Mapping of Igk-V genes using backcrossed laboratory and wild mice

In the mouse, the genes coding for the Ly-2 antigen, the chain of the T-cell receptor, and the immunoglobulin kappa light chain have been located on chromosome 6. Although a tentative order has been proposed for these genes, very few data have been reported concerning their genetic distance. To address this question, we have produced backcross mice between SJL and MAI (a wild-derived strain belonging to the Mus musculus), since these mice segregate for the Ly-2 and Igk-C proteins and for the Igk-V24, Igk-V21, Igk-V10, Igk-V8, and Igk-V4 genes. Twelve recombinants were obtained from 163 backcross mice studied. Two mice showed a recombination between the (Igk-V24, Igk-V10, Igk-V8, Igk-V4) and the (Ly-2, Igk-C, Igk-V21) groups, and ten mice displayed a recombination between the (Igk-V24, Igk-V10, Igk-V8, Igk-V4) group and the Tcrb-C loci. These data imply the following gene order: Tcrb-C .... (Igk-V24, Igk-V10, Igk-V8, Igk-V4) .... (Igk-V21, Igk-C, Ly-2). They indicate a distance of 6.1 cM between Tcrb-C and (Igk-V24, Igk-V10, Igk-V8, Igk-V4) and 1.2 cM between Igk-V24, Igk-V10, Igk-V8, Igk-V4 and the (Igk-V21, Igk-C, Ly-2) groups.     

Size, activity and catabolic diversity of the soil microbial biomass in a wetland complex invaded by reed canary grass

Reed canary grass (Phalaris arundinacea, L.) invasion of wetlands is an ecological issue that has received attention, but its impact on soil microbial diversity is not well documented. The present study assessed the size (substrate-induced respiration), catabolic diversity (CLPP, community level physiological profiles) and composition (selective inhibition) of the soil microbial community in invaded (>95% P. arundinacea cover) and in non-invaded areas of a wetland occupied by native species grown either as a mixed assemblage (22 species) or as quasi-monotypic stands of Scirpus cyperinus (74% cover). The study also tested the hypothesis that decomposition of lignin- and phenolics-rich plant tissues would be fastest in soils exhibiting high catabolic diversity. Results showed that soil respiration, microbial biomass and diversity were significantly higher (P?<?0.03; 1.5 to 3 fold) in P. arundinacea-invaded soils than in soils supporting native plant species. Fungal to bacterial ratios were also higher in invaded (0.6) than in non-invaded (0.4) plots. Further, canonical discriminant analysis of CLPP data showed distinct communities of soil decomposers associated with each plant community. However, these differences in microbial attributes had no effect on decomposition of plant biomass which was primarily controlled by its chemical composition. While P. arundinacea invasion has substantially reduced plant diversity, this study found no parallel decline in the size and diversity of the soil microbial community in the invaded areas.     

New insights into the effect of medium-chain-length lactones on yeast membranes. Importance of the culture medium

In hydrophobic compounds biotechnology, medium-chain-length metabolites often perturb cell activity. Their effect is usually studied in model conditions of growth in glucose media. Here, we study whether culture on lipids has an impact on the resistance of Yarrowia lipolytica to such compounds: Cells were cultured on glucose or oleate and submitted to γ-dodecalactone. After a 60-min exposure to 3 g L⁻¹, about 80% of the glucose-grown cells (yeast extract peptone dextrose (YPD) cells) had lost their cultivability, 38% their membrane integrity, and 31% their reducing capacity as shown with propidium iodide and methylene blue, respectively. For oleate-grown cells, treatment at 6 g L⁻¹ did not alter cultivability despite some transient loss of membrane integrity from 3 g L⁻¹. It was shown with diphenylhexatriene and 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene that oleate-grown cells had membranes more fluid and less sensitive to the lactone-induced fluidization. Analyses revealed also higher contents of ergosterol but, for YPD- and minimum-oleate-grown cells (YNBO cells), the addition of lactone provoked a decrease in the concentration of ergosterol in a way similar to the depletion by methyl-β-cyclodextrin and an important membrane fluidization. Ergosterol depletion or incorporation increased or decreased, respectively, cell sensitivity to lactone. This study shows that the embedment of oleate moieties into membranes as well as higher concentrations of sterol play a role in the higher resistance to lactone of oleate-grown cells (YPO cells). Similar oleate-induced increase in resistance was also observed for Rhodotorula and Candida strains able to grow on oleate as the sole carbon source whereas Saccharomyces and Sporidiobolus cells were more sensitive after induction.     

Cell size and water permeability as determining factors for cell viability after freezing at different cooling rates

This work studied the viabilities of five types of cells (two yeast cells, Saccharomyces cerevisiae CBS 1171 and Candida utilis; two bacterial strains, Escherichia coli and Lactobacillus plantarum; and one human leukemia K562 cell) as a function of cooling rate during freezing. The range of investigated cooling rates extended from 5 to 30,000 degrees C/min. Cell viability was classified into three ranges: (i) high viability for low cooling rates (5 to 180 degrees C/min), which allow cell water outflow to occur completely and do not allow any intracellular crystallization; (ii) low viability for rapid cooling rates (180 to 5,000 degrees C/min), which allow the heat flow to prevail over water outflow (in this case, cell water crystallization would occur as water was flowing out of the cell); (iii) high viability for very high cooling rates (>5,000 degrees C/min), which allow the heat flow to be very rapid and induce intracellular crystallization and/or vitrification before any water outflow from the cell. Finally, an assumption relating cell death to the cell water crystallization as water is flowing out of the cell is made. In addition, this general cell behavior is different for each type of cell and seems to be moderated by the cell size, the water permeability properties, and the presence of a cell wall.     

The Hsp90 co-chaperones Cdc37 and Sti1 interact physically and genetically

Cdc37 associates with the heat-shock protein 90 (Hsp90) molecular chaperone as one of several auxiliary proteins that are collectively referred to as Hsp90 co-chaperones. Cdc37 has been proposed to be a specificity factor for Hsp90, directing it notably towards kinases. It is not known whether Cdc37 is essential for viability in the budding yeast Saccharomyces cerevisiae because of Hsp90-dependent or -independent functions or both. Sti1 and Cpr7 are non-essential Hsp90 co-chaperones that bind to a common surface on Hsp90 through tetratricopeptide repeats (TPR). We have found that Sti1 is specifically retained from yeast extracts by immobilized Cdc37. Similarly, the endogenous proteins are also found in a complex. Moreover, purified recombinant Sti1 and Cdc37 interact in the complete absence of Hsp90. Complexes between Cdc37 and Sti1 are not unique to this TPR protein since endogenous Cdc37 can be co-purified with exogenously expressed Cpr7 fused to glutathione-S-transferase. The heterogeneity of Cdc37 complexes, both with and without Hsp90, may expand the functional diversity of Cdc37. Here we show that the combination of cdc37 and sti1 mutations is synthetically lethal, suggesting that direct contacts between Cdc37 and Sti1 may at least contribute to vital functions in yeast.     

An extended HLA-DQ-DR haplotype rather than DRB1 alone contributes to RA predisposition

 In the present study, we tested our hypothesis on the role of a DQ-DR haplotype in rheumatoid arthritis (RA) predisposition. Using two groups of patients and controls, one from The Netherlands and one from Switzerland, we found that DQA1*0301-homozygous and DQA1*0301//DQA1*0101/04-heterozygous individuals are highly predisposed to RA in both populations, while DQA1*0101/04-homozygous are not. The DQA1*0301-DRB1*0403/06/07 and DQA1*0301-DRB1*0901 haplotypes are not associated with RA by themselves but strongly increase the risk of developing disease in DQA1*0301- and DQA1*0101/04-heterozygous. DRB1 alleles carrying the motif DERAA in their third hypervariable region, i.e., *0103, *0402, *1102, *1103, *1301, and *1302, provide a long-lasting protection against RA in DQA1*0101/04- but not in DQA1*0301-positive individuals. These data show that considering both DQ and DR gives a better distinction between patients and controls than the shared epitope hypothesis. Received: 5 March 1998 / Revised: 21 April     

A profound alteration of blood TCRB repertoire allows prediction of cerebral malaria

Cerebral malaria (CM) is one of the severe complications of Plasmodium infection. In murine models of CM, Talphabeta cells have been implicated in the neuropathogenesis. To obtain insights into the TCRB repertoire during CM, we used high throughput CDR3 spectratyping and set up new methods and software tools to analyze data. We compared PBL and spleen repertoires of mice infected with Plasmodium berghei ANKA that developed CM (CM(+)) or not (CM(-)) to evidence modifications of the TCRB repertoire associated with neuropathology. Using distinct statistical multivariate methods, the PBL repertoires of CM(+) mice were found to be specifically altered. This alteration is partly due to recurrently expanded T cell clones. Strikingly, alteration of the PBL repertoire can be used to distinguish between CM(+) and CM(-). This study provides the first ex vivo demonstration of modifications of Talphabeta cell compartment during CM. Finally, our original approach for deciphering lymphocyte repertoires can be transposed to various pathological conditions.     

Quantitative gas chromatography/mass spectrometry determination of C-mannosylation of tryptophan residues in glycoproteins

C-mannosylation of Trp residue is one of the most recently discovered types of glycosylation, but the identification of these mannosylated residues in proteins is rather tedious. In a previous paper, it was reported that the complete analysis of all constituents of glycoproteins (sialic acids, monosaccharides, and amino acids) could be determined on the same sample in three different steps of gas chromatography/mass spectrometry of heptafluorobutyrate derivatives. It was observed that during the acid-catalyzed methanolysis step used for liberation of monosaccharide from classical O- and N-glycans, Trp and His were quantitatively transformed by the addition of a methanol molecule on their indole and imidazole groups, respectively. These derivatives were stable to acid hydrolysis used for the liberation of amino acids. Since monosaccharide derivatives were also stabilized as heptafluorobutyrate derivatives of O-methyl-glycosides, it was suggested that C-mannosides of Trp residues could quantitatively be recovered. Based on the analyses of standard compounds, peptides and RNase 2 from human urine, we report that C((2))-mannosylated Trp could be quantitatively recovered and identified during the step of amino acid analysis. Analyses of different samples indicated that this type of glycosylation is absent in bacteria and yeasts.     

Depth-related ecological zonation of a carboniferous carbonate ramp: Upper Viséan of Béchar Basin,Western Algeria

Summary Following the demise of the stromatoporoid-coral reef community in Late Frasnian time, Lower Carboniferous carbonate shelf profiles possessed a ramp geometry, with major organic buildups represented by mud-rich mounds. Microfacies petrography of the exceptionally well-preserved Upper Viséan (Lower Carboniferous) carbonate ramp of the Béchar Basin, Algerian Sahara, may well contribute significantly to our understanding of the paleoecological zonation of Carboniferous non-rimmed platforms, and of the still enigmatic mounds commonly referred to as Waulsortian banks or mounds. Facies are grouped into two broad groups: (a) a mound facies group which comprises sponge wackestone-bafflestone, sponge-fenestellid bafflestone-wackestone, crinoid wackestone-packstone, and bedded flanks of intraclastic wackestone-packstone, all four facies composing the actual mud-rich mounds, and (b) a supramound facies group composed of coral-microbial framestone, crinoid packstone-grainstone, algal-foraminiferal grainstone and oolite grainstone. Calcareous algae are important bathymetric indicators and are used to delineate three bathymetric zones based on light penetration: the aphotic zone, which contains no calcareous algae; the dysphotic zone, where there is little ambient light, and which is characterized by the presence of red algae (Fasciella, Ungdarella, Stacheia, Epistacheoides) and absence of green algae; and the euphotic zone, which receives the full spectrum of sunlight, and is characterized by the occurrence of both green algae (Koninckopora, Kamaenella, Kamaena, Palaeoberesella, Calcisphaera, Anthracoporellopsis, Issinella, Exvotarisella) and red algae. Integration of algal zonation, distribution of the other biota, and recurrence of distinct assemblages, enable recognition of seven depth-related benthic assemblages. Together with the physical properties of the facies, the benthic assemblages were used to define seven bathymetric zones, from upper to lower ramp: (1) algal assemblage (upper ramp); (2) crinoid-ramose bryozoan assemblage (mid-ramp); and (3) productid brachiopod assemblage, (4) colonial rugose coral-microbial encruster assemblage, (5) crinoid-fenestellid assemblage, (6) sponge-fenestellid, and (7) sponge assemblage (lower ramp). The vertical zonation of the mud-rich mounds and associated facies differ from that reported from the classical Upper Tournaisian-Lower Viséan Waulsortian mound-bearing successions.