Biomedical subjects

Guy Vernet

Publications and source records attributed to Guy Vernet.

18 recordsLinked to original sources

PubMed2006

Molecular epidemiology of hepatitis B virus in Dakar, Sénégal.

Using DNA chip technology and real-time quantitative PCR, molecular profile of HBV strains infecting blood donors and patients in Dakar, Sénégal was studied. All HBsAg-positive blood donors (n = 175) and all patients who presented with chronic hepatitis B (n = 29) between 1st June 2003 and 31st July 2003 were studied. One patient, a blood donor, was coinfected by HCV, and nine patients had anti-HDV antibodies. Few persons in either group were HBeAg-positive. Viral load values were relatively low but correlated with biochemical abnormalities. Patients were infected mainly by genotype E (72%). Patients infected by genotype A (28%) tended to be younger than other patients. There was no significant difference between the blood donors and the patients with hepatitis B as regards virological markers, including viral load, when the HBV genotype was taken into account. The BCP A1762T and G1764A mutations were found in four patients and one patient, respectively; the two mutations were never found in the same patient. The W28* mutation at position 1896 of the core was detected in 19 of the 32 genotyped patients, 18 (83%) of whom had genotype E infection. ALT levels were not influenced by HBV mutations. This study shows a low frequency of clinical signs in HBsAg-positive blood donors, a relatively low level of viral replication, and a high frequency of pre-core mutants in this West African population. These results underline the importance of molecular characterization of HBV infection as specific treatments become available in this region.

Adult↗

PubMed2005

Effects of the herbicide isoproturon on metallothioneins, growth, and antioxidative defenses in the aquatic worm Tubifex tubifex (Oligochaeta, Tubificidae).

Metallothioneins (MTs) are low molecular weight proteins, mainly implicated in metal ion detoxification. Increase in MT contents is considered to be a specific biomarker of metal exposure. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth, and antioxidative defenses. Therefore, the induction of MTs as biomarkers of exposure to the pesticide isoproturon has been investigated in the aquatic worms Tubifex tubifex. MT levels in exposed worms increased significantly (p < 0.05) after 2, 4, and 7 days of exposure to different concentrations of isoproturon (maximum increase compared to unexposed controls: +148.56% for 10 mg l(-1) after 4 days of exposure). In response to isoproturon, the activity of glutathione-S-transferase (max. +52%), glutathione-reductase (max. +100%), and catalase (max. +117%) increased, demonstrating the occurrence of an oxidative stress response to the herbicide. Thus, the increase in MT contents caused by isoproturon was interpreted as a defense response towards increased oxidative stress generated by the herbicide. Residues of isoproturon and its metabolites, 1-(4-isopropylphenyl)-3-methylurea, 1-(4-isopropylphenyl) urea, and 4-isopropylanilin were detected in the worm growth medium. Half-life of the herbicide was shorter at a low (0.1 mg l(-1)) initial concentration. The herbicide accumulated in T. tubifex but no metabolite could be detected.

Animals↗

PubMed2005

Determination of photosynthetic and enzymatic biomarkers sensitivity used to evaluate toxic effects of copper and fludioxonil in alga Scenedesmus obliquus.

Modulated PAM fluorometry and Plant Efficiency Analyser methods were used to investigate photosynthetic fluorescence parameters of alga Scenedesmus obliquus exposed to inhibitory effect of fungicides copper sulphate and fludioxonil (N-(4-nitrophenyl)-N'-propyl-uree). The change of those parameters were studied when alga S. obliquus have been exposed during 48 h to different concentrations of fungicides (1, 2 and 3 mgl(-1)). Under the same condition, enzymatic activities of catalase, ascorbate peroxidase, glutathione reductase and glutathione S-transferase were investigated to evaluate antioxidative response to fungicides effects. The change of sensitivity of those parameters was dependent to the mode of fungicide action, their concentration and time of exposure. For copper effects, the most indicative photosynthetic biomarkers were parameters Q(N) as non-photochemical fluorescence quenching, Q(Emax) as the proton induced fluorescence quenching and ABS/RC as the antenna size per photosystem II reaction center. Copper induced oxidative stress was indicated by increased activity of catalase serving as the most sensitive and valuable enzymatic biomarker. On the other hand, fludioxonil effect on photosynthetic parameters was very negligible and consequently not very useful as biomarkers. However, fludioxonil induced strong antioxidative activities associated with cytosol enzymes, as we found for catalase, ascorbate peroxidase and glutathione S-transferase activities. By obtained results, we may suggest for the activation of those enzymes to be sensitive and valuable biomarkers of oxidative stress induced by fludioxonil. Determination of biomarkers sensitivity may offer advantages in providing real criteria to use them for ecotoxicological diagnostic studies.

Ascorbate Peroxidases↗

PubMed2005

High-density DNA probe arrays for identification of staphylococci to the species level.

Rapid and accurate identification and speciation of staphylococci clinical isolates is important for predicting medical pathology. We evaluated the ability of a high-density DNA probe array based on 16S rDNA sequences to identify Staphylococcus species. Correct identification was observed for 185 out of the 201 strains (92%). Of the 33 tested species, the array was able to correctly identify 30 of them. The total time required for identification of 4 isolates was 5 h. Such a tool represents a powerful method for routine microbiological diagnostic as well as for epidemiological studies.

Animals↗

PubMed2005

Metallothionein induction, antioxidative responses, glycogen and growth changes in Tubifex tubifex (Oligochaete) exposed to the fungicide, fenhexamid.

Laboratory studies were conducted to determine the effects of different concentrations of fenhexamid (0.1, 1, and 10 mg L(-1)) on growth, oxidative stress, protein, glycogen, and metallothionein (MT) contents in Tubifex tubifex after an exposure of 2, 4, and 7 days. In addition, residues of the fungicide were followed in water and in the worms. In water, fenhexamid concentration decreased slowly (maximum -2 +/- 0.03% after 2 days for 1 mg L(-1)). In the worms, it increased after 4 days and decreased thereafter, confirming that the worms were exposed to the fungicide and not to a degradation product. LC50 values were between 95.22 +/- 5.36 and 32.11 +/- 1.8 mg L(-1) depending on exposure time. Exposure to fenhexamid had a negative effect on T. tubifex growth (maximum effect -12.2 +/- 0.8% after 7 days with 10 mg L(-1)) demonstrating the toxic effect of the pesticide. This growth rate decrease was accompanied by a reduction in protein and glycogen contents. The activity of catalase (CAT), and glutathione reductase (GR) increased in response to the fungicide demonstrating an oxidative stress in the worms. In contrast glutathion-S-transferase activity (GST) decreased. Exposure to fenhexamid also induced synthesis of MT (maximum +78 +/- 8% after 2 days for 10 mg L(-1)). The specificity of MT concentration increase in response to metals is discussed.

Amides↗

PubMed2005

Evaluation of a high-density oligonucleotide array for characterization of grlA, grlB, gyrA and gyrB mutations in fluoroquinolone resistant Staphylococcus aureus isolates.

Using high-density oligonucleotide array technology, 30 Staphylococcus aureus strains were studied for the presence of mutations in genes involved in fluoroquinolone resistance: grlA, gyrA, grlB and gyrB. For the two most important genes, gyrA and grlA, correlation with sequencing reached 95.1%. If all genes were considered, correlation was 88.8%.

Codon↗

PubMed2005

The DNA-Chip technology as a new molecular tool for the detection of HBV mutants.

Genetic variability of the Hepatitis B virus (HBV) strongly impacts the natural history of infection and the efficiency of diagnosis, vaccination and treatment. A genotypc assay able to provide genetic information on all HBV genes would be a very important tool for clinicians and epidemiologists. The DNA-Chip technology has proved to be powerful and convenient for the diagnosis of infectious diseases due to bacteria and viruses. We have designed a genotyping assay based on polymerase chain reaction and DNA-Chip for HBV Starting from a unique specimen, analysis of polymorphisms at 150 positions along the genome and 383 mutations is possible as well as the determination of the genotype. Preliminary experiments with plasmas from infected patients show that results obtained with this reagent are strongly correlated with those obtained with sequencing.

Genetic Techniques↗

PubMed2005

DNA probe array for the simultaneous identification of herpesviruses, enteroviruses, and flaviviruses.

Viral infections of the central nervous system (CNS) are caused by a variety of viruses, namely, herpesviruses, enteroviruses, and flaviviruses. The similar clinical signs provoked by these viruses make the diagnosis difficult. We report on the simultaneous detection of these major CNS pathogens using amplification by PCR and detection of amplified products using DNA microarray technology. Consensus primers were used for the amplification of all members of each genus. Sequences specific for the identification of each virus species were selected from the sequence alignments of each target gene and were synthesized on a high-density microarray. The amplified products were pooled, labeled, and cleaved, followed by hybridization on a single array. This method was successfully used to identify herpesviruses, namely, herpes simplex virus type 1 (HSV-1), HSV-2, and cytomegalovirus; all serotypes of human enteroviruses; and five flaviviruses (West Nile virus, dengue viruses, and Langat virus). This approach, which used highly conserved consensus primers for amplification and specific sequences for identification, would be extremely useful for the detection of variants and would probably help solve some unexplained cases of encephalitis. The analytical sensitivity of the method was shown to be 500 genome equivalents ml(-1) for HSV-1, 0.3 50% tissue culture infectious doses (TCID50s) ml(-1) for the enterovirus coxsackievirus A9, and 200 TCID50s ml(-1) for West Nile virus. The clinical sensitivity of this method must now be evaluated.

Base Sequence↗

PubMed2005

Characterization of 26 isolates of Staphylococcus aureus, predominantly from dairy sheep, using four different techniques of molecular epidemiology.

Little information is available regarding the molecular epidemiology of Staphylococcus aureus-induced mastitis in dairy sheep. In this study, 4 different typing techniques were compared in typing 26 S. aureus isolates, predominantly from cases of subclinical mastitis in dairy ewes. The 4 techniques were pulsed-field gel electrophoresis (PFGE), restriction fragment length polymorphism (RFLP) on 2 genes (coagulase and clumping factor B), randomly amplified polymorphic DNA-polymerase chain reaction (PCR) (RAPD-PCR), and multilocus sequence typing (MLST). On the basis of discriminatory power as the key parameter of typing systems, MLST and PFGE were found to be the most powerful techniques. The MLST and PFGE could contribute to epidemiological surveillance and evaluation of mastitis control programs, by documenting prevalence and dissemination of endemic clones in infected populations. The results of this study show that a single clone of S. aureus is widely distributed in infected ewe mammary glands.

Animals↗

PubMed2004

Humoral immune factors modulated by copper and chitosan in healthy or parasitised carp (Cyprinus carpio L.) by Ptychobothrium sp. (Cestoda).

As an environmental protection point of view, the potential toxicity of chitosan on aquatic animal health, alone or associated with copper must be investigated. Fish possess defence mechanisms to counteract the impact of toxics. The non-cellular and non-specific immune defences (total immunoglobulin, ceruloplasmin, lysozyme and potential killing activity of phagocytic cells) can be modulated by the potential environmental pollutants but also by natural stimulants such as bacteria, viruses or parasites. In this study, we investigate the potential toxicity of copper (0.1 and 0.25 mg/L) or chitosan (75 and 150 mg/L) and the combination copper and chitosan (0.1 and 75 mg/L, respectively) on two groups of carp: healthy or parasitised by Ptychobothrium sp. Fish exposed to water-soluble chitosan for 96 h had significantly high levels of natural antibodies in plasma. Moreover, activities of lysozyme and ceruloplasmin were also increased in plasma after the same treatment. The exposition of fish to copper have shown apparently contradictory effects on the immune parameters measured but, significant increase of this bacteriolytic activity was observed, particularly in head kidney after 4 days of treatment of fish with copper. The two products may induce separately an acute, short and local inflammatory acute phase response by stimulating some components of the innate immune response of healthy fish. The mixture seems to reduce the impact of the each product due to the physical and chemical properties of chitosan to complex with copper. The responses of humoral immune factors of treated carp was modulated by the presence of the parasite, as shown by the high elevation of lysozyme activity observed in parasitised carps after exposition to copper and by increases in natural antibodies levels observed in parasitised carp treated with the copper-chitosan mixture. This could indicate an additive effect on the stress response mediated by parasite. It occurred a greater stress response in the parasitised group than healthy group exposed to the same treatment evoking an additive effect. So, it is important to specify the health status of organisms to understand responses of immunological markers in fish.

Animals↗

PubMed2004

Modulation in hepatic and head kidney parameters of carp (Cyprinus carpio L.) induced by copper and chitosan.

Copper is used in treatment mixtures to control fungal diseases in vineyards plants. High concentrations of copper are inducing antioxidant stress in some aquatic ecosystems, and potential bioaccumulation in aquatic organisms has prompted the demand for alternative use of low toxic molecules in culture treatments. Chitosan is a biomolecule with antifungal and heavy metal ion chelating properties that may be used as a biopesticide. In this study, we investigate the potential toxicity of chitosan for aquatic animal health, alone or associated with copper. Carp (Cyprinus carpio L.) were exposed to different chitosan concentrations (from 37.5 to 375 mg/l) or to two sublethal copper concentrations (0.1 and 0.25 mg/l) or to chitosan and copper (75 and 0.1 mg/l, respectively). Antioxidant enzyme activities were enhanced in chitosan treated fish after 4 days and depressed after 8 days. This phenomenon indicated a non-negligible toxicity of chitosan in fish physiology. However, the mixture copper-chitosan seems to induce a lower degree of oxidative stress than each fungicide alone. These observations show that chitosan is a potentially noxious molecule for some fish and any industrial and/or agricultural uses of this compound will have to address this problem.

Animals↗

PubMed2004

Molecular diagnostics in virology.

Molecular biology has significantly improved diagnosis in the field of clinical virology. Virus discovery and rapid implementation of diagnostic tests for newly discovered viruses has strongly beneficiated from the development of molecular techniques. Viral load and antiviral resistance or subtyping assays are now part of the biological monitoring of patients chronically infected by human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV) and CMV. It will be important to add to this panel assays for other viruses of the herpesviridae family. Qualitative assays for the detection of blood-borne viruses have increased safety of blood donation and organ transplantation. Screening of other blood-borne viruses (parvovirus B19, HAV), multiplexing of detection and test automation to improve practicability and reduce costs will be the next steps. A major evolution in the near future will be the generalization of NAT for the diagnosis of viral etiology in patients, mostly with respiratory, CNS or gastro-intestinal diseases. Major technical improvements have been made to avoid obstacles that still limit this generalization, i.e. genetic variability of viruses, multiplex detection, contamination risk. Commercial offers already exist but menus must be extended to limit the validation and documentation work associated with home-brew assays. Real-time amplification has allowed the development of new NAT platforms but automation and integration of all steps of the reaction are still required to reduce hands-on-time, time-to-result and costs, and to increase throughput.

Antibodies, Viral↗

PubMed2003

Acute and sublethal effects of two insecticides on earthworms (Lumbricus terrestris L.) under laboratory conditions.

Earthworms (Lumbricus terrestris L.) were exposed to commercial formulations of endosulfan and aldicarb for 2, 7, and 15 days, and the LC(10), LC(25), and LC(50) were determined. Worms were then exposed to LC(10), LC(25), and LC(50) concentrations of endosulfan and LC(10) and LC(25) concentrations of aldicarb. The growth rate and total protein content were determined and related to endosulfan and aldicarb residues in soil and earthworms. Aldicarb was more toxic than endosulfan under the experimental conditions. The residues of endosulfan and aldicarb caused a significant reduction in the growth rate and total protein content of earthworms. The residues of endosulfan and aldicarb were monitored in soil and earthworms after 2, 7, and 15 days of exposure. The residues remaining in the soil after the experiments ranged between 37.75% and 68.54% of the applied concentration for endosulfan and between 10.13% and 67.71% of the applied concentration for aldicarb. Small amounts of both insecticides were detected in worms, and accumulation was more important for endosulfan. This study proposes the use of growth rate and total protein content as biomarkers for contamination by endosulfan and aldicarb.

Aldicarb↗

PubMed2003

The F684/F735 chlorophyll fluorescence ratio: a potential tool for rapid detection and determination of herbicide phytotoxicity in algae.

The use of herbicides constitutes the principal method of weed control but the introduction of these compounds into the aquatic environment (primarily through runoff) may have severe consequences for non-target plants. In this study, we describe a sensitive and inexpensive method for detection of photosynthesis-inhibiting herbicides, based on chlorophyll (Chl) fluorescence emission. Algae exhibited a Chl fluorescence signature with two maxima around 684 and 735 nm, correlated with the total Chl content of the algal suspension. The ratio of these two maxima (i.e. F684/F735) can be used as an indicator of stress in the photosynthetic apparatus, and thus represents a very simple method for in vivo evaluation of the health status of algae. Determination of the F684/F735 fluorescence ratio revealed the presence and phytotoxicity of atrazine, metribuzin, terbuthylazine, diuron, DCPMU, DCPU and paraquat. The toxic effect of these pollutants was estimated by monitoring the increase in the F684/F735 value, which reflects photosystem II and photosystem I photochemistry. We observed a drastic increase in the magnitude of this ratio, correlating quantitatively with herbicide concentration and corresponding to a decline in algal photosynthetic activity. For the tested herbicides affecting photosynthetic electron transport, the magnitude of the effect was as follows: diuron= DCPMU > metribuzin > atrazine > terbuthylazine > paraquat > DCPU. The F684/F735 Chl fluorescence ratio thus gives toxicity responses which compare favourably with tests such as the algal growth inhibition test, and could therefore be used to detect the presence and phytotoxicity of herbicides in aquatic environments.

Chlorophyll↗

PubMed2003

Pigments as biomarkers of exposure to the vineyard herbicide flazasulfuron in freshwater algae.

Weed control in Champagne vineyards has long relied on the use of diuron and substituted triazines; these compounds are now being replaced by flazasulfuron, a sulfonylurea that is used at a much lower dosage. The vineyards of Champagne are planted on steep slopes and runoff is important, and even though low doses of these herbicides are used, they may present some potential risk for freshwater ecosystems. Therefore, the effects of the sulfonylurea herbicide, flazasulfuron (formulated as Katana) was investigated on the unicellular green alga Scenedesmus obliquus. The pigment content of the algal suspensions was followed as a biomarker of exposure to the herbicide. The results demonstrate that flazasulfuron induced a reduction in chlorophyll content at concentrations of 10 microg/L, while the increase of pigment content in the culture was reduced with the lowest concentration tested (0.1 microg/L). Among the three pigments tested, chlorophyll a appeared to be the most sensitive biomarker. In the algal medium, flazasulfuron was slowly degraded (DT(50) approximately 8 days) in a compound that was tentatively identified. The toxicity of this herbicide for the algae was comparable to that of older herbicides which are used at a much higher rate. Therefore, we may speculate that even if flazasulfuron comes into contact with freshwater ecosystems, its effects on algae will be less deleterious than that of traditional herbicides.

Agriculture↗

PubMed2003

Effects of methanol on photosynthetic processes and growth of Lemna gibba.

Effects of methanol on growth and photosynthetic activity of Lemna gibba exposed under continuous illumination were examined. As a higher plant, L. gibba appeared to be much more sensitive to methanol inhibitory effect compared with some algae (Theodoridou et al. [2002] Biochim. Biophys. Acta, 1573, 189-198). We found that stimulatory or inhibitory effects were strongly dependent on the methanol concentration and the time of exposure. When the exposure was up to 0.2% methanol, the growth rate of biomass was improved by 50%. However, stimulatory effect of methanol appeared to be smaller when plants were exposed for 48 h compared with 24 h. Increase in biomass induced by methanol was not based on the increase in primary photosynthetic process but rather on accommodation of energy dissipation during photosynthesis. Inhibitory effect on the growth of L. gibba already observed for 0.5% methanol was strongly associated with the increase in the nonphotochemical energy dissipation. The ratio between biomass and methanol concentration appeared to determine the stimulatory or the inhibitory effect. Suggested explanations for the stimulatory and the inhibitory effects are presented.

Methanol↗

PubMed2002

In vivo and in vitro modulation of carp (Cyprinus carpio L.) phagocyte oxidative burst activity by gallium.

Since gallium is a metal ion used in semiconductor industry, the toxicological effects were previously evaluated in mammals but the ecotoxicological impacts remain unknown. In term of ecotoxicological risk assessment, the median lethal concentration (LC50 for 96 h of gallium to carp (Cyprinus carpio L.) and the oxidative response of carp phagocytes after the fish were exposed to sublethal levels of gallium were determined. The LC50 of gallium on C. carpio at 96 h was estimated as 96.25 +/- 14.3 mg/L. To determine the effect in vivo of gallium on the phagocyte response, fish were exposed for 96 h to 5 or 50 mg Ga(3+)/L. Carp maintained for 48 or 96 h in water containing 50 mg/L gallium had a significant fall in phagocyte oxidative burst activity in comparison with controls, as well as decreased leukocyte number in blood and increased cytotoxicity. To determine the effect in vitro of gallium on the phagocyte response, isolated phagocytes were exposed for 5 or 15 min to 50 nM, 500 nM, or 5, 50, 100, or 200 microM of Ga(3+). The oxidative burst was increased after in vitro incubation of phagocytes with 50 or 500 nM gallium for 15 min or with 500 nM gallium for 5 min. Moreover, for 50, 100, or 200 microM gallium, the oxidative burst activity of carp phagocytes was significantly decreased. Results indicate that the lethal toxicity of gallium for carp of gallium is not as high as for other metal ions. However, gallium was immunosuppressive for carp at the highest concentrations used (from 50 microM) in vivo and in vitro. At low concentrations, it could be an immunostimulant as observed in vitro.

Animals↗

PubMedDate not supplied

Metallothionein induction related to hepatic structural perturbations and antioxidative defences in roach (Rutilus rutilus) exposed to the fungicide procymidone.

A variety of stresses, hormones, glucocorticoids and cytokines are known to induce metallothioneins (MTs) in animals. The aim of this study was to investigate the effects of chemical stress induced by the dicarboximide fungicide procymidone on hepatic structure, MT content and antioxidative defences (catalase and glutathione reductase activities and glutathione content) in the common fish Rutilus rutilus. Catalase and glutathione reductase activities remained stable throughout the experiment. Four days of exposure to 0.2 or 0.4 mg l(-1) of procymidone induced an obvious increase in liver MT content, perturbation of metal MT contents, and an increase in hepatic glutathione content. After 14 days' exposure, obvious and large structural alterations of the hepatic parenchyma occurred simultaneously with a decrease in MT and glutathione content. These events were interpreted as degeneration of the liver. Fish exposed for 14 days to procymidone and then placed for 14 days in clean water showed nearly complete decontamination of the liver, but MT concentrations remained high. The toxicological significance of these events is discussed.

Animals↗