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Biomedical subjects

G Vernet

Publications and source records attributed to G Vernet.

At least 37 records · Page 2Linked to original sources

Biochemical and (ultra)structural hepatic perturbations of Brachydanio rerio (Teleostei, Cyprinidae) exposed to two sublethal concentrations of copper sulfate.

The aim of this study was to compare biochemical and (ultra)structural perturbations induced by Cu in the liver of Brachydanio rerio exposed for 14 days to sublethal concentrations of copper and then replaced for 14 days to clean water. Toxicity of Cu was clearly demonstrated: simultaneously to Cu accumulation, the liver developed large lysed areas and hepatocytic alterations. However, the majority of the parenchyma was composed of functional dark hepatocytes displaying typical feature of increased metabolism: development of rough reticulum, increase in size of nucleus and nucleolus, glycogenic depletion. Increase in hepatic protein content and of anti-oxidative defences (glutathione content, catalase and glutathione-S-transferase (GST) activities) indicated that the overall response of the liver was adaptative. In all hepatocytes a cord-like structure of the nucleoli was suspected to be associated to metal deposition. After 14 days depuration, the liver still contained high Cu concentrations and the hepatic alterations were not reversed. Such complementary studies are necessary for a better understanding of the deleterious effects of pollutants and for the development of biomarkers for metal toxicity.

Journal Article↗

Toxic effects and bioaccumulation of cadmium in the aquatic oligochaete Tubifex tubifex.

Although Tubifex tubifex (Oligochaeta, Tubificida) has been proposed as a test organism for ecotoxocological studies, very few data concerning sublethal toxicity and bioaccumulation are available on this worm. The aims of this work were to assess the toxicity of cadmium, one of the most toxic metals frequently encountered in polluted areas, on T. tubifex and the ability of the worm to accumulate this metal. Acute toxicity was analyzed by measurement of the 96-h LC(50) and daily survival rates. Results indicated that T. tubifex undergoes an adaptation period to Cd, the duration of which decreases with increasing Cd concentration. The various parameters affecting toxicity are discussed. Sublethal toxicity was studied by scanning electron microscopy. Observations revealed that Cd induced autotomy of the caudal region and mucus production. Autotomy is proposed as a criterion for sublethal toxicity. The results of bioaccumulation studies revealed that Cd is highly and rapidly taken up by the worm, suggesting involvement of efficient detoxification mechanisms. Consequently, the ability to accumulate large amounts of Cd may represent a potential toxicological risk to predators of the worm if Cd is accumulated in bioavailable forms.

Animals↗

Metallothioneins in liver of Rutilus rutilus exposed to Cu2+. Analysis by metal summation, SH determination and spectrofluorimetry.

Metallothioneins (MTs) have important roles in the homeostasis of essential metals and in the detoxication of heavy metals. They also represent a potential indicator of aquatic contamination by metals. Routine methods are needed for MTs quantification in ecotoxicological studies. This paper investigates the possibility to use the spectrofluorescent properties of Cu-MTs for MTs quantification. Cu displacement of metals coordinated to MTs and spectrofluorimetric determination of the obtained Cu-MTs was tested with commercial MTs and Cu2+-induced MTs in roach liver (Rutilus rutilus). Results of this original and simple spectrofluorimetric quantification of MTs presented a good correlation with data obtained with SH quantification, but not with metal summation evaluation of MTs (analysis of Zn, Cu and Cd coordinated to MTs). The three methods showed an clear induction of MTs in roach liver after 7 days of Cu2+ exposure. After 14 days of contamination, a reduction of hepatic MTs content was observable and not correlated to liver recovery. Results show that this low cost spectrofluorimetric method is useful to quantify MTs.

Animals↗

Phytotoxicity of diuron alone and in combination with copper or folpet on duckweed (Lemna minor).

The photosystem II-herbicide diuron is widely used for weed control in Champagne's vineyards. Its important use and its relative persistence make it of particular interest for ecotoxicological studies. Toxicity of diuron was assessed on Lemna minor L., a representative aquatic macrophyte regularly used for toxicological studies. Toxicity assessments were based on inhibition of growth and total chlorophyll content of L. minor cultures after 7 days. Growth was inhibited and IC(50) and IC(90) were, respectively, 25 and 60 microg l(-1), but chlorophyll content of L. minor increased in response to the herbicide. When diuron was combined with copper, growth inhibition of L. minor depended on the concentrations of both chemicals. For some concentrations, combination of these chemicals resulted in a slight (but non-significant) antagonism. Additivity was observed for all other mixtures. When diuron was combined with folpet, growth and chlorophyll content of L. minor only depended on the concentration of the herbicide. Diuron was also found to prevent the copper-induced decrease of chlorophyll content when it was combined with this metal. A multifactorial model was found more appropriate to characterize interactions between pesticides than Abott's model.

Journal Article↗

Toxic responses and catalase activity of Lemna minor L. Exposed to folpet, copper, and their combination.

Toxicity of copper and folpet--two fungicides widely used on grape--was evaluated on Lemna minor L., a sensitive aquatic weed regularly used for (eco)toxicological studies. Toxicity assessments were based on inhibition of growth and chlorophyll content of L. minor cultures after 7 days. IC10, IC50, and IC90 were determined for both compounds alone and were respectively, 0.03, 0.16, and 0.95 mg liter-1 for copper and 1.20, 7.50, and > 40 mg liter-1 for folpet. When both compounds were combined, the response of L. minor depended on the initial folpet concentration. Indeed, a slight synergy was observed for 5 mg liter-1 folpet, while at folpet concentrations of 20 to 35 mg liter-1, the two fungicides were antagonists. The antagonism was positively correlated with folpet concentration. Antagonism between Cu and folpet could not be explained by a reduced bioavailability of Cu since concentration of free copper in the mixture did not depend on the presence of folpet. One physiological defense response elicited by copper in plants is an increase in catalase activity. Copper and folpet stimulated catalase activity and changes in the activity of the enzyme could not account for the synergy but possibly for the antagonism. Nevertheless, catalase activity increase significantly after a 24-h exposure to 25 micrograms liter-1 of copper. The use of this property as a rapid and sensitive biomarker to monitor the toxicity of xenobiotics alone or in combination and of environmental water is discussed.

Catalase↗

Determination of anti-Toxoplasma gondii immunoglobulin G avidity: adaptation to the Vidas system (bioMérieux).

The determination of specific anti-Toxoplasma gondii IgG avidity has been proposed to improve the determination of the date of toxoplasmic seroconversion in pregnant women. In this study, we adapted this serological technique to the Vidas system (bioMérieux) using 6 M urea as the dissociating agent. We studied 356 sera, including 42 sequential sera from sero-conversions in pregnant women. Our results show that the test is easy to use, and that an avidity index higher than 0.300 allows the exclusion of a recent infection acquired less than 4 months before serum sampling.

Animals↗

Effects of platinum (Pt4+) on Lumbriculus variegatus Müller (Annelida, Oligochaetae): acute toxicity and bioaccumulation.

The acute toxicity and bioaccumulation (rates, kinetic) of tetravalent platinum in Lumbriculus variegatus under different physicochemical conditions (temperature and total water hardness) were investigated. Increased Pt4+ concentration (from 0.05 to 50 mg/L), exposure (up to 30 days), temperature (from 4 to 20 degrees C) and decreasing water hardness (from 300 to 0 mg/L CaCO3) increased Pt toxicity. The metal accumulated at a constant rate that was concentration, temperature and time dependent. The median lethal concentration (96h LC50) varied greatly from 0.397 mg/L in distilled water to 30 mg/L in the hard water from Champagne. Thus, L. var. can tolerate high levels of Pt. As a result, L. var. can be used in the laboratory to analyze mechanisms of adaptation to the induced stress and in the field as an indicator of Pt pollution.

Animals↗

Prevalence and persistence of antibody titers to recombinant HIV-1 core and matrix proteins in HIV-1 infection.

Numerous studies have established the correlation between antibodies to the core protein p24 of HIV-1 and the progression of the acquired immunodeficiency syndrome. In this study, we analyzed the immune response to two recombinant gag proteins, p24 and p17, in order to evaluate their diagnostic or prognostic significance. Immune response to the immunodominant domain of the transmembrane glycoprotein gp41 was used as a reference. Sera collected from individuals from France and Burundi (Central Africa) at various CDC stages of HIV-1 infection were tested using three sandwich enzyme-linked immunoassays developed with a synthetic peptide corresponding to the immunodominant domain of gp41, SP gp41, or recombinant p24 and p17 cloned and expressed in Escherichia coli. These assays allowed detection of titer antibodies to the three cited antigens. Antibodies to SP gp41 were detected in every HIV-1-positive patient from France and Burundi, generally at a high and stable level. Results obtained with p24 confirmed the value of antibodies to p24 as a prognostic marker only in European and North American populations, since the African population had very high levels of these antibodies even at an advanced stage of the disease. They also confirmed that initial antibody response to p24 is more predictive of outcome than antibody titer change over time. Although antibodies to p17 decline during progression to AIDS, they are frequently absent in French patients at early, asymptomatic stages and therefore could not be used as a prognostic marker. In contrast, antibodies to p17 are significantly less common in African patients with AIDS when compared with symptomless HIV-1-infected African individuals.(ABSTRACT TRUNCATED AT 250 WORDS)

Acquired Immunodeficiency Syndrome↗

Cellular immunity in an annelid (Nereis diversicolor, Polychaeta): production of melanin by a subpopulation of granulocytes.

We attempted to identify the nature and origin of the pigment produced by the marine worm Nereis diversicolor in order to isolate, in inert brown capsules, foreign objects introduced into its body cavity. This brown pigment, characterized by cytochemical techniques, could be a melanin. The activity of the enzyme phenoloxidase responsible for melanin biosynthesis was detected by enzyme cytochemistry techniques in vacuoles and the Golgi apparatus of coelomocytes activated by the presence of foreign bodies. Morphological techniques combined with a monoclonal immunological probe enabled us to establish that the "G2" granulocytes contain both the precursor of the pigment in dense bodies and the capacity for phenoloxidase synthesis when activated to encapsulate foreign bodies. The "G2" granulocyte may therefore be compared to a melanocyte in which melanin is not stored as in mammals, but immediately extruded following synthesis in the form of a thick fluid.

Animals↗

Molecular cloning and immunolocalization of two variants of the major basic nuclear protein (HCc) from the histone-less eukaryote Crypthecodinium cohnii (Pyrrhophyta).

Two clones that encode variants (HCc1 and HCc2) of the major basic nuclear protein of the dinoflagellate Crypthecodinium cohnii, were identified by immunoscreening of a cDNA expression library. The first clone carries a full-length cDNA with an open reading frame (HCc1) encoding 113 amino acids. The cDNA from the second clone lacks some of the 5' end, and the coding sequence is only 102 residues. The two proteins display 77% sequence similarity and their NH2-ends are homologous to the NH2-peptide of the HCc protein determined by P. Rizzo. The amino acid composition, which confirms the basic nature of lysine-rich HCc proteins, differs markedly from other known DNA-binding proteins such as histones, HMGs or prokaryotic histone-like proteins. No convincing homology was found with other proteins. HCc antigens were localized on C. cohnii by immunofluorescence, and by electron microscopy (EM) with immunogold labelling. HCc proteins are mainly detected at the periphery of the permanently condensed chromosomes, where active chromatin is located, as well as in the nucleolar organizing region (NOR). This suggests that these basic, non-histone proteins, with a moderate affinity for DNA, are involved at some level in the regulation of gene expression.

Amino Acid Sequence↗

Basic nuclear proteins of the histone-less eukaryote Crypthecodinium cohnii (Pyrrhophyta): two-dimensional electrophoresis and DNA-binding properties.

Unlike typical eukaryotes, the Dinoflagellate Crypthecodinium cohnii does not contain histones but six major basic, low molecular weight nuclear proteins which represent only 10% of the DNA mass and differ from histones in their electrophoretic and DNA-binding properties. These proteins are resolved in two-dimensional electrophoresis (AUT-PAGE x SDS-PAGE). Three proteins with an apparent molecular mass of 16, 16.5 and 17 kDa (p16, p16.5 and p17) are present in addition to the major 14 kDa basic nuclear component (HCc). HCc itself is resolved in three proteins (alpha, beta and gamma). When the proteins are not reduced with 2-mercaptoethanol before 2D-PAGE, the migration of HCc alpha, beta and gamma is modified in a way which suggests the formation of both inter- and intramolecular disulfide bridges and thus, the presence of at least two cysteines. The amino-acid analysis of HCc proteins resolved in 2D gels confirms that they are lysine-rich. HCc alpha, beta and gamma as well as p16, p16.5 and p17 are removed from isolated chromatin with 0.6 M NaCl, indicating that their affinity for DNA in vivo is lower than that of core histones. Furthermore, in vitro, they bind more tightly to single-stranded than to double-stranded DNA.

Animals↗

[Chronic cryptogenic pneumonia].

We report a case of a 72 year old man with the appropriate criteria for the recently identified chronic cryptogenic pneumonia: dyspnoea, cough, low general state, fever, raised sedimentation rate, localised opacities on the chest x-ray; no cause has been identified; the patient will improve on steroid therapy even though antibiotics are without effect, but relapses on stopping the steroids. A lung biopsy shows a predominant intra-alveolar fibrosis. A definitive cure can be obtained by 12 weeks of steroid therapy.

Aged↗

[Familial polycythemia caused by high oxygen affinity hemoglobin. 2nd world case of hemoglobin J Providence].

The cas observed in Auvergne and reported here raises the aetiological problem of polycythaemias. Young subjects with polycythaemia should be investigated for congenital anomaly of oxygen transport by measuring P50 and 2,3-DPG, which provides information on the oxygen-carrying capacity of haemoglobin. When confronted with familial polycythaemia due to high oxygen affinity haemoglobin, clinicians must know that a cause-effect relationship is not always demonstrable since other factors, such as tobacco-smoking in this particular case, may intervene.

Adult↗

Study of pp60v-src protein kinase activity in synchronized chicken embryo fibroblasts infected with Rous sarcoma virus.

Chicken embryo fibroblast (CEF) cultures, synchronized by the addition of serum to stationary cells, were exposed to Schmidt-Ruppin strain of Rous Sarcoma Virus (SR-RSV) and the appearance of pp60v-src protein kinase activity was examined through the cell cycle. In cells infected either at the beginning or at the end of G1, the onset of pp60v-src protein kinase activity was coincidental, closely following mitosis, with a delay between the infection of cells with SR-RSV and the appearance of protein kinase activity of about 20 and 16 h, respectively. In cells infected during the S phase this delay was 16 h, as observed for late G1 cells. These experiments show that the activity of pp60v-src protein kinase, which cannot be detected before the first mitosis following infection does not depend on G1. The aphidicolin prevented protein kinase activity if added before or at the beginning of S phase, but not if added later, which is presumably related to the inhibition of S phase, required for provirus integration. The use of colcemid, which suppresses cell division, did not inhibit but delayed the appearance of protein kinase activity. These results show that the synthesis of an active oncogene product, such as pp60v-src protein kinase, depends on both S phase and mitosis.

Animals↗

Immunological study of a cellular 35K phosphorylated polypeptide detected in Rous sarcoma virus transformed cells.

We have immunoprecipitated a phosphoprotein of 35K daltons (35K) common to RSV-transformed chick embryo fibroblasts (CEF) and rodent cells. The phosphorylation of this antigen depends on the expression of the v-src gene and contains phosphotyrosine. The pre-existing 35K protein, of CEF infected de novo, was further shown to become phosphorylated shortly after the appearance of active pp60v-src, and about 1 day before morphological transformation. The experiments with RSV-transformed rodent cells have shown that the 35K phosphoprotein is associated with the cellular framework. Another phosphoprotein of 37K was found in the RSV-transformed rodent cells, but not in the transformed CEF. However, this protein was not phosphorylated at tyrosine residues and its phosphorylation persisted, at the restrictive temperature, in cells transformed by a ts mutant of RSV.

Animals↗