CO2 efflux from a Mediterranean semi-arid forest soil. I. Seasonality and effects of stoniness

We studied the seasonality of total soil CO₂efflux and labeled C-CO₂ released from ¹⁴Clabeled straw incubated in the H horizon of asemi-arid Mediterranean forest soil. Fieldmeasurements were carried out over 520 days in aseries of reconstructed soil profiles with and withouta gravel layer below the H horizon. We monitored soilclimate and related this to soil CO₂ efflux.Seasonal variations in soil CO₂ efflux in asemiarid Mediterranean forest were mainly related tochanges in soil temperature. In spite of drought, highrespiration rates were observed in mid summer. Highsoil CO₂ efflux in hot and dry episodes wasattributed to increases in soil biological activity.The minimum soil CO₂ efflux occurred in latesummer also under dry conditions, probably related toa decrease in soil biological activity in deephorizons. Biological activity in organic layers waslimited by water potential () in summer and bytemperature in winter. Rewetting a dry soil resultedin large increases in soil CO₂ efflux only at hightemperatures. These large increases represented asignificant contribution to the decomposition oforganic matter in the uppermost horizons. Soilbiological activity in the uppermost horizons was moresensitive to changes in soil and hence tosummer rainstorms than the bulk soil microbialactivity. The presence of a layer of gravel improvedboth moisture and temperature conditions for thedecomposition of organic matter. As a result, soilCO₂ efflux increased in soils containing rockfragments. These effects were especially large for theorganic layers.     

Complete exon sequencing of all known Usher syndrome genes greatly improves molecular diagnosis

Background

Usher syndrome (USH) combines sensorineural deafness with blindness. It is inherited in an autosomal recessive mode. Early diagnosis is critical for adapted educational and patient management choices, and for genetic counseling. To date, nine causative genes have been identified for the three clinical subtypes (USH1, USH2 and USH3). Current diagnostic strategies make use of a genotyping microarray that is based on the previously reported mutations. The purpose of this study was to design a more accurate molecular diagnosis tool.

Methods

We sequenced the 366 coding exons and flanking regions of the nine known USH genes, in 54 USH patients (27 USH1, 21 USH2 and 6 USH3).

Results

Biallelic mutations were detected in 39 patients (72%) and monoallelic mutations in an additional 10 patients (18.5%). In addition to biallelic mutations in one of the USH genes, presumably pathogenic mutations in another USH gene were detected in seven patients (13%), and another patient carried monoallelic mutations in three different USH genes. Notably, none of the USH3 patients carried detectable mutations in the only known USH3 gene, whereas they all carried mutations in USH2 genes. Most importantly, the currently used microarray would have detected only 30 of the 81 different mutations that we found, of which 39 (48%) were novel.

Conclusions

Based on these results, complete exon sequencing of the currently known USH genes stands as a definite improvement for molecular diagnosis of this disease, which is of utmost importance in the perspective of gene therapy.     

High variation in foliage and leaf litter chemistry among 45 tree species of a neotropical rainforest community

Distinct ecosystem level carbon : nitrogen : phosphorus (C : N : P) stoichiometries in forest foliage have been suggested to reflect ecosystem-scale selection for physiological strategies in plant nutrient use. Here, this hypothesis was explored in a nutrient-poor lowland rainforest in French Guiana. Variation in C, N and P concentrations was evaluated in leaf litter and foliage from neighbour trees of 45 different species, and the litter concentrations of major C fractions were also measured. Litter C ranged from 45.3 to 52.4%, litter N varied threefold (0.68-2.01%), and litter P varied seven-fold (0.009-0.062%) among species. Compared with foliage, mean litter N and P concentrations decreased by 30% and 65%, respectively. Accordingly, the range in mass-based N : P shifted from 14 to 55 in foliage to 26 to 105 in litter. Resorption proficiencies indicated maximum P withdrawal in most species, but with a substantial increase in variation in litter P compared with foliage. These data suggest that constrained ecosystem-level C : N : P ratios do not preclude the evolution of highly diversified strategies of nutrient use and conservation among tropical rainforest tree species. The resulting large variation in litter quality will influence stoichiometric constraints within the decomposer food web, with potentially far-ranging consequences on nutrient dynamics and plant-soil feedbacks.     

αB-Crystallin Interacts with Cytoplasmic Intermediate Filament Bundles during Mitosis

The small heat-shock protein αB-crystallin interacts with intermediate filament proteins. Using cosedimentation assay, we showed previously that in vitro binding of αB-crystallin to peripherin and vimentin was temperature-dependent. Furthermore, when NIH 3T3 cells were submitted to different stress conditions a dynamic reorganization of the intermediate filament network was observed concomitantly with the recruitment of αB-crystallins on the intermediate filament proteins. Thus, the intracellular state of αB-crystallin correlated directly with the remodeling of the intermediate filament network in response to stress. Here, we show data suggesting that αB-crystallin is implicated in remodeling of intermediate filaments during cell division. We investigated the intracellular distribution of αB-crystallin in naturally occurring mitotic NIH 3T3 cells and in neuroblastoma N2a and N1E115 cells. In NIH 3T3 cells, αB-crystallin remained diffused throughout the cell cycle. Subcellular fractionation of αB-crystallin showed that αB-crystallin remained in the cytosolic compartment during mitosis. Furthermore, αB-crystallin accumulated in mitotically arrested NIH 3T3 cells. This increased level of αB-crystallin protein was due to an increased level of αB-crystallin mRNA in mitotic NIH 3T3 cells. In the neuroblastoma cells, the intermediate filaments were rearranged into thick cable-like structures and αB-crystallin was recruited onto them. In neuroblastoma N2a cells the level of expression did not change during the cell cycle. However, a small fraction of αB-crystallin switched onto the insoluble fraction in mitotically arrested N2a cells. Our results suggested that depending on the state of rearrangement of the intermediate filament network during mitosis αB-crystallin was either recruited onto the intermediate filaments or upregulated in the cytosolic compartment.     

CO2 efflux from a Mediterranean semi-arid forest soil. II. Effects of soil fauna and surface stoniness

Many forest soils in the Mediterranean basin areshallow and contain high amounts of gravel in theorganic layers. Recent studies on soil organic matteraccumulation have shown high amounts of organic matteroccurring mainly in soils with high levels ofstoniness at the soil surface. The gravel layer mayaffect the microclimatic conditions of the soilsurface and probably the distribution and activity ofsoil fauna.In order to quantify the combined effects soil fauna(epigeic macrofauna and earthworms) and stoniness onthe release of soil CO₂, we performed a threefactor field experiment by using a series ofreconstructed soil profiles. Factors 1 and 2 consistedof the exclusion/presence of soil epigeic macrofaunaand earthworms, and factor 3 of the presence/absenceof a gravel layer intermingled with the H horizon. Weincubated ¹⁴C straw in the H horizon and carriedout three 40 mm rainfall simulations.Soil respiration primarily depended on the season. Theeffects of soil fauna were generally small and did notcoincide with periods of high faunal activity. Thelargest effects of both earthworms and soil epigeicfauna were found after wetting the soil in summer. Theeffects of the earthworms were concentrated in themineral soil while the effects of the epigeic faunawere concentrated in the H horizon and mainly arosetowards the end of the experiment. This suggests thatthe effects of epigeic fauna may have beenunderestimated due to the length of the experiment.The gravel layer increased the effect of faunaprobably by creating more favorable microclimaticconditions. The accumulation of organic matter insoils with high levels of stoniness cannot beexplained by the effect of gravel on soil microclimatenor by its effect on the activity of soil fauna.     

Genetic variability of the length of postpartum anoestrus in Charolais cows and its relationship with age at puberty

Fertility records (n = 1 802) were collected from 615 Charolais primiparous and multiparous cows managed in an experimental herd over an 11-year period. The objectives of the study were to describe the genetic variability of the re-establishment of postpartum reproductive activity and the relationship with body weight (BW) and body condition score (BCS) at calving and age at puberty. The length of postpartum anoestrus was estimated based on weekly blood progesterone assays and on twice daily detection of oestrus behaviour. The first oestrus behaviour was observed 69 days (± 25 days s.d.) post-calving and the first positive progesterone measurement (≥ 1 ng mL^-1) was observed at 66 days (± 22 days s.d.) for the group of easy-calving multiparous suckling cows. Estimates of heritability and repeatability were h² = 0.12 and r = 0.38 respectively, for the interval from calving to first oestrus (ICO). Corresponding values were h² = 0.35 and r = 0.60 for the interval from calving to the first positive progesterone test (ICP). The genetic correlation between both criteria was high (r_g = 0.98). The genetic relationships between postpartum intervals and BW and BCS of the female at calving were negative: the genetic aptitude to be heavier at calving and to have high body reserves was related to shorter postpartum intervals. A favourable genetic correlation between age at puberty and postpartum intervals was found (r_g between 0.45 and 0.70). The heifers which were genetically younger at puberty also had shorter postpartum intervals.     

THE CHITINOUS NATURE OF FILAMENTS EJECTED BY PHAEOCYSTIS (PRYMNESIOPHYCEAE)1

Filaments ejected by Phaeocystis globosa Scherffel, organized in star-like structures, were observed and analyzed before and after their discharge from cells. Ultrastructural observations obtained after cryofixation and cryosubstitution led to a model for their storage within the cell and for their ejection from the cell. Electron diffraction analysis on the ejected filaments demonstrated their chitinous composition. This technique indicated without ambiguity that each filament was in fact a whisker-like α-chitin crystal, with the axes of the corresponding polymer chains aligned with the filament's axis. X-ray microanalysis of the mats of filaments indicated that the silica content suggested by earlier workers was an artifact resulting from the filtration procedure.     

Crystallization,preliminary X-ray diffraction study,and crystal packing of a complex between anti-Hen lysozyme antibody F9.13.7 and Guinea-fowl lysozyme

The complex formed between the Fab fragment of a murine monoclonal anti-hen egg lysozyme antibody F9.13.7 and the het-erologous antigen Guinea-fowl egg lysozyme has been crystallized by the hanging drop technique. The crystals, which diffract X-rays to 3 Å resolution, belong to the monoclinic space group P2₁, with a = 83.7 Å, b = 195.5 Å, c = 50.2 Å, β = 108.5° and have two molecules of the complex in the asymmetric unit The three-dimensional structure has been determined from a preliminary data set to 4 Å using molecular replacement techniques. The lysozyme–Fab complexes are arranged with their long molecular axes approximately parallel to the crystallo-graphic unique axis. Fab F9.13.7 binds an anti-genie determinant that partially overlaps the epitope recognized by antilysozyme antibody HyHEL10. © 1993 Wiley-Liss, Inc.