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Biomedical subjects

A Girard

Publications and source records attributed to A Girard.

At least 109 records · Page 6Linked to original sources

[Effects of aminoglycosides on phosphoinositide metabolism in renal proximal tubules. In vivo and in vitro studies].

Phosphoinositides play a major role in the interaction between aminoglycosides and renal epithelial cells. We therefore investigated both in vivo and in vitro, in isolated proximal tubules and in various parts of the rabbit kidney, the influence of neomycin, gentamicin and amikacin on the phosphoinositide metabolism. This was carried out by measuring the 32p incorporation into phosphatidylinositol 4,5-bisphosphates (PI-P2) and phosphatidylinositol 4-phosphates (PI-P). In aminoglycoside-treated animals, compared to untreated controls, the 32 p-PI-P2 and 32p-PI-P levels were decreased and increased respectively in proximal tubules and in cortical areas but these levels were unchanged in papilla and in medulla. Similar variations could be obtained after exogenous addition of aminoglycosides. PI-P2 is a key membrane component in the regulation of both the Ca2+ transport and the intracellular Ca2+ mobilization. Thus, the aminoglycoside-induced modifications of the phosphoinositide metabolism may be considered as a primary event in the drug toxicity which may result from alterations in the cell calcium homeostasis.

Amikacin↗

[Phosphoinositide metabolism in essential and experimental hypertension].

The metabolism of phosphoinositides, a class of membrane lipids that appears to be intimately involved in the regulation by the membrane of the intracellular Ca2+ level, has been reported to be modified in the erythrocyte of the spontaneously hypertensive rat (SHR and SHR/SP). In order to elucidate the link between the phosphoinositide alteration and hypertension, the metabolism of phosphoinositides was studied in human essential hypertension and in Sabra rats under various patho-physiological conditions. Experiments were performed in vitro on isolated ghost membranes by measuring the radioactivity incorporated into triphosphoinositides (PI-P2) and diphosphoinositides (PI-P) following the incubation of membranes with [gamma 32P]-ATP. 32P-PI-P2, in moderate untreated essential hypertensive controls (n = 31) was higher than in normotensives (n = 30) (1.18 +/- 0.06 vs 0.92 +/- 0.04, 32P nmol/15 min/mg prot, p less than 0.005); 32P-PI-P2 and 32P-PI-P in hypertensive patients treated with beta-blocking agents (n = 20) did not differ from the values observed in untreated hypertensives. In Sabra rats, 32P-PI-P2 values were 0.79 +/- 0.03 and 1.32 +/- 0.08 for SbN and SbH, respectively (8-11 animals per group); difference was significant. 32P-PI-P values varied similarly. Both 32P-PI-P2 and 32P-PI-P did not change significantly when animals were fed a high sodium diet or were injected with DOCA, though such treatments rose the blood pressure. Our data indicate that the modification of phosphoinositide metabolism that we observed both in rat and human hypertension is not a consequence of the blood pressure elevation, but may be considered as an intrinsic membrane defect. Changes in the phosphoinositide metabolism may therefore be associated with the functional and structural alterations concerning the transmembrane Na+ and Ca2+ fluxes which may be of pathogenic importance.

Adult↗

Polyacrylamide gel electrophoresis in quantification of high-density lipoprotein cholesterol.

We evaluated a method for quantifying high-density lipoprotein cholesterol in plasma, based on electrophoretic migration of the prestained (with Sudan Black III) sample through a discontinuous polyacrylamide++ gel and densitometric integration of the stain associated with each class of lipoprotein. With this method, operations can be carried out on all types of lipoproteins over a broad range of concentrations. Overloading with very-low and low-density lipoproteins did not affect reliability within a wide range of HDL concentrations (0.45 to 16.60 mmol/L). Results for 22 individual plasma samples from normal and dyslipemic subjects correlated well with those by ultracentrifugal analysis (r=0.96; Student's t= 0.90, p > 0.30). We conclude that this method is reliable, sensitive, and accurate, It may be used for simultaneously typing dyslipoproteinemias and assaying HDL cholesterol.

Cholesterol, HDL↗

[Not Available].

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France↗

Triphosphoinositide breakdown and dense body release as the earliest events in thrombin-induced activation of human platelets.

The activation by thrombin of human platelets prelabelled with 32P induced a 30-40% decrease in 32P-triphosphoinositides (TPI) in the first 10 sec; the decrease in the other 32P-labelled phosphoinositides occurred by 20-30 sec. At 10 sec., the intensity of these effects was maximum with 0.2-0.4 U/ml thrombin. Under these conditions, 53, 20 and 15% of the dense granule, alpha-granule and lysosome constituents, respectively were released and thromboxane B2 synthesis reached only 10% of its maximum. Together with experiments carried out with chlorpromazine - or PGE1 - treated platelets, our results suggest the existence of a close relationship between TPI-breakdown and dense body release which appear to be the earliest events resulting from the activation of human platelets by thrombin.

Blood Platelets↗

Cells with neuronal properties in permanent cultures of quail embryo neuroretinas infected with Rous sarcoma virus.

Neuroretina cells from 7-day quail embryos infected 'in vitro' with the mutant ts NY-68 of Rous sarcoma virus, were established into permanent cultures. An initial stage of cellular proliferation was followed by a period of minimal multiplication. After recovery from this crisis, cell proliferation resumed. About 30% of the cells had binding sites for tetanus toxin and the monoclonal antibody A2B5 which seem to be specific for neurons, and an ultrastructural study suggested the presence of neurons and Müller (astroglial) cells. The specific activity of glutamic acid decarboxylase, the enzyme responsible for the synthesis of the neurotransmitter gamma-aminobutyric acid was high (10-30 nmol CO2/h/mg of protein) and electrophysiology showed that some cells had 'active' membranes. After about 18 months in culture, approximately 20% of the cells were able to respond to electrical stimulation by producing action potentials which were inhibited by 10(-7) M tetrodotoxin. These electrophysiological properties are stable: they have been repeatedly found at regular intervals throughout a 20 months period. Furthermore, a clone in which all tested cells are excitable, has been derived from the mass culture. Quail embryo neuroretina cells with typical neuronal properties can thus be established into permanent cultures after infection with Rous sarcoma virus.

Action Potentials↗

A neuronal clone derived from a Rous sarcoma virus-transformed quail embryo neuroretina established culture.

Neuroretina (NR) is an evagination of the central nervous system (CNS) which is composed of photoreceptors, glial (Müller) cells and horizontal, bipolar, amacrine and ganglion neuronal cells. We describe here the usefulness of Rous sarcoma virus (RSV) in the establishment of a neuronal clone from quail embryo neuroretina. When primary cultures of chick and quail embryo neuroretina cells are transformed by RSV, neuronal markers such as ribbon synapses, choline acetyltransferase (CAT) and glutamic acid decarboxylase (GAD) specific activity are present. These RSV-transformed primary cultures can be established into permanent cell lines from which neuronal clones have been isolated. One of them, clone QNR/D, can generate tetrodotoxin(TTX)-inhibitable action potentials on electrical stimulation, has a high GAD activity and binds monoclonal antibodies raised against chick embryo neuroretina. The presence of these neuronal markers suggests that the QNR/D clone is derived from cells of the amacrine or ganglionic lineage. This is the first time that a neuronal cell clone of defined origin has been obtained from the CNS. The neuronal markers of the QNR/D clone are expressed at both the permissive and the non-permissive temperatures for transformation.

Action Potentials↗

Impairment of membrane phosphoinositide metabolism by aminoglycoside antibiotics: streptomycin, amikacin, kanamycin, dibekacin, gentamicin and neomycin.

Like many amphiphilic cationic drugs, aminoglycosides are able to produce phospholipidosis, mainly by inhibiting enzymes involved in phospholipid metabolism. Phosphoinositides have been suggested to function as receptors for aminoglycosides. Therefore, we investigated the influence of these drugs upon phosphoinositide metabolism by measuring the 32P-incorporation into the polyphosphoinositides, using the rat erythrocyte membrane as a model. Depending upon the experimental conditions, neomycin induced a decrease and/or an increase in the 32P-labeling of triphosphoinositides (TPI) and of diphosphoinositides (DPI), respectively. These variations were rapid and depended upon the drug concentration. At 0.3 mM, neomycin reversed the distribution of radioactivities associated with DPI and TPI without modifying the total radioactivity incorporated. This drug concentration altered neither the Mg++-activated TPI-specific phosphomonoesterase activity nor the Ca++-activated polyphosphoinositide phosphodiesterase activity. It appears likely that the drug inhibits the DPI-kinase activity, by interacting with DPI and thereby lowering the substrate availability. Over the range of concentrations studied (up to 1-2 mM), gentamicin, kanamycin and dibekacin behave as neomycin. However, their effects could be observed only at drug concentrations higher than those of neomycin. By contrast, streptomycin and amikacin did not alter the 32P-labeling of TPI and of DPI. The order of potency of aminoglycosides for the impairment of the phosphoinositide interconversion was neomycin, gentamicin, dibekacin, kanamycin. A possible relationship between the toxicity of aminoglycosides and their capacity to impair the phosphoinositide metabolism is discussed.

Amikacin↗

Seroepidemiological survey of maedi-visna virus infection in sheep and goat flocks in Quebec.

Maedi-visna, a chronic viral disease of adult sheep characterized by progressive dyspnoea or neurological manifestations, was first recognized and described clinically in Canada in 1970. Seroepidemiological study was conducted in sheep and goats in various areas of Quebec. Sera of 10% of the animals of selected flocks were collected and specific antibodies against maedi-visna virus were tested by a modified direct complement fixation test. Results show seropositive rate of 67.6% for Sherbrooke sheep; of 40.5, 41.1 and 47.1% for Quebec, Saint-Hyacinthe and Nicolet sheep respectively and only 29.2 and 20.0% positive sera in l'Assomption and Rimouski animals. Prevalence rate of positive goats varied according to geographic areas (0 to 36.8%). Statistical analysis of various factors, e.g. age, breed, mode of raising, origin and size of flock showed no relation between these factors and the geographic areas. But, some clinical problems in the sheep flocks such as cough, rapid breathing, mortality and abortion were associated with high infection rate (greater than or equal to 50%) to maedi-visna virus. In goats, no correlation was demonstrated between these clinical signs and serological results. Our results suggest that it is important to consider this disease in an adequate program of preventive medicine in Quebec.

Animals↗

[Immunoreactive neuropeptides in cell lines derived from nervous and non nervous tissues].

Immunoreactive neuropeptides have been found in cells of permanent lines originating from nervous (cerebellum, retina) and non nervous (mesenchyme, kidney, bladder, colon, lung, melanocyte) tissues. The pattern of neuropeptides varied from one cell type to another, but all the cell lines studied contained immunoreactive alpha-endorphin and methionin-enkephalin.

Animals↗

Expression of neuronal markers in chick and quail embryo neuroretina cultures infected with Rous sarcoma virus.

Cultures of neuroretina (NR) cells from 7-day chick and quail embryos were infected with ts NY-68, a thermosensitive mutant of Rous sarcoma virus (RSV) which transformed NR cells at 36 degrees C. The following differentiation markers for neurones were studied: tetanus toxin-binding sites at the cell surfaces, presence of synapses, and the specific activity of the enzymes choline acetyltransferase (CAT) and glutamic acid decarboxylase (GAD). Appearance of synapses and expression of CAT were similar in control and transformed cultures. Tetanus toxin-binding cells were observed in transformed primary cultures and also in quail NR subcultures. GAD-specific activity was markedly stimulated in chick and quail primary cultures transformed by ts NY-68 and further increased in subcultures of ts NY-68-transformed quail NR cells. Stimulation of GAD activity is controlled by the transforming (src) gene of RSV since it was not observed in cultures infected with RAV-1, a leukosis virus which lacks the src gene. These data show that infection of chick and quail NR cultures with RSV results in the transformation of cells with neuronal markers.

Animals↗

[Demonstration of neuropeptides in cultured neuroretinal cells of chick embryo].

Immunoreactive neuropeptides have been found in cultures of neuroretina cells obtained from 7 day Chick embryos. Methionine-enkephalin and thyroliberin (TRH) were present in a large number of neurons, while somatostatin, beta-melanotropin (MSH) and alpha-endorphin were occasionally seen in a few neurons. In cultures devoid of neurons and probably made of Müller (astroglial) cells, methionine-enkephalin, alpha-MSH and TRH were each detected in 60 to 80% of the cells. Immunoreactive methionine enkephalin was also localized in Müller cells in retina sections of 10 and 20 day old embryos.

Animals↗

Gamma ray inactivation of some animal viruses.

Twenty samples of animal viruses comprising 14 different viruses in 12 families were subjected to varying doses of gamma irradiation from a 60Co source in a Gamma Cell 220 (Atomic Energy of Canada Limited) to determine lethal dose levels. The dose responses appeared linear throughout inactivation. The D10 values, that is the dose necessary to reduce infectivity by one log10, ranged from less than 0.20 Megarads to approximately 0.55 Megarads. There was not a complete inverse correlation between the target size (virion core) and the D10 value.

Dose-Response Relationship, Radiation↗

[Hydroxy 9-methyl 2-ellipticinium acetate (NSC 264-137). Toxicologic study and therapeutic effect in 100 cancers (author's transl)].

A new derivative of ellipticine, hydroxy-9-methyl-2-ellipticinium acetate, was found to be a useful anti-tumor drug in advanced cancers which could not be treated any longer successfully by any other procedure. In our series of 100 patients, the best results were obtained with bone metastases from breast carcinomas and with anaplastic thyroid carcinomas. Most patients usually received a weekly perfusion of 80 mg/m2. The main characteristic of this drug is its lack of hematologic, and hepatic toxicity. No renal trouble was observed during the first year, but 2 deaths from renal insufficiency occured during the 18th and 15th month of treatment. The most frequent side effect consists of digestive troubles (nausea, vomiting) which rarely compelled to stop the treatment (4 times in 100 patients).

Alkaloids↗

Studies on intercellular adhesion. III. Adhesion of cells to isotypic or heterotypic collecting particles and monolayers.

Adhesion between cells of different tissues from chick embryos was studied by the modified collecting particle (Pessac et al., '77a) and stationary monolayer assays. These assays measure the number of 3H labeled cells that adhere to the surface of cell aggregates and tissue fragments or on top of cell monolayers. The numbers of cells from a given suspension that adhere to identical or different cells can be compared. Intercellular adhesion may be considered as tissue specific if, in identical conditions, more cells adhere to identical cells than to cells from different tissues. Neural retina, cerebrum, optic tectum, liver and kidney cells from 9-day-old chick embryos showed no tissue specificity of adhesion, while heart cells adhered preferentially to collecting heart fragments.

Animals↗