Activin promotes oocyte development in ovine preantral follicles <Emphasis Type="Italic">in vitro</Emphasis>

Activins have been implicated as important regulating factors for many reproductive processes. The aim of this study was to determine the effect of activin A on the development of ovine preantral follicles in vitro. Mechanically isolated preantral follicles (161 ± 2 microm) were cultured for 6 days in the presence of human recombinant activin A (0, 10 and 100 ng/ml). Half of the medium was replaced every second day and follicle diameters were measured. Conditioned medium was subsequently analysed for oestradiol content using a delayed enhancement lanthanide fluorometric immunoassay (DELFIA). At the end of the culture period, follicles were fixed and processed for histology, after which oocyte diameter and granulosa cell death were measured. There was significant follicle growth over 6 days in all groups (p < 0.001). Activin, at both concentrations, increased follicle growth over control levels by Day 6 (p < 0.05). Oocyte diameters were also significantly increased by Day 6 of culture in all groups (p < 0.05), with 100 ng/ml activin increasing oocyte diameter over control levels (p < 0.05). Activin, at both concentrations, increased oestradiol production on Day 2 of culture, but this increase was not sustained during the culture period. Moreover, activin did not have any effect on antrum formation or follicle survival. In conclusion, activin promoted ovine preantral follicle and oocyte growth in vitro, but did not accelerate follicle differentiation over a six-day culture period. These results support a paracrine role for activin A during early oocyte and follicular development.     

Predicting spatially heterogeneous invasive spread: Pyracantha angustifolia invading a dry Andean valley in northern Argentina

Biological Invasions - Understanding the drivers of invasive species spread is key to designing optimal management programmes for controlling them. Population models, parameterized from demographic...     

Targeting the role of a key conserved motif for substrate selection and catalysis by 3-deoxy-D-manno-octulosonate 8-phosphate synthase

3-Deoxy-D-manno-octulosonate 8-phosphate synthase (KDO8PS) catalyzes the reaction between three-carbon phosphoenolpyruvate (PEP) and five-carbon d-arabinose 5-phosphate (A5P), generating KDO8P, a key intermediate in the biosynthetic pathway to 3-deoxy-D-manno-octulosonate, a component of the lipopolysaccharide of the Gram-negative bacterial cell wall. Both metal-dependent and metal-independent forms of KDO8PS have been characterized. KDO8PS is evolutionarily and mechanistically related to the first enzyme of the shikimate pathway, the obligately divalent metal ion-dependent 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAH7PS) that couples PEP and four-carbon D-erythrose 4-phosphate (E4P) to give DAH7P. In KDO8PS, an absolutely conserved KANRS motif forms part of the A5P binding site, whereas in DAH7PS, an absolutely conserved KPR(S/T) motif accommodates E4P. Here, we have characterized four mutants of this motif (AANRS, KAARS, KARS, and KPRS) in metal-dependent KDO8PS from Acidithiobacillus ferrooxidans and metal-independent KDO8PS from Neisseria meningitidis to test the roles of the universal Lys and the Ala-Asn portion of the KANRS motif. The X-ray structures, determined for the N. meningitidis KDO8PS mutants, indicated no gross structural penalty resulting from mutation, but the subtle changes observed in the active sites of these mutant proteins correlated with their altered catalytic function. (1) The AANRS mutations destroyed catalytic activity. (2) The KAARS mutations lowered substrate selectivity, as well as activity. (3) Replacing KANRS with KARS or KPRS destroyed KDO8PS activity but did not produce a functional DAH7PS. Thus, Lys is critical to catalysis, and other changes are necessary to switch substrate specificity for both the metal-independent and metal-dependent forms of these enzymes.     

LOCAL CYTONUCLEAR EXTINCTION OF THE GOLDEN-WINGED WARBLER

I compared the mtDNA compositions of two adjacent populations of Vermivora chrysoptera (golden-winged warbler) at different stages of transient hybridization with its sister species V. pinus (blue-winged warbler). Pinus mtDNA introgresses asymmetrically and perhaps rapidly into chrysoptera phenotypes without comparable reverse introgression of chrysoptera mtDNA into replacing pinus populations. Pinus mtDNA was virtually fixed (98%) in an actively hybridizing lowland population with varied phenotypes. Pinus mtDNA increased from 27% (n = 11) in 1988 to 70% (n = 10) in 1992 in successive samples of a highland population in the initial stages of hybridization. This population comprised mostly pure and slightly introgressed chrysoptera phenotypes. The rapid pace of asymmetrical introgression may be the result of initial invasion of chrysoptera populations by pioneering female pinus and/or an unknown competitive advantage of pinus females and their daughters over chrysoptera females.     

Synthetic prions generated in vitro are similar to a newly identified subpopulation of PrPSc from sporadic Creutzfeldt-Jakob Disease

In recent studies, the amyloid form of recombinant prion protein (PrP) encompassing residues 89-230 (rPrP 89-230) produced in vitro induced transmissible prion disease in mice. These studies showed that unlike "classical" PrP(Sc) produced in vivo, the amyloid fibrils generated in vitro were more proteinase-K sensitive. Here we demonstrate that the amyloid form contains a proteinase K-resistant core composed only of residues 152/153-230 and 162-230. The PK-resistant fragments of the amyloid form are similar to those observed upon PK digestion of a minor subpopulation of PrP(Sc) recently identified in patients with sporadic Creutzfeldt-Jakob disease (CJD). Remarkably, this core is sufficient for self-propagating activity in vitro and preserves a beta-sheet-rich fibrillar structure. Full-length recombinant PrP 23-230, however, generates two subpopulations of amyloid in vitro: One is similar to the minor subpopulation of PrP(Sc), and the other to classical PrP(Sc). Since no cellular factors or templates were used for generation of the amyloid fibrils in vitro, we speculate that formation of the subpopulation of PrP(Sc) with a short PK-resistant C-terminal region reflects an intrinsic property of PrP rather than the influence of cellular environments and/or cofactors. Our work significantly increases our understanding of the biochemical nature of prion infectious agents and provides a fundamental insight into the mechanisms of prions biogenesis.     

THE MORPHOLOGY AND DEVELOPMENT OF SOME PROMINENTLY STALKED SOUTHERN AUSTRALIAN HALYMENIACEAE (CRYPTONEMIALES,RHODOPHYTA). II. THE SPONGE-ASSOCIATED GENERA THAMNOCLONIUM KUETZING AND CODIOPHYLLUM GRAY1

Three members of the red algal family Halymeniaceae (Thamnoclonium dichotomum (J. Ag.) J. Ag., Codiophyllum flabelliforme (Sond.) Schmitz, and C. decipiens (J. Ag.) Schmitz) are investigated. All are endemic to southern and southwestern Australia, possess basal stalks of substantial size and firmness, and are consistently associated with specific sponge taxa. In each case, the sponges are bonded by collagen-like fibrils to the host cuticle without modifying the algal tissue at the ultrastructural level. Secondary cortication and prominent growth rings occur in the stalks of all three species, and in each the pit plugs between cells become wider, more convoluted and less electron dense with increasing distance from the surface. Such pit plugs are apparently a unique attribute of the stalked Halymeniaceae. The three species share pit plug, sponge association and stalk morphological features but are not otherwise closely related, as they actually represent three distinct genera.     

GacS sensor domains pertinent to the regulation of exoproduct formation and to the biocontrol potential of Pseudomonas fluorescens CHA0

In the root-colonizing biocontrol strain CHA0 of Pseudomonas fluorescens, cell density-dependent synthesis of extracellular, plant-beneficial secondary metabolites and enzymes is positively regulated by the GacS/GacA two-component system. Mutational analysis of the GacS sensor kinase using improved single-copy vectors showed that inactivation of each of the three conserved phosphate acceptor sites caused an exoproduct null phenotype (GacS-), whereas deletion of the periplasmic loop domain had no significant effect on the expression of exoproduct genes. Strain CHA0 is known to synthesize a solvent-extractable extracellular signal that advances and enhances the expression of exoproduct genes during the transition from exponential to stationary growth phase when maximal exoproduct formation occurs. Mutational inactivation of either GacS or its cognate response regulator GacA abolished the strain's response to added signal. Deletion of the linker domain of the GacS sensor kinase caused signal-independent, strongly elevated expression of exoproduct genes at low cell densities. In contrast to the wild-type strain CHA0, the gacS linker mutant and a gacS null mutant were unable to protect tomato plants from crown and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici in a soil-less microcosm, indicating that, at least in this plant-pathogen system, there is no advantage in using a signal-independent biocontrol strain.     

Explicit Mentalizing Mechanisms and Their Adaptive Role in Memory Conformity

Memory conformity occurs when an individual endorses what other individuals remember about past events. Research on memory conformity is currently dominated by a ‘forensic’ perspective, which views the phenomenon as inherently undesirable. This is because conformity not only distorts the accuracy of an individual''s memory, but also produces false corroboration between individuals, effects that act to undermine criminal justice systems. There is growing awareness, however, that memory conformity may be interpreted more generally as an adaptive social behavior regulated by explicit mentalizing mechanisms. Here, we provide novel evidence in support of this emerging alternative theoretical perspective. We carried out a memory conformity experiment which revealed that explicit belief-simulation (i.e. using one''s own beliefs to model what other people believe) systematically biases conformity towards like-minded individuals, even when there is no objective evidence that they have a more accurate memory than dissimilar individuals. We suggest that this bias is functional, i.e. adaptive, to the extent that it fosters trust, and hence cooperation, between in-group versus out-group individuals. We conclude that memory conformity is, in more fundamental terms, a highly desirable product of explicit mentalizing mechanisms that promote adaptive forms of social learning and cooperation.