Biomedical subjects

M Raymond

Publications and source records attributed to M Raymond.

At least 55 records · Page 3Linked to original sources

PubMed1997

Aedes polynesiensis in the Society Islands: environmental correlates of isoenzyme differentiation.

Isoenzyme genetic differentiation of Aedes polynesiensis mosquitoes in Raiatea island, French Polynesia, was evaluated by two models of population structure based on seven gene-enzyme systems: Ak, Est, Got, Gpi, Hk, Mdh and Pgm. The ecological model tested whether genetic differentiation is congruent with habitat differences. The isolation model evaluated whether genetic differentiation is proportional to geographical distribution. The ecological model found no significant differentiation between populations of Ae.polynesiensis from beach and forest ecotopes, whereas the isolation model was consistent with the data. However, incipient speciation is opposed by the source-sink system of population dynamics in such small neighbouring islands, where Ae.polynensiensis extinction is readily followed by reinvasion causing considerable gene flow between island populations.

Aedes↗

PubMed1997

Analysis of culture filtrate and cell wall-associated antigens of Mycobacterium paratuberculosis with monoclonal antibodies.

Proteins secreted by Mycobacterium species have been suggested as major immune targets in the early phase of infection. In this study, we sought to identify specific antigens in culture filtrates and in soluble cell extracts of Mycobacterium paratuberculosis. The release of antigens into the culture medium during growth of the bacilli and the distribution of specific epitopes within the Mycobacterium species were investigated by immunoblot analysis with monoclonal antibodies (MAbs) raised against M. paratuberculosis antigens. MAb B6A interacted with a cellular antigen with an apparent molecular mass of 34.5 kDa in lysates of M. paratuberculosis. MAb B6A did not interact with lysates from any other mycobacterial species, suggesting recognition of an M. paratuberculosis species-specific epitope. MAb FL1-A1 reacted with an antigen of 44.3 kDa in M. paratuberculosis and a 9-kDa antigen in Mycobacterium kansasii. MAb PII-B1 reacted with concanavalin A (ConA)-binding cellular and filtrate molecules of M. paratuberculosis and with lysates of Mycobacterium kansasii and Mycobacterium avium 18. The affinity-purified glycosylated antigens migrated as a diffuse band of between 35 and 45.6 kDa and reacted strongly with ovine and bovine paratuberculosis serum and polyclonal serum against M. tuberculosis lipoarabinomannan antigens. These glycoconjugates were the earliest antigens detected in culture filtrates of M. paratuberculosis. Deglycosylation of the ConA-binding molecules with alpha-mannosidase enzyme abolished the reaction with MAb PII-B1 and with bovine but not ovine paratuberculosis serum, suggesting selective immunogenicity in the different animal species.

Animals↗

PubMed1997

The Candida albicans CDR3 gene codes for an opaque-phase ABC transporter.

We report the cloning and functional analysis of a third member of the CDR gene family in Candida albicans, named CDR3. This gene codes for an ABC (ATP-binding cassette) transporter of 1,501 amino acids highly homologous to Cdr1p and Cdr2p (56 and 55% amino acid sequence identity, respectively), two transporters involved in fluconazole resistance in C. albicans. The predicted structure of Cdr3p is typical of the PDR/CDR family, with two similar halves, each comprising an N-terminal hydrophilic domain with consensus sequences for ATP binding and a C-terminal hydrophobic domain with six predicted transmembrane segments. Northern analysis showed that CDR3 expression is regulated in a cell-type-specific manner, with low levels of CDR3 mRNA in CAI4 yeast and hyphal cells, high levels in WO-1 opaque cells, and undetectable levels in WO-1 white cells. Disruption of both alleles of CDR3 in CAI4 resulted in no obvious changes in cell morphology, growth rate, or susceptibility to fluconazole. Overexpression of Cdr3p in C. albicans did not result in increased cellular resistance to fluconazole, cycloheximide, and 4-nitroquinoline-N-oxide, which are known substrates for different transporters of the PDR/CDR family. These results indicate that despite a high degree of sequence conservation with C. albicans Cdr1p and Cdr2p, Cdr3p does not appear to be involved in drug resistance, at least to the compounds tested which include the clinically relevant antifungal agent fluconazole. Rather, the high level of Cdr3p expression in WO-1 opaque cells suggests an opaque-phase-associated biological function which remains to be identified.

ATP-Binding Cassette Transporters↗

PubMed1997

Esterase gene amplification in Culex pipiens.

In the mosquito Culex pipiens one of the major resistance mechanisms to organophosphorous pesticides (OPs) is increased detoxification of insecticide. This resistance is the consequence of overproduction of two types of esterases, esterases A and B, coded at two loci, Est-3 (A esterase) and Est-2 (B esterase). We have analysed the genomic structure of these genes in different strains resistant to OPs and have attempted to characterize the different types of mutations leading to the resistant phenotypes. It is shown that, concerning the more frequent resistant phenotypes, mutations leading to resistance are of two main types. First, overproduction of one A esterase present in Southern France results from a regulatory mechanism. The second type of mutation is gene amplification which involves events that have initially generated the duplication of both the A and B esterase or only the B esterase locus. We report the point that the most frequent esterase overproductions are the results of eight different mutations and that, given the range of distribution of these genotypes, mutation leading to an efficient resistance gene is one of the most limiting factors for the evolution toward resistance in Culex pipiens.

Animals↗

PubMed1996

Frequency-dependent maintenance of left handedness in humans.

The percentage (10-13%) of left handedness in human has apparently not changed since the Neolithic. Left handedness is heritable and appears to be repeatedly associated with some reduced fitness components; the persistence of left handedness implies that left handers have a fitness advantage in some situations. We propose that left handers have a frequency-dependent advantage in fights and for that reason a fitness advantage. To test this hypothesis, left handedness frequencies in the general population and in sporting individuals (both students and the sporting elite) have been compared, as sporting performance is likely to be a good indicator of fighting abilities. The higher proportion of left-handed individuals in interactive sports (reflecting some fighting elements), reaching 50% in some sports categories, but not in noninteractive sports, is consistent with the fighting hypothesis. The greater frequency of left handedness in males than in females is also consistent with this hypothesis, as male-male fights are universally more frequent than other combinations. The frequency-dependent advantage in fights of left handers might explain the stability of left handedness.

Adolescent↗

PubMed1996

The influence of time of administration on the pharmacokinetics of a once-a-day diltiazem formulation: morning against bedtime.

Twenty-three young, healthy, male volunteers received, in a randomized crossover design, 240 mg of a once-a-day diltiazem formulation at 08:00 (AM) or 22:00 (HS) for 6 days. A 7 day washout period was observed between the two modes of administration. Diltiazem plasma concentrations were monitored every hour for 24 h and at 30, 36, and 48 h after the last dose. Differences were found between AM and HS dosing for Cmin (mean (SD) = 47 center dot 2 (25 center dot 8) against 39 center dot 6 (21 center dot 1) ng mL-1, p = 0 center dot 038), AUC0-24 (2008 (814) against 1754 (714) ng h mL-1, p = 0 center dot 024), and AUC0-48 (2662 (1244) against 2395 (238) ng h mL-1, p = 0 center dot 034). Overall the two modes of administration did not produce bioequivalent pharmacokinetic profiles. Also HS dosing gave significantly higher plasma concentrations of diltiazem in the early morning hours when the incidence of cardiovascular events is higher. If one assumes a strong correlation between plasma concentrations and myocardial protection then HS dosing should be recommended for QD formulation of diltiazem. Clinical studies should be performed to confirm this theoretical pharmacokinetic advantage.

Adult↗

PubMed1996

Determination of Ace.1 Genotypes in Single Mosquitoes: Toward an Ecumenical Biochemical Test

The occurrence of two acetylcholinesterases, AChE1 and AChE2, in the mosquito Culex pipiens has been recently documented. Resistance to organophosphates and carbamates due to target insensitivity is the result of a qualitative change of only AChE1, encoded by the Ace.1 gene. Because AChE1 and AChE2 differ in their sensitivity to inhibitors, Ace.1 genotypes can be misclassified by previous tests. We describe a new rapid microplate test that allows unambiguous identification of Ace.1 genotypes. This test involves comparing AChE activities in the absence of insecticide and in the presence of two propoxur concentrations: a low concentration that inhibits only the sensitive AChE1 and a higher concentration that inhibits also AChE2 but not the insensitive AChE1 responsible of insecticide resistance. This comparison allows the identification of the three Ace.1 genotypes: resistant (Ace.1RR), susceptible (Ace.1SS) homozygotes, and heterozygotes (Ace.1RS). The similarity of propoxur sensitivity of modified AChE1s found in various resistant strains from the United States, Europe, and Africa indicates that this test is probably suitable for all the Ace.1 alleles described so far in C. pipiens.

Journal Article↗

PubMed1996

Duplication of the Ace.1 locus in Culex pipiens mosquitoes from the Caribbean.

In Culex pipiens mosquitoes, AChE1 encoded by the locus Ace.1 is the target of organophosphorus and carbamate insecticides. In several resistant strains homozygous for Ace.1RR, insensitive AChE1 is exclusively found. An unusual situation occurs in two Caribbean resistant strains where each mosquito, at each generation, displays a mixture of sensitive and insensitive AChE1. These mosquitoes are not heterozygotes, Ace.1RS, as preimaginal mortalities cannot account for the lethality of both homozygous classes. This situation is best explained by the existence of two Ace.1 loci, coding, respectively, a sensitive and an insensitive AChE1. Thus, we suggest that in the Caribbean a duplication of the Ace.1 locus occurred before the appearance of insecticide resistance at one of the two copies.

Acetylcholinesterase↗

PubMed1996

Lithium poisoning treated by high-performance continuous arteriovenous and venovenous hemodiafiltration.

Intermittent hemodialysis is considered the modality of choice when enhanced lithium removal is indicated. However, postdialysis rebound in serum lithium concentration is frequently observed after the dialysis sessions and results from incomplete intracellular removal. Continuous renal replacement therapy could provide a more gradual and complete lithium removal since it is performed over longer time periods, thus avoiding rebound following therapy. Seven patients presenting with symptomatic lithium intoxication were treated by continuous renal replacement therapy (continuous arteriovenous and venovenous hemodiafiltration [CAVHDF and CVVHDF]). For CAVHDF, the dialysate flow rate was increased to 4 L/hr to optimize solute clearances. Five intoxicated patients (four acute and one chronic) were treated by high dialysate flow rate (HDFR) (4 L/hr) CAVHDF and two patients with chronic poisoning were treated by CVVHDF, one with a dialysate flow rate of 1 L/hr and one with a dialysate flow rate of 2 L/hr. Serum lithium concentrations for the four acute poisoning cases were 4.0, 4.6, 4.4, and 3.2 mEq/L, at initiation of HDFR CAVHDF, and decreased respectively to 1.2, 0.8, 1.2, and 1.1 mEq/L after 15, 19, 35, and 21 hours of treatment. No lithium rebound was observed over 24 to 36 hours following CAVHDF. For the three chronic intoxication cases, serum lithium concentrations dropped from 1.7, 2.2, and 3.8 mEq/L to 0.7, 0.17, and 0.4 mEq/L, respectively, after 18, 42, and 44 hours of HDFR CAVHDF or CVVHDF. The chronic case treated for only 18 hours presented a slight rebound in lithium level (0.3 mEq/L), whereas no significant rebound was observed for the two other cases treated for longer periods. Mean +/- SEM dialyser urea, lithium, and creatinine clearance during HDFR CAVHDF were 50.5 +/- 5.0, 41.4 +/- 4.6, and 37.6 +/- 3.7 mL/min, respectively (number of measurements = 41). Dialyser lithium clearance during CVVHDF was 48.4 +/- 1.4 mL/min (n = 10) and 61.9 +/- 2.3 mL/min (n = 7), with dialysate flow rates of 1 and 2 L/hr, respectively. Mean dialyzer lithium removal for the seven cases was 106.4 mEq, while mean renal lithium removal was 21.5 mEq during the same period. We conclude that HDFR CAVHDF and CVVHDF are effective alternatives to intermittent hemodialysis for treatment of lithium poisoning. They provide excellent lithium clearances (60 to 85 L/d); in addition, because of their continuous nature, they prevent posttherapy lithium rebound by allowing a more gradual and complete removal from intracellular compartments, and they may be particularly useful in chronic poisoning in which intracellular lithium accumulation is more extensive.

Acute Disease↗

PubMed1996

Esterase polymorphism in insecticide susceptible populations of the mosquito Culex pipiens.

Gene amplification involving a particular haplotype has been found at the esterase B locus of mosquitoes from various countries. This similarity has been explained by a unique amplification event followed by migration and selection by organophosphate (OP) insecticides. This assumes that the polymorphism of non-amplified esterase haplotypes is so large that the chance of independent amplification in two distinct populations is negligible. In order to test this assumption, three susceptible populations from northern Europe were sampled and analysed for esterase and haplotype polymorphism. At the protein level, 18 and 16 alleles were found for esterase A and B respectively in one French population (n = 74), and 16 and 14 in an English one (n = 50). At the DNA level, 24 alleles at the esterase B locus were detected in a sample of 72 mosquitoes from one population, with the use of only one restriction enzyme (EcoR V). Restriction maps of two non-amplified haplotypes randomly sampled from a single breeding site in Belgium were built with six restriction enzymes. 60% of all restriction sites were different among the two maps. The huge polymorphism found in northern Europe requires specific explanations for its stability, but it considerably strengthens the hypothesis of migration of amplified haplotypes.

Animals↗

PubMed1996

Testing the unique amplification event and the worldwide migration hypothesis of insecticide resistance genes with sequence data.

In the mosquito Culex pipiens, over-production of esterases is a common insecticide resistance mechanism. Different alleles at the esterase A and B loci are known to occur in natural populations, as shown by enzyme electrophoresis and RFLP studies on the esterase B locus. Here we analyse the variability of the esterase A locus at the nucleotide level in mosquitoes possessing or lacking over-produced esterase A. A surprisingly high level of nucleotide polymorphism is found in coding and noncoding regions, and the extent of polymorphism detected is higher than that previously described for the most polymorphic loci in Drosophila. We also show that eight strains from different localities (Africa, America, Asia) that possess the over-produced esterase A2 share the same nucleotide sequence at the esterase A locus, strongly supporting the evolutionary scenario of a unique event of amplification of this gene followed by dispersal through migration rather than the occurrence of multiple independent mutational events of the gene.

Amino Acid Sequence↗

PubMed1996

Existence of two acetylcholinesterases in the mosquito Culex pipiens (Diptera:Culicidae).

Two acetylcholinesterases (AChEs), AChE1 and AChE2, differing in substrate specificity and in some aspects of inhibitor sensitivity, have been characterized in the mosquito Culex pipiens. The results of ultracentrifugation in sucrose gradients and nondenaturing gel electrophoresis of AChE activity peak fractions show that each AChE is present as two molecular forms: one amphiphilic dimer possessing a glycolipid anchor and one hydrophilic dimer that does not interact with nondenaturing detergents. Treatment by phosphatidylinositol-specific phospholipase C converts each type of amphiphilic dimer into the corresponding hydrophilic dimer. Molecular forms of AChE1 have a lower electrophoretic mobility than those of AChE2. However, amphiphilic dimers and hydrophilic dimers have similar sedimentation coefficients (5.5S and 6.5S, respectively). AChE1 and AChE2 dimers, amphiphilic or hydrophilic, resist dithiothreitol reduction under conditions that allow reduction of Drosophila AChE dimers. In the insecticide-susceptible strain S-LAB, AChE1 is inhibited by 5 x 10(-4) M propoxur (a carbamate insecticide), whereas AChE2 is resistant. All animals are killed by this concentration of propoxur, indicating that only AChE1 fulfills the physiological function of neurotransmitter hydrolysis at synapses. In the insecticide-resistant strain, MSE, there is no mortality after exposure to 5 x 10(-4) M propoxur: AChE2 sensitivity to propoxur is unchanged, whereas AChE1 is now resistant to 5 x 10(-4) M propoxur. The possibility that AChE1 and AChE2 are products of tissue-specific posttranslational modifications of a single gene is discussed, but we suggest, based on recent results obtained at the molecular level in mosquitoes, that they are encoded by two different genes.

Acetylcholinesterase↗

PubMed1996

Dominance of insecticide resistance presents a plastic response.

Dominance level of insecticide resistance provided by one major gene (an insensitive acetylcholinesterase) in the mosquito Culex pipiens was studied in two distinct environments. Dominance level was found to be very different environments, varying from almost complete dominance to almost recessive when either propoxur (a carbamate insecticide) or chlorpyrifos (an organophosphorus insecticide) was used. To better understand this plastic response, three environmental parameters were manipulated and their interactions studied. For chlorpyrifos, each parameter had a small effect, but when all parameters were changed, the dominance level was greatly affected. For propoxur, one environmental parameter had a large effect by itself. It was further studied to understand the causal relationship of this plasticity. Recessivity of resistance was associated with more demanding environments. These results are discussed in the context of the various theories of the evolution of dominance. It appears that dominance of insecticide resistance cannot be directly predicted by Wright's physiological theory.

Animals↗

PubMed1996

Testing differentiation in diploid populations.

We examine the power of different exact tests of differentiation for diploid populations. Since there is not necessarily random mating within populations, the appropriate hypothesis to construct exact tests is that of independent sampling of genotypes. There are two categories of tests, FST-estimator tests and goodness of fit tests. In this latter category, we distinguish "allelic statistics", which account for the nature of alleles within genotypes, from "genotypic statistics" that do not. We show that the power of FST-estimator tests and of allelic goodness of fit tests are similar when sampling is balanced, and higher than the power of genotypic goodness of fit tests. When sampling is unbalanced, the most powerful tests are shown to belong to the allelic goodness of fit group.

Alleles↗

PubMed1996

An insensitive acetylcholinesterase in Culex pipiens (Diptera:Culicidae) from Portugal.

Resistance mechanisms of a strain (PRAIAS) of northern house mosquito, Culex pipiens L., collected in Portugal in 1993, and highly resistant to organophosphates and carbamates, were investigated by comparing the resistance characteristics to 3 organophosphorous (temephos, chlorpyrifos, malathion) and 1 carbamate (propoxur) insecticides in the presence or absence of synergists; and by determining the possible occurrence of overproduced esterases or insensitive acetylcholinesterase (AChE). The reference strain MSE from southern France, with an insensitive AChE, was included in all analyses for comparison. For organophosphorous insecticides, resistance in PRAIAS was caused by an insensitive AChE and an increase in oxidative metabolism, although the 2nd mechanism has only a marginal effect. For propoxur, the insensitive AChE was the only resistance mechanism detected. Biochemical properties of both the French and Portuguese insensitive AChEs were similar. We cannot exclude the possibility that PRAIAS and MSE strains possess exactly the same insensitive AChE allele.

Acetylcholinesterase↗

PubMed1996

Coamplification of esterase A and B genes as a single unit in Culex pipiens mosquitoes.

In Culex pipiens mosquitoes, resistance to organophosphorous insecticides often results from increased detoxification by two types of esterases, A and B, which are closely linked. Overproduction of all esterase B so far investigated (B1, B2, B4, B5 and B6) is from gene amplification. An esterase A gene (esterase A2) has recently been cloned from mosquitoes with the overproduced esterases A2 and B2, and amplification of this gene has also been reported. We describe the cDNA sequences of three additional esterase genes from insecticide-resistant strains of Culex pipiens originating from France and California which show at least 93 per cent homology with the esterase A2 gene sequence. Restriction enzyme mapping shows that the esterase A gene lies within 2.2 kb of the esterase B gene. In mosquitoes with overproduced esterases A2 and B2, the amplification level of esterase A is equal to that of esterase B suggesting that the genes are coamplified. Furthermore, we show that in one strain with an overproduced A esterase (A1), gene amplification cannot account for the increased protein level. This indicates that overproduction of esterases A can be achieved through two different mechanisms: gene amplification and a regulatory mechanism--the nature of which remains to be identified.

Animals↗

PubMed1996

[Resistance of insects to insecticides. Molecular mechanisms and epidemiology].

The aim of this paper is to indicate the main strengths which are involved in the spread of insecticide resistance genes. These forces are more or less well known in population genetics, there are: mutations which are involved in the creation of new alleles well adapted to insecticides; migration which is responsible for the geographical extension of resistance and selection which screened among the various alleles those which are best adapted in the geographical and ecological context. The biological model described in this paper is the mosquito Culex pipiens which is the best known model at that time. Many works have been done on the resistance to insecticides in this species. On the other hand, the molecular mechanisms involved in insecticide resistance have been described from various invertebrates.

Animals↗

PubMed1995

Commensalism, adaptation and gene flow: mosquitoes of the Culex pipiens complex in different habitats.

Two ecotypes have been described for Culex pipiens mosquitoes of the temperate zone: a human commensal type and a feral type, but their degree of evolutionary differentiation and taxonomic status are still unclear. The commensal form is characterized by life-history traits probably adaptive to underground man-made environments. This situation has sometimes been considered as an example of recent speciation although the existence of intermediate forms indicates that the balance between gene flow and disruptive selection should first be assessed. The present study was concerned with (1) the determination of biological traits involved in adaptation to commensalism, and (2) the pattern of gene flow within and between ecotypes in a restricted area. It was found that (1) significant differences in biological traits exist between mosquitoes from different habitats, (2) characteristics of the commensal type are not universal in mosquitoes from underground man-made habitats, (3) allozyme markers do not clearly differentiate ecotypes and (4) insecticide resistance genes, which reveal recent migration, occur in each ecotype. These results are discussed in the context of possible speciation due to commensalism.

Adaptation, Physiological↗