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1.
Infect Genet Evol ; 10(5): 629-37, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20381646

ABSTRACT

Biting midges of the Culicoides obsoletus Meigen species complex (Diptera: Ceratopogonidae) are increasingly suspected as vectors of the recent emergence of bluetongue virus in Europe. Within this complex, identification of the C. obsoletus and Culicoides scoticus females is considered as difficult or sometimes not possible while the identification of males is easy, based on genitalia observation. Nolan et al. (2007) concluded that the distinction of C. obsoletus and C. scoticus females is not possible according to morphology but require molecular analyses. In 2010, the identification of biting midges is done under a stereomicroscope without specific identification within the C. obsoletus species complex. However, such a specific identification distinguishing C. obsoletus s. str. and C. scoticus s. str. is crucial to identify the European competent vectors of the virus, their relative abundances and then accurately assess the risk. We performed morphometric analyses of head, genitalia and thorax of females combined with sequencing of the cytochrome oxidase I barcode fragment of mitochondrial DNA on 88 specimens in order to have a molecular identification of our sampled species. As we knew the actual species of individuals thanks to molecular results, we explored the discriminant power of 15 morphometric variables to distinguish the females according to their species. Multivariate analyses were performed on the morphometric measurements to identify and validate a combination of variables leading to an accurate species identification. It appears that females of C. obsoletus and C. scoticus can be accurately distinguished based on only four variables: width between chitinous plates, length and width of spermathecae1 and length of spermatheca2. This approach should improve the accuracy of morphologically-based species identification.


Subject(s)
Bluetongue/transmission , Ceratopogonidae/anatomy & histology , Ceratopogonidae/classification , Electron Transport Complex IV/genetics , Insect Vectors/anatomy & histology , Insect Vectors/classification , Mitochondria/enzymology , Protein Subunits/genetics , Animals , Bluetongue/epidemiology , Bluetongue virus , Ceratopogonidae/enzymology , Ceratopogonidae/genetics , DNA, Mitochondrial/analysis , Female , Insect Vectors/enzymology , Insect Vectors/genetics , Male , Mitochondria/genetics , Molecular Sequence Data , Phylogeny , Principal Component Analysis
2.
J Helminthol ; 83(2): 181-9, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19356266

ABSTRACT

The prevalence of human cercarial dermatitis (HCD) caused by bird schistosomes appears to be increasing in France, in light of the impact of tourism combined with high densities of wild aquatic hosts in freshwater areas. The present work expands our knowledge of schistosome systematics by including samples of bird schistosomes collected from their natural hosts in France. Heads (318) and viscera (81) of aquatic birds belonging to 16 species from five orders, collecting during the hunting seasons or found dead, were autopsied for nasal and visceral schistosomes. Eggs and/or adults were analysed by molecular methods using the D2 domain and the second internal transcribed spacer (ITS-2) region of rDNA to determine species. Even if nasal eggs were polymorphic according to the host, all haplotypes were similar to that of Trichobilharzia regenti. Marked diversity of visceral species was observed. Final hosts under natural conditions were reported. For the first time, Trichobilharzia franki is reported in its natural bird hosts, Anas platyrhynchos, Anas crecca, Aythya fuligula and Cygnus olor. We also identified T. szidati in A. crecca and Anas clypeata. Bilharziella polonica was found in six species of aquatic birds, including Grus grus. This finding is the first record of bird schistosomes in this aquatic bird. Three new taxa of visceral schistosomes in Anser anser are strongly suspected according to their haplotypes. Futhermore, a new haplotype of visceral schistosomes isolated in Cygnus olor and similar to Allobilharzia visceralis was identified.


Subject(s)
Bird Diseases/parasitology , Schistosoma/isolation & purification , Schistosomiasis/veterinary , Animals , Animals, Wild/genetics , Animals, Wild/parasitology , Bird Diseases/epidemiology , Bird Diseases/genetics , Birds/parasitology , DNA, Helminth/genetics , DNA, Ribosomal Spacer/genetics , France/epidemiology , Fresh Water , Molecular Sequence Data , Schistosoma/genetics , Schistosomiasis/epidemiology , Sequence Analysis, DNA
3.
Bull Soc Pathol Exot ; 100(4): 291-5, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17982863

ABSTRACT

A comparative morphological and molecular study was carried out on 11 different populations of Phlebotomus (Paraphlebotomus) caucasicus Marzinovsky 1917 caught in 7 provinces in Iran (2004-2005). Differences in the implantation level of the two distal spines of the style, the number of setae of the basal lobe of coxite, and the length of the third antennal segment, revealed the existence of two morphotypes within P. (Pa) caucasicus, a species having a confused history if we take into account an unclear synonymisation with Phlebotomus (Paraphlebotomus) grimmi Porchinsky 1874. Sequencing of mtDNA (a fragment of cytchromeB gene, tRNA for serine gene and a fragment of NADH1 gene) and Neighbour-Joining analysis showed a partial correlation between morphotypes and haplotypes. We also found a correlation between the latter and the geographical origin of the specimens. These results need further studies in order to appreciate the role of each morphotype/haplotype in the transmission of Leishmania major.


Subject(s)
Phlebotomus/classification , AT Rich Sequence/genetics , Animals , Cytochromes b/genetics , Genitalia, Male/anatomy & histology , Haplotypes , Head/anatomy & histology , Insect Vectors , Iran , Male , NAD/genetics , Phlebotomus/anatomy & histology , Phlebotomus/genetics , Serine/genetics , Topography, Medical
4.
Clin Res Cardiol ; 96(6): 347-58, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17393058

ABSTRACT

UNLABELLED: The aim of this study was to evaluate subjective noise perception and objective parameters of circulation in the vicinity of the Frankfurt airport. Two areas were selected in which aircraft noise was the predominant source of noise (and was) created by planes induced by take off but not during landing. Data of residents living in the two areas were observed over a period of twelve weeks, one area being exposed to air traffic noise for three quarters of the given time, the other for one quarter of the time. METHODS: Fifty three volunteers (age 50-52 +/- 15 y) monitored their blood pressure and heart rate over a period of three months by using an automatic device with digitized readings. They also protocolled their own subjective perception of noise and sleep quality. Thirty one probands were living West of the airport (West group) and were exposed to a nocturnal equivalent continuous air traffic noise level of L(eq(3)) = 50 dB(A) outside, during flight direction 25 to the West. Twenty two probands were living East of the airport (East group) and were exposed to L(eq(3)) = 50 dB (A) during flight direction 07 to the East. During the opposite flight directions air craft noise corresponded to L(eq(3)) = 40 dB(A) in both areas. Frankfurt airport operates direction 25 for about 75% of the time on average and direction 07 for 25% of the time. RESULTS: The average blood pressure was significantly higher in the West group with higher noise exposure. Morning systolic blood pressure was 10 mmHg and diastolic pressure 8 mmHg higher in the West group. Throughout the observation period, the East group showed a parallel between daily changes in noise and subjective noise perception. In the West group such a parallel did not appear. This reaction was considered to be the consequence of the high noise exposure of the West group. CONCLUSIONS: It is concluded that a population exposed to a nocturnal equivalent continuous air traffic noise level of L(eq(3)) = 50 dB(A) for three quarters of a given time has a higher average blood pressure compared to a population exposed to the same equal energy noise level for only one quarter of the time. Within the East group a parallel between noise exposure and noise perception was observed, while in the West group this parallel did not appear. The difference is considered to be the consequence of higher noise stress levels in the West group. The data are in accordance with recent epidemiological studies and indicate that a nocturnal aircraft noise of L(eq(3)) = 50 dB(A) can have negative effects on subjective noise perception and on objective parameters of circulation.


Subject(s)
Aircraft , Attitude , Blood Pressure , Heart Rate , Noise, Transportation/adverse effects , Urban Health , Adolescent , Adult , Aged , Circadian Rhythm , Female , Germany , Humans , Male , Middle Aged , Seasons , Sleep
6.
Int J Pharm ; 229(1-2): 147-53, 2001 Oct 23.
Article in English | MEDLINE | ID: mdl-11604267

ABSTRACT

The purpose of this study was to determine the pharmacokinetics of gacyclidine, a non-competitive NMDA antagonist, in plasma and spinal cord extracellular fluid (ECF) after IV administration of single enantiomers in rats. After implantation of microdialysis probes in spinal cord, concentrations in plasma and ECF dialysates were determined by a chiral GC/MS assay over 5 h after administration of either (+)-gacyclidine or (-)-gacyclidine (1.25 mg/kg). Plasma protein binding was estimated in vitro by equilibrium dialysis. Plasma concentrations decayed in parallel in a biphasic manner (t(1/2)alpha approximately 9 min; t(1/2)beta approximately 90 min) with no significant difference between the two enantiomers. Clearance of (+)-gacyclidine and (-)-gacyclidine (291 versus 275 ml/min per kg, respectively), volume of distribution (Vdbeta: 38 versus 40 l/kg), and protein binding (90 versus 89%) were not stereoselective. Both gacyclidine enantiomers were quantifiable in spinal cord ECF 10 min after drug administration and their concentrations remained stable over the duration of the experiment in spite of changing blood concentrations. Penetration of the two enantiomers in spinal cord ECF was similar although highly variable between animals. Exposure of spinal cord ECF was comparable for both enantiomers, and not correlated with plasma AUCs. This study showed the absence of any pharmacokinetic difference between the two enantiomers when administered individually, and no enantiomeric inversion. Both gacyclidine enantiomers penetrate rapidly and extensively into spinal cord ECF, and their distribution may involve an active transport system.


Subject(s)
Cyclohexanes/pharmacokinetics , Excitatory Amino Acid Antagonists/pharmacokinetics , Piperidines/pharmacokinetics , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Spinal Cord/metabolism , Animals , Blood Proteins/metabolism , Calibration , Cyclohexanes/chemistry , Cyclohexenes , Excitatory Amino Acid Antagonists/chemistry , Extracellular Space/metabolism , Half-Life , Injections, Intravenous , Male , Microdialysis , Piperidines/chemistry , Rats , Rats, Wistar , Stereoisomerism
7.
J Pharm Sci ; 90(1): 70-8, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11064380

ABSTRACT

The pharmacokinetics of gacyclidine enantiomers, a noncompetitive N-methyl-D-aspartate (NMDA) antagonist, were studied in plasma and spinal cord extracellular fluid (ECF) after experimental spinal cord injury in rats. Spinal cord trauma was produced by introducing an inflatable balloon in the dorsal subdural space. Upon implantation of microdialysis probes in spinal cord (T9) and intravenous (iv) bolus administration of (+/-)-gacyclidine (2.5 mg/kg), concentrations in plasma and ECF were monitored over 5 h and analyzed by a stereospecific gas chromatography-mass spectrometry (GC-MS) assay. In plasma, concentrations of (+)-gacyclidine were approximately 25% higher than those of (-)-gacyclidine over the duration of the experiment and decayed in parallel (t(1/2 alpha) approximately 7 min; t(1/2 beta) approximately 90 min) with no significant difference between the two enantiomers. Clearance (CL) and volume of distribution (Vd) of (-)-gacyclidine were approximately 20% higher than those of its optical antipode (CL: 285 versus 236 mL. kg(-1). min(-1); Vd(beta): 39.3 versus 31.2 l/kg). Protein binding (approximately 91%) was not stereoselective. In spinal cord ECF, both enantiomers were quantifiable within 10 min after drug administration, and their concentration remained stable over the duration of the experiment in spite of changing blood concentrations. Repeated iv bolus injections of gacyclidine did not modify these profiles. Areas under the curves (AUCs) of concentration in ECF versus time were similar for both enantiomers and not correlated with AUCs in plasma. Penetration of (-)-gacyclidine was, however, significantly higher (approximately 30%) than that of (+)-gacyclidine. In summary, the disposition of gacyclidine enantiomers is stereoselective. Both enantiomers exhibit a high affinity for spinal cord tissue, and the drug exchange between plasma and spinal cord ECF involves an active transport system. These findings contribute to the explanation of the discrepancy between drug concentrations in plasma and spinal cord ECF.


Subject(s)
Cyclohexanes/pharmacokinetics , Neuroprotective Agents/pharmacokinetics , Piperidines/pharmacokinetics , Spinal Cord Injuries/metabolism , Animals , Biological Transport, Active , Calibration , Cyclohexenes , Disease Models, Animal , Dose-Response Relationship, Drug , Male , Protein Binding/drug effects , Rats , Rats, Wistar , Stereoisomerism , Tissue Distribution
8.
Am J Cardiol ; 85(2): 172-7, 2000 Jan 15.
Article in English | MEDLINE | ID: mdl-10955372

ABSTRACT

Earlier studies have reported on the potentiated muscarinic vasoconstriction of intracoronary acetylcholine after metoprolol application in patients with coronary artery disease. The present study investigated the effect of celiprolol, atenolol, and placebo on acetylcholine-induced vasomotion in patients with coronary artery disease. Furthermore, direct effects on coronary vasomotion and on hemodynamics were evaluated. Acetylcholine (intracoronary concentrations of 6.3x10(-7), 2.0x10(-6), and 6.3x10(-6) M) was given before and after double-blind celiprolol (0.30 mg/kg IV), atenolol (0.15 mg/kg IV), or placebo in 3x12 patients. Vasomotion was investigated by quantitative coronary angiography in proximal and distal segments of epicardial coronary arteries, and by the determination of the coronary resistance index based on Doppler-flow measurements. The investigated drugs had no direct affect on the diameter of the epicardial coronary arteries. However, celiprolol, in contrast to atenolol, significantly reduced systemic vascular resistance (change after atenolol: from 1,855+/-308 to 2,161+/-550 dyne s cm(-5); celiprolol: 1,691+/-435 to 1,411+/-343 dyne s cm(-5); and placebo: 1,722+/-215 to 1,710+/-213 dyne s cm(-5), p<0.001) and the coronary resistance index (change after atenolol: 2.52+/-3.58 to 2.86+/-4.24; celiprolol: 2.70+/-1.55 to 2.49+/-2.26; and placebo: 1.97+/-1.35 to 1.92+/-1.25, p<0.01). Celiprolol, atenolol, and placebo did not have different effects on acetylcholine-induced coronary vasomotion of epicardial conductance vessels (diminution of proximal lumen diameter before/after atenolol: 0.42+/-0.39/0.44+/-0.39 mm; celiprolol: 0.32+/-0.26/0.30+/-0.24 mm; and placebo: 0.36+/-0.29/0.43+/-0.40 mm) and of coronary resistance vessels (reduction of coronary resistance index before/after atenolol: 1.95 +/-4.74/ 1.92+/-3.74; celiprolol: 0.98+/-0.73/1.41+/-1.50; and placebo: 1.16+/-1.29/1.16+/-1.04). In contrast to atenolol, celiprolol possesses vasodilative properties in systemic and coronary resistance vessels. There was no direct effect on the diameter of conductance vessels. Acetylcholine-induced coronary vasomotion both in conductance and resistance vessels was not influenced by the beta blockers that were studied. This suggests that atenolol and celiprolol do not influence endothelium-dependent, nitric oxide related vasomotion.


Subject(s)
Acetylcholine/pharmacology , Adrenergic beta-Antagonists/pharmacology , Atenolol/pharmacology , Celiprolol/pharmacology , Coronary Disease/physiopathology , Coronary Vessels/drug effects , Coronary Vessels/physiopathology , Vasodilator Agents/pharmacology , Double-Blind Method , Humans , Male , Middle Aged
9.
Eur J Cancer ; 36(12): 1578-84, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10930807

ABSTRACT

The influence of the administration schedule (intravenous (i.v.) bolus versus i.v. infusion) on the pharmacokinetics of methotrexate (MTX) in plasma and extracellular fluid (ECF) of a brain C6-glioma was investigated in rats. MTX concentrations were determined by high performance liquid chromatography (HPLC)-ultraviolet radiation (UV). MTX (50 mg/kg) was administered by i.v. bolus or i.v. infusion (4 h). Concentration-time profiles were fitted to a two-compartment open model. Maximum MTX concentrations ranged between 178 and 294 microgram/ml (i.v. bolus), and between 11 and 24 microgram/ml (i.v. infusion) in plasma. MTX rapidly entered the tumour tissue although its concentrations in the ECF were much lower than those observed in plasma for both modes of administration. In spite of an important interindividual variability, AUC(ECF) was approximately 5-fold higher and mean MTX penetration in tumour ECF (AUC(ECF)/AUC(Plasma)) was approximately 3-fold higher after i.v. bolus than after i.v. infusion administration. These results indicate that i.v. bolus administration schedules promote MTX delivery in brain tumour tissue.


Subject(s)
Antimetabolites, Antineoplastic/therapeutic use , Brain Neoplasms/drug therapy , Methotrexate/therapeutic use , Animals , Antimetabolites, Antineoplastic/pharmacokinetics , Drug Administration Schedule , Infusions, Intravenous , Male , Methotrexate/pharmacokinetics , Rats , Rats, Wistar , Tumor Cells, Cultured
10.
Herz ; 25(2): 143-50, 2000 Mar.
Article in German | MEDLINE | ID: mdl-10829254

ABSTRACT

Since 1995, the German Society for Cardiac Angiography and Interventions in Private Practice (BNK) has been intensely involved in quality assurance; since 1996, it has registered relevant data for diagnostic left heart catheterization (DIAG) and coronary interventions (PTCA). The following data are based on a total of 72,777 cardiac catheters, of which 54,513 were DIAG and 18,264 were PTCA (DIAG in 1996: 10,316; in 1997: 15,692 and in 1998: 28,505. PTCA in 1996: 2,597; in 1997: 5,600 and in 1998: 10,067). The mean patient age was 61.9 years for males and 65.3 years for females (31%). DIAG was performed on an out-patient basis in 60% of the cases. The proportion of self-referrals is circa one-third and has not increased over time. A s/p PTCA was present in 21% of the DIAG patients; those after coronary bypass surgery showed a steadily increasing trend towards a follow-up DIAG (8% of the patients). There was a distinctive decrease in the use of contrast medium from an average of 164 ml (1996) to 138 ml (1998). The symptoms leading to DIAG did not substantially change in the years from 1996 to 1998; the angiographic range of the coronary disease and the degree of an impaired myocardial function have remained virtually unchanged. The treatment recommendations appear to tend towards medical therapy with decreasing indications for PTCA (20%) and open heart surgery (16%). The mean duration of a PTCA procedure decreased from 57 +/- 8 (1996) to 44.6 +/- 25 (1998) minutes. The success rates remained unchanged for coronary stenoses as well as for recanalization of chronic total occlusions. PTCA complications: the incidence of abrupt coronary occlusions was 2% and of emergency bypass surgery 0.4%; mortality was 0.1%.


Subject(s)
Angioplasty, Balloon, Coronary/standards , Cardiac Catheterization/standards , Coronary Angiography/standards , Quality Assurance, Health Care , Aged , Female , Germany , Humans , Male , Middle Aged , Practice Guidelines as Topic , Private Practice , Societies, Medical , Software
11.
Pharm Res ; 17(2): 148-53, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10751028

ABSTRACT

PURPOSE: Determination of the pharmacokinetics of gacyclidine enantiomers, a non-competitive NMDA antagonist, in plasma and spinal cord extracellular fluid (ECF) of rats. METHODS: Implantation of microdialysis probes in spinal cord (T9). Serial collection of plasma samples and ECF dialysates over 5 hours after IV bolus administration of (+/-)-gacyclidine (2.5 mg/kg). Plasma protein binding determined in vivo by equilibrium dialysis. Chiral GC/ MS assay. RESULTS: Plasma concentrations of (+)-gacyclidine were approximately 25% higher than those of (-)-gacyclidine over the duration of the experiment in all animals. Plasma concentrations decayed in parallel in a biphasic manner (t1/2alpha approximately 9 min; t1/2beta approximately 90 min) with no significant difference between enantiomers. Clearance and volume of distribution of (-)-gacyclidine were approximately 20% higher than those of its optical antipode (CL: 248 vs 197 ml.kg(-1)x min(-1); Vdbeta: 31.6 vs 23.5 1/kg). Protein binding (approximately 90%) was not stereoselective. Both gacyclidine enantiomers were quantifiable in spinal cord ECF 10 min after drug administration and remained stable over the duration of the experiment in spite of changing blood concentrations. Penetration of (-)-gacyclidine was significantly higher (approximately 40%) than that of (+)-gacyclidine in all animals. Yet, exposure of spinal cord ECF was similar for both enantiomers, and not correlated with plasma AUCs. CONCLUSIONS: The disposition of gacyclidine enantiomers is stereoselective. Both enantiomers exhibit a high affinity for spinal cord tissue and their distribution may involve a stereoselective and active transport system. This hypothesis could also explain the discrepancy between drug concentrations in plasma and spinal cord ECE


Subject(s)
Cyclohexanes/pharmacokinetics , Excitatory Amino Acid Antagonists/pharmacokinetics , Piperidines/pharmacokinetics , Spinal Cord/metabolism , Animals , Blood Proteins/metabolism , Catheterization , Cyclohexanes/chemistry , Cyclohexenes , Excitatory Amino Acid Antagonists/chemistry , Extracellular Space/metabolism , Injections, Intravenous , Male , Microdialysis , Piperidines/chemistry , Protein Binding , Rats , Rats, Wistar , Stereoisomerism
12.
Gesundheitswesen ; 62(2): 78-85, 2000 Feb.
Article in German | MEDLINE | ID: mdl-10740355

ABSTRACT

1956 7th grade students of high schools and secondary modern schools in three cities with 110,000 to 180,000 inhabitants were asked about cigarette, alcohol and drug consumption using questionnaires. Subsequently 1080 of the students took part in a newly invented non-smoking curriculum which was held during 8 lessons within 4 weeks (intervention group (I)). The lessons were performed by non-smoking physicians who had passed a training programme before. 876 students served as control (C) group. 2 years later the questioning was repeated. The number of students who started smoking within the two years was significantly lower in the intervention group. In the high schools a significant effect was seen in boys (I: 13.0%, K: 22.4%; p < 0.01) and girls (I: 21.4%, K: 28.4%; p < 0.05) whereas in secondary modern schools a significant effect was seen only in boys (I: 17.4%, K: 25.2%; p < 0.05) not in girls (I: 18.0%, K: 22.0%; n.s.). First-time consumers of drugs like cannabis, marihuana and organic solvents were found less often in the intervention group among boys in high schools than in the control group (I: 14.9%; K: 23.6%; p < 0.05). No effects of the intervention were seen in girls of high schools and in students of secondary modern schools. No effects of the intervention were seen on alcohol consumption. Students who drank beer, wine or liquor several times a week were found at the same rate in the intervention and control groups. We conclude that a non-smoking intervention of 8 lessons in the 7th grade of high schools and secondary modern schools may lower the rate of first-time consumers of cigarettes but not of alcohol and drugs.


Subject(s)
Health Promotion/methods , Smoking Prevention , Adolescent , Female , Germany , Health Knowledge, Attitudes, Practice , Humans , Male , Prospective Studies , Smoking/adverse effects , Urban Population
13.
FEBS Lett ; 458(2): 97-102, 1999 Sep 17.
Article in English | MEDLINE | ID: mdl-10481044

ABSTRACT

Vinblastine and vincristine are two medically important bisindole alkaloids from Catharanthus roseus (Madagascar periwinkle). Attempts at production in cell cultures failed because a part of the complex pathway was not active, i.e. from tabersonine to vindoline. It starts with tabersonine 16-hydroxylase (T16H), a cytochrome P450-dependent enzyme. We now show that T16H is induced in the suspension culture by light and we report the cloning of the cDNA. The enzyme was expressed in Escherichia coli as translational fusion with the P450 reductase from C. roseus, and the reaction product was identified by mass spectrometry. The protein (CYP71D12) shares 47-52% identity with other members of the CYP71D subfamily with unknown function. The induction by light was strongly enhanced by a nutritional downshift (transfer into 8% aqueous sucrose). We discuss the possibility that the entire pathway to bisindoles can be expressed in suspension cultures.


Subject(s)
Alkaloids/biosynthesis , Alkaloids/chemistry , Cytochrome P-450 Enzyme System/chemistry , Indole Alkaloids , Indoles , Mixed Function Oxygenases , Plants/enzymology , Quinolines , Cells, Cultured , Cloning, Molecular , Cytochrome P-450 Enzyme System/biosynthesis , Cytochrome P-450 Enzyme System/genetics , DNA, Complementary/isolation & purification , Enzyme Induction , Escherichia coli/enzymology , Escherichia coli/genetics , Light , Molecular Sequence Data , Plant Proteins/biosynthesis , Plant Proteins/chemistry , Plant Proteins/genetics , Plants/genetics , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/genetics
14.
Plant J ; 19(2): 183-93, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10476065

ABSTRACT

We investigated the P450 dependent flavonoid hydroxylase from the ornamental plant Catharanthus roseus. cDNAs were obtained by heterologous screening with the CYP75 Hf1 cDNA from Petunia hybrida. The C. roseus protein shared 68-78% identity with other CYP75s, and genomic blots suggested one or two genes. The protein was expressed in Escherichia coli as translational fusion with the P450 reductase from C. roseus. Enzyme assays showed that it was a flavonoid 3', 5'-hydroxylase, but 3'-hydroxylated products were also detected. The substrate specificity was investigated with the C. roseus enzyme and a fusion protein of the Petunia hybrida CYP75 with the C. roseus P450 reductase. Both enzymes accepted flavanones as well as flavones, dihydroflavonols and flavonols, and both performed 3'- as well as 3'5'-hydroxylation. Kinetics with C. roseus cultures on the level of enzyme activity, protein and RNA showed that the F3'5'H was present in dark-grown cells and was induced by irradiation. The same results were obtained for cinnamic acid 4-hydroxylase and flavanone 3beta-hydroxylase. In contrast, CHS expression was strictly dependent on light, although CHS is necessary in the synthesis of the F3'5'H substrates. Immunohistochemical localization of F3'5'H had not been performed before. A comparison of CHS and F3'5'H in cotyledons and flower buds from C. roseus identified CHS expression preferentially in the epidermis, while F3'5'H was only detected in the phloem. The cell-type specific expression suggests that intercellular transport may play an important role in the compartmentation of the pathways to the different flavonoids.


Subject(s)
Cytochrome P-450 Enzyme System/genetics , Mixed Function Oxygenases/genetics , Plants/genetics , Acyltransferases/genetics , Acyltransferases/metabolism , Amino Acid Sequence , Base Sequence , Blotting, Southern , Cells, Cultured , Cytochrome P-450 Enzyme System/metabolism , DNA, Complementary/chemistry , DNA, Complementary/genetics , Escherichia coli/genetics , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Plant , Immunohistochemistry , Mixed Function Oxygenases/metabolism , Molecular Sequence Data , Plant Cells , Plants/enzymology , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Substrate Specificity , Tissue Distribution
15.
Pharm Res ; 16(8): 1219-25, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10468023

ABSTRACT

PURPOSE: Establishment of the pharmacokinetic profile of methotrexate (MTX) in the extracellular fluid (ECF) of a brain C6-glioma in rats. METHODS: Serial collection of plasma samples and ECF dialysates after i.v. infusion of MTX (50 or 100 mg/kg) for 4 h. HPLC assay. RESULTS: Histological studies revealed the presence of inflammation, edema, necrosis, and hemorrhage in most animals. In vivo recovery (reverse dialysis) was 10.8 +/- 5.3%. MTX concentrations in tumor ECF represented about 1-2% of the plasma concentrations. Rapid equilibration between MTX levels in brain tumor ECF and plasma. ECF concentrations almost reached steady-state by the end of the infusion (4 h), then decayed in parallel with those in plasma. Doubling of the dose did not modify MTX pharmacokinetic parameters (t1/2alpha, t1/2beta, MRT, fb, Vd, and CL(T)), except for a 1.7-fold increase of AUC(Plasma) and a 3.8-fold increase in AUC(ECF), which resulted in a 2.3-fold increase in penetration (AUC(ECF)/AUC(Plasma)). In spite of an important interindividual variability, a relationship between MTX concentrations in plasma and tumor ECF could be established from mean pharmacokinetic parameters. CONCLUSIONS: High plasma concentrations promote the penetration of MTX into brain tissue. However, free MTX concentrations in tumor ECF remain difficult to predict consistently.


Subject(s)
Antimetabolites, Antineoplastic/pharmacokinetics , Brain Neoplasms/metabolism , Extracellular Space/metabolism , Glioma/metabolism , Methotrexate/pharmacokinetics , Animals , Antimetabolites, Antineoplastic/blood , Brain Neoplasms/pathology , Capillary Permeability , Computer Simulation , Glioma/pathology , Infusions, Intravenous , Male , Methotrexate/blood , Microdialysis , Neoplasm Transplantation , Rats , Rats, Wistar , Tissue Distribution , Tumor Cells, Cultured
16.
Alcohol Clin Exp Res ; 23(5): 885-90, 1999 May.
Article in English | MEDLINE | ID: mdl-10371410

ABSTRACT

The pharmacokinetics of toloxatone and ethanol were determined in plasma and cerebrospinal fluid of conscious rabbits. According to a cross-over design, rabbits (n = 5) randomly received on three separate occasions either toloxatone (5 mg/kg), ethanol (1 g/kg), or toloxatone and ethanol (5 mg/kg and 1 g/kg, respectively) by intravenous injection. Toloxatone and ethanol concentrations were measured by HPLC with UV detection and GC with flame ionization detection, respectively. Ethanol concentration profiles in plasma were characterized by a rapid decline occurring within the first 30 min after administration followed by a linear (zero-order) phase that persisted for the length of the experiment. The maximum ethanol elimination rate was 0.31+/-0.20 g/h x kg. Toloxatone concentrations in plasma and cerebrospinal fluid were characterized by a multiexponential decay with effective half-lives of 0.39+/-0.06 and 0.56+/-0.07 hr, respectively. Toloxatone passage through the blood brain barrier was rapid and important. Our results also demonstrate that acute ethanol administration had no effect on toloxatone pharmacokinetics and that toloxatone administration had no effect on ethanol pharmacokinetics.


Subject(s)
Ethanol/pharmacology , Monoamine Oxidase Inhibitors/pharmacokinetics , Oxazoles/pharmacokinetics , Oxazolidinones , Animals , Cross-Over Studies , Drug Interactions , Ethanol/metabolism , Ethanol/pharmacokinetics , Male , Monoamine Oxidase Inhibitors/metabolism , Monoamine Oxidase Inhibitors/pharmacology , Oxazoles/metabolism , Oxazoles/pharmacology , Rabbits , Random Allocation
17.
J Invasive Cardiol ; 11 Suppl B: 14B-18B, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10745612

ABSTRACT

UNLABELLED: One-hundred patients undergoing routine diagnostic or interventional catheterization were randomly assigned to receive either percutaneously applied collagen (group A; n = 50) or conventional pressure dressing (group B; n = 50) for sealing of the femoral artery. Clinical variables were comparable in both groups. The heparin dose was 100 IU/kg in 30 patients and 200 IU/kg in 20 patients of either group. The average compression time was 4.3 minutes in group A and 42.3 minutes in group B (p < 0.001). Bleeding was not observed in group A but was observed in 6/50 patients in group B. The time to ambulation was 6.4 hours (range: 4-12 hours) in group A and 21.6 hours (range: 10-48 hours) in group B (p < 0.001). Hematomas with a diameter of > 6 cm developed in 4/50 patients in group A and in 11/50 patients in group B (p < 0.05). Blood transfusion or surgical interventions were not required and there was no loss of ankle pulses in either group. CONCLUSION: Percutaneously applied collagen reduced compression time and duration of bedrest after diagnostic catheterization and PTCA. Despite earlier ambulation, the incidence of bleeding was lower with collagen than with conventional pressure dressing.


Subject(s)
Angioplasty, Balloon, Coronary/adverse effects , Cardiac Catheterization/adverse effects , Hemostatic Techniques , Needlestick Injuries/therapy , Postoperative Hemorrhage/prevention & control , Aged , Bandages , Collagen/therapeutic use , Equipment and Supplies , Female , Femoral Artery/injuries , Hemostatics/therapeutic use , Humans , Male , Middle Aged , Needlestick Injuries/complications , Postoperative Hemorrhage/etiology , Pressure , Treatment Outcome
18.
C R Acad Sci III ; 321(10): 849-55, 1998 Oct.
Article in French | MEDLINE | ID: mdl-9835021

ABSTRACT

Phylogenetic relationships among Phlebotominae were inferred through a pilot study using parsimony analysis of the D2 domain of ribosomal DNA sequences: 455 pairs of bases were sequenced in nine species of Phlebotomine sandflies which belong to the genera Lutzomyia, Phlebotomus and Sergentomyia. Two taxa are used as outgroups: Psychoda sp. and Nemapalpus flavus which is the sister group of the Phlebotominae. The South American genus Lutzomyia appears to be monophyletic. The Mediterranean species Sergentomyia dentata is its sister group and is not clustered with the Old World genus Phlebotomus. The latter is a paraphyletic genus with an early individualization of the branch including the closely related subgenera Phlebotomus and Paraphlebotomus, and a late individualization of the subgenus Larroussius. These results have some consequences on the biogeography of the leishmaniasis in the Old World.


Subject(s)
Evolution, Molecular , Phlebotomus/genetics , Phylogeny , Psychodidae/genetics , Animals , Base Sequence , DNA, Ribosomal/genetics , Molecular Sequence Data , Pilot Projects , Sequence Homology, Nucleic Acid
20.
Alcohol ; 15(4): 319-25, 1998 May.
Article in English | MEDLINE | ID: mdl-9590518

ABSTRACT

Arterio-venous ethanol concentrations in both whole blood and plasma were determined as a function of time in the rabbit. Following i.v. injection of 1.0 g/kg, both arterial and venous ethanol concentrations showed an abrupt decline occurring immediately after the end of the administration, followed by a pseudolinear phase that persisted for the length of the experiment. This work substantiates the arterio-venous ethanol concentration differences reported in the literature. It illustrates that equal arterial and venous ethanol concentrations may not be achieved readily after rapid i.v. injection. Moreover, it demonstrates a faster decay of ethanol concentrations in arterial than in venous plasma.


Subject(s)
Ethanol/administration & dosage , Ethanol/blood , Animals , Arteries , Injections, Intravenous , Male , Osmolar Concentration , Plasma/metabolism , Rabbits , Time Factors , Veins
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