Functional and evolutionary consequences of cranial fenestration in birds

Ostrich‐like birds (Palaeognathae) show very little taxonomic diversity while their sister taxon (Neognathae) contains roughly 10,000 species. The main anatomical differences between the two taxa are in the crania. Palaeognaths lack an element in the bill called the lateral bar that is present in both ancestral theropods and modern neognaths, and have thin zones in the bones of the bill, and robust bony elements on the ventral surface of their crania. Here we use a combination of modeling and developmental experiments to investigate the processes that might have led to these differences. Engineering‐based finite element analyses indicate that removing the lateral bars from a neognath increases mechanical stress in the upper bill and the ventral elements of the skull, regions that are either more robust or more flexible in palaeognaths. Surgically removing the lateral bar from neognath hatchlings led to similar changes. These results indicate that the lateral bar is load‐bearing and suggest that this function was transferred to other bony elements when it was lost in palaeognaths. It is possible that the loss of the load‐bearing lateral bar might have constrained diversification of skull morphology in palaeognaths and thus limited taxonomic diversity within the group.     

Isolation of bovine follicular dendritic cells allows the demonstration of a particular cellular prion protein

As interaction of cellular prion protein (PrPc) and the infectious agent (PrPres) appears to be a crucial pathogenic step promoted by homology, variation in PrPc isoforms on bovine immune cells may explain the absence of infectivity in most bovine lymph organs. In this study, we examined PrPc expression in bovine lymph organs (tonsils and lymph nodes) and on isolated follicular dendritic cells (FDCs). We used a panel of different monoclonal antibodies (MoAbs) raised against different epitopes of prion protein. Two MoAbs recognise amino acids 79-92 (SAF 34 and SAF 32 Mo-Abs); the 6H4 antibody reacts with a specific peptide comprising the 144-152 amino acids, and the 12F10 MoAb recognises the sequence 142-160. After immunolabelling of frozen sections of lymph organs with 6H4 or 12F10 MoAbs, we detected cellular prion protein in germinal centres. However, using the SAF 34 or SAF 32 antibodies, PrPc was revealed outside the lymphoid tissues. No PrPc was observed in the germinal centres. Therefore, we adapted the method of FDC isolation, making it suitable for the study of PrPc expression on their surface. Using electron microscopy, the presence of PrPc on the surface of FDCs was demonstrated only with 6H4 MoAb. These results suggest that bovine follicular dendritic cells express a particular form of prion protein. Either the N-terminal part of PrPc is cleaved or the accessibility of the specific epitope (79-92) of SAF 34 MoAb is abolished by interaction with other molecules. This particular isoform of PrPc on bovine FDCs might be related to the apparent absence of infectivity in lymph organs in cattle affected by bovine spongiform encephalopathy.     

Spatial and temporal diversity in hyperparasitoid communities of Cotesia glomerata on garlic mustard,Alliaria petiolata

1. Interactions between two trophic levels can be very intimate, often making species dependent on each other, something that increases with specialisation. Some specialised multivoltine herbivores may depend on multiple plant species for their survival over the course of a growing season, especially if their food plants are short‐lived and grow at different times. Later generations may exploit different plant species from those exploited by previous generations. 2. Multivoltine parasitoids as well as their natural enemies must also find their hosts on different food plants in different habitats across the season. Secondary hyperparasitoid communities have been studied on cocoons of the primary parasitoid, Cotesia glomerata (Hymenoptera: Braconidae), on black mustard (Brassica nigra) – a major food plant of its host, the large cabbage white (Pieris brassicae) – which grows in mid‐summer. 3. Here, hyperparasitoid communities on C. glomerata pupal clusters were studied on an early‐season host, garlic mustard, Alliaria petiolata, over ‘time’ (one season, April–July) in six closely located ‘populations’ (c. 2 km apart), and within two different ‘areas’ at greater separation (c. 100 km apart). At the plant level, spatial effects of pupal ‘location’ (canopy or bottom) on the plant were tested. 4. Although large‐scale separation (area) did not influence hyperparasitism, sampling time and small‐scale separation (population) affected hyperparasitism levels and composition of hyperparasitoid communities. Location on the plant strongly increased proportions of winged species in the canopy and proportions of wingless species in bottom‐located pupae. 5. These results show that hyperparasitism varies considerably at the local level, but that differences in hyperparasitoid communities do not increase with spatial distance.     

Characteristics and properties of a caldo-active bacterium producing extracellular enzymes and two related strains

Summary The caldo-active strain YT-P was found to produce a variety of extracellular enzymes, including an amylase and a protease, which were further examined. With azo-casein as a substrate, optimum conditions with respect to enzyme and substrate concentration were determined for the protease. The optimum temperature was found to be 70°C, with a sharp decline to both lower and higher temperatures. The enzyme was found to be extremely heat-stabile, with unaltered activity after 8 hours at 80°C.Optimum conditions for the amylase were also examined. This enzyme was shown to be less heat-stabile, though the temperature optimum was again at 70°C. The activity or stability was not influenced by absence or presence of Ca-ions. The main activity of the amylase was found in the 20–40% ammonium sulfate fraction, which also contained the bulk of the proteolytic enzyme.This strain growth optimally on a variety of carbon sources at 72°C. Typical submicroscopical features are the double-layered cell wall, and a cytoplasmic membrane with a varying number of small dots and dot-free patches.Furthermore the nutritional requirements and submicroscopical features of two other strains, YT-G and YT-F, are described and compared to strain YT-P.Based on the fatty acid composition of the three spore forming caldo-active strains we suggest that they belong to the genus Bacillus, and propose the names B. caldolyticus for strain YT-P, B. caldovelox for strain YT-F, and B. caldotenax for strain YT-G.     

Attachment of the soluble complement regulator factor H to cell and tissue surfaces: relevance for pathology

Complement is a central element of innate immunity and this vital defense system initiates and coordinates immediate immune reactions which attack and eliminate microbes, foreign particles and altered self cells. Newly generated activation products are extremely toxic and consequently, activation is highly restricted in terms of time and space. The initial activation of the alternative complement pathway occurs continuously and the early phase acts indiscriminatoryl and forms on any surface. However, the system discriminates between self and foreign, and therefore allows activation on foreign surfaces e.g. microbes, and restricts activation on host cells. Consequently, self cells and tissues are protected from the harmful activation products. This protection is mediated by specific regulators or inhibitors, which exist in the fluid phase and/or in membrane-bound forms. Here we review a novel mechanism, i.e. the attachment of the soluble complement regulator factor H to the surface of self cells. This attachment, which is demonstrated experimentally by means of immunofluorescense microscopy and by flow cytometry, increases the inhibitory potential at the cell surface and mediates protection by reducing the local formation of toxic inflammatory products. This attachment is highly relevant and has pathophysiological consequences in several human diseases, including Factor H-associated hemolytic uremic syndrome (FH-HUS), membrano-proliferative glomerulonephritis type II, recurrent microbial infections and chronic inflammation, e.g. rheumatoid arthritis and immune evasion of tumor cells. Defects of this safeguard activity have been recently understood in patients with FH-HUS. Point mutations in the Factor H gene occurring in the C-terminus of the protein result in impaired cell binding capacity of Factor H and, consequently, during an inflammatory insult endothelial cells are not properly protected and are damaged.     

Comparative studies on the nature and distribution of pigments from two thermophilic bacteria

Summary The pigments of two strains of extremely thermo-resistant bacteria were extracted and examined spectrophotometrically. Both strains contain -carotene as the main pigment according to the spectral characteristics of the pigment in various solvents. According to quantitative data Thermus aquaticus contains more than one hundred times as much of the pigment than is found in strain YT-G, representing almost 80% of the total lipid. The carotene of strain YT-G is destroyed by hot saponification, while that of T. aquaticus resists that procedure without alterations. T. aquaticus contains a small amount of a second pigment with absorption maxima at 370, 400 and 420 m.Solubilization of the membrane of spheroplasts derived from acetate-C¹⁴-labelled cells of T. aquaticus, followed by separation of the crude membrane fraction, showed that about 50% of the C¹⁴ is associated with the membrane fraction. More than 90% of the C¹⁴ is recovered in the yellow supernatant of the acetone-extracted crude membrane fraction, showing that a rapid incorporation of the label into the pigment occurs. The assumption that the pigment is associated with the membrane is supported by submicroscopical observations: T. aquaticus has a well-defined cytoplasmic membrane, and on top of this a sandwiched layer, which is embedded in a very electron-dense material, that we assume to consist of the pigment. Strain YT-G which contains only minute quantities of the same pigment, does not possess this kind of an outer membrane, but shows only a cytoplasmic membrane envelopped by the cell wall.The main experiments were done while on leave to the Ames Research Center, as a recipient of a Senior Post-doctoral Resident Research Associateship of the National Research Council, Washington, D.C., from the Dept. of Exobiology, University of Nijmegen, Driehuizerweg 200, Nijmegen, The Netherlands.     

Silicium-Stoffwechsel bei Mikroorganismen

Zusammenfassung Die Verteilung von Kiselsäurekomponenten auf verschiedene Zellfraktionen von Proteus mirabilis wurde nach Inkubation der Bakterien in verschiedenen Silicat-haltigen Medien bestimmt. Die Si-Konzentration sowohl der Zellwände und des Extraktes als auch der aus dem Extrakt erhaltenen Fraktionen (Protein-Fraktion und Alkohol-Äther-lösliche Fraktion) ist abhängig von den Konzentrationsbedingungen: Bei Inkubation der Bakterien in Si-Lösungen mit Glucose oder Glucose, Nitrat und Phosphat ergeben sich für die Zellwände viel höhere Si-Konzentrationen als beim Fehlen der Glucose im Inkubationsmedium. Werden abgetötete Zellen verwendet, so findet sich keine Kieselsäure in den Zell-wänden, wohl aber in beiden Fraktionen des zell-freien ExtraktesDie Protein-Fraktion enthält wasserlösliche Kieselsäurekomponenten, die durch Hydrolyse in molybdat-aktive Kieselsäure und Hexosen zerfallen. Der Hauptanteil der Hexosesilicat-Ester wird in den ersten 5 min der nach 20 min beendeten Reaktion hydrolysiert, wobei sowohl reduzierende Verbindungen als auch Furfurol-bildende Kohlenhydrate auftreten. Aufgrund der Zunahme der freien Kieselsäure während der Hydrolyse liegen KH-Kieselsäure-Ester auch in der EES-Fraktion vor. Signifikante Mengen dieser Kieselsäure-Ester werden nur dann gebildet, wenn die Bakterien in Si-Lösungen inkubiert werden, die zugleich auch Glucose enthalten.

---

Summary The distribution of silicon compounds in several cell fractions of Proteus mirabilis has been followed after incubation of the bacteria in different silicate containing solutions. The concentration in the cell walls, cell extracts as well as the ethanol-ether soluble (EES) and the protein fraction, both derived from the cell-free extract, depends on the kind of media used for incubation:After incubation of the bacteria in silicate solutions containing glucose alone, or glucose, nitrogen and phosphate compounds, the incorporation of Si into the cell walls is much higher than in the absence of glucose. Inactivated cells fail to incorporate the element into the cell walls but accumulate it in both fractions of the cell-free extract. The alcohol-ether insoluble fraction contains water soluble silicon compounds, which upon hydrolysis release molybdate-active silicic acid and hexoses. The main portion of these hexose-silicate esters are split during the first five minutes, while it takes about 20 min to complete the hydrolysis. Both furfurolforming sugars and reducing substances have been shown to appear during this reaction. Silicon linked to carbohydrates has also been shown to be present in the EES fraction, according to the increase of free silicic acid during hydrolysis. These esters are formed in significant amounts only after incubating the bacteria in silicate solutions containing glucose, but not in glucose-free media.

     

Lotus corniculatus nodulation specificity is changed by the presence of a soybean lectin gene

Plant lectins have been implicated as playing an important role in mediating recognition and specificity in the Rhizobium-legume nitrogen-fixing symbiosis. To test this hypothesis, we introduced the soybean lectin gene Le1 either behind its own promoter or behind the cauliflower mosaic virus 35S promoter into Lotus corniculatus, which is nodulated by R. loti. We found that nodulelike outgrowths developed on transgenic L. corniculatus plant roots in response to Bradyrhizobium japonicum, which nodulates soybean and not Lotus spp. Soybean lectin was properly targeted to L. corniculatus root hairs, and although infection threads formed, they aborted in epidermal or hypodermal cells. Mutation of the lectin sugar binding site abolished infection thread formation and nodulation. Incubation of bradyrhizobia in the nodulation (nod) gene-inducing flavonoid genistein increased the number of nodulelike outgrowths on transgenic L. corniculatus roots. Studies of bacterial mutants, however, suggest that a component of the exopolysaccharide surface of B. japonicum, rather than Nod factor, is required for extension of host range to the transgenic L. corniculatus plants.