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Biomedical subjects

P Mangin

Publications and source records attributed to P Mangin.

At least 19 recordsLinked to original sources

Testing human hair for flunitrazepam and 7-amino-flunitrazepam by GC/MS-NCI.

To validate information on flunitrazepam use, we investigated human hair for flunitrazepam and its major metabolite 7-amino-flunitrazepam by gas chromatography coupled to mass spectrometry in negative chemical ionization mode of detection. Samples were twice decontaminated with methylene chloride, pulverized in a ball mill and 50 mg of powdered hair were incubated in Soerensen buffer (pH 7.6) in the presence of diazepam-d5 used as internal standard. After liquid-liquid extraction of the incubation medium with diethylether-chloroform (80:20, by vol.), the organic phase was evaporated and the dry extract was derivatizated with heptafluorobutyric anhydride. Benzodiazepines were separated on a 30 m capillary column and detected using single ion monitoring. Among 40 hair samples tested (obtained from drug addicts deceased by heroin overdose), 14 were positive for both flunitrazepam and 7-amino-flunitrazepam and 12 for 7-amino-flunitrazepam only. Concentrations ranged from 31 to 129 pg/mg (mean: 60 pg/mg) and from 3 to 161 pg/mg (mean: 46 pg/mg) for flunitrazepam (14 cases) and 7-amino-flunitrazepam (26 cases), respectively. This first report described the detection of flunitrazepam and 7-amino-flunitrazepam in hair of chronic abusers. Due to the low concentrations observed, negative chemical ionization appears to be the alternative to test flunitrazepam and other benzodiazepines in hair.

Flunitrazepam

HPLC/MS determination of buprenorphine and norbuprenorphine in biological fluids and hair samples.

An original method, based upon HPLC (high performance liquid chromatography)/Ionspray-MS, has been developed for the identification of buprenorphine (BUP) and norbuprenorphine (norBUP) in biological fluids and hair samples. Biological fluids (2 mL) are extracted at pH 8.4 by CHCl3/2-propanol/n-heptane (25:10:65, v/v) after addition of deuterated BUP (BUP-d4, 10 ng). Hair samples (40 mg) are extracted in the same conditions after decontamination by CH2Cl2, mechanical pulverization, addition of BUP-d4 (1 ng), acidic incubation (1 mL 0.1 N HCl, 56 degrees C overnight), then neutralization by NaOH. Analytes are separated on a 4-microns NovaPak C18 (Waters) column (150 by 2.0 mm, ID) with a mobile phase of acetonitrile/2 mM NH4COOH buffer, pH 3.0 (80:20, v/v; flow rate 200 microL/min; post column split 1:3). Detection is done by a Perkin-Elmer Sciex API-100 mass analyzer equipped with an ISP interface (nebulizing and curtain gas:99.95-% N2; main settings: orifice + 50 V, electron multiplier + 2400 V). The mean retention times for BUP, BUP-d4, and norBUP are 5.84, 5.79, and 4.42 min, respectively. For all compounds, mass spectra exhibit a unique, protonated molecular ion [M + H]+ at m/z 414 (norBUP), 468 (BUP), and 472 (BUP-d4), without any significant fragmentation. The lower limits of detection are 0.10 and 0.05 ng/mL blood, and 4 and 2 pg/mg hair for BUP and norBUP, respectively. BUP and norBUP concentrations measured in hair from six addicts under substitutive therapy by BUP ranged from 4 to 140 pg/mg, and from nondetected to 67 pg/mg, respectively. The good performances of this method in terms of both sensitivity and specificity make it a convenient alternative to HPLC/coulometry and GC/MS for the separate analysis of BUP and norBUP in biological samples.

Adult

Simultaneous high-performance liquid chromatographic analysis of flunitrazepam and four metabolites in serum.

A high-performance liquid chromatographic method for the simultaneous determination of flunitrazepam and four metabolites, desmethylflunitrazepam (DMF), 7-aminodesmethylflunitrazepam (7-NH2DMF), 7-aminoflunitrazepam (7-NH2F) and 3-hydroxyflunitrazepam (3-OHF), in serum is described. The method involves a simple extraction from alkalinized plasma (pH 9.5) into diethyl ether-chloroform (80:20, v/v). Prazepam was used as an internal standard for the quantification of the five compounds. Separation was achieved with a 10 microns RSil CN column (300 x 3.9 mn I.D.). The detection wavelength was set at 242 nm. The limits of detection ranged from 2.5 to 5 micrograms/l with a limit of quantification of 10 micrograms/l for all analytes.

Anti-Anxiety Agents

Headspace GC/MS testing for chlorodifluoromethane in two fatal cases.

Two cases of lethal poisoning due to chlorodifluoromethane (Freon 22) inhalation are described. The fluorocarbon was determined in biological tissues by headspace gas chromatography/mass spectrometry. Ions monitored were m/z 67, 86 and 51, the latter being used for quantification. Blood concentrations were 26.0 and 37.1 microliters/ml. In both cases, the drug was also identified in urine, vitreous humor and bile, but in much lower concentrations.

Accidents, Occupational

Hair analysis for nordiazepam and oxazepam by gas chromatography--negative-ion chemical ionization mass spectrometry.

A procedure is presented for the identification of nordiazepam and its metabolite, oxazepam, in human hair. The method involves decontamination of hair with dichloromethane, incubation in phosphate buffer (pH 7.6) in the presence of deuterated internal standards, liquid-liquid extraction, derivatization with N,O-bis(trimethylsilyl)trifluoroacetamide plus 1% trimethylchlorosilane and gas chromatography-mass spectrometry using negative-ion chemical ionization with methane. Among thirty samples obtained from polydrug abusers, thirteen tested positive for nordiazepam, in the range of 0.25-18.87 ng/mg. Five samples were also positive for oxazepam, in the range 0.11-0.50 ng/mg.

Adult

High-performance liquid chromatography coupled to ion spray mass spectrometry for the determination of colchicine at ppb levels in human biofluids.

An original method based upon high-performance liquid chromatography coupled to ion spray mass spectrometry (HPLC-ISP-MS) has been developed for the identification and quantification of colchicine (COL) in human blood, plasma or urine. After single-step liquid-liquid extraction by dichloromethane at pH 8.0 using tofisopam (TOF) as an internal standard, solutes are separated on a 5-microns C18 Microbore (Alltech) column (250 x 1.0 mm, I.D.), using acetonitrile-2 mM NH4COOH, pH 3 buffer (75: 25, v/v) as the mobile phase (flow-rate 50 microliters/min). Detection is done by a Perkin-Elmer Sciex API-100 mass analyzer equipped with a ISP interface (nebulizing and curtain gas: N2, quality U; main settings: ISP, +4.0 kV; OR, +50 V; Q0, -10 V; Q1, -13 V; electron multiplier, +2.2 kV); MS data are collected as either total ion current (TIC, m/z 100-500 or 380-405), or selected ion monitoring (SIM) at m/z 400 and 383 for COL and TOF, respectively. COL mass spectrum shows a prominent molecular ion [M + H]+ at m/z 400. Increasing OR potential fails to provide a significant fragmentation. Retention times are 2.70 and 4.53 min for COL and TOF, respectively. The quantification method shows a good linearity (r = 0.998) over a concentration range from 5 to 200 ng/ml. The lower limit of detection in SIM mode is 0.6 ng/ml COL, making the method convenient for both clinical and forensic purposes.

Anti-Anxiety Agents

Water-jet dissection in renal surgery: experimental study of a new device in the pig.

The pig model was used for experiments with a new type of water-jet dissector, which produces high-pressure water by application of a gas and maintains the water fully sterile in a single-use delivery apparatus. The experiment was conducted ex vivo (14 kidneys) and in vivo to compare electric cautery section with water-jet dissection (5 vs 11 partial nephrectomies). Ex vivo study confirmed sparing of blood vessels and pelvicaliceal system. In vivo study did not show significant differences in blood loss but, the water-jet allowed precise dissection and tight closure of the excretory system. More frequent haemorrhages were noted on histological examination of the WJ group, but no coagulation necrosis. These are preliminary findings and further studies of long-term results may confirm the benefits of the absence of parenchymal necrosis and the definite advantage of precise closure of the pelvicaliceal system, as morbidity of partial nephrectomy is often related to secondary haemorrhages and urinary fistulae.

Animals

French Caucasian population data obtained from fluorescently detected HUMvWFA31/A and HUMF13A01 short tandem repeat loci.

Allele and phenotype frequencies for two tetranucleotide STR (short tandem repeat or microsatellite) systems, HUMvWFA31/A and HUMF13A01, were obtained from a sample of approximately 240 unrelated individuals randomly selected from the French Caucasian population. PCR (polymerase chain reaction) products were analysed on 6% polyacrylamide denaturing gels and visualized using fluorescently labelled primers on the automated 373A ABI DNA sequencer (Applied Biosystems Inc.). French Caucasian allele frequencies were compared to other published Caucasian data. Conditions were optimised for the quadruplex PCR amplification of these two STR loci together with the HUMFESFPS and HUMTH01 loci and the quadruplex PCR was also performed on various forensic DNA samples.

Alleles

Trichloroethanol is not a metabolite of alpha chloralose.

Head space capillary gas chromatography was used to detect alpha chloralose and its potent metabolite, trichloroethanol in clinical and forensic cases. Although alpha chloralose was identified in blood and urine in all cases, trichloroethanol was never detected. In a fatal case the alpha chloralose concentration in blood was 151.3 mg/l. It was concluded that trichloroethanol is not a metabolite of alpha chloralose.

Adult

Detection and quantification of lorazepam in human hair by GC-MS/NCI in a case of traffic accident.

A traffic accident caused by a man who declared that he was driving under influence of drugs (Temesta), led our laboratory to develop a procedure for the detection and the quantification of lorazepam in human hair. The method involves decontamination of hair with dichloromethane, incubation in Soerensen buffer (pH 7.6) in the presence of lorazepam-d4, liquid-liquid extraction with diethylether-chloroform (80:20, v/v) at pH 8.4, derivatization by silylation and detection by GC-MS/NCI. The increasing concentrations of lorazepam from the end to the roots of a 16-cm-long hair strand (i.e. 31 pg/mg, 40 pg/mg and 49 pg/mg) proved that the driver had taken the drug over a long period of time.

Accidents, Traffic

Nephrogenic adenoma of the bladder in renal transplant recipients: a report of 9 cases with assessment of deoxyribonucleic acid ploidy and long-term followup.

PURPOSE: We evaluated the outcome of nephrogenic adenoma, a benign tumor rarely encountered in renal transplant recipients. MATERIALS AND METHODS: Between 1985 and 1993, 9 renal transplant recipients with a nephrogenic bladder adenoma removed by endoscopic resection were followed for 24 to 88 months (mean 40). Tumor deoxyribonucleic acid ploidy was assessed by flow cytometry at diagnosis and/or relapse. RESULTS: The relapse rate was 88%. The tumors were diploid and of low proliferating potential, and showed no malignant transformation. CONCLUSIONS: Our study confirms the lack of premalignant potential of nephrogenic adenomas. However, since transplant recipients might be at increased risk for bladder cancer, they should be followed closely.

Adenoma

Computer-controlled monitoring of bladder pressure in the prevention of 'TUR syndrome': a randomized study of 53 cases.

OBJECTIVE: To evaluate the performance of a computer-controlled monitor of bladder pressure in the prevention of transurethral resection (TUR) syndrome. PATIENTS AND METHODS: The in vitro pressure loss in catheters and endoscopes of different size was measured for irrigant flow rates of 0-500 mL/min to calibrate them before surgery. The calibrations were used in a computerized monitoring system designed to control bladder pressure during TUR of the prostate (TURP). The performance of the system was assessed in a randomized study of 53 patients with a prostate adenoma or carcinoma (Group A, 27 unmonitored patients; Group B, 26 monitored patients). The primary criterion for evaluating the absorption of irrigating fluid was the level of glycine in the blood. RESULTS: When patients with capsule perforation were included in the analysis, there was no statistically significant difference in mean glycine absorption between the groups, although glycine levels were highest in Group A, particularly in those cases with perforation (four in Group A; two in Group B). When the results for patients with capsule perforation were excluded from the analysis, there was a significant difference in the incidence of irrigant absorption between the groups. The extent of absorption was not related to the duration of operation (which was shorter in Group B) nor to the weight of resection chippings. CONCLUSION: The continuous computerized monitoring of bladder pressure during TURP effectively reduced the absorption of irrigant fluid, making the procedure safer for the patient and easier for the surgeon.

Adenoma

Sweat testing in opioid users with a sweat patch.

For many years, toxicologists have detected the presence of drugs of abuse in biological materials using blood or urine. In recent years, remarkable advances in sensitive analytical techniques have enabled the analysis of drugs in unconventional samples such as sweat. In a study conducted in a detoxification center, sweat patches were applied to 20 known heroin abusers. Subjects wore the patch with minimal discomfort for five days. During the same period, two urine specimens were also collected. Target drugs analyzed either by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-mass spectrometry (LC-MS) included opiates (heroin, 6-monoacetylmorphine, morphine, codeine), cocaine (cocaine, benzoylecgonine, ecgonine methyl ester), delta 9-tetrahydrocannabinol, benzodiazepines (nordiazepam, oxazepam), amphetamines (amphetamine, methamphetamine, methylenedioxyamphetamine [MDA], methylenedioxymethamphetamine [MDMA], methylenedioxyethylamphetamine [MDEA]), and buprenorphine. Patches were positive for opiates in 12 cases. Heroin (37-175 ng/patch) and/or 6-acetylmorphine (60-2386 ng/patch) were identified in eight cases, and codeine exposure (67-4018 ng/patch) was determined in four cases. When detected, heroin was always present in lower concentrations than 6-acetylmorphine, which was the major analyte found in sweat. Cocaine (324 ng/patch) and metabolites were found in only one case. delta 9-Tetrahydrocannabinol (4-38 ng/patch) was identified in nine cases. Benzodiazepine concentrations were very low, ranging from 2 to 44 and from 2 to 15 ng/patch for nordiazepam and oxazepam, respectively. MDEA (121 ng/patch) and its metabolite, MDA (22 ng/patch), were detected in one case. Buprenorphine, which was administered as therapy under close medical supervision, was detected in the range 1.3-153.2 ng/patch with no apparent relationship between the daily dose and amount excreted in sweat. All the urine tests were consistent with the sweat findings, but to identify the same drugs it was necessary to test two urine specimens along with only one sweat specimen. It was concluded that sweat testing appears to offer the advantage of being a relatively noninvasive means of obtaining a cumulative estimate of drug exposure over the period of a week. This new technology may find useful applications in the treatment and monitoring of substance abusers, as the patch provides a long-term continuous monitor of drug exposure or noncompliance.

Amphetamines

First isolation of tandemly repeated DNA sequences in New World vultures and phylogenetic implications.

A highly repeated DNA sequence composed of closely related subunits that ranged from 171 to 176 base pairs has been cloned and characterized in the king vulture (Sarcoramphus papa). Related sequences were also isolated in the black vulture (Coragyps atratus). This new family of avian repetitive DNA elements is here termed the "HaeIII family." Genomic DNAs from a number of avian species were probed with one of the king vulture restriction fragments. In the cathartids, the hybridization patterns showed no individual or sexual variations. A strong HaeIII ladder was present in the two aforementioned species as well as in the Andean condor (Vultur gryphus), but in the black vulture the bands of the ladder alternated in intensity. Weaker hybridization signals were obtained in two ciconids, the jabiru stork (Jabiru mycteria) and the white stork (Ciconia ciconia). The HaeIII repeat was not detected in accipitrid birds of prey, a Polyborinae falconid, pelecanids, and psittacids.

Animals

[Nephronogenic nephroma: a benign kidney tumor in adults. Two case reports].

The authors report 2 cases of a benign and uncommon type of renal tumour in adult, the nephronogenic nephroma. The pathology is discussed and compared to other groups of solid renal tumour. The confusion with malign tumour is usual. According to radiological investigations and pathological analysis, aspects of surgical treatment are presented and discussed.

Aged

Continuous river monitoring of the diatoms in the diagnosis of drowning.

The diagnosis of drowning is one of the most difficult in forensic pathology. Diatom analysis has been proposed to provide supportive evidence of drowning but the reliability and applicability of quantitative and qualitative diatom analysis in the diagnosis of drowning is still disputed in the literature. In order to further examine such cases, the authors report the development of a water monitoring system based on algae performed each month at three aquatic locations where drowning victims are frequently found. Water samples and stones were taken from the surface and from the bed of the river. This protocol was performed during 1993 with analyses both on water samples and human tissue samples (30 bodies). The diatom profile of the drowning sites was compared with the tissue analysis. The extraction of diatoms from the tissues was performed with an enzymatic digestion method using Proteinase K. Results indicate that the monitoring of river diatom populations is an accurate method of generating profiles of the river flora, which can then be compared with the diatom genera found in tissues.

Diatoms

[The PROGENE study, the French project of genetic analysis of familial prostatic cancer: recruitment and analysis].

OBJECTIVES: To initiate a genetic linkage study in order to localize one or several predisposition gene(s) for hereditary prostatic cancer (PC), as various epidemiological studies have demonstrated a possible family aggregation in about 25% of cases. A family segregation study [14] has also shown that a genetic predisposition, with autosomal dominant transmission and high penetrance (88% at 85 years) could be responsible for 9% of all PC. METHODS: A national collection of families with at least 2 cases of PC allowed: 1) identification of families with hereditary forms of PC, 2) creation of a constitutional DNA bank after collecting blood samples from subjects belonging to these families, and 3) a simulation study of genetic linkage analysis prior to microsatellite genotyping. RESULTS: From July 1994 to September 1995, we included 67 families (180 cases of PC). Another 45 families are currently being included. 24 of these 67 families (89 PC, 54 survivors) satisfied at least one of the criteria defined in the study by CARTER et al. for hereditary forms of familial PC. Two families were also included as the 3 patients with PC were second degree relatives. A total of 26 families therefore presented a hereditary form, 18 of which (73 PC, 46 survivors) were considered to be informative for a genetic linkage study (lod score = 4 for theta = 0.001 with an 8 allele marker). The constitutional DNA of 271 individuals of these informative families was extracted from circulating cells obtained from blood samples, immortalized lymphocytes, and the genotyping was initiated for 216 microsatellite markers distributed throughout the genome, an average of every 20 cM. CONCLUSION: Although the recruitment allowed us to identify many informative families for an inherited risk of PC, the predictive study suggested a high probability for localization of a predisposition gene by genetic linkage analysis. It would therefore be possible to identify, within the families concerned, the subjects carrying the genetic anomaly and consequently at high-risk of PC. Finally, the demonstration of the locus would allow cloning and identification of the gene (s) involved.

Female