Heliobacteria Reveal Fermentation As a Key Pathway for Mercury Reduction in Anoxic Environments.

The accumulation of mercury (Hg) in rice, a dietary staple for over half of the world's population, is rapidly becoming a global food safety issue. Rice paddies support the anaerobic production of toxic methylmercury that accumulates in plant tissue, however the microbial controls of Hg cycling in anoxic environments remain poorly understood. In this study, we reveal a novel reductive Hg metabolism in a representative of the family Heliobacteria ( Heliobacterium modesticaldum Ice1) that we confirm in model chemotrophic anaerobes. Heliobacteria served as our initial model because they are a family of spore-forming fermentative photoheterotrophs commonly isolated from terrestrial environments. We observed that H. modesticaldum reduced up to 75% of HgII under phototrophic or fermentative conditions. Fermentative HgII reduction relied on the ability of cells to oxidize pyruvate whereas phototrophic HgII reduction could be supported even in the absence of a carbon source. Inhibiting pyruvate fermentation eliminated HgII reduction in all chemotrophic strains tested, whereas phototrophic cells remained unaffected. Here we propose a non mer-operon dependent mechanism for Hg0 production in anoxic environments devoid of light where external electron acceptors are limited. These mechanistic details provide the foundation for novel bioremediation strategies to limit the negative impacts of Hg pollution.

Rainfall-induced runoff from exposed streambed sediments: an important source of water pollution.

When surface water levels decline, exposed streambed sediments can be mobilized and washed into the water course when subjected to erosive rainfall. In this study, rainfall simulations were conducted over exposed sediments along stream banks at four distinct locations in an agriculturally dominated river basin with the objective of quantifying the potential for contaminant loading from these often overlooked runoff source areas. At each location, simulations were performed at three different sites. Nitrogen, phosphorus, sediment, fecal indicator bacteria, pathogenic bacteria, and microbial source tracking (MST) markers were examined in both prerainfall sediments and rainfall-induced runoff water. Runoff generation and sediment mobilization occurred quickly (10-150 s) after rainfall initiation. Temporal trends in runoff concentrations were highly variable within and between locations. Total runoff event loads were considered large for many pollutants considered. For instance, the maximum observed total phosphorus runoff load was on the order of 1.5 kg ha. Results also demonstrate that runoff from exposed sediments can be a source of pathogenic bacteria. spp. and spp. were present in runoff from one and three locations, respectively. Ruminant MST markers were also present in runoff from two locations, one of which hosted pasturing cattle with stream access. Overall, this study demonstrated that rainfall-induced runoff from exposed streambed sediments can be an important source of surface water pollution.

A novel experimental setup for simultaneous adsorption and induced deformation measurements in microporous materials.

A new experimental setup is presented allowing the simultaneous measurement of adsorption isotherms and adsorption-induced deformations. It is composed of a manometric technique coupled with a digital image correlation setup for full-field displacement measurements. The manometric part is validated by comparing adsorption isotherms with those obtained by a gravimetric method. The principles and methods of both adsorption isotherm and induced deformation measurements are presented in detail. As a first application of this new apparatus, the coupling between adsorption and induced deformation is characterised for a microporous media (activated carbon) saturated by pure CO2 (318.15 K, [0-60] bars) and pure CH4 (303.15 K, [0-130] bars). For this very homogeneous porous material, the induced deformation is characteristic of a pure volumetric swelling but the full-field setup may allow the characterisation of the localised pattern of deformation for heterogenous or cracked microporous media.

Bovine leukemia virus, a versatile agent with various pathogenic effects in various animal species.

The bovine leukemia virus is the etiological agent of a chronic lymphatic leukemia in cows, sheep, and goats. The same virus seems to induce a kind of wasting disease in experimentally infected rabbits. Antibodies to highly purified bovine leukemia viral Mr 51,000 glycoprotein and Mr 24,000 protein cross-react with human T-lymphotropic virus III/lymphadenopathy-associated virus antigens present in cultured lymphocytes of African patients suffering from acquired immune deficiency syndrome. Bovine leukemia virus has many structural and functional characteristics in common with the human T-lymphotropic viruses. The most striking feature of these retroviruses is the existence of a long open reading frame located at the 3' side of the provirus between the right end of the 3' side of env gene and the left end of the long terminal repeat. It is believed that the long open reading frame protein product acts in trans upon a number of genes to account for the biological effects of the virus.

Experimental infection of sheep and goat with bovine leukemia virus: localization of proviral information on the target cells.

Bovine leukemia virus (BLV) proviral integration was studied in the DNA from circulating leucocytes or tumor cells of sheep and goats experimentally infected with BLV. Southern blot analysis of infected cell DNA for BLV proviral sequences indicate that: (1) the provirus may be found as unintegrated molecules in the circulating leucocytes of infected sheep; (2) the provirus is integrated at many sites in the genome of the leucocytes of infected goats and occasionally in infected sheep; (3) the provirus is present at only a few sites in the DNA of sheep or goat tumor cell clones. A second case of goat lymphosarcoma is also reported.

Synthesis of catalytically active choline acetyltransferase in Xenopus oocytes injected with messenger RNA from rat central nervous system.

Choline acetyltransferase (ChAT) mRNA and ChAT enzymatic activity have been compared in different regions of the rat central nervous system. mRNA was assayed by exploiting the Xenopus oocyte system which was first tested by measuring the electrophysiological response to glycine after injection of mRNA derived from the ventral part of the spinal cord (VSC). This tissue was found to contain the highest ChAT mRNA level. The striatum, which yielded the maximal enzymatic activity, contained 10 times less ChAT mRNA than the VSC. These results are discussed in terms of the neuroanatomical differences between the two structures.

The conformation of purified Toxoplasma gondii SAG1 antigen, secreted from engineered Pichia pastoris, is adequate for serorecognition and cell proliferation.

A truncated form of SAG1, the immunodominant surface antigen of Toxoplasma gondii, has been produced in the methylotrophic yeast, Pichia pastoris. By construction, the recombinant protein lacks C-terminal residues 308-336 which, in native SAG1, encompass the glycosylphosphatidylinositol anchorage site. Secretion of anchor-less SAG1 proceeded via the yeast prepro alpha-mating factor signal peptide and yielded two immunoreactive protein species having apparent molecular masses of 31.5 and 34.5 kDa, respectively, and differing only by N-glycosylation of the single Asn-X-Ser site present in the molecule. Purification of the anchor-less SAG1 was achieved by a combination of ion-exchange and size-exclusion chromatographies. N-terminal amino acid sequencing of the products indicated the presence of additional residues glutamic acid--alanine at the N-terminal end of the products. Despite incomplete processing and unnatural glycosylation, anchor-less SAG1 proteins apparently adopted a suitable conformation recognized by monoclonal and human serum-derived antibodies, specific for the native SAG1. In addition, the recombinant anchor-less SAG1 proved competent for inducing proliferation, in vitro, of mononuclear cells from seropositive individuals. Finally, properly adjuvanted anchor-less SAG1 was able to induce protection of mice against a lethal challenge with T. gondii tachyzoites.

Platinum group element enrichments and possible chondritic Ru:Ir across the Frasnian-Famennian boundary, western New York State.

The Frasnian-Famennian boundary is recognized as the culmination of a global mass extinction in the Late Devonian. In western New York State the boundary is a distinct horizon within a pyritic black shale bed of the upper Hanover Shale defined by the first occurrence of Palmatolepis triangularis in the absence of Frasnian conodonts. The boundary is characterized by a minor disconformity marked by a lag concentration of conodonts. Iridium at the boundary is 0.11-0.24 ng/g, two to five times background levels of <0.05 ng/g; other Ir enrichments of 0.38 ng/g and 0.49 ng/g occur within 50 cm of the conodont-constrained boundary. Numerous Ir enrichments in the boundary interval suggest extraterrestrial accretion and platinum group element (PGE) concentration at disconformities, or mobilization and concentration in organic-rich/pyritic-rich laminations from cosmic or terrestrial sources. PGE ratios of Pt/Pd and Ku/Ir at the boundary horizon approximate chondritic ratios and are suggestive of an unaltered extraterrestrial source. These values do not conclusively establish a single extraterrestrial impact as the ultimate cause of the Frasnian-Famennian mass extinction, especially given the presence of similar Ir enrichments elsewhere in the section and the absence at the boundary of microtektites and shocked mineral grains.

Separation of platinum and palladium with silicone-rubber foam treated with dimethylglyoxime.

Open-pore silicone-rubber foam is shown to be a good inert solid support for column separations. Foams treated with dimethylglyoxime were studied for their adsorption of platinum and palladium from solution. The separation of platinum and palladium was achieved in solutions containing as little as 1.0 ppm.

Bovine leukaemia virus and enzootic bovine leukosis.

Infection of bovines with bovine leukaemia virus (BLV) manifests itself in either of two ways: 30-70% of carriers develop persistent lymphocytosis (PL), with the viral genome integrated at a large number of different sites in the DNA of the affected B-lymphocytes, without causing any chromosomal abnormalities. Only 0,1-10% of carriers develop lymphoid tumours, which also consist of B-lymphocytes. In contrast to PL, however, they are of mono- or oligoclonal origin in terms of the integration site, which is characteristic for each tumour. All cells contain one or more copies of the viral genome, chromosomal aberrations are common and if deletions are present they are invariably found in the 5'-half of the virus DNA sequence. In both types of affected cells transcription is repressed in vivo, but transient virus production can be induced in vitro and detected by means of syncytia induction or haemagglutination. In vivo production of virus in some unknown cell is suggested by the presence of high antibody titres in infected animals, especially against the envelope glycoprotein gp51. This can be detected by various techniques such as immunodiffusion, radioimmune assay or ELISA. Monoclonal antibodies against gp51 have revealed 8 epitopes, 3 of which are recognized by neutralizing antibodies and one by a cytolytic antibody. The BLV genome, about 9 kb in size, have been cloned, and some of the information obtained on its molecular structure and function is discussed. It codes for at least 4 non-glycosylated and 2 glycoproteins. Of special interest is the recently discovered serological relationship between some of the non-glycosylated proteins and those of the human T-cell leukaemia virus. The functional role of BLV in leukaemogenesis is largely unknown. The presence of the viral genome seems to be necessary for the maintenance of the transformed state, but not its continuous expression nor an LTR-mediated promotion of transcription of cellular genes. No oncogene is carried by the virus. Although bovine leukosis is not of major economic importance, its eradication is desirable and feasible in countries with a relatively low incidence, by means of testing and elimination. For endemic situations vaccination would be preferable, and distinct possibilities exist for the development of gp51 based vaccines.

Cascade ultrafiltration and competing ligand exchange for kinetic speciation of aluminium, iron, and nickel in fresh water.

Kinetic speciation of nickel, aluminium, and iron in fresh water has been investigated by cascade ultrafiltration followed by competing ligand exchange of the ultrafiltered fractions. Graphite furnace atomic absorption spectrometry was used to measure the kinetics of metal complex dissociation. Dissolved metal species were fractionated by cascade ultrafiltration. Metal speciation in each ultrafiltered fraction was then characterized as free metal ions, "labile" metal complexes (with dissociation rate constants >/=10(-3) s(-1)), "slowly labile" metal complexes (with dissociation rate constants >10(-6) s(-1)), and "inert" metal complexes (with dissociation rate constants <10(-6) s(-1)). The experimental results were compared with the predictions of a computer-based equilibrium speciation model, the Windermere humic aqueous model (WHAM) V. Cascade ultrafiltration coupled with kinetic speciation of the metal species in each molecular weight cut-off (MWCO) fraction provided a more comprehensive picture and insight into the physical and the chemical characteristics of the metal species than either ultrafiltration or measurement of dissociation kinetics alone.

Reproductive and therapeutic cloning, germline therapy, and purchase of gametes and embryos: comments on Canadian legislation governing reproduction technologies.

In Canada, the Assisted Human Reproduction Act received royal assent on 29 March 2004. The approach proposed by the federal government responds to Canadians' strong desire for an enforceable legislative framework in the field of reproduction technologies through criminal law. As a result of the widening gap between the rapid pace of technological change and governing legislation, a distinct need was perceived to create a regulatory framework to guide decisions regarding reproductive technologies. In this article the three main topics covered in the new legislation are commented on: cloning, germline therapy, and purchase of gametes and embryos. Some important issues also covered in the new legislation, such as privacy and access to information, data protection, identity of donors, and inspection, will not be addressed.

High level of genetic diversity among spelt germplasm revealed by microsatellite markers.

The genetic diversity of spelt (Triticum aestivum (L.) Thell. subsp. spelta (L.) Thell.) cultivated presently is very narrow. Although the germplasm collections of spelt are extensive, the related genetic knowledge is often lacking and makes their use for genetic improvement difficult. The genetic diversity and structure of the spelt gene pool held in gene banks was determined using 19 simple sequence repeat (SSR) markers applied to 170 spelt accessions collected from 27 countries and 4 continents. The genetic distances (1 - proportion of shared alleles) were calculated and an unweighted pair-group method with arithmetic averaging (UPGMA)-based dendrogram was generated. The genetic diversity was high: 259 alleles were found and the mean interaccession genetic distance was 0.782 +/- 0.141. The dendrogram demonstrated the much higher genetic diversity of spelt held in germplasm collections than in the currently used genotypes. Accessions with the same geographical origin often tended to cluster together. Those from the Middle East were isolated first. All but one of the Spanish accessions were found in a unique subcluster. Most accessions from eastern Europe clustered together, while those from northwestern Europe were divided into two subclusters. The accessions from Africa and North America were not separated from the European ones. This analysis demonstrates the extent of genetic diversity of spelts held in germplasm collections and should help to widen the genetic basis of cultivated spelt in future breeding programs.

Role of the 3' long open reading frame region of bovine leukemia virus in the maintenance of cell transformation.

Viral RNA expression was studied by dot blot hybridization with polyadenylated RNAs extracted from a bovine (YR-1) and an ovine (YR-2) tumor cell clone. Both clones were derived from in vivo bovine leukemia virus-induced tumors. The probes used were either the bovine leukemia virus information or only the long open reading frame sequences. No viral RNA corresponding to the bovine leukemia virus long open reading frame region was detected in YR-2, and a very limited amount of bovine leukemia virus messages was unraveled in YR-1. These results strongly suggest that viral expression, even in the long open reading frame region, is not required to maintain transformation of at least some tumor cells.

[Treatment of respiratory distress caused by laryngeal obstruction]. / Traitement des détresses respiratoires par obstacle laryngé

Respiratory distress due to laryngeal obstruction produces a more or less dramatic syndrome depending on the cause of the obstruction and its evolution in time. Treatment should be immediate in order to avoid death or cerebral and cardiac sequelae. It consists, above all, of restoring the patency of the respiratory tract, the short-circuit obtained by intubation or tracheotomy should, in no case, leave any laryngeal sequelae. The authors present their personal series based on a study of respiratory distress due to laryngeal obstruction in the child. These observations were obtained from a large series of 480 cases of laryngeal dyspnea observed over the last 5 years on the E.N.T. unit of the Edouard Herriot Hospital in Lyons. They contrast a very small number of sub-glottic laryngitis cases requiring intubation or tracheotomy to a very large number of cases of subglottic laryngitis, even with respiratory distress, which recovered with well designed medical treatment. Foreign bodies, of which they report eleven cases, all required emergency surgery. They emphasize the severity of this cause. As far as papillomas and angiomas are concerned, they require emergency treatment, either straightaway or after a trial of medical treatment.

[Emergency treatment of tracheal and bronchial foreign bodies]. / Traitement des corps étrangers trachéaux et bronchiques en urgence

Basing their report on personal experience of more than 300 tracheal or bronchial foreign bodies, the authors emphasize: -the difficulty of extraction, even in very expert hands. -the absolute necessity of a fully trained operator, and experienced anaesthetist and appropriate material. Apart from acute respiratory distress, they consider preferable to defer operation for a few hours, under strict supervision, in order to make available all these optimal conditions.

Determination of trace elements in granites by inductively coupled plasma mass spectrometry.

Proposed is a simple and reliable method for the dissolution of granite and the determination of 38 elements by inductively coupled plasma mass spectrometry. One hundred milligrams of sample are digested with 1 ml of HF and 0.5 ml of HNO(3) in screw top PTFE-lined stainless steel bombs at 190 degrees C for 12 h. Insoluble residues are dissolved using 8 ml of 40% HNO(3) (v/v) heated to 110 degrees C for 3 h. Six granite standard reference materials (GSR-1, JG-2, G-2, NIM-G, SG-3, SG-1a) were studied. Analytical calibration was accomplished using aqueous standard solutions. Rhodium was used as an internal standard to correct for matrix effects and instrument drift. We report data for: Li, Be, Sc, V, Cr, Co, Ni, Cu, Zn, Ga, Rb, Sr, Y, Zr, Nb, Mo, Cs, Ba, Hf, Ta, W, Pb, Th, U and 14 of the rare earth elements. The recoveries for most of these elements in granite ranged from 90 to 110%.

Comparison of thermostable macromolecular antigens from leprosy-associated bacteria.

Three types of bacteria are associated with leprosy: Mycobacterium leprae, leprosy-derived corynebacteria (LDC), and armadillo-derived mycobacteria (ADM). The immunological relationships between these three types of bacteria and Mycobacterium bovis BCG, used as a reference, were determined by cross-immunoelectrophoresis. When compared with the reference, cross-reactions were observed with a variable number of antigens: 2 in the case of strain LDC 15, 4 with M. leprae, and from 1 to 10 in the case of the ADM, depending on their subgroup. Next, thermostable macromolecular antigens (TMAs), the major cross-reactive antigens of leprosy-associated bacteria, were compared by anti-TMA antibody ELISA tests. The LDC TMAs displayed high cross-reactivity between the subgroups and lower cross-reactivity with the TMAs of M. bovis BCG. Evidence for the presence of a species-specific moiety in TMA of the different LDC was obtained by using depleted anti-TMA antisera. Western blot analysis revealed the presence of many proteins in the TMAs of LDC and M. bovis BCG, some of them being species-specific and other cross-reactive.