Indian Ocean. The sequence consists of nannofossil chalk and is discontinuous across the boundary; missing is an estimated 0.3-m.y. late Maestrichtian and early Danian interval. Nonetheless, because calcareous nannofossil Zones NP1 and NP2 are well-developed, micropaleontological studies of the sequence have yielded a detailed record of Danian high-latitude microplankton evolution. In addition, stable carbon isotope analyses of planktonic and benthic foraminifer and bulk samples provide a record of late Maestrichtian and early Danian surface- and deep-water carbon isotope variations. Together, the carbon isotope and carbonate accumulation records serve as an index of regional marine net productivity across the boundary. Earliest Danian nannoplankton assemblages consisted mainly of persistent genera that were generally rare or absent in the Upper Cretaceous at Hole 750A. However, by 0.5-0.6 m.y. after the boundary, newly evolving Danian taxa became dominant. The turnover in nannofossil assemblages was accompanied by significant changes in rates of net productivity as gauged by carbon isotope distributions and carbonate accumulation rates. During the period dominated by persistent taxa, net productivity was extremely low, as reflected by the absence of vertical delta13C gradients and reduced carbonate accumulation rates. Later in the Danian, as new species evolved and flourished, vertical delta13C gradients reappeared and carbonate accumulation rates increased, signaling partial recovery of net productivity in this region. The absolute timing and magnitude of late Maestrichtian and early Danian biotic and geochemical changes in the southern Indian Ocean were similar to those recorded in other pelagic K/P boundary sequences from low- and mid-latitude Atlantic and Pacific sites, indicating that these events were ubiquitous.

Planktonic foraminifers in DSDP Holes 30-289 and 33-315A

The work is based on samples from Deep-Sea Drilling in the Pacific Ocean and from natural sections in its continental setting. Species composition of planktonic foraminifera from Maastrichtian sediments of the Pacific and South Atlantic oceans, as well as from marginal seas of Australia and New Zealand and epicontinental basins of the northern hemisphere has been analysed. Two main issues: reconstruction of Maastrichtian climatic zonality, and reconstruction of Maastrichtian paleodepths. Four bipolar climatic zones have been distinguished. According to preservation of planktonic foraminifera and composition of their complexes three levels of dissolution have been identified.

Planktonic foraminifera in the Maastrichtian of Hor-Hahar

The lengthy warm, stable climate of the Cretaceous terminated in the Campanian with a cooling trend, interrupted in the early and latest Maastrichtian by two events of global warming, at ~70-68 Ma and at 65.78-65.57 Ma. These climatic oscillations had a profound effect on pelagic ecosystems, especially on planktic foraminiferal populations. Here we compare biotic responses in the tropical-subtropical (Tethyan) open ocean and mesotrophic (Zin Valley, Israel) and oligotrophic (Tunisia) slopes, which correlate directly with global warming and cooling. The two warming events coincide with blooms of Guembelitria, an extreme opportunist genus best known as the main survivor of the Cretaceous-Paleogene (K-Pg) catastrophe. In the Maastrichtian, Guembelitria bloomed in the uppermost surface water above shelf and slope environments but failed to reach the open ocean as it did at K-Pg. The coldest interval of the late Maastrichtian (~68-65.78 Ma) is marked by an acme of the otherwise rare species Gansserina gansseri, a deep-dwelling keeled globotruncanid. The G. gansseri acme event can be traced from the deep ocean even onto the Tethyan slope, marking copious production and circulation of cold intermediate water. This acme is abruptly terminated by extinction of the species, a dramatic reversal attributed to a short-term global warming episode. This extinction corresponds precisely with the second bloom of Guembelitria that began ~300 kyr prior to the K-Pg event. The antithetical relationship between blooming of Guembelitria and the G. gansseri acme reflects planktic foraminiferal sensitivity to warm-cool-warm-cool climatic oscillations marking the end of the Cretaceous.

Biostratigraphy of benthic and planktonic foraminifera in Paleocene samples of DSDP HOle 93-605

Paleocene benthic and planktonic foraminifers occur throughout a long interval of the sedimentary succession cored at Site 605. A biostratigraphic zonation based on planktonic foraminifers is proposed for this Paleocene section. Zones identified are Subbotina pseudobulloides Zone, Morozovella trinidadensis Zone, M. uncinata Zone, M. pusilla pusilla Zone, Planorotalites pseudomenardii Zone, and M. velascoensis Zone. Fluctuations in the sedimentation rate occurred at Site 605. Rates of deposition were high during the M. pusilla pusilla and P. pseudomenardii zones, and a depositional hiatus may occur at the base of the M. velascoensis Zone. Qualitative and quantitative analysis of benthic foraminiferal assemblages suggests that the Paleocene sediments of Site 605 were deposited near the upper limit of Nuttallides truempyi, that is, approximately in the middle bathyal zone (600 m or more).

Campanian-Maestrichtian planktonic foraminifera of the southern South Atlantic

Campanian-Maestrichtian planktonic foraminifers were examined from Sites 698 (2128 m water depth) and 700 (3611 m water depth) on the Northeast Georgia Rise (southern South Atlantic, 51°S). Site 698 penetrated 72.5 m of Campanian-Maestrichtian chalk and limestone with only 18.2% recovery, whereas Site 700 recovered 66.8% of a 152.7-m section of Coniacian-Maestrichtian limestone. Preservation of planktonic foraminifers from both sites is moderate in Maestrichtian samples, but worsens with increasing depth in the Campanian. The Northeast Georgia Rise planktonic foraminifers are typical of Late Cretaceous Austral Province faunas described from other southern high-latitude sites; species diversity is low and the assemblages are dominated by species of Heterohelix, Globigerinelloides, Hedbergella, and Archaeoglobigerina. Five species, including Globigerinelloides impensus Sliter, Archaeoglobigerina australis Huber, Archaeoglobigerina mateola Huber, Hedbergella sliteri Huber, and Rugotruncana circumnodifer (Finlay), are considered to be endemic to the Austral Province. Formation of a cool temperate water mass in the circum-Antarctic region, resulting from the final breakup of the Gondwana continents, may have led to increased provincialism of the Austral Province planktonic foraminifers during Campanian-Maestrichtian time. Magnetobiostratigraphic correlation of eight planktonic foraminifer datum events at Hole 700B with ages determined for datums at ODP Leg 113 Holes 689B and 690C (Maud Rise, 65°S) demonstrates regional synchroneity of first and last occurrences within the Austral Province. As was observed at the Maud Rise, several keeled and nonkeeled species previously thought to have been restricted to warmer low-latitude regions first occur later at the Northeast Georgia Rise than at the low-latitude sites. The causes for high-latitude diachroneity among these immigrant species are not clear; neither oxygen and carbon isotope data from the Maud Rise sites nor calcareous nannoplankton distributions for the southern South Atlantic region show conspicuous changes that correlate to the delayed first occurrences.

Maastrichtian planktic foraminifera of the South Atlantic Ocean

Stratigraphic, faunal and isotopic analyses of the Maastrichtian at DSDP sites 525A and 21 in the South Atlantic reveal a planktic foraminiferal fauna characterized by two major events, an early late Maastrichtian diversification and end-Maastrichtian mass extinction. Both events are accompanied by major changes in climate and productivity. The diversification event which occurred in two steps between 70.5 and 69.1 Ma increased species richness by a total of 43% and coincided with the onset of major cooling in surface and bottom waters and increased surface productivity. The onset of the terminal decline in Maastrichtian species richness began at 67.5 Ma and the first significant decline in surface productivity occurred at 66.2 Ma, coincident maximum cooling to 13°C in surface waters and the reduction of the surface-to-deep temperature gradient to less than 5°C. Major climatic and moderate productivity changes mark the mass extinction and the last 500 kyr of the Maastrichtian. Between 200 and 400 kyr before the K-T boundary surface and deep waters warmed rapidly by 3-4°C and cooled again during the last 100 kyr of the Maastrichtian. Surface productivity decreased only moderately across the K-T boundary. Species richness began to decline during the late Maastrichtian cooling and by K-T boundary time, the mass extinction had claimed 66% of the species. Viewed within the context of Maastrichtian climate and productivity changes, the K-T mass extinction could have resulted from extreme environmental stress even without the addition of an extraterrestrial impact.