Plant cyclotides: A unique family of cyclic and knotted proteins that defines the cyclic cystine knot structural motif

Several macrocyclic peptides (similar to 30 amino acids), with diverse biological activities, have been isolated from the Rubiaceae and Violaceae plant families over recent years. We have significantly expanded the range of known macrocyclic peptides with the discovery of 16 novel peptides from extracts of Viola hederaceae, Viola odorata and Oldenlandia affinis. The Viola plants had not previously been examined for these peptides and thus represent novel species in which these unusual macrocyclic peptides are produced. Further, we have determined the three-dimensional struc ture of one of these novel peptides, cycloviolacin O1, using H-1 NMR spectroscopy. The structure consists of a distorted triple-stranded beta-sheet and a cystine-knot arrangement of the disulfide bonds. This structure is similar to kalata B1 and circulin A, the only two macrocyclic peptides for which a structure was available, suggesting that despite the sequence variation throughout the peptides they form a family in which the overall fold is conserved. We refer to these peptides as the cyclotide family and their embedded topology as the cyclic cystine knot (CCK) motif. The unique cyclic and knotted nature of these molecules makes them a fascinating example of topologically complex proteins. Examination of the sequences reveals they can be separated into two subfamilies, one of which tends to contain a larger number of positively charged residues and has a bracelet-like circularization of the backbone. The second subfamily contains a backbone twist due to a cis-Pro peptide bond and may conceptually be regarded as a molecular Moebius strip. Here we define the structural features of the two apparent subfamilies of the CCK peptides which may be significant for the likely defense related role of these peptides within plants. (C) 1999 Academic Press.

Expression of EphA5 during development of the olfactory nerve pathway in rat

The olfactory neuroepithelium is a highly plastic region of the nervous system that undergoes continual turnover of primary olfactory neurons throughout life. The mechanisms responsible for persistent growth and guidance of primary olfactory axons along the olfactory nerve are unknown. In the present study, we used antibodies against the Eph-related receptor, EphA5, to localise EphA5, and recombinant EDhA5-IgG fusion protein to localise its ligands. We found that although both EphA5 and its ligands were both expressed by primary olfactory neurons within the embryonic olfactory nerve pathway, there was no graded or complementary expression pattern. In contrast, the expression patterns altered postnatally such that primary olfactory neurons expressed the ligands, whereas the second-order olfactory neurons, the mitral cells, expressed EphA5. The role of EphA5 was analysed by blocking EphA5-ligand interactions in explant cultures of olfactory neuroepithelium using anti-EphA5 antibodies and recombinant EphA5. These perturbations reduced neurite outgrowth from explant cultures and suggest that intrafascicular axon repulsion may serve to limit adhesion and optimise conditions for axon growth. (C) 2000 Wiley-Liss, Inc.

A review of the Arhythmacanthidae (Acanthocephala) with a description of Heterosentis hirsutus n. sp. from Cnidoglanis macrocephala (Plotosidae) in Australia

Heterosentis hirsutus n. sp. is described from Cnidoglanis macrocephalo (Siluriformes: Plotosidae) from the Swan Estuary, Western Australia. It is distinguished by having 14 longitudinal rows of 6-7 hooks per row on the proboscis, a trunk armed anteriorly and posteriorly (=genital spines) with minute spines and lemnisci that may extend to the poster;or margin of the proboscis receptacle The new species also has prominent fragmented nuclei in its trunk well. New information is given for Heterosentis plotosi Yamoguti, 1935 from Plotosus lineatus (Siluriformes: Plotosidae) and H. poraplagusiarum (Nickol, 1972) Amin, 1985 from Paraplogusia guttata (Pleuronectiformes: Cynoglossidoe), both from Queensland. A key to the species of Heterosentis Van Cleave, 1931 is provided. The Arhythmacanthidae subfamilies are reviewed: there is little utility in the recognition of these taxa because of the small number of genera involved and the validity/ of the characters on which they ore based is in doubt, particularly whether trunk spines are present or absent. Only Acanthocephaloides Meyer, 1932, Breizocanthus Golvon, 1969, Euzetocanthus Golvan & Houin, 1964, Heterosentis, Hypoechinorhynchus Yamaguti, 1939 and Paracanthocepholoides Golvan, 1969 of the Arhythmacanthidae are considered valid. A key to these genera is provided. The monotypic genus Neocanthocepholoides Cable & Quick, 1954 is considered a new synonym of Acanthocephaloides thus creating Acanthocephaloides spinicaudatus (Cable & Quick, 1954) n. comb. Arhythmocanthus Yamaguti, 1935 is maintained as a synonym of Heterosentis because the distinction between two and three hook types is made equivocal when the transition between the opical and subapical hooks is gradual.

Effects of habitat modification in mangroves on the structure of mollusc and crab assemblages

The abundance and species richness of mollusc and crab assemblages were examined in a subtropical mangrove forest in Moreton Bay, Queensland, Australia, which has been disturbed and damaged by the construction of a wooden boardwalk and a path. Sections of the forest immediately adjacent to the boardwalk and path were compared with reference areas to determine whether changes to the small-scale structural complexity within the forest affected the benthic fauna. The disturbed area was characterised by having 65-80% fewer pneumatophores, significantly fewer species and individuals of molluscs, but significantly more species and individuals of crabs than the reference areas. The abundance of mangrove pneumatophores and the attached epiphytic algae were manipulated at two sites to determine whether observed differences in these features could account for the differences in the assemblage of molluscs in the disturbed area of the forest compared with reference areas. Five experimental treatments were used: undisturbed controls, pneumatophore removals (abundance reduced by ca. 65%), epiphytic algal removals (algae removed from ca. 65% of pneumatophores), pneumatophore disturbance controls and algal disturbance controls. The experimental reduction of the abundance of mangrove pneumatophores and the associated epiphytic algae led to significant declines (by as much as 83%) in the number of molluscs utilising the substratum in the modified plots. There was no significant difference in the abundance of molluscs in the pneumatophore and algal removal plots suggesting any effect was primarily related to removal of the epiphytic algae from the surface of the pneumatophores. The responses by the biota to the changes in the physical environment demonstrate that even relatively small-scale modifications to the physical structure of subtropical mangrove forests can lead to significant effects on the diversity and abundance of macrobenthic organisms in these habitats. Such modifications have the potential to cause cascading effects at higher trophic levels with a deterioration in the value of these habitats as nursery and feeding grounds. Future efforts at conservation of these estuarine environments must focus on the prevention or reduction of modifications to the physical structure and integrity of the system, rather than just on the prevention of loss of entire patches of habitat. (C) 2000 Elsevier Science B.V. All rights reserved.

Structure of the tegument and ectocommensal microorganisms of Gyliauchen nahaensis (Digenea : Gyliauchenidae), an inhabitant of herbivorous fish of the Great Barrier Reef, Australia

The ultrastructure of the tegument and tegument-associated microorganisms of the gyliauchenid digenean Gyliauchen nahaensis is described by transmission and scanning electron microscopy. The tegument is devoid of surface spines and is characterized by a moderately folded apical membrane, abundant vesicles, basal mitochondria, a folded basal plasma membrane, and a thick basal matrix. Microorganisms form a dense biofilm on the tegument of the posterodorsal surface and the excretory papilla. At least 7 microbial morphotypes were identified, including eubacteria, spirochaetes, and nanobacteria.

Modeling the optical properties of AlSb, GaSb, and InSb

Optical constants of AlSb, GaSb, and InSb are modeled in the 1-6 eV spectral range. We employ an extension of Adachi's model of the optical constants of semiconductors. The model takes into account transitions at E-0, E-0 + Delta(0), E-1, and E-1 + Delta(1) critical points, as well as higher-lying transitions which are modeled with three damped harmonic oscillators. We do not consider indirect transitions contribution, since it represents a second-order perturbation and its strength should be low. Also, we do not take into account excitonic effects at E-1, E-1 + Delta(1) critical points, since we model the room temperature data. In spite of fewer contributions to the dielectric function compared to previous calculations involving Adachi's model, our calculations show significantly improved agreement with the experimental data. This is due to the two main distinguishing features of calculations presented here: use of adjustable line broadening instead of the conventional Lorentzian one, and employment of a global optimization routine for model parameter determination.

Approach to designing rotating drum bioreactors for solid-state fermentation on the basis of dimensionless design factors

The development of large-scale solid-stale fermentation (SSF) processes is hampered by the lack of simple tools for the design of SSF bioreactors. The use of semifundamental mathematical models to design and operate SSF bioreactors can be complex. In this work, dimensionless design factors are used to predict the effects of scale and of operational variables on the performance of rotating drum bioreactors. The dimensionless design factor (DDF) is a ratio of the rate of heat generation to the rate of heat removal at the time of peak heat production. It can be used to predict maximum temperatures reached within the substrate bed for given operational variables. Alternatively, given the maximum temperature that can be tolerated during the fermentation, it can be used to explore the combinations of operating variables that prevent that temperature from being exceeded. Comparison of the predictions of the DDF approach with literature data for operation of rotating drums suggests that the DDF is a useful tool. The DDF approach was used to explore the consequences of three scale-up strategies on the required air flow rates and maximum temperatures achieved in the substrate bed as the bioreactor size was increased on the basis of geometric similarity. The first of these strategies was to maintain the superficial flow rate of the process air through the drum constant. The second was to maintain the ratio of volumes of air per volume of bioreactor constant. The third strategy was to adjust the air flow rate with increase in scale in such a manner as to maintain constant the maximum temperature attained in the substrate bed during the fermentation. (C) 2000 John Wiley & Sons, Inc.

Social identity and self-categorization processes in organizational contexts

Although aspects of social identity theory are familiar to organizational psychologists, its elaboration, through self-categorization theory, of how social categorization and prototype-based depersonalization actually produce social identity effects is less well known. We describe these processes, relate self-categorization theory to social identity theory, describe new theoretical developments in detail, and show how these developments can address a: range of organizational phenomena. We discuss cohesion and deviance, leadership, subgroup and sociodemographic structure, and mergers and acquisitions.

The politics and ethics of relocated art

At the end of Word War II, Soviet occupation forces removed countless art objects from German soil. Some of them were returned during the 1950s, but most either disappeared for good or were stored away secretly in cellars of Soviet museums. The Cold War then covered the issue with silence. After the collapse of the Soviet Union, museums in St Petersburg and Moscow started to exhibit some of the relocated art for the first time in half a century. The unusual quality of the paintings-mostly impressionist masterpieces-not only attracted the attention of the international art community, but also triggered a diplomatic row between Russia and Germany. Both governments advanced moral and legal claims to ownership. To make things even more complicated, many of the paintings once belonged to private collectors, some of whom were Jews. Their descendants also entered the dispute. The basic premise of this article is that the political and ethical dimensions of relocated art can be understood most adequately by eschewing a single authorial standpoint. Various positions, sometimes incommensurable ones, are thus explored in an attempt to outline possibilities for an ethics of representation and a dialogical solution to the international problem that relocated art has become.

Subgroup relations: A comparison of mutual intergroup differentiation and common ingroup identity models of prejudice reduction

Two studies examined relations between groups (humanities and math-science students) that implicitly or explicitly share a common superordinate category (university student). In Experiment 1, 178 participants performed a noninteractive decision-making task during which category salience was manipulated in a 2 (superordinate category salience) x 2 (subordinate category salience) between-groups design. Consistent with the mutual intergroup differentiation model, participants for whom both categories were salient exhibited the lowest levels of bias, whereas bias was strongest when the superordinate category alone was made salient. This pattern of results was replicated in Experiment 2 (N = 135). In addition, Experiment 2 demonstrated that members of subgroups that are nested within a superordinate category are more sensitive to how the superordinate category is represented than are members of subgroups that extend beyond the boundaries of the superordinate category.

Systemic coadministration of chloramphenicol with intravenous but not intracerebroventricular morphine markedly increases morphine antinociception and delays development of antinociceptive tolerance in rats

Chloramphenicol, an in vitro inhibitor of the glucuronidation of morphine to its putative antianalgesic metabolite, morphine-3-glucuronide (M3G), was coadministered with morphine in adult male Sprague-Dawley rats to determine whether it inhibited the in vivo metabolism of morphine to M3G, thereby enhancing morphine antinociception and/or delaying the development of antinociceptive tolerance. Parenteral chloramphenicol was given acutely (3-h studies) or chronically (48-h studies). Morphine was administered by the i.v. or i.c.v. route. Control rats received chloramphenicol and/or vehicle. Antinociception was quantified using the hotplate latency test. Coadministration of chloramphenicol with i.v. but not i.cv. morphine increased the extent and duration of morphine antinociception by approximate to 5.5-fold relative to rats that received i.v. morphine alone. Thus, the mechanism through which chloramphenicol enhances i.v. morphine antinociception in the rat does not directly involve supraspinal opioid receptors. Acutely, parenteral coadministration of chloramphenicol and morphine resulted in an approximate to 75% increase in the mean area under the serum morphine concentration-time curve but for chronic dosing there was no significant change in this curve, indicating that factors other than morphine concentrations contribute significantly to antinociception. Antinociceptive tolerance to morphine developed more slowly in rats coadministered chloramphenicol, consistent with our proposal that in vivo inhibition of M3G formation would result in increased antinociception and delayed development of tolerance. However, our data also indicate that chloramphenicol inhibited the biliary secretion of M3G. Whether chloramphenicol altered the passage of M3G and morphine across the blood-brain barrier remains to be investigated.

Hypoechinorhynchus robustus sp n. from Notolabrus parilus (Labridae) from Western Australia with a discussion on the validity of the hypoechinorhynchidae (Acanthocephala : Palaeacanthocephala)

Hypoechinorhynchus robustus sp. n. is described from Notolabrus parilus (Richardson) (Labridae) from Pt Peron, Western Australia. It has a proboscis with 30 hooks arranged in ten longitudinal rows: 5 rows of a small apical spine, a large anterior hook and a small posterior spine, 5 rows of a large anterior hook, a middle spine and a posterior spine. The new species is distinguished from other species of the genus by having a set of 5 small apical spines anterior to the large hooks on the proboscis, by having lemnisci that barely extend beyond the proboscis receptacle and testes which are more adjacent than tandem. H. robustus also has robust trunk spines anteriorly. Re-examination of Hypoechinorhynchus alaeopis Yamaguti, 1939 (type species) revealed trunk spines that had been overlooked previously. The Hypoechinorhynchidae is made a junior synonym of Arhythmacanthidae because there is considerable overlap between the two family diagnoses, particularly in that both families have a proboscis armature that changes abruptly from small basal spines to large apical (or subapical if present) hooks. The genus Hypoechinorhynchus is placed in the subfamily Arhythmacanthinae because it has trunk spines and a spherical proboscis with few hooks (relative to other arhythmacanthid genera). It is also proposed that Heterosentis magellanicus (Szidat, 1950) be returned to the genus Hypoechinorhynchus since it was transferred to Heterosentis primarily because it had trunk spines. The other hypoechinorhynchid genus contained only Bolborhynchoides exiguus (Achmerov et Dombrowskaja-Achmerova, 1941) Achmerov, 1959 and is relegated to incertae sedis.

Litterfall from two eucalypt woodlands in central Queensland

The mean annual litterfall at two dry woodland sites in central Queensland was 1129 kg ha(-1) for an open E. populnea F. Muell. woodland (n = 2 years), and 2318 kg ha(-1) for a woodland dominated by E. cambageana Maiden (n = 1 year). Leaves formed the largest component of total litterfall, which varied seasonally with a spring-summer maximum. Annual litterfall at these sites conformed with a pattern of decreasing litter production with declining annual rainfall, consistent with a range of eucalypt-dominated communities.

Conformations of cyclic octapeptides and the influence of heterocyclic ring constraints upon calcium binding

A comparison is made between the structures and calcium binding properties of four cyclic octapeptides that differ in the number of heterocyclic thiazole and oxazoline ring constraints. The conformations of the naturally occurring cyclic octapeptides ascidiacyclamide 1 and patellamide D 2, which each contain two oxazoline and two thiazole rings, are compared by H-1 NMR spectroscopy with the analogues cyclo(Thr-D-Val(Thz)-Ile)(2) 3 with just two thiazoles, and cyclo(Thr-D-Val-alpha Abu-Ile)(2) 4, with no 5-membered rings. The conformations observed in the solid state for ascidiacyclamide (saddle) and patellamide D (twisted figure of eight) were retained in solution, whilst peptide 3 was found to have a chair shape and peptide 4 displayed a range of conformations. The solid state structure of 4 revealed that the peptide takes a relatively planar conformation with a number of transannular hydrogen bonds, which are apparently retained in solution. Complexation studies utilising H-1 NMR and CD spectroscopy yielded 1∶1 calcium-peptide binding constants (log K) for the four peptides (2.9 (1), 2.8 (2), 4.0 (3) and 5.5 (4)) as well as a 1 : 2 metal-peptide binding constant for 3 (log K = 4.5). The affinity for Ca2+ thus decreases with increasing number of 5-membered ring constraints in the macrocycle (4 > 3 > 2 approximate to 1).

Implications of specimen preparation and of surface contamination for the measurement of the grain boundary carbon concentration of steels using x-ray microanalysis in an UHVFESTEM

The purpose of the present investigation was to gain an understanding of the nature of the carbon contamination on the surface of standard steel transmission electron spectroscopy (TEM) specimens, the effect of exposure of a clean specimen to normal laboratory air, and the efficacy of plasma-cleaning treatments. This knowledge is a necessary prerequisite to the development of appropriate specimen preparation and/or specimen cleaning methods. X-ray photoelectron spectroscopy in combination with argon ion beam profiling was used to characterize the specimen surfaces of X65 steel and 316 stainless steel. The only clean carbon-free surface obtained was that during argon etching of the sample in the surface analysis chamber. Any exposure of a previously cleaned sample to laboratory air resulted in a rapid carbon (hydrocarbon) contamination of the sample surface and the development of surface oxidation, Plasma cleaning with subsequent exposure of the specimen to the laboratory air also resulted in a carbon-contaminated surface. This suggests that procedures of preparation of TEM specimens of steels outside an ultrahigh vacuum chamber are unlikely to result in the lowering of contamination rates on specimens to levels where measurements for carbon in the grain boundaries are possible. What is needed is a cleaning system as an integral part of the specimen insertion system into the field-emission scanning transmission electron microscope. This cleaning could be carried out by argon ion etching. Copyright (C) 2000 John Wiley & Sons, Ltd.