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1.
Ann Biol Clin (Paris) ; 77(6): 669-680, 2019 12 01.
Article in French | MEDLINE | ID: mdl-31859644

ABSTRACT

Although the use of EDTA-containing collection tubes is known to stabilize the complement analytes and to make the results more reliable, no external quality assessment (EQA) scheme based on EDTA plasma samples is available to date in France. Consequently, a number of clinical laboratories currently participate to EQA program on samples whose matrix is different from their routine practice. The aim of this work was to offer a new external quality assessment scheme, as an inter-laboratory exchange (ILE). The ILE samples come from pooled EDTA plasmas of healthy subjects and are diluted to obtain distinct control levels. The protocol has been validated on CH50, C3, C4 and C1-inhibitor measurements, through: (i) a stability study of post-centrifugation storage of EDTA plasma samples at room temperature, 4̊C and -20̊C; (ii) the demonstration of the linearity of the dilution steps; and (iii) a stability study of the diluted samples. Our results demonstrate a four-weeks stability of the ILE samples prepared and stored according to our protocol. Those results are compatible with the ILE implementation constraints, and the program has been implemented in January 2018. The one-year ILE implementation experience is also presented. The newly implemented ILE will be useful for the accreditation of the complement activity of French laboratories using EDTA plasma samples.


Subject(s)
Blood Chemical Analysis/methods , Blood Specimen Collection , Complement System Proteins/analysis , Edetic Acid/chemistry , Plasma/chemistry , Blood Chemical Analysis/standards , Blood Preservation/methods , Blood Preservation/standards , Blood Specimen Collection/methods , Blood Specimen Collection/standards , Complement System Proteins/drug effects , Complement System Proteins/metabolism , Edetic Acid/pharmacology , Excipients/chemistry , Excipients/pharmacology , Humans , Laboratory Proficiency Testing , Plasma/drug effects , Plasma/metabolism , Protein Stability/drug effects , Quality Assurance, Health Care/methods , Quality Control , Time Factors , Transportation/standards
2.
Psychopharmacology (Berl) ; 214(1): 197-208, 2011 Mar.
Article in English | MEDLINE | ID: mdl-20101392

ABSTRACT

RATIONALE: In rats, prenatal restraint stress (PRS) induces persistent behavioral and neurobiological alterations leading to a greater consumption of psychostimulants during adulthood. However, little is known about alcohol vulnerability in this animal model. OBJECTIVES: We examined in adolescent and adult male Sprague Dawley rats the long-lasting impact of PRS exposure on alcohol consumption. METHODS: PRS rats were subjected to a prenatal stress (three daily 45-min sessions of restraint stress to the mothers during the last 10 days of pregnancy). Alcohol preference was assessed in a two-bottle choice paradigm (alcohol 2.5%, 5%, or 10% versus water), in both naïve adolescent rats and adult rats previously exposed to a chronic alcohol treatment. Behavioral indices associated with incentive motivation for alcohol were investigated. Finally, plasma levels of transaminases (marker of hepatic damages) and ΔFosB levels in the nucleus accumbens (a potential molecular switch for addiction) were evaluated following the chronic alcohol exposure. RESULTS: Alcohol preference was not affected by PRS. Contrary to our expectations, stressed and unstressed rats did not display signs of compulsive alcohol consumption. The consequences of the alcohol exposure on locomotor reactivity and on transaminase levels were more prominent in PRS group. Similarly, PRS potentiated alcohol-induced ΔFosB levels in the nucleus accumbens. CONCLUSION: Our data suggest that negative events occurring in utero do not modulate alcohol preference in male rats but potentiate chronic alcohol-induced molecular neuroadaptation in the brain reward circuitry. Further studies are needed to determine whether the exacerbated ΔFosB upregulation in PRS rats could be extended to other reinforcing stimuli.


Subject(s)
Alcohol Drinking/epidemiology , Ethanol/administration & dosage , Prenatal Exposure Delayed Effects , Stress, Psychological/complications , Animals , Disease Models, Animal , Female , Male , Motor Activity/drug effects , Nucleus Accumbens/drug effects , Nucleus Accumbens/metabolism , Pregnancy , Proto-Oncogene Proteins c-fos/drug effects , Proto-Oncogene Proteins c-fos/metabolism , Rats , Rats, Sprague-Dawley , Restraint, Physical , Reward , Transaminases/blood
3.
Alcohol Clin Exp Res ; 33(8): 1346-54, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19413649

ABSTRACT

BACKGROUND: Although it is generally believed that chronic ethanol consumption impairs learning and memory, results obtained in experimental animals are not univocal, and there are conditions in which ethanol paradoxically improves cognitive functions. In the present work, we investigated the effects of prenatal stress and of chronic ethanol exposure during adulthood on spatial memory in rats. METHODS: Rats were subjected to a prenatal stress delivered as 3 daily 45-minute sections of restraint stress to the mothers during the last 10 days of pregnancy (PRS rats). After 7 months of ethanol exposure (ethanol 10%, oral intake), memory performances were evaluated in a spatial discrimination test in control and PRS male rats. Then, the oxidative damages and the expression of metabotropic glutamate (mGlu) receptors were assessed in their hippocampus. RESULTS: Chronic ethanol exposure resulted in a reduced performance in a spatial recognition task in control animals. Unexpectedly, however, the same treatment attenuated spatial memory deficits in rats that had been subjected to prenatal stress. This paradigm of ethanol administration did not produce detectable signs of oxidative damage in the hippocampus in either unstressed or PRS rats. Interestingly, ethanol intake resulted in differential effects in the expression of mGlu receptor subtypes implicated in mechanisms of learning and memory. In control rats, ethanol intake reduced mGlu2/3 and mGlu5 receptor levels in the hippocampus; in PRS rats, which exhibited a constitutive reduction in the levels of these mGlu receptor subtypes, ethanol increased the expression of mGlu1a receptors but did not change the expression of mGlu2/3 or mGlu5 receptors. CONCLUSION: Our findings support the idea that stress-related events occurring before birth have long-lasting effects on brain function and behavior, and suggest that the impact of ethanol on cognition is not only dose- and duration-dependent, but also critically influenced by early life experiences.


Subject(s)
Ethanol/administration & dosage , Hippocampus/metabolism , Memory Disorders/prevention & control , Prenatal Exposure Delayed Effects/metabolism , Receptors, Metabotropic Glutamate/biosynthesis , Stress, Psychological/metabolism , Animals , Female , Gene Expression Regulation, Developmental/drug effects , Gene Expression Regulation, Developmental/physiology , Hippocampus/drug effects , Male , Memory Disorders/metabolism , Pregnancy , Rats , Rats, Sprague-Dawley , Spatial Behavior/drug effects , Spatial Behavior/physiology , Up-Regulation/drug effects
4.
Brain Res ; 1191: 55-62, 2008 Jan 29.
Article in English | MEDLINE | ID: mdl-18096141

ABSTRACT

Prenatal restraint stress (PRS) in rats is associated with hippocampal dysfunctions and several behavioural and endocrine disorders related to this brain area. Recently, we have reported that the PRS modifies the hypothalamic-pituitary-adrenal (HPA) response to an ethanol challenge in adolescent animals. Since hippocampus is particularly sensitive to the deleterious effects of ethanol during adolescence, we investigated in this study the combined effects of PRS and ethanol administration on the oxidative status in the hippocampus of 28-day-old male rats. Thirty minutes after an intraperitoneal (i.p.) injection of ethanol (1.5 g/kg), the activities of several antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) but also non-enzymatic antioxidant (reduced glutathione) were assayed. Thiobarbituric acid reactive substances (TBARS) levels were also measured as a marker of lipid peroxidation. Ethanol enhanced superoxide dismutase activity in control rats but not in PRS rats. At basal level, catalase activity was lower in PRS rats than in control rats, indicating a potentially higher sensitivity to oxidative damages after this early stress. However, the hippocampal TBARS levels were not significantly affected by the ethanol administration, showing that an acute ethanol exposure does not induce oxidative damage in adolescent male rats. In conclusion, our data suggest that PRS affects both basal antioxidant status in the hippocampus and antioxidant response after an acute ethanol exposure. These findings extend previous works showing that PRS leads to hippocampal dysfunctions and raise the question of the potential increase of the hippocampal oxidative damage in PRS rats after repeated exposure to ethanol.


Subject(s)
Alcoholic Intoxication/enzymology , Hippocampus/enzymology , Oxidative Stress/drug effects , Oxidoreductases/metabolism , Prenatal Exposure Delayed Effects/enzymology , Stress, Psychological/enzymology , Age Factors , Alcoholic Intoxication/complications , Animals , Antioxidants/metabolism , Catalase/drug effects , Catalase/metabolism , Disease Models, Animal , Ethanol/pharmacology , Female , Glutathione/drug effects , Glutathione/metabolism , Glutathione Peroxidase/drug effects , Glutathione Peroxidase/metabolism , Hippocampus/drug effects , Hippocampus/growth & development , Male , Oxidoreductases/drug effects , Pregnancy , Rats , Restraint, Physical , Stress, Psychological/complications , Superoxide Dismutase/drug effects , Superoxide Dismutase/metabolism , Thiobarbituric Acid Reactive Substances/metabolism
5.
Eur J Neurosci ; 24(4): 1193-200, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16925589

ABSTRACT

The period of adolescence and environmental factors, such as stress, are important in determining ethanol vulnerability in both humans and rats. Ethanol is a powerful activator of the hypothalamic-pituitary-adrenal (HPA) axis but attenuated responses of the HPA axis to ethanol have been described in populations with a high risk of ethanol abuse. In rats, prenatal stress leads to prolonged stress-induced corticosterone secretion and increases the vulnerability to drugs of abuse, such as amphetamine and nicotine in adulthood and 3,4-methylenedioxymethamphetamine in adolescent rats. The aim of the present study was to assess the impact of a prenatal stress on HPA axis responsiveness to a moderate dose of ethanol (1.5 g/kg i.p.) in adolescent male rats (28 days old). The parameters evaluated were plasma adrenocorticotropic hormone, plasma corticosterone and mRNA expression of HPA axis central markers (mineralocorticoid receptor, glucocorticoid receptor, corticotropin-releasing hormone and pro-opiomelanocortin). Contrary to prior expectations, our results demonstrate that prenatal stress blunts the HPA axis responsiveness to a moderate dose of ethanol in adolescent rats in spite of similar blood ethanol levels. These data suggest that prenatal stress may have the opposite effect on the response to stress depending on the attributes of the stressor stimulus. They thus raise questions about the possible impact of prenatal stress on the further development of ethanol vulnerability.


Subject(s)
Ethanol/pharmacology , Hypothalamo-Hypophyseal System/drug effects , Pituitary-Adrenal System/drug effects , Stress, Psychological , Adrenocorticotropic Hormone/blood , Animals , Body Weight , Corticosterone/blood , Corticotropin-Releasing Hormone/genetics , Corticotropin-Releasing Hormone/metabolism , Dentate Gyrus/cytology , Dentate Gyrus/metabolism , Female , Humans , Hypothalamo-Hypophyseal System/physiology , Male , Paraventricular Hypothalamic Nucleus/cytology , Paraventricular Hypothalamic Nucleus/metabolism , Pituitary Gland/cytology , Pituitary Gland/metabolism , Pituitary-Adrenal System/physiology , Pregnancy , Pro-Opiomelanocortin/genetics , Pro-Opiomelanocortin/metabolism , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Receptors, Glucocorticoid/genetics , Receptors, Glucocorticoid/metabolism , Receptors, Mineralocorticoid/genetics , Receptors, Mineralocorticoid/metabolism
6.
J Anal Toxicol ; 30(2): 86-90, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16620537

ABSTRACT

The purpose of this work is to characterize chemical compounds added to an ingested soda by (1)H nuclear magnetic resonance ((1)H NMR) spectroscopy and by gas chromatography-mass spectrometry in the electron impact mode. A second point was to highlight possible metabolic disturbances by considering urinary profile. Without any pretreatment, dimethylphtalate, 2-butanone, and 2,2,4-trimethylpentanediol diisobutyrate were found in the adulterated soda. Quantitative analysis was performed by relative integration of peak areas. Huge quantities of 2,2,4-trimethylpentanediol diisobutyrate and dimethylphtalate were found in the oily layer. 2-Butanone, which is miscible in water, was found in the two phases as well as small quantities of dimethylphtalate. The urine sample was collected on hospital admission and was also analyzed by (1)H NMR spectroscopy. The major abnormal compound found was 1,2-propanediol. Other disturbances concerned endogenous metabolites such as 2-ketoglutaric acid, lactic acid, and betaine.


Subject(s)
Carbonated Beverages/analysis , Food Contamination/analysis , Foodborne Diseases/diagnosis , Gas Chromatography-Mass Spectrometry/methods , Magnetic Resonance Spectroscopy/methods , Xenobiotics/poisoning , Butanones/urine , Female , Foodborne Diseases/urine , Forensic Medicine/methods , Humans , Middle Aged , Octanes/urine , Phthalic Acids/urine , Propylene Glycol/urine , Workplace , Xenobiotics/urine
7.
Ther Drug Monit ; 25(4): 421-32, 2003 Aug.
Article in English | MEDLINE | ID: mdl-12883224

ABSTRACT

Recently, interest in hair analysis in such fields as drug abuse, driving, or for clinical purposes (determination of drug-exposed neonates especially) has grown because of the highly sensitive method of detection (GC-MS) that can now be applied. Neonates born to drug-addicted mothers can suffer from neonatal withdrawal syndrome (NWS), which requires morphine treatment in its severe forms. To assess and measure toxicologic factors predicting the appearance and the severity of this syndrome, matrices such as urine, meconium, and hair are necessary. Cannabinoids, opiates, cocaine (and its metabolites), and methadone in particular were determined in the various matrices collected in 17 mother/neonate pairs. An immunologic screening method was used, and quantification was achieved with GC-MS. In spite of some bias (color, length, race) that might hinder an accurate interpretation, the results of hair analysis makes it possible to confirm a fetal drug exposure and to reinforce the diagnosis of the NWS observed, particularly when results obtained in other matrices are negative. Hair analysis contributes to our ability to predict a NWS.


Subject(s)
Hair/chemistry , Neonatal Abstinence Syndrome/diagnosis , Substance Abuse Detection/methods , Cannabinoids/adverse effects , Cannabinoids/analysis , Cannabinoids/urine , Cocaine/adverse effects , Cocaine/analysis , Cocaine/urine , Female , Gas Chromatography-Mass Spectrometry , Humans , Infant, Newborn , Meconium/chemistry , Methadone/adverse effects , Methadone/analysis , Methadone/urine , Narcotics/adverse effects , Narcotics/analysis , Narcotics/urine , Pregnancy
8.
Forensic Sci Int ; 133(1-2): 57-62, 2003 Apr 23.
Article in English | MEDLINE | ID: mdl-12742690

ABSTRACT

The increasing interest in toxicological hair analysis as a marker of human exposure to xenobiotics such as illicit substances or therapeutic drugs, has been made feasible by the extension of mass spectrometry, a highly sensitive method of detection. A newborn exposed to drugs in utero can suffer from a varying degree of withdrawal syndrome, a few days after birth. If of opiate origin, the withdrawal syndrome can be treated with morphine, among other therapeutics, but it is not easy to diagnose because of atypical symptoms presented by neonates and especially when maternal drug addiction has not been revealed. To assess and measure toxicological factors linked with the appearance and the severity of this syndrome, maternal and neonatal matrices such as urine, meconium and hair were collected during a protocol approved by the ethical committee. Opiates in particular were measured with GC-MS and potential combined dependences (cannabis, cocaine, amphetamine, LSD and benzodiazepines) and/or substitutive therapeutics (methadone or buprenorphine) were also assessed in 17 mother/neonate couples. Gestational opiate exposure profiles were drawn up and linked with the observed withdrawal syndromes. A withdrawal syndrome seems to appear more frequently after foetal exposure to an association of opiates/substitutive molecules (8 out of 10 withdrawal syndromes observed in this study), although the impact of cocaine and benzodiazepines must also be taken into account. The results obtained in neonatal hair make it possible to affirm foetal drug exposure and are in accordance, for the majority, with the appearance of a neonatal withdrawal syndrome (NWS). Neonatal hair analysis could contribute to assess in utero exposure to opiates, particularly when results in urine and meconium are negative or when these matrices are not available.


Subject(s)
Hair/chemistry , Maternal-Fetal Exchange , Narcotics/analysis , Female , Gas Chromatography-Mass Spectrometry , Humans , Immunoassay , Infant, Newborn , Meconium/chemistry , Pregnancy , Substance Abuse Detection/methods
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