Characterization of native and reconstituted exosome complexes from the hyperthermophilic archaeon Sulfolobus solfataricus

The eukaryotic exosome is a protein complex with essential functions in processing and degradation of RNA. Exosome-like complexes were recently found in Archaea. Here we characterize the exosome of Sulfolobus solfataricus. Two exosome fractions can be discriminated by density gradient centrifugation. We show that the Cdc48 protein is associated with the exosome from the 30S-50S fraction but not with the exosome of the 11.3S fraction. While only some complexes contain Cdc48, the archaeal DnaG-like protein was found to be a core exosome subunit in addition to Rrp4, Rrp41, Rrp42 and Csl4. Assays with depleted extracts revealed that the exosome is responsible for major ribonucleolytic activity in S. solfataricus. Various complexes consisting of the Rrp41-Rrp42 hexameric ring and Rrp4, Csl4 and DnaG were reconstituted. Dependent on their composition, different complexes showed variations in RNase activity indicating functional interdependence of the subunits. The catalytic activity of these complexes and of the native exosome can be ascribed to the Rrp41-Rrp42 ring, which degrades RNA phosphorolytically. Rrp4 and Csl4 do not exhibit any hydrolytic RNase activity, either when assayed alone or in context of the complex, but influence the activity of the archaeal exosome.     

Investigating damselfly populations at springs in Banff National Park, Canada, with special focus on Argia vivida, Amphiagrion abbreviatum, and Ischnura cervula (Odonata: Coenagrionidae)

The objective of this study was to estimate Argia vivida (Odonata: Coenagrionidae) populations, identify breeding habitat, and investigate movement of adults within Banff National Park, Alberta, Canada, during the summer of 2003. Mark-recapture techniques and standardized dip-net surveys were used to monitor Argia vivida at various life stages. A reproductive index identified which sites Argia vivida recognized as suitable breeding habitat, and exuvia surveys confirmed breeding sites. The basic structure of emergent and surrounding vegetation was measured to investigate the importance of available ovipositing or roosting sites and the condition of the matrix habitat. Data was recorded for Amphiagrion abbreviatum and Ischnura cervula (both Odonata: Coenagrionidae) to determine if these spring-associated damselflies were successfully breeding within Banff National Park. Comparisons were made between the highly protected Middle Springs and the heavily altered Cave & Basin Springs. Additional surveys at the Vermilion Lake cool spring and Middle Springs Bog investigated their use as breeding habitat for Amphiagrion abbreviatum and Argia vivida, respectively. Results suggest the ecological value of thermal springs extends beyond their origin to outflows and downstream pools. Conservation of Argia vivida must recognize the value of unobstructed thermal outflows, and consider the condition of the forested habitat surrounding springs with regard to its potential use as nocturnal roosts and dispersal corridors. Amphiagrion abbreviatum was confirmed breeding within Banff National Park, while no sign of breeding activity was recorded for Ischnura cervula.     

Evolution of resistance during clonal expansion

Acquired drug resistance is a major limitation for cancer therapy. Often, one genetic alteration suffices to confer resistance to an otherwise successful therapy. However, little is known about the dynamics of the emergence of resistant tumor cells. In this article, we consider an exponentially growing population starting from one cancer cell that is sensitive to therapy. Sensitive cancer cells can mutate into resistant ones, which have relative fitness alpha prior to therapy. In the special case of no cell death, our model converges to the one investigated by Luria and Delbrück. We calculate the probability of resistance and the mean number of resistant cells once the cancer has reached detection size M. The probability of resistance is an increasing function of the detection size M times the mutation rate u. If Mu < 1, then the expected number of resistant cells in cancers with resistance is independent of the mutation rate u and increases with M in proportion to M(1-1/alpha) for advantageous mutants with relative fitness alpha>1, to l nM for neutral mutants (alpha = 1), but converges to an upper limit for deleterious mutants (alpha<1). Further, the probability of resistance and the average number of resistant cells increase with the number of cell divisions in the history of the tumor. Hence a tumor subject to high rates of apoptosis will show a higher incidence of resistance than expected on its detection size only.     

Juvenile ornithopod (Dinosauria: Rhabdodontidae) remains from the Upper Cretaceous (Lower Campanian, Gosau Group) of Muthmannsdorf (Lower Austria)

The fragmentary remains of a juvenile rhabdodontid ornithopod from the Coal-bearing Complex of the Gosau Group (Lower Campanian, Grünbach syncline) at Muthmannsdorf near Wiener Neustadt, Lower Austria are revised. The material, probably belonging to a single individual, includes a right dentary (lectotype of Iguanodon suessi Bunzel, 1871, designated herein), teeth, a fragmentary parietal, fragments of scapula, ?radius, femur, tibia, two vertebrae (lost) and a manual ungual.The lectotype dentary does not provide clear autapomorphies or sufficient diagnostic features to determine its position within the Rhabdodontidae at generic level. By this “Iguanodon suessi” Bunzel, 1871 and the genus “Mochlodon” Seeley, 1881, to which it was latter referred as type species, cannot be characterized sufficiently by differential diagnosis and these are best considered nomina dubia. Based upon combined character comparisons (mainly postcranial features) the Muthmannsdorf ornithopod is referred herein to Zalmoxes Weishampel, Jianu, Csiki and Norman, 2003, a genus so far known from the late Maastrichtian of Romania. It probably but not evidently represents a yet unnamed species, most closely related to Zalmoxes shqiperorum Weishampel, Jianu, Csiki and Norman, 2003. At the present state of knowledge the Austrian material is not further diagnostic at the species level and kept in open nomenclature as Zalmoxes sp.     

Drivers of litter mass loss and faunal composition of detritus patches change over time

Decomposition of vegetal detritus is one of the most fundamental ecosystem processes. In complex landscapes, the fate of litter of terrestrial plants may depend on whether it ends up decomposing in terrestrial or aquatic conditions. However, (1) to what extent decomposition rates are controlled by environmental conditions or by detritus type, and (2) how important the composition of the detritivorous fauna is in mediating decomposition in different habitats, remain as unanswered questions. We incubated two contrasting detritus types in three distinct habitat types in Coastal Georgia, USA, to test the hypotheses that (1) the litter fauna composition depends on the habitat and the litter type available, and (2) litter mass loss (as a proxy for decomposition) depends on environmental conditions (habitat) and the litter type. We found that the abundance of most taxa of the litter fauna depends primarily on habitat. Litter type became a stronger driver for some taxa over time, but the overall faunal composition was only weakly affected by litter type. Decomposition also depends strongly on habitat, with up to ca. 80% of the initial detrital mass lost over 25 months in the marsh and forest habitats, but less than 50% lost in the creek bank habitat. Mass loss rates of oak versus pine litter differed initially but converged within habitat types within 12 months. We conclude that, although the habitat type is the principle driver of the community composition of the litter fauna, litter type is a significant driver of litter mass loss in the early stages of the decomposition process. With time, however, litter types become more and more similar, and habitat becomes the dominating factor in determining decomposition of older litter. Thus, the major driver of litter mass loss changes over time from being the litter type in the early stages to the habitat (environmental conditions) in later stages.     

Novel Humanized and Highly Efficient Bispecific Antibodies Mediate Killing of Prostate Stem Cell Antigen-Expressing Tumor Cells by CD8+ and CD4+ T Cells

Prostate cancer is the most common noncutaneous malignancy in men. The prostate stem cell Ag (PSCA) is a promising target for immunotherapy of advanced disease. Based on a novel mAb directed to PSCA, we established and compared a series of murine and humanized anti-CD3-anti-PSCA single-chain bispecific Abs. Their capability to redirect T cells for killing of tumor cells was analyzed. During these studies, we identified a novel bispecific humanized Ab that efficiently retargets T cells to tumor cells in a strictly Ag-dependent manner and at femtomolar concentrations. T cell activation, cytokine release, and lysis of target cells depend on a cross-linkage of redirected T cells with tumor cells, whereas binding of the anti-CD3 domain alone does not lead to an activation or cytokine release. Interestingly, both CD8(+) and CD4(+) T cells are activated in parallel and can efficiently mediate the lysis of tumor cells. However, the onset of killing via CD4(+) T cells is delayed. Furthermore, redirecting T cells via the novel humanized bispecific Abs results in a delay of tumor growth in xenografted nude mice.     

Naturally occurring fragments from two distinct regions of the prostatic acid phosphatase form amyloidogenic enhancers of HIV infection

Semen is the major vector for HIV-1 transmission. We previously isolated C-proximal fragments of the prostatic acid phosphatase (PAP) from semen which formed amyloid fibrils that potently enhanced HIV infection. Here, we used the same methodology and identified another amyloidogenic peptide. Surprisingly, this peptide is derived from an N-proximal fragment of PAP (PAP85-120) and forms, similar to the C-proximal fragments, positively charged fibrillar structures that increase virion attachment to cells. Our results provide a first example for amyloid formation by fragments of distinct regions of the same precursor and further emphasize the possible importance of amyloidogenic peptides in HIV transmission.