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Biomedical subjects

S Noël

Publications and source records attributed to S Noël.

At least 19 recordsLinked to original sources

Molecular identification of two species of myiasis-causing Cuterebra by multiplex PCR and RFLP.

The myiasis-causing flies Cuterebra grisea (Coquillet) and Cuterebra fontinella (Clark) (Diptera: Oestridae) are normally parasites of mice, predominantly of the genus Peromyscus. The morphological similarities of these species and the existence of intermediate morphotypes bearing characters of both species make the identification of adults problematic; furthermore the identification of larvae is apparently not possible. This study presents two molecular approaches to discriminate between these species using specific band patterns: (i) species-specific primers designed in the cytochrome oxidase II (COII) region used in multiplex polymerase chain reaction (PCR) and (ii) restriction fragment length polymorphism (RFLP) on amplified segments of cytochrome oxidase I (COI) gene. Both methods were tested on Cuterebra larvae and on adult museum specimens. The two techniques showed a clear difference between C. grisea and C. fontinella, although species-specific primers were more successful than RFLP for degraded DNA. No intraspecific variation in RFLP and species-specific amplifications were detected for the two species of Cuterebra. The results exhibit discrepancies between molecular and morphological identification, suggesting that some of the adults were misidentified.

Animals↗

The heterogeneity of sprout inhibitor application with chlorpropham.

The CIPC or chlorpropham is used on potatoes as "sprouting inhibitor". These lasts years, some set of belgian potatoes treated by CIPC exceeded the Maximum Residue Limit (MRL), fixed at 5 mg/kg. The heterogeneity of sprout inhibitor application would be one of the causes of over-dosage. In order to estimate the distribution of CIPC between potatoes, according to the formulation used (DP, EC and HN), a research project financed by "le fond budgétaire des matières premieres" has been set up. In order to evaluate the distribution of CIPC into the pile of potatoes, the efficiency of the different formulations as well as the residues caused by their application on tubers, some tests have been placed in the storage hall. These tests consist in taking off some samples at different places into the pile of potatoes during storage and destocking. The content of CIPC is analysed by capillary gas chromatography with detection by mass spectrometry detection (GC-MS) or nitrogen specific detection (GC-NPD). To estimate the efficiency, the sprouting are observed every month and samples of potatoes are taken off on the top of the pile to control the evolution of CIPC content. Considering the results of the tests, the inhibitor treatment with the formulation DP+HN seems to be the more efficient. As for the residues, the formulation DP leads to a higher content of CIPC than HN, that has a very low concentration. After treatment, the quantity really applied on tubers depends on the formulation used. In fact, 50% of CIPC applied by DP formulation are found on tubers but hardly 10%, after gas application.

Belgium↗

[Neurologic and muscular mechanisms of fatigue].

Fatigue of voluntary muscular effort is a complex phenomenon, influenced by peripheral and central nervous system factors. Peripheral mechanisms of fatigue include impairment in neurotransmission and impulse propagation down the sarcolemna, dysfunction within the sarcoplasmic reticulum involving calcium release and uptake, impairment in the actin-myosin cross-bridge interactions and substrate depletion or accumulation of metabolites. The central fatigue is associated with reduction of motor cortical excitability, failure to drive the motoneurons adequately, changes in neurotransmitter activity, etc. Is fatigue an unexplained disabling symptom or a protective mechanism to prevent muscle fiber degeneration?

Diagnosis, Differential↗

Study of potatoes' sprout inhibitor treatments with chlorprophame.

Studies carried out in 1999 by the University of Ghent showed that 36% of potatoes' samples contained Chlorprophame (CIPC) residues and that 7.9% of them exceeded the maximal limit of residues (RML), fixed at 5 ppm. The heterogeneity of sprout inhibitor application would be one of the causes of over-dosage. However, this heterogeneity would also cause under-dosages leading to problems when controlling the sprouting in potatoes stored over 6 degrees C. This study aims at determining some technical causes of the heterogeneity of CIPC sprout inhibitor treatments when storing potatoes. The study concerns two treatment techniques: dusting and spraying. To draw up an inventory of mechanical treatments in Belgium, a survey has been conducted among 28 farmers throughout Belgium. 35 samples have been taken at random in the different storage rooms to analyse the content of CIPC residue. In order to do so, a method of analysis: the gas chromatography in capillary phase with detection by mass spectrophotometry, has been developed. Tests have been carried out by changing several parameters such as the material, the product or the place in the storage line, in order to assess the CIPC application techniques. The survey made it possible to analyse qualitatively, from the declarations of farmers, the causes of heterogeneity linked to treatment techniques. An almost systematically over-dosage of the CIPC quantity has been noticed. However, out of the 35 samples analysed, only 2 had residue contents higher than the RML. The comparative analysis of the quantities applied and the residues contained in the samples made it possible to quantify the heterogeneity of the applications depending on the techniques. The tests carried out show in a general way that mechanical dusting, even though having a less constant flowrate than sprayers, leads to less important variation of the residue between samples. In testing conditions, the heterogeneity of the antigerminative treatment decreases when applied by means of a mechanical duster. In practise, these results are distorted by topical applications of CIPC. The combination of this practise with a too high heterogeneity of the treatment are to be avoided in order to have a good preservation and meet the residues standards.

Belgium↗

Molecular detection of Y chromosome microdeletions: an Irish study.

The region of the Y chromosome most critical for male fertility is called the azoospermia factor (AZF) region and it is located within subintervals five and six on the long arm of the Y chromosome. Several genes, all residing here, contribute to spermatogenesis and deletions in these genes are thought to be pathogenetically involved in some cases of male infertility associated with azoospermia or oligozoospermia. The aim of this study was to establish the prevalence of microdeletions in the AZF region of the Y chromosome in an Irish male population undergoing fertility treatment. To do this, we applied and compared two independent polymerase chain reaction (PCR) based screening methods, namely, a PCR protocol using several sequence-tagged site (STS) primer sets and a recently published multiplex PCR Y chromosome screening protocol. A total of 78 patients, attending the IVF unit at University College Hospital, Galway, were included in this study. Of them, 56 suffered from idiopathic azoospermic/oligozoospermic infertility. The remaining 22 patients had various conditions, which may have contributed to their infertility. A total of 50 age-matched normospermic men were included as controls. Two microdeletions were found; one in the AZFa region and one in AZFb region. These deletions were observed among the truly idiopathic cases. Further analysis was performed to study the extent of the deletions and it was confirmed that each deletion encompassed the respective AZF region including the AZF candidate gene.

Adult↗

Trypanosoma cruzi: infectivity of clonal genotype infections in acute and chronic phases in mice.

Eight Trypanosoma cruzi stocks pertaining to the clonal genotypes 19/20, 32, and 39 have been characterized for three experimental parameters of infectivity in Balb/c mice: (i) percentage of mice with a patent parasitemia (% MPP), (ii) maximum parasitemia (MP), and (iii) percentage of mice with positive hemoculture (% MPH). By order of decreasing values, the values recorded for the clonal genotypes ranked as follows: 19/20, 32, and 39, except for the % MPP parameter, for which 19/20 and 32 were not statistically different. The rate of successful reisolation after infection in mice, analyzed by multilocus enzyme electrophoresis and random amplified polymorphic DNA typing, was statistically different according to the clonal genotype and was different for uniclonal infections and for mixed infections by two different clonal genotypes. These results confirm that T. cruzi clonal genotypes differ significantly in their infectivity in mice.

Acute Disease↗

Kinetic evidence for the formation of a Michaelis-Menten-like complex between horseradish peroxidase compound II and di-(N-acetyl-L-tyrosine).

The formation of a reversible adsorption complex between a dimer of N-acetyl-L-tyrosine [di-(N-acetyl-L-tyrosine), (NAT)2] and horseradish peroxidase (HRP) compound II (CII) was demonstrated using a kinetic approach. A specific KIIm value (0.58 mM) was deduced for this step from stopped-flow measurements. The dimerization of the dipeptide Gly-Tyr was analysed at the steady state and compared with (NAT)2 dimerization [(NAT)2-->(NAT)4]. A saturation of the enzyme was observed for both substrates within their range of solubility. In each case the rate of dimerization reflected the rate-limiting step of compound II reduction to the native HRP (E) (kappcat/Kappm approximately kII-->E). The kappcat values for (Gly-Tyr)2 and (NAT)4 formation were 254 s-1 and 3.6 s-1 respectively. The KappM value of Gly-Tyr was 24 mM. It was observed that the value (0.7 mM) for (NAT)2 was close both to its specific KIIm value for the second step of reduction (CII-->E) and to its thermodynamic dissociation constant (Kd=0.7 mM) with the resting form of the enzyme. As (NAT)2 was a tighter ligand but a poorer substrate than Gly-Tyr, a steady-state kinetic study was performed in the presence of both substrates. A kinetic model which includes an enzyme-substrate adsorption prior to each of the two steps of reduction was derived. This one agreed reasonably well with the experimental data.

Adsorption↗

Release of deuterated nonenal during beer aging from labeled precursors synthesized in the boiling kettle.

The use of labeled nonenal enabled the demonstration that the appearance of the cardboard flavor in finished beer comes from lipid auto-oxidation during wort boiling and not from lipoxygenasic activity during mashing. Free trans-2-nonenal produced by linoleic acid auto-oxidation in the kettle disappears, owing to retention by wort amino acids and proteins. This binding linkage protects trans-2-nonenal from yeast reduction but is reversible, allowing release of the compound at lower pH during aging. Labeled trans-2-nonenal is detected after aging when deuterated precursors form in the boiling kettle. The amount of alkenal released correlates with the concentration of reversible associations in the pitching wort. This work brings new illumination to the formation of trans-2-nonenal and overturns many previous hypotheses. It also explains why a reduction in the beer pH intensifies the cardboard flavor.

Aldehydes↗

A phylogenetic analysis by multilocus enzyme electrophoresis and multiprimer random amplified polymorphic DNA fingerprinting of the Leishmania genome project Friedlin reference strain.

We have assessed the phylogenetic status of the Leishmania genome project Friedlin reference strain by MLEE and multiprimer RAPD including a set of 9 stocks representative of the main Leishmania species and of the whole genetic diversity of the Leishmania genus. To our knowledge, the detailed genetic characterization of the Friedlin strain has never been published before. As previously recorded (Tibayrenc et al. 1993), MLEE and RAPD data gave congruent phylogenetic results. The Friedlin reference strain was definitely attributed to Leishmania (Leishmania) major Yakimoff et Schokhor, 1914. Five specific RAPD patterns made it possible to distinguish between the Friedlin strain and the 2 other L. (L.) major stocks included in the study. Various specific MLEE and RAPD characters permitted to distinguish between the Leishmania species included in the study. All these characters are usable to detect accidental laboratory mix-ups involving the Friedlin reference strain. In confirmation with previous studies involving a more limited set of genetic markers, the general genetic diversity of the Leishmania genus proved to be considerable. It must be made clear that only one strain cannot be considered as representative of the whole genetic variability of the genus Leishmania. In the future, it is therefore advisable to complement the results obtained in the framework of the Leishmania genome project with data from other strains that should be selected on a criterion of important genetic differences with the Friedlin strain.

Animals↗

A phylogenetic analysis of the Trypanosoma cruzi genome project CL Brener reference strain by multilocus enzyme electrophoresis and multiprimer random amplified polymorphic DNA fingerprinting.

We have assessed the phylogenetic status of the Trypanosoma cruzi Genome Project CL Brener reference strain by multilocus enzyme electrophoresis (MLEE) and multiprimer random amplified polymorphic DNA (RAPD) including a set of cloned stocks representative of the whole genetic diversity of T. cruzi. MLEE and RAPD data gave congruent phylogenetic results. The CL Brener reference strain fell into the second major phylogenetic subdivision of T. cruzi, and was genetically very close to the Tulahuen reference strain. No reliable RAPD character and only one MLEE character permitted us to distinguish between the CL Brener and Tulahuen reference strains. In contrast, many RAPD and MLEE characters were able to distinguish between the CL Brener reference strain and the other T. cruzi genotypes analyzed here, in particular the formerly described principal zymodemes I, II and III. It is suspected that both CL Brener and Tulahuen are hybrid genotypes, a fact that should be taken into account when interpreting sequence data. Moreover, our study confirms that the species T. cruzi is genetically very heterogeneous. We recommend future comparison of sequencing data from the CL Brener reference strain with those of at least one radically distinct T. cruzi genotype, belonging to the other major phylogenetic subdivision of this species.

Animals↗

Trypanosoma cruzi: impact of clonal evolution of the parasite on its biological and medical properties.

Trypanosoma cruzi populations are subdivided into natural clones that can exhibit considerable genetic differences. It has been proposed that T. cruzi clonal structure has a major impact on this parasite's biological properties. The present work aims at testing this hypothesis. Twenty-one stocks isolated from various ecological cycles, places, and hosts were characterized by multilocus enzyme electrophoresis (MLEE) with 22 genetic loci and random amplification of polymorphic DNA (RAPD) with 10 primers on the one hand and by 14 different biological parameters on the other hand. These parameters were related to: (i) growth kinetics of epimastigotes and amastigotes; (ii) infection of culture cells by amastigotes; (iii) viability of extracellular trypomastigotes; or (iv) sensitivity of epimastigotes, trypomastigotes, and amastigotes to Benznidazole and Nifurtimox. MLEE and RAPD results exhibited parity to each other, as previously noted (M. Tibayrenc, K. Neubauer, C. Barnabé, F. Guerrini, D. Skarecky, and F. J. Ayala, 1993, Proceedings of the National Academy of Sciences of the USA 90, 1335-1339), and showed that the 21 stocks were distributed into three main genetic groups, 19/20, 32, and 39, corresponding to the major clones 19, 20, 32, and 39 previously described on the basis of 15 isozyme loci. Most biological parameters showed a strong correlation to the genetic distances evaluated from either MLEE or RAPD, which favors the working hypothesis. The only exception came from drug sensitivity estimated on trypomastigote forms. The overall results made it possible to firmly reject the null hypothesis that there is no relationships between evolutionary distances and biological differences in T. cruzi natural clones.

Animals↗

Trypanosoma cruzi: compared vectorial transmissibility of three major clonal genotypes by Triatoma infestans.

Twenty Trypanosoma cruzi stocks attributed to the 19/20, 32, and 39 major clones (Tibayrenc et al. 1986) were used to infect experimentally third instar larvae of Triatoma infestans. Three variables were considered: (i) percentage of infected insects; (ii) number of flagellates per insect (NFI); and (iii) percentage of metacyclic trypomastigotes per insect. Differences between the genotypes under study for all parameters considered were detected. These differences were statistically significant (P < 10(-3)), except between the 39 and 32 clonal genotypes for the NFI parameter. The correlation coefficient between the genetic distance and the biological parameters determined by the nonparametric Mantel's test was strongly significant (P < 10(-4)). Data obtained suggest clearly that populations of parasites belonging to the 19/20 genotype are more efficiently transmitted (high transmissibility genotype) by the vector than the 32 genotype (low transmissibility genotype), while the 39 genotype presents intermediary characteristic. Results confirm the working hypothesis that the subdivision of T. cruzi into discrete clonal lineages has an impact on the vectorial competence of T. infestans, the most important vector of the chagasic infection in South America, and that different clonal lineages do not exhibit the same vectorial transmissibility. This fact is relevant both for Chagas' disease epidemiology and for the use of xenodiagnosis.

Animals↗

Compared vectorial transmissibility of pure and mixed clonal genotypes of Trypanosoma cruzi in Triatoma infestans.

A total of 15 mixtures involving 9 different stocks attributed to the 19/20, 32 and 39 major clonal genotypes of Trypanosoma cruzi were used to infect third-instar nymphs of Triatoma infestans via an artificial feeding device. Three biological parameters were considered: (1) the percentage of infected insects (%II), (2) the number of flagellates per insect (NFI), and (3) the percentage of trypomastigotes per insect (%DIF). Genetic characterization by both multilocus enzyme electrophoresis (MLEE) and random amplification of polymorphic DNA (RAPD) indicated that in almost all cases (87%), mixtures remained present after completion of the whole cycle in the insect vector. Two lines of comparison were performed: (1) pure clonal genotypes versus corresponding mixed clonal genotypes and (2) the actual behavior of mixed clonal genotypes versus the expected behavior of the theoretical mixture (i.e. the arithmetic mean of the results observed for each of the two clonal genotypes taken separately). Statistical analyses of the variables were made difficult because of the presence of large standard deviations. Nevertheless, in several cases, mixtures differed significantly from pure clonal genotypes, and in one case the actual mixture differed significantly from the theoretical mixture. In some cases, interaction (either potentialization or reciprocal inhibition) could be suspected.

Animals↗

The gp63 gene locus, a target for genetic characterization of Leishmania belonging to subgenus Viannia.

In the present study the gp63 gene locus was used as a target for genetic characterization of Leishmania parasites by 2 methods: (i) RFLP analysis with several restriction enzymes (gp63-RFLP), and (ii) intra-genic PCR amplification coupled with restriction analysis (PCR-RFLP). Both methods were applied to a large number of natural isolates belonging to 4 species of the subgenus Viannia, namely L. (V.) braziliensis, L. (V.) peruviana, L. (V.) guyanensis and L. (V.) lainsoni; reference stocks of subgenus Leishmania were included as outgroups. Multilocus isoenzyme typing (MLEE) was used as a reference. On the one hand gp63-RFLP evidenced an extensive polymorphism and revealed specific markers for subgenus, species and geographical populations: congruence with MLEE was demonstrated statistically. The particular interest of gp63-RFLP was illustrated by infra-specific polymorphism, because of the possible relationship with phenotype diversity. On the other hand intra-genic amplification was less resolutive than gp63-RFLP, but also allowed discrimination of the 2 subgenera (PCR alone) and all the species tested in the subgenus Viannia (PCR-RFLP). PCR-RFLP presents an important operational advantage as it allows genetic characterization of minute amounts of parasites, using Leishmania specific primers. The polymorphism revealed by gp63-RFLP and PCR-RFLP illustrates the very high genomic and genetic plasticity of gp63 genes.

Animals↗

Movement disorders due to cerebral Toxoplasma gondii infection in patients with the acquired immunodeficiency syndrome (AIDS).

Hemichorea and parkinsonism are unusual manifestations of cerebral toxoplasmosis in patients with AIDS. We here describe two such cases and we reviewed extensively the literature (through computer searches using MEDLINE) for other reported instances. In our patients, unlike the other neurological symptoms, the response of the movement disorders to anti-toxoplasmosis therapy was delayed and only partial. We demonstrate that tetrabenazine is a valuable additional symptomatic treatment for choreic movements in one of our patients. We emphasize that, among patients suffering from AIDS, particularly in countries with high prevalence of toxoplasmosis, the occurrence of movement disorders should first suggest the diagnosis of cerebral toxoplasmosis.

Acquired Immunodeficiency Syndrome↗