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Biomedical subjects

G Vernet

Publications and source records attributed to G Vernet.

At least 19 recordsLinked to original sources
PubMed2006

European multicenter evaluation of high-density DNA probe arrays for detection of hepatitis B virus resistance mutations and identification of genotypes.

Polymorphisms along the hepatitis B virus (HBV) genome have an impact on disease outcome, sensitivity to antiviral treatment, escape from vaccination, and laboratory diagnosis. We have designed a diagnostic tool based on duplex amplification of the whole HBV genome and a high-density DNA chip designed to detect 245 mutations, 20 deletions, and 2 insertions at 151 positions and to determine the genotype of the virus in serum. Assay performances were evaluated with 170 samples, characterized by determination of viral load and sequencing of the Pol, S, and precore genes and the basal core promoter. One hundred fifty-three samples (90%) could be amplified and analyzed by the chip. Only two samples with more than 10(3) genome copies/ml could not be analyzed. Genotype had no impact on analytical sensitivity. Reproducibility studies showed no difference between repeats for codon and genotype determination. Genotype determination by sequencing and the chip were concordant in 148 of 151 samples. Twelve thousand one hundred sixty-one codons were analyzed by both techniques. Only 89.4% could be determined by sequencing, and among the remaining 11,335 codons, 92.8% were identical by sequencing and the chip. Failures to identify an amino acid by the chip were mainly due to reduced hybridization efficiency attributed to unexpected polymorphisms. Optimization of the chip-based reagent for the analysis of the HBV genome is ongoing. This first evaluation showed that DNA chip technology can provide important information in relation to the clinical management of chronic hepatitis B.

DNA, Viral↗
PubMed2004

Metallothionein induction in aquatic oligochaete tubifex tubifex exposed to herbicide isoproturon.

Metallothioneins (MTs) are low-molecular-weight proteins mainly involved in metal ion detoxification. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth and antioxidative defenses. Moreover, pesticides can induce their synthesis. The aim of the current work was to determine the effects of isoproturon, either pure or formulated as Matin (suspension containing an isoproturon concentration of 500 g. L(-1)), on the metallothionein and total protein contents of the aquatic worm Tubifex tubifex. MT levels in exposed worms increased significantly after 7 and 15 days of exposure to a concentration of the herbicide of 50 mg. L(-1). Isoproturon reduced the metal (Cu, Zn, and Cd) content of metallothioneins, and it also increased the total protein content of the worms. These results suggest that MT induction may not be considered a specific biomarker of metal exposure but that it can be used as a nonspecific biomarker of the effect of isoproturon effect in aquatic worms.

Animals↗
PubMed2004

Diagnosis of zoonotic viral encephalitis.

Human infections by zoonotic encephalitis viruses are usually asymptomatic or symptoms are not specific to these viruses. Some of them have high mortality and morbidity rates and most often no specific treatment exist. This emphasizes the need for a precise identification of arboviruses in clinical specimens from humans and animals. Because these diseases are frequent in developing countries and tend to emerge or re-emerge in others, diagnostic tools must detect the broadest possible range of viruses with a high sensitivity and this is a key factor for surveillance, control of transmission and prevention through vaccination. In countries with limited diagnostic infrastructures, low-cost and easy-to-use tests are required. The diagnosis of arboviral encephalitis has been significantly improved in the recent years. Sensitive ELISA assay to detect antibodies against many arboviruses in serum or CSF are commercially available and can be used to detect early infections. Immunochromatographic rapid tests for the detection of specific IgM that could be used on fingertip blood would be valuable tools in developing countries. A limitation of these serologic assays is their lack of specificity as many arboviruses are antigenically related. Virus isolation or molecular assays from different human or animal tissues are also important diagnostic tools. Molecular assays have been extensively described in the recent years. They are very sensitive and have the advantage over cell culture that specimen transportation is less critical. Real-time detection has even improved sensitivity and reduced time-to-result. Although the utility of molecular assays for the detection of arboviruses in mosquito pools has been demonstrated, an extensive validation of their pertinence in clinical settings is still required. The use of DNA-microarrays may further extend the range of viruses that can be detected in a single test and allow isolates typing for epidemiology purposes.

Agglutination Tests↗
PubMed2004

Immunology-related perturbations induced by copper and chitosan in carp (Cyprinus carpio L.).

Copper is used in treatment mixtures to control fungal diseases in vineyards. Its concentrations are relatively high in some aquatic ecosystems, and the main problem observed in this study was the antioxidant stress induced by this heavy metal. Copper toxicologic effects in aquatic organisms have prompted the demand for alternative use of low-toxicity molecules in culture treatments. Chitosan is a polymer with antifungal property similar to copper and may be an interesting biopesticide. Thus, it is necessary to investigate the potential toxicity of chitosan for aquatic animal health, either alone or in conjunction with copper. In this study, carp were exposed to two sublethal chitosan concentrations (75 and 150 mg/L) or to two sublethal copper concentrations (0.1 and 0.25 mg/L) or to a mixture of chitosan plus copper (75 mg/L and 0.1 mg/L, respectively). The results of the present study show that exposure to copper at environmentally realistic levels or to chitosan at sublethal concentrations may significantly stimulate various aspects of immune functions in carp such as nonspecific cellular immunity, represented by total immunoglobulin level, ceruloplasmin activity, and oxidative activity of phagocytes. This acute-phase inflammatory response induced separately by the two treatments was not observed, especially on phagocyte oxidative activity, when carp were exposed to the copper-chitosan mixture. This fact could be explained by a possible chelation of copper by chitosan decreasing the biodisponibility of the two products for immune cells. Thus, the immunotoxicologic impact of copper and chitosan on fish immune response would be less pronounced with the combined treatments than with separate treatments in an aquatic environment.

Animals↗
PubMed2004

Detection of human immunodeficiency virus type 1 antiretroviral resistance mutations by high-density DNA probe arrays.

Genotypic resistance testing has become an important tool in the clinical management of patients infected with human immunodeficiency virus type 1 (HIV-1). Standard sequencing methodology and hybridization-based technology are the two principal methods used for HIV-1 genotyping. This report describes an evaluation of a new hybridization-based HIV-1 genotypic test of 99 clinical samples from patients infected mostly with HIV-1 subtype B and receiving treatment. This test combines RNA extraction with magnetic silica particles, amplification by nested reverse transcriptase PCR, and detection with high-density probe arrays designed to detect 204 antiretroviral resistance mutations simultaneously in Gag cleavage sites, protease, reverse transcriptase, integrase, and gp41. The nested reverse transcriptase PCR success rates at viral loads exceeding 1,000 copies/ml were 98% for the 2.1-kb amplicon that covers the Gag cleavage sites and the protease and reverse transcriptase genes, 92% for the gp41 amplicon, and 100% for the integrase amplicon. We analyzed 4,465 relevant codons with the HIV-1 DNA chip genotyping assay and the classic sequence-based method. Key resistance mutations in protease and reverse transcriptase were identified correctly 95 and 92% of the time, respectively. This test should be a valuable alternative to the standard sequence-based system for HIV-1 drug resistance monitoring and a useful diagnostic tool for simultaneous multiple genetic analyses.

Anti-HIV Agents↗
PubMed2003

Chitosan improves development, and protects Vitis vinifera L. against Botrytis cinerea.

We evaluated the potential of chitosan both to stimulate plant development and to induce protection from Botrytis cinerea in Vitis vinifera L. plantlets. The presence of 1.75% (v/v) chitogel in the culture medium was the optimal concentration for in vitro grapevine plantlet growth, as determined by measurements on enhancement of root and shoot biomass. Photosynthesis and related parameters were also stimulated in chitogel-treated plantlets. Chitogel reduced the development of Botrytis cinerea and induced cytological alterations to the pathogen. When challenged with the fungus, a significant decrease in disease incidence was observed in plants growing on medium supplemented with chitogel. Furthermore, exogenous foliar applications of chitogel to plantlets growing on chitogel-free medium sensitized them so as to be protected against Botrytis cinerea attack. Our results indicate that chitogel can be used in the vineyard as a means to attain protection against Botrytis cinerea and that its application may counteract the wide use of chemical pesticides.

Botrytis↗
PubMed2003

Oxyfluorfen toxic effect on S. obliquus evaluated by different photosynthetic and enzymatic biomarkers.

The effect of oxyfluorfen was investigated when alga Scenedesmus obliquus has been exposed to different concentrations (7.5, 15, and 22.5 microg x L(-1)) at 12, 24, and 48 hours of exposure. Toxicity test was done by using 13 biomarkers concerning growth rate, chlorophyll content and indicators of photosynthetic and antioxidant enzyme activities. The change of the 13 parameters showed a great variation of sensitivity indicating differences in parameters' suitability to be used as biomarkers when alga culture was exposed to oxyfluorfen toxicity. The order of sensitivity between those biomarkers was: Antenna size (ABS/RC) > Chlorophyll content > Catalase (CAT) > Operational PSII quantum yield (phiS(PSII)) > Glutathione S-transferase (GST) > Functional plastoquinone pool (Q(PQ)) > Glutathione reductase (GR) > Growth rate > Nonphotochemical quenching (QN) > Proton gradient quenching (Q(Emax)) > Ascorbate peroxidase (APX) > Photochemical quenching (Q(p)) > Maximum PSII quantum yield (Phi(PSII)). The effect of oxyfluorfen on the changes of those parameters was interpreted as a result of herbicide mode of action at molecular level of alga cellular system. This study indicated for some photosynthetic and enzymatic biomarkers to be useful indicators of toxicity effect induced in non-target alga species. Determination of biomarkers' sensitivity order may facilitate their selection to be used in environmental risk assessment of polluted water.

Biomarkers↗
PubMed2003

Metallothionein induction in the aquatic oligochaete Tubifex tubifex exposed to the herbicide isoproturon.

Metallothioneins (MTs), are low molecular weight proteins, mainly involved in metal ion detoxification. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth, and anti-oxidative defenses. Moreover, pesticides can induce their synthesis. The aim of the current work was to determine the effects of isoproturon either pure or formulated as Matin (suspension containing 500 g x l(-1) isoproturon) on the metallothionein and total protein content of the aquatic worm Tubifex tubifex. MT levels in exposed worms increased significantly after 7 and 15 days of exposure to 50 mg x l(-1) of herbicide. Isoproturon reduced metal (Cu, Zn, and Cd) content of metallothioneins, and it also increased total protein content of the worms. The results suggest that MT induction may not be considered as a specific biomarker of metal exposure but it can be used as a non specific biomarker of isoproturon effect in the worm.

Animals↗
PubMed2002

Antioxidant response modulated by copper in healthy or parasitized carp (Cyprinus carpio L.) by Ptychobothrium sp. (Cestoda).

An increased antioxidant response (catalase, glutathione S-transferase (GST) and glutathione reductase (GRd) activities in liver and GST activity in head kidney) was observed in carp parasitized by Ptychobothrium sp. compared to healthy fish. In case of a copper contamination of these fish, the decrease in enzymatic activities observed was less pronounced in parasitized than in healthy carp.

Animals↗
PubMed2002

Photosynthetic responses of Lemna minor exposed to xenobiotics, copper, and their combinations.

The effects on the photosynthetic process of copper and pesticides, used in vineyards, and their combinations, were investigated by measuring different chlorophyll fluorescence parameters in Lemna minor. Cu and flumioxazin had a severe impact on duckweed since a decrease in their photosynthetic capacity was detected after 24h of exposure to 200 and 1 microg.L(-1), respectively. However, fungicides used to control Botrytis cinerea (procymidone, pyrimethanil, and fludioxonil) seem to have no marked effects on duckweed even at very high concentrations (50 mg.L(-1)). Analysis of the combinations between copper (200 microg.L(-1)) and pesticides revealed different patterns of response: a synergistic effect was observed when Cu(2+) was added to flumioxazin (1 microg.L(-1)). In contrast, an antagonism was detected when duckweed was exposed to a mixture of Cu(2+) and fludioxonil or procymidone. However, these interactions always tended toward additivity when pesticide concentrations increased. Additivity was also observed for the Cu(2+)-pyrimethanil mixture at each fungicide concentration.

Araceae↗
PubMed2002

Biological effects of two insecticides on earthworms (Lumbricus terrestris L.) under laboratory conditions.

Earthworms (Lumbricus terrestris L.) were exposed to endosulfan and aldicarb for 2, 7, and 15 days and lethal concentrations were determined. Worms were then exposed to these concentrations (LC10, LC25, and LC50) of endosulfan and (LC10 and LC25) of aldicarb. Growth rate, total protein content, and insecticide residues were determined. Aldicarb was more toxic than endosulfan and both insecticides caused a significant reduction of growth rate and total protein content of earthworms. Residues remaining in the soil after 2 to 15 days ranged between 37.75% and 68.54% of applied concentration for endosulfan and between 10.13% and 67.71% of applied concentration for aldicarb. Less than 1% of soil concentration was detected in worms and accumulation was more important in the case of endosulfan. This study proposes the use of growth rate and total protein content as biomarkers for contamination by endosulfan and aldicarb.

Aldicarb↗
PubMed2002

Fate and sublethal effects of isoproturon on mature earthworm (Lumbricus terrestris L.).

This study was conducted to investigate the effects of isoproturon in mature earthworm (Lumbricus terrestris L.) under laboratory condition. Earthworms were exposed to soils contaminated with different concentrations for various duration. Residues were monitored in soil and earthworms after 7, 15, 30, 45, and 60 days of exposure to different isoproturon concentrations. Acute toxicity of isoproturon was determined together with growth rate and total soluble protein content of worms. These parameters were related to isoproturon concentration in soil and earthworms. No lethal effect of isoproturon was observed even at the highest concentration tested (1.4 g/kg soil) after 60 days after treatment. Residues of isoproturon have caused a significant reduction of the growth rate. Additionally a reduction of total soluble protein was observed in all treated worms. Decrease of isoproturon concentration in soil was slow and depended on herbicide initial concentration. In the worms, it increased during the first 15 days and decreased thereafter. This study is suggesting the use of the growth rate of earthworms as biomarker of exposure to isoproturon.

Animals↗
PubMed2001

Characterisation of the yeast Pichia membranifaciens and its possible use in the biological control of Botrytis cinerea, causing the grey mould disease of grapevine.

Pichia membranifaciens strain FY-101, isolated from grape skins, was found to be antagonistic to Botrytis cinerea, the causal organism of the grey mould disease of the grapevine. When grown together on solid as well as liquid media, the yeast brings about the inhibition of this parasitic fungus, coagulation and leakage of its cytoplasm, and suppression of its ability to produce the characteristic grey mould symptoms on the grapevine plantlets. In vitro experiments confirm that this yeast can be used as a biological control organism against B. cinerea. An account of the molecular characterisation of P. membranifaciens (complete sequence of the ITS region of its ribosomal DNA, GenBank accession No. AF 270935), as well as the interaction between B. cinerea and the yeast, are given here.

Antibiosis↗
PubMed2001

Immunocytochemical localization of hydroxyindole-o-methyltransferase (HIOMT) in the brain of Myoisophagos lacteus (Nemertea: Heteronemertea: Lineidae).

In an attempt to identify the brain structures that synthesize melatonin and that probably mediate the photoperiodic response of the heteronemertean Myoisophagos lacteus, we utilized immunocytochemical techniques and employed immunoglobulins directed against hydroxyindole-O-methyltransferase (HIOMT, EC 2.1.1.4). This enzyme catalyzes the last step of melatonin biosynthesis. In immunocytochemically treated head sections of Myoisophagos lacteus, antibodies labelled a few cells in the dorsal region of the dorsal cerebral ganglia. Previous studies have shown that melatonin is present both in the brain and eyes of this nemertean species and that melatonin is involved in control of the worm reproduction. Other studies have demonstrated the presence of photoreceptor-like cells in the same region of the worm brain that showed HIOMT immunostaining. Therefore, anatomical findings of the present study, coupled with results of previous works, provide strong evidence that this region of the worm brain houses a photoperiodic receptor involved in melatonin biosynthesis. J. Exp. Zool. 290:156-162, 2001.

Acetylserotonin O-Methyltransferase↗
PubMed2001

Effects of procymidone, fludioxonil and pyrimethanil on two non-target aquatic plants.

Procymidone, fludioxonil, and pyrimethanil are widely used to control the pathogenic fungus Botrytis cinerea in Champagne's vineyards. These fungicides may end up in surface waters and present potential risks for aquatic vascular plants and algae. Therefore, their toxicity was evaluated on Lemna minor and Scenedesmus acutus in six-day or 48-h tests, respectively. Based on growth and chlorophyll (Chl) content of L. minor and S. acutus cultures, the results showed that the alga was the most sensitive to the fungicides. Among the fungicides, pyrimethanil was the most toxic for L. minor, its nominal IC50 was 46.16 mg l(-1) and that of the other two was >100 mg l(-1). In contrast, pyrimethanil appeared the least toxic for S. acutus at low concentration, nominal IC50 were 22.81, 4.85, and 4.55 mg l(-1) for pyrimethanil, fludioxonil, and procymidone, respectively. Fate of the fungicides in the media was also investigated and acute toxicity of the agrochemicals is discussed in regard to concentration in the culture media. Poor solubility of procymidone and fludioxonil appeared to be partly responsible for the low toxicity of these fungicides. Based on these toxicity data and the concentrations found in ponds collecting vineyard runoff water, these pesticides should not impair the establishment of pioneer plants.

Agriculture↗