Water quality trends in the last decade for ten watersheds dominated by diffuse pollution in Québec (Canada).

The aim of this work is to evaluate and discuss river water quality trends over the last decade in ten watersheds where diffuse pollution represents more than half of the annual load of phosphorus (P) and nitrogen (N). Trend analyses taking into account flow data indicate a significant reduction of total P in eight rivers, of ammonia N in five rivers, of nitrate + nitrite in four rivers, of total filtered N in three rivers and of suspended solids in two rivers. An increase of turbidity was observed in four rivers and, for fecal coliforms, no trends. P decrease can be explained by reduced mineral P inputs on cropped lands related to means such as agro-environmental fertilization plans and addition of phytase in pig and poultry feed. However, for seven of them, median P concentrations remain at least two times greater than the Québec water quality guideline for protection of rivers against eutrophication. Concentrations of other parameters remain problematic in some rivers too. These results indicate the need to continue the efforts for further diffuse pollution reduction. Future work should better quantify actions taken at the watershed scale to reduce diffuse pollution.

Suppression of valid inferences and knowledge structures: the curious effect of producing alternative antecedents on reasoning with causal conditionals.

These studies looked at the difficulty that reasoners have in accepting conditional ("If P then Q") major premises that are not necessarily true empirically, as a basis for deductive reasoning. Preliminary results have shown that when reasoners are asked to produce possible alternate antecedents to the major premise ("If A then Q"), they paradoxically tend to deny the modus ponens (MP) inference ("If P is true, then Q is true"). Three studies further explored these results. The first study gave university students paper-and-pencil tests in which instructions to "suppose that the major premise is true" was followed by a request to determine the next number in a sequence, to retrieve information unrelated to the premises, or to retrieve a possible case of "If A then Q." Relative to a control group, reasoners asked to produce an alternative antecedent showed a significant tendency to deny the MP inference, whereas no such tendency was observed for the two other tasks used. A second study compared performance on a condition in which reasoners were asked to produce an alternative antecedent with that when they were given an explicit alternative. Premises used in this study were such that the latter alternative antecedent was also spontaneously produced by over 70% of reasoners. Results showed that the tendency to refuse the MP premise could not be accounted for by the specific nature of the alternative produced. A third study found that the tendency to refuse the MP inference after producing an alternative antecedent was affected by the number of "disabling conditions" (i.e., conditions that allow "P to be true" and "Q to be false") available for the major premise. These results are interpreted as being consistent with a model that supposes that logical reasoning requires selective inhibition of real-world knowledge.

Mechanisms of action of antimalarials in inflammation: induction of apoptosis in human endothelial cells.

Antimalarials are beneficial therapeutic agents in systemic lupus and rheumatoid arthritis. These autoimmune diseases have abnormally low apoptosis of inflammatory cells. Both disorders have an abnormal angiogenesis. In the present report, antimalarials were demonstrated to selectively increase apoptosis of HUVECs in vitro. A 24-h exposure to 50 or 150 microM of the drugs was associated with a significant loss of substrate-adherent cells. Chloroquine exhibited an inhibitory effect on HUVEC proliferation over 7 days. Programmed cell death in HUVECs rendered nonadherent by chloroquine was confirmed by the induction of DNA fragmentation in floating cells. Northern blot analysis revealed a rapidly increased expression of the bcl-x(s) gene without any change in the expression of the bcl-2 gene, indicating that HUVECs under chloroquine were undergoing apoptosis. The onset of the apoptotic cascade in HUVECs appeared shortly after the addition of chloroquine. The effect of chloroquine on apoptosis was distinct from acute cell lysis and was restricted to HUVECs. Antimalarials also induced IL-1alpha production. In parallel, chloroquine alone did not increase the expression of IL-6. Anti-IL-1alpha Ab or IL-1Ra only marginally reversed chloroquine-induced depression of proliferation for the low drug concentration, but not the massive cell death effect at and above 50 microM. Taken together, these data may indicate that antimalarials repress angiogenesis. The autocrine mechanism involving IL-1alpha accounts only for a minor fraction of the full antiendothelial effect of chloroquine, which is mainly dependent on apoptosis.

Equilibrium model in an in vitro poly(ADP-ribose) turnover system.

Poly(ADP-ribose) metabolism plays an important role in numerous DNA-related functions. This homopolymer is synthesized by poly(ADP-ribose) polymerase and is degraded mainly by the poly(ADP-ribose) glycohydrolase. The activities of these two enzymes in the nucleus are closely coordinated. To better understand the interactions between these enzymes, we designed an in vitro system in which both enzymes are present at the same time. In this work, we report a model describing the synthesis and degradation of the poly(ADP-ribose) in turnover conditions. Because the half-life of the polymer in the cell is close to 1 min, we studied the very early kinetic interactions of these two enzymes.

The US-1 element from the gene encoding rat poly(ADP-ribose) polymerase binds the transcription factor Sp1.

By comparing the upstream DNA sequence of the rat and human genes encoding poly(ADP-ribose) polymerase (PARP), we have defined a 16-bp conserved region and designated it as US-1 for 'upstream sequence 1'. This element is homologous to the recently described binding site for the transcription factor Sp1 in the promoter sequence of the mouse p12 gene which encodes a protease inhibitor. Analyses in gel mobility shift assays revealed that a nuclear protein, produced by all tissue-culture cells tested, specifically binds the US-1 element. The pattern of shifted DNA protein complexes obtained was strikingly similar to that for Sp1, which is supported by the positive displacement of these complexes by an oligomer containing the Sp1 binding site in gel shift competition experiments. Replacement of the Sp1 binding site from the basal promoter of the mouse p12 gene by the rPARP US-1 element did not result in any significant variations in the level of expression of the chloramphenicol acetyltransferase (CAT) reporter gene upon transient transfection of tissue-culture cells. However, when point mutations are introduced in the US-1 element in a similar substitution experiment, a significant reduction in CAT gene expression could be observed. These data are consistent with Sp1 interacting with the US1 element. Results from DNase I footprinting experiments clearly indicated that purified Sp1 not only binds to the US-1 element but also to four other closely located cis-acting sites scattered in the promoter of the rat PARP gene, therefore suggesting that Sp1 is likely to modulate strongly the expression of that gene in different tissues.

Expression in Escherichia coli of the 36 kDa domain of poly(ADP-ribose) polymerase and investigation of its DNA binding properties.

We have expressed in Escherichia coli the 36 kDa domain of the human poly(ADP-ribose) polymerase. This polypeptide comprises the C-terminal part of the DNA binding domain, as well as the automodification region of the enzyme, but lacks the zinc-finger motifs of the N-terminal region and the C-terminal catalytic domain. By probing the crude E. coli protein extracts with radioactive DNA probes (South-Western blots), we have shown that the 36 kDa domain binds a DNA probe of 222 bp but does not bind a shorter probe of 66 bp. This interaction is stronger when the polypeptide is fused to the 55 kDa catalytic domain of the enzyme.

Structural analysis of the putative regulatory region of the rat gene encoding poly(ADP-ribose) polymerase.

A lambda EMBL3 clone containing the first three exons along with part of the 4th exon of the rat poly(ADP-ribose) polymerase gene was isolated from a genomic DNA library. This clone also contains 6.6 kbp of upstream sequences. Nucleotide sequence analysis of the proximal 5' 670 nucleotides flanking the major RNA start site of the rat gene does not reveal significant global homology with the same region of the human gene, but a series of short sequences are identical. Among these sequences are found two putative Sp1 binding sites along with a decanucleotide sequence responsible for the attachment of the transcription factor AP-2.

Pattern of cadmium contamination in the liver and kidneys of moose and white-tailed deer in Québec.

Samples of moose (N = 431) and white-tailed deer (N = 225) liver and kidneys were collected during the 1985 hunting season from 14 zones south of the 50 degree latitude in Québec. Regional differences in cadmium level in the liver were detected and three homogeneous areas were delineated for each species. Uptake was greater for moose than for deer: in the liver, mean concentrations were 2.9-15.9 mg kg-1 (dry weight) for moose and 0.8-2.6 for deer, depending on the area and sex; in kidneys, means ranged between 31.8-100.5 and 20.9-39.0 mg kg-1, respectively. Female moose had lower levels than bulls. Less affected moose, in eastern Québec, contained cadmium concentrations comparable to the highest values measured in Scandinavia. Cadmium uptake in deer was on the same level or higher than in the United States. Our results indicate a widespread presence of this heavy metal in the environment that may be linked to acid precipitation. We do not recommend consuming wild cervid liver or kidneys in Québec for the moment. Further research is needed on the overall mechanisms involved in the cadmium contamination of the environment and on the actual intake of this metal in the human diet.