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Biomedical subjects

M Gauthier

Publications and source records attributed to M Gauthier.

At least 37 records · Page 2Linked to original sources
PubMed2001

Activation of human neutrophils by technical toxaphene.

Toxaphene is a persistent organic pollutant (POP) known to be composed of numerous congeners. Toxaphene technical mixture applied as a pesticide consists of over 800 congeners. Among these, T(2) and T(12) are the two environmentally prevalent forms found in humans. Although toxaphene is known to exert some toxic effects, including potential proinflammatory properties, little is known concerning its action on cells of the human immune system, especially neutrophils. In the present study, we found that toxaphene was not necrotic for human neutrophils incubated for up to 24 h with concentrations ranging from 0.1 to 50 microg/ml. Toxaphene was found to induce neutrophil superoxide production (O(-)(2)) in a concentration-dependent manner. The potency and the kinetics of toxaphene-induced O(-)(2) by neutrophils were found to be similar to that of the classical neutrophil agonists phorbol 12-myristate 13-acetate (PMA). Furthermore, the use of various transduction signal inhibitors (genistein, pertussis toxin, staurosporine, H-7, and HA-1077), suggests that, as for PMA, toxaphene mediates its effect primarily via PKCs and, to a lesser extend, via tyrosine kinases. In this respect, staurosporine, H-7, and genistein were found to inhibit toxaphene- and PMA-induced O(-)(2) production by 52, 72, and 31% and by 63, 62, and 23%, respectively. Toxaphene was also found to significantly enhance neutrophil phagocytosis of opsonized sheep red blood cells and to induce neutrophil apoptosis. The induction of neutrophil apoptosis was paralleled with a decrease in CD16 expression. T(2) and T(12), the two prevalent congeners found in humans, were also found to significantly increase the O(-)(2) production in neutrophils at a concentration of 5 microg/ml. We conclude that neutrophils are important targets for toxaphene, as this POP can activate O(-)(2) production by a PKC- and tyrosine kinase-dependent mechanism, induce phagocytosis, and accelerate the apoptotic rate. This is the first study that focuses on toxaphene/human neutrophil interactions.

Apoptosis↗
PubMed2001

Contrasting effects of imidacloprid on habituation in 7- and 8-day-old honeybees (Apis mellifera).

We examined the effects of sublethal doses (0.1, 1, and 10 ng per animal) of a new neonicotinoid insecticide, Imidacloprid, on habituation of the proboscis extension reflex (PER) in honeybees (Apis mellifera) reared under laboratory conditions. In untreated honeybees, the habituation of the proboscis extension reflex is age-dependent and there is a significant increase in the number of trials required for habituation in older bees (8-10 days old) as compared to very young bees (4-7 days old). Imidacloprid alters the number of trials needed to habituate the honeybee response to multiple sucrose stimulation. In 7-day-old bees, treatment with Imidacloprid leads to an increase in the number of trials necessary to abolish the response, whereas in 8-day-old bees, it leads to a reduction in the number of trials for habituation (15 min and 1 h after treatment), and to an increase 4 h after treatment. The temporal effects of Imidacloprid in both 7- and 8-day-old bees suggest that 4h after treatment the observed effects are due to a metabolite of Imidacloprid, rather than to Imidacloprid itself. Our results suggest the existence of two distinct subtypes of nicotinic receptors in the honeybee that have different affinities to Imidacloprid and are differentially expressed in 7- and 8-day-old individuals.

Age Factors↗
PubMed2001

Memory impairment induced by cholinergic antagonists injected into the mushroom bodies of the honeybee.

The role of honeybee central brain structures, suspected to be cholinergic, has been studied in learning and memory. The nicotinic antagonist mecamylamine and the muscarinic antagonist scopolamine were locally injected into the calyces and the alpha-lobes of mushroom bodies, and their effects on memory acquisition and retrieval were investigated using one-trial olfactory conditioning of the proboscis extension reflex. A strong impairment of the olfactory learning was noticed following mecamylamine injection into the mushroom body calyces. Mecamylamine and scopolamine disturbed retrieval processes when injected into the alpha-lobes of mushroom bodies but remain without effect on these processes when injected into the mushroom body calyces. These results emphasise the role of the cholinergic networks of the mushroom bodies in the formation and recall of memory in the honeybee. They suggest that the role of the brain structures in these processes is sequential. Mushroom body calyces involved in the associative process of olfactory learning could be relayed by the alpha-lobes for information retrieval.

Animals↗
PubMed2001

Sexual maturation and fertility of male Nigerian Dwarf goat (Capra hircus) clones produced by somatic cell nuclear transfer.

Three, genetically identical, Nigerian Dwarf bucks produced by somatic cell nuclear transfer (NT) of fetal fibroblasts were monitored for sexual maturation and fertility. Starting at four months of age, these male clones were trained to serve an artificial vagina (AV). Average age of the NT-derived bucks at first semen collection was 20 weeks, which was not different from that of other young bucks of this breed (average age at first collection = 20 weeks). Average sperm production at 5 months of age for the NT-derived bucks was 5.0 x 10(8) spermatozoa, which was comparable to that of dwarf bucks of similar age (3.4 x 10(8) spermatozoa). At seven months of age, semen collected from two NT-derived bucks was used to artificially inseminate six females (three does per buck). Five does were confirmed pregnant by ultrasound at day 42. Nine healthy kids, four males and five females, were born in March and April 2000. Viable spermatozoa were collected from one of the F1 males at 28 weeks of age. These results demonstrated that NT-derived bucks and one of their male offspring developed sexually within the normal timeframe for their breed and that the clones were fertile.

Animals↗
PubMed2001

Activation of human neutrophils by chlordane: induction of superoxide production and phagocytosis but not chemotaxis or apoptosis.

We have recently reported that human neutrophils are important targets of different xenobiotics, including chemicals of environmental concern. In the present study, we found that chlordane was not toxic for human neutrophils incubated for up to 24 h with concentrations ranging from 0.1 to 50 microg/ml. Chlordane was found to induce neutrophil superoxide production (O2-) in a concentration-dependent fashion and its potency to induce this response was found to be similar to phorbol 12-myristate 13-acetate (PMA), a classical neutrophil agonist. The use of different transduction signal inhibitors (genistein, pertussis toxin, staurosporine, and calphostin C) indicates that, as for PMA, chlordane induces O2- production via protein kinase C (PKC). In this respect, staurosporine and calphostin C were found to inhibit chlordane- and PMA-induced O2- production by 65% and 72%, and by 83% and 85%, respectively. Chlordane was also found to significantly enhance neutrophil phagocytosis of opsonized sheep red blood cells (SRBCs). Despite these effects, chlordane did not alter neutrophil apoptosis as assessed by cytology (Diff-Quick staining) and by flow cytometry (CD-16 expression). In addition, chlordane did not alter neutrophil chemotaxis (48-well Boyden chamber). Cells were, however, responsive as they were activated by the well-characterized interleukin (IL)-8 chemokine. We conclude that chlordane can activate O2- production by a PKC-dependent mechanism and induce phagocytosis without altering chemotaxis and apoptosis. Chlordane must be added to a growing list of environmental contaminants that share some pro-inflammatory properties.

Apoptosis↗
PubMed2001

Activation of human neutrophils in vitro and dieldrin-induced neutrophilic inflammation in vivo.

Many chemicals of environmental concern are known to alter the immune system and are considered toxic molecules because they affect immune cell functions. Inflammation related to environmental chemical exposure, however, is poorly documented, except that from air pollutants. In this study, we found that the organochlorine insecticide dieldrin could not alter the ability of human neutrophils to phagocytose opsonized sheep red blood cells at nonnecrotic concentrations (0.1, 1, 10, and 50 microM). However, dieldrin was found to increase human neutrophil superoxide production, RNA synthesis, and proinflammatory cytokine interleukin-8 production. The normal apoptotic rate of neutrophils evaluated by both cytology and flow cytometry (CD-16 staining) was not altered by dieldrin treatments, and this was correlated with its inability to inhibit spreading of neutrophils onto glass. Using the murine air pouch model, we found that dieldrin induces a neutrophilic inflammation. Taken together, these results demonstrated that dieldrin is a proinflammatory contaminant. To our knowledge, this is the first report establishing that dieldrin is a contaminant exhibiting proinflammatory properties. In addition, it is the first time that the murine air pouch model has been successfully used to confirm that a chemical of environmental concern can induce an inflammatory response in vivo.

Animals↗
PubMed2000

Damping mechanism in dynamic force microscopy.

A general theory is presented which describes the damping in dynamic force microscopy due to the proximity of the surface, consistently with resonant frequency shift effects. Orders of magnitude for the experimentally measured "dissipation" and image corrugation are reproduced. It is suggested that the damping does not mainly result from energy dissipation, but arises because not all solutions of the microlever equation of motion are accessible. The damping is related to the multivalued nature of the analytical resonance curve, which appears at some critical tip-surface separation.

Biopolymers↗
PubMed2000

Functional cytochrome oxidase histochemistry in the honeybee brain.

The variations of neural metabolism induced by surgical and chemical treatments were studied in the honeybee brain by the means of cytochrome oxidase (CO) histochemistry. CO staining is considerably reduced in the alpha-lobe by antennal input deprivation. Chemical stimulation (50 mM K(+)) was linked to an increase of CO staining in antennal lobes (AL) and to a decrease in the basal ring of calyces (Cal). Application of the nicotinic ligand imidacloprid (10(-4) M) resulted in increased CO labelling within 30 min in all the structures analysed. Treatment with a lower concentration (10(-8) M) resulted in reduced staining in Cal and central body (CB). We conclude that CO histochemistry can be used to identify the target structures of cholinergic ligands in the honeybee brain.

Acetylcholine↗
PubMed2000

Frequency of Escherichia coli strains producing the cytotoxic necrotizing factor (CNF1) in nosocomial urinary tract infections.

The presence of cytotoxic necrotizing factor 1 (CNF1), together with various associated virulence factors (alpha-haemolysin, P-, S- and A-fimbriae), was screened in 175 uropathogenic Escherichia coli strains isolated from hospitalized adult patients. The cnf1 gene was detected in 30% of the selected strains independently of the severity of the clinical urinary infection. A significant association between CNF1, haemolytic activity and the products of the pap/sfa genes was found. However, CNF1 appeared not to play a major role in nosocomial E. coli urinary tract infections.

Adhesins, Bacterial↗
PubMed1999

Voluntary and reflex control of human back muscles during induced pain.

1. Back pain is known to change motor patterns of the trunk. The purpose of this study was to examine the motor output of the erector spinae (ES) muscles during pain in the lumbar region. First, their voluntary activation was assessed during flexion and re-extension of the trunk. Second, effects of cutaneous and muscle pain on the ES stretch reflex were measured, since increased stretch reflex gain has been suggested to underlie increased muscle tone in painful muscles. 2. The trunk movement and electromyographical (EMG) signals from the right and left ES during pain were compared with values before pain. Controlled muscle pain was induced by infusion of 5 % saline into the right lumbar ES. Cutaneous pain was elicited by mechanical or electrical stimulation of the dorsal lumbar skin. The stretch reflex was evoked by rapidly indenting the right lumbar ES with a servo-motor prodder. 3. The results from the voluntary task show that muscle pain decreased the modulation depth of ES EMG activity. This pattern was associated with a decreased range and velocity of motion of the painful body segment, which would normally serve to avoid further injury. Interestingly, when subjects overcame this guarding tendency and made exactly the same movements during pain as before pain, the EMG modulation depth was still reduced. The results seem to reconcile the controversy of previous studies, in which both hyper- and hypoactivity of back muscles in pain have been reported. 4. In the tapped muscle, the EMG response consisted of two peaks (latency 19.3 +/- 2.1 and 44.6 +/- 2.5 ms, respectively) followed by a trough. On the contralateral side the first response was a trough (26.2 +/- 3.2 ms) while the second (46.4 +/- 4.3 ms) was a peak, similar to the second peak on the tapped side. Cutaneous pain had no effect on the short-latency response but significantly increased the second response on the tapped side. Surprisingly, deep muscle pain had no effect on the stretch reflex. A short-latency reciprocal inhibition exists between the right and left human ES. 5. It is concluded that deep back pain does not influence the stretch reflexes in the back muscles but modulates the voluntary activation of these muscles.

Adult↗
PubMed1999

The 5' region of cnf1 harbours a translational regulatory mechanism for CNF1 synthesis and encodes the cell-binding domain of the toxin.

The Escherichia coli cytotoxic necrotizing factor 1 (CNF1) is organized into three functional domains: the N-terminal part containing the cell-binding domain, a putative central membrane-spanning region, and a C-terminal catalytic region. On the basis of competition assays between CNF1 and GST-recombinant proteins containing different N-terminal fragments, and point mutations, we restricted the binding region to the first 190 amino acids. Hydrophilic amino acids 53-75 are strictly necessary to cell receptor recognition. Using different cnf1-lacZ translational fusions, we demonstrated that the mRNA corresponding to the first 48 codons of cnf1 is involved in the translational regulation of CNF1 synthesis. This regulation consists of both a positive and a negative control. The positive control is exerted by codons 6-20, including a putative downstream box that enhances the translational expression of cnf1. The negative control depends on codons 45-48. In this region, an anti-Shine-Dalgarno sequence, highly homologous to the core of the internal complementary sequence already reported for growth rate-regulated metabolic genes, has been detected. To some extent, the inner structural organization of CNF1 would thus suggest the compiling of several functions in a single mRNA protein system.

5' Untranslated Regions↗
PubMed1999

A novel mechanism for benign essential blepharospasm.

PURPOSE: The purpose of this study is to test the hypothesis that the photophobia of benign essential blepharospasm (BEB) is caused by sympathetically maintained pain. METHODS: Nineteen patients with photophobia and BEB were enrolled in an unblinded prospective treatment trial. The intervention was blockade of the superior sympathetic ganglion with local anesthetic. Outcome measures included the patient's subjective report of ocular surface dryness, foreign body sensation, and eyelid spasm. We also obtained video recordings of eyelid movements. RESULTS: Of the 19 patients, 13 reported subjective improvement in BEB symptoms after cervical sympathetic blockade (CSB). Thirteen of 19 patients also had objective evidence of decreased light-induced eyelid spasm after CSB. Ocular surface disease was present in 18 of 19 patients. CONCLUSION: These data support the hypothesis that in many patients with BEB there is a sympathetically maintained pain syndrome associated with external ocular disease. We speculate on a neurologic circuit that may explain these findings.

Anesthetics, Local↗
PubMed1999

Ribavirin in ventilated respiratory syncytial virus bronchiolitis. A randomized, placebo-controlled trial.

We undertook a prospective, double-blind, placebo-controlled trial to resolve the question of the clinical effectiveness of ribavirin in previously well infants who require ventilation for respiratory distress secondary to respiratory syncytial virus (RSV) bronchiolitis. Aerosol ribavirin or NaCl 0.9% was administered within 24 h of initiation of ventilation, 18 h/d, for a maximum of 7 d or until extubation. From March 1994 to March 1997, 42 children were randomized and 41 patients were retained for analysis. Baseline characteristics of each group-ribavirin and placebo (20:21)-were not significantly different with respect to age (62.5 +/- 35.9 versus 62.7 +/- 30.9 d), sex, weight, and length of ventilation pre-aerosol. "Intent to treat" outcome analysis found no significant differences in the length of the following: ventilation (102.16 +/- 65.26 versus 126.28 +/- 78.72 h; p = 0.29), aerosol therapy, stay in the intensive care unit, total oxygen therapy, and hospitalization. The aerosols were well tolerated and no deaths occurred. This trial demonstrates the lack of effectiveness of aerosolized ribavirin in reducing the length of ventilation and course of illness in infants with no underlying illness ventilated for respiratory distress secondary to RSV bronchiolitis.

Administration, Inhalation↗
PubMed1998

Purification of a recombinant protein expressed in yeast: optimization of analytical and preparative chromatography.

The industrial production of recombinant proteins requires control of both fermentation and purification steps. For the serodiagnosis of toxoplasmosis, the main antigen is a membrane protein of 30 kDa (P30). The P30 gene was cloned and expressed in Schizosaccharomyces pombe at 0.7 microg/ml in culture medium. Batch fermentation was optimized by the specific choice of peptones, which enabled optimum growth and protein expression without reducing the efficacy of the purification step. Analytical purification was then carried out using cation-exchange chromatography. For larger volumes, scaling up was performed on expanded mode by using a Streamline system (Pharmacia). This purification step allowed us to obtain a 67.5% recovery with a purification factor greater than 27-fold. Expanded bed adsorption technology is a convenient and effective technique for protein capture directly from feedstock, and the eluted fraction is ready for a second affinity chromatography step. This second step is performed with a yield of 40% and provides a final purification factor of 2000-fold.

Adsorption↗
PubMed1998

Effects of the muscarinic antagonists atropine and pirenzepine on olfactory conditioning in the honeybee.

One-trial conditioning of the proboscis extension reflex (PER) in honeybees was used to examine the qualitative effects of two muscarinic antagonists, atropine and pirenzepine, on the acquisition and retrieval of memory following intracranial injection. The main result of this study is that atropine, at a relatively high concentration of 10(-2) M, impairs memory retrieval but not acquisition of memory after a single olfactory conditioning trial (at this concentration, there is no effect of atropine on the sensorimotor components of the PER). This result is in agreement with the effects of scopolamine, reported in a previously published article. Pirenzepine, at the same concentration as atropine, had no effect on either acquisition or retrieval of memory. These results suggest that blockade of muscarinic-like receptors, except those that bind to pirenzepine, induces solely an impairment of memory retrieval.

Animals↗
PubMed1998

A new attempt to assess the effect of learning processes on the cholinergic system: studies on fruitflies and honeybees.

The effect of training on the functioning of the cholinergic system was investigated in fruitflies and in honeybees. Drosophila were submitted to a passive avoidance conditioning of the proboscis extension response (PER). Flies had to learn to suppress the sugar-induced PER to avoid an aversive quinine reinforcement. In a yoked control group, the punishment was administered with no relation to the response displayed. Honeybees underwent a five-trial olfactory conditioning of the PER elicited by an antennal gustatory stimulation. In the control group, olfactory and gustatory stimulations were unpaired to prevent a learning process from developing. Immediately at the end of the learning session, acetylcholinesterase (AChE) activity was individually measured on the whole animal for Drosophila and on the head for the honeybee in experimental and in control groups. In fruitflies and honeybees, the AChE rate did not differ between the experimental group and its respective control group. Moreover, no significant correlation could be found individually between the learning performance and the AChE rate in either Drosophila or in honeybees. This experiment did not reveal any modulatory effect of the learning acquisition level on the AChE activity in insects as was previously reported in honeybees.

Acetylcholinesterase↗
PubMed1998

Bacterial toxins and the Rho GTP-binding protein: what microbes teach us about cell regulation.

In the present review activities of two bacterial toxins, Clostridium botulinum exoenzyme C3 and Escherichia coli CNF1, both acting on the GTP-binding protein Rho are analyzed. Proteins belonging to the Rho family regulate the actin cytoskeleton and act as molecular switches in a number of signal transduction pathways. C3 and CNF1 have opposite effects on Rho thus representing useful tools for studies on cell division, cell differentiation and apoptosis.

ADP Ribose Transferases↗