ALBERT

Notes: . In vitro excystation of Spironucleus muris cysts, purified by sequential sucrose and Percoll gradients from mouse feces, was studied. Three in vitro excystation procedures, used for Giardia, were assessed to determine the most useful method. Excystation was monitored by light microscopy and subsequently characterized by transmission and scanning electron microscopy. Spironucleus muris excysted routinely at a level greater than 90% when induced in Hanks’ balanced salt solution containing sodium bicarbonate at pH 2.0 and transferred to Tyrodes’ salt solution as an excystation medium. Similarly, high rates of excystation were recorded after induction of S. muris cysts in 0.1 M potassium phosphate buffer (pH 7.0) with sodium bicarbonate and excystation in trypticase-yeast extract-iron medium (TYI medium) or phosphate-buffered saline. A lower rate and percentage of excystation were observed after induction of S. muris cysts in an aqueous hydrochloric acid solution (pH 2.0) followed by excystation in TYI medium. All excystation methods produced extremely active S. muris trophozoites with normal morphology. Nonexcysting S. muris cysts have a wall composed of an outer fibrous and an inner membranous portion. Following induction, numerous vesicles appeared in the peritrophic space. Excystation began by the cyst wall opening at one pole, and the anterior part of the trophozoite protruding from the cyst wall. The trophozoite emerged progressively from the cyst wall and the empty cyst wall appeared to collapse. Excysted trophozoites exhibited normal morphological features of S. muris trophozoites isolated from the mouse intestine.

Notes: In ciliates, development of the polyploid somatic macronucleus after sexual events involves extensive and reproducible rearrangements of the germ-line genome, including chromosome fragmentation and precise excision of numerous internal sequence elements. In Paramecium aurelia, alternative macronuclear versions of the same germ-line genome can be maternally inherited across sexual generations, showing that rearrangement patterns are not strictly determined by the germ-line sequence. Homology-dependent maternal effects can be evidenced by transformation of the vegetative macronucleus with cloned macronuclear sequences: new fragmentation patterns or internal deletions are specifically induced during differentiation of a new macronucleus, in sexual progeny of transformed clones. Furthermore, transformation of the maternal macronucleus with germ-line sequences containing internal eliminated sequences (short single-copy elements) can result in a specific inhibition of the excision of the same elements in the zygotic macronucleus. These experiments show that the processing of many germ-line sequences in the developing macronucleus is sensitive to the structure and copy number of homologous sequences in the maternal macronucleus. The generality and sequence specificity of this trans-nuclear, epigenetic regulation of rearrangements suggest that it is mediated by pairing interactions between germ-line sequences and sequences imported from the maternal macronucleus.

Notes: An analysis was conducted of multidimensional subsurface moisture flow in a hypothetical low-level radioactive waste disposal facility. The key design feature examined in the analysis was a sloping sand/gravel capillary barrier designed to route natural infiltration around a concrete vault. Three barrier slopes (1:25, 1:10, and 1:5) and three sand/gravel property combinations were considered. The slopes and material property combinations were selected to represent a range in barrier effectiveness. The porous media flow code used for this analysis was VAM3DCG, a three-dimensional, finite-element code which was able to use nonorthogonal grid discretizations and employed robust, efficient numerical techniques. Three-dimensional modeling demonstrated that flow in the hypothetical design exhibited cross-slope flow because of the pressure gradient produced in the third, cross-slope dimension. Barrier effectiveness was shown to be highly sensitive to the sand/ gravel material properties. Barrier slope was less important, especially for the effective material combinations. The presence of three-dimensional flow could be important in a performance assessment if the quantity of water predicted to breach the capillary barrier by a two-dimensional model were different from that predicted by a three-dimensional model. Comparative modeling demonstrated that a two-dimensional analysis resulted in underestimation of barrier effectiveness. For the considered design, two-dimensional modeling is a conservative, yet reasonable, approach in a performance assessment application.

Notes: Spatial mid temporal variations in the distribution of photosynthesis over the leaf area were investigated during induction upon illumination of Rosa rubiginosa L. leaves. Gas exchange and maps of relative photosynthetie electron transport activity computed from chlorophyll fluorescence images were simultaneously monitored. In air, after 15 h of dark adaptation, linear electron transport was heterogeneously distributed over the leaf area during the induction. This patchy induction was explained by asynchronous metabolism activation for the first 10 min of illumination, concomitant asynchronous limitation by intrinsic metabolism and stomatal apertures (10–30 min) and finally by only stomatal limitation beyond 30 min. A brief transition to non-photorespiratory conditions after 20 min of illumination under subsaturating irradiance revealed a marked heterogeneity of CO2 assimilation, presumably as a result of heterogeneous stomatal apertures. The frequency distribution of CO2 assimilation was unimodal. During the induction, heterogeneity gradually decreased and photosynthesis was uniform at steady-state. After 10 min of dark adaptation, heterogeneity of linear electron transport activity occurred during the first 15 min of a second induction and mainly resulted from metabolic limitation.

Notes: We present geological and morphological observations at different scales to constrain rates of faulting and the distribution of deformation in the seismically active Aegean region. We focus first on the 130 km long Corinth Rift, an asymmetric graben where a flight of terraces of marine origin are uplifted. We show that the edges of the terraces lie in the footwall of the normal fault bounding the Corinth Rift and correspond to sea-level highstands of laic Pleistocene age. Using a detailed analysis of aerial and SPOT imagery supported by field observations, we have mapped 10 terrace platforms and strandlines ranging in elevation from 10 to 400 m over distances of 2 to 20 km from the fault. The elevation of the terraces' inner edges was estimated at 172 sites with an error of ±5m. This data set contains a precise description of the uplift and flexure of 10 different palaeohorizontal lines with respect to the present sea level. To date the deformation, we correlate the Corinth terraces with late Pleistocene oxygen-isotope stages of high sea-level stands and with global sea-level fluctuations. Using a thick elastic plate model consistent with our current understanding of the earthquake cycle and a boundary-element technique we reproduce the geometry of the shorelines to constrain both mechanical parameters and the slip on the fault. We show that the seismogenic layer behaves over the long term as if its elastic modulus were reduced by a factor of about 1000. All the terraces are fitted for fault slip increasing in proportion to terrace age, and the component of regional uplift is found to be less than 0.3 mm yr−1. The best fits give a slip rate of 11±3 mm yr−1 on the main rift-bounding fault over the last 350 kyr. Other geological and morphologic information allows us to estimate the total age of the main fault (∼1 Ma) and to examine the mechanical evolution of the Corinth Rift. The minimum observed sediment thickness in the Gulf places an extreme check on the results of the modelling and a lower bound on slip rate of 6–7 mm yr−1 (40 per cent less than estimated with modelling). Even this slip rate is nearly 10 times higher than for comparable features in most of the Aegean and elsewhere in the world.At a larger scale, the spacing and asymmetry of the rift systems in the Aegean suggest strain localization in the upper mantle, with slow extension starting 15 Myr ago or earlier. The more recent (1 Myr), rapid phase of rifting in Corinth partly reactivated this earlier phase of extension. The younger faulting in Corinth appears to result from its present location in the inhomogeneous stress field (process zone) of the south-westward propagating tip of the southern branch of the North Anatolian Fault. We extend these relations to propose a mechanical model for the Late Cenozoic evolution of the Aegean. As the Arabia/Europe collision progressed in eastern Turkey it caused Anatolia to move to the west and the North Anatolian Fault to propagate into the Aegean, where the early slow extension started to be modified about 5 Ma ago. The process of propagation dramatically increased the activity of some but not all of the earlier rifts. The model we present is compatible with tectonic observations, as well as with the seismicity, the palaeomagnetic rotations and the displacement field now observed with GPS and SLR.

Notes: Field work carried out in Gansu province and complemented with analysis of SPOT panchromatic scenes allows us to characterize the deformations along the eastern segment of the Altyn Tagh fault and to place bounds on its Holocene left slip rate. East of 96°E, the long-term, left-lateral offset of stream channels, alluvial fans, and terrace edges is about 50 m. These offsets are most probably of Holocene age (12 ± 2 ka) and imply that the corresponding derived slip rate is 4 ± 2 mm yr−1. This observation is consistent with a north-eastward along-strike decreasing slip rate on the Altyn fault due to partitioning of slip on multiple, more easterly trending splays.

Notes: We characterized a novel mutant phenotype (tetrapac, tpc) of Neisseria gonorrhoeae (Ngo) associated with a distinctive rough-colony morphology and bacterial growth in clusters of four. This phenotype, suggesting a defect in cell division, was isolated from a mutant library of Ngo MS11 generated with the phoA minitransposon TnMax4. The tpc mutant shows a 30% reduction in the overall murein hydrolase activity using Escherichia coli murein as substrate. Tetrapacs can be resolved by co-cultivation with wild-type Ngo, indicating that Tpc is a diffusible protein. Interestingly, Tpc is absolutely required for the natural transformation competence of piliated Ngo. Mutants in tpc grow normally, but show a ∼ 10-fold reduction in their ability to invade human epithelial cells. The tpc sequence reveals an open reading frame of ∼1 kb encoding a protein (Tpc) of 37kDa. The primary gene product exhibits an N-terminal leader sequence typical of lipoproteins, but palmitoylation of Tpc could not be demonstrated. The ribosomal binding site of tpc is immediately downstream of the translational stop codon of the folC gene coding for an enzyme involved in folic acid biosynthesis and one-carbon metabolism. The tpc gene is probably co-transcribed from the folC promoter and a promoter located within the folC gene. The latter promoter sequence shares significant homology with E. coli gearbox consensus promoters. All three mutant phenotypes, i.e. the cell separation defect, the transformation deficiency and the defect in cell invasion can be restored by complementation of the mutant with an intact tpc gene. To some extent the tcp phenotype is reminiscent of iap in Listeria, lytA in Streptococcus pneumoniae and lyt in Bacillus subtilis, all of which are considered to represent murein hydrolase defects.