Specific adipocytokines profiles in patients with hyperactive and/or binge/purge form of anorexia nervosa.

OBJECTIVE: The aim of our study was to determine whether eating behaviors and/or physical activity level may explain contradicting results in adipocytokines levels in anorexia nervosa (AN). SUBJECTS/METHODS: Fasting levels of circulating adipocytokines (adiponectin, resistin and leptin), insulin, glucose, C-reactive protein, cytokines (tumor necrosis factor-alpha and interleukin (IL)-1beta), body composition and resting energy expenditure were measured in 24 women AN patients and 14 women controls. These parameters were compared according to AN subtypes: 15 patients with restrictive (R-AN) form versus 9 patients with binge/purge (BP-AN) form; 15 patients with hyperactive (H-AN) form versus 9 patients with nonhyperactive (NH-AN) form. RESULTS: BP-AN patients had significantly higher serum adiponectin levels compared with R-AN patients (P<0.05), and H-AN patients had higher serum leptin and lower serum resistin levels compared with NH-AN patients (P<0.05 for both). CONCLUSIONS: Our study shows specific adipocytokines profiles depending on the subtype of AN: restrictive versus binge/purge and hyperactive versus Nonhyperactive forms. We suggest that these biological signatures could interfere with the outcome of the disease.

T-cell activation during exacerbations: a longitudinal study in refractory asthma.

BACKGROUND: Asthma exacerbations represent the main source of costs and morbidity in asthma care, and drugs specifically designed to prevent exacerbations are needed. A prerequisite is to dispose of a precise knowledge of inflammatory events leading to exacerbations. OBJECTIVE: To study T-cell activation during exacerbations from severe refractory asthmatics. METHODS: Proportions of blood T-cell interleukin (IL)-13, interferon-gamma, IL-4, IL-5, IL-10 production and of CD4+CD25+(high)CD62L+CD45RO+ [T regulatory (Treg)] cells were determined by flow cytometry. Blood cytokine mRNA was studied by reverse transcription-polymerase chain reaction and the respective protein levels were determined by cytokine beads array. Depletion of Treg cells was performed to study their activation. T-cell cytokines were detected in parallel in induced sputum. RESULTS: At baseline, T helper 2 (Th2) cells were increased in asthmatics, whereas T helper 1 (Th1) and Treg T cells were decreased. T helper 2 cells increased before exacerbations, followed by Th1 cells, in blood and induced sputum, albeit Treg cells decreased in parallel with IL-10-producing T cells. Concordant results were found at the mRNA level. The suppressive activity of Treg cells was impaired during exacerbations compared to baseline. CONCLUSIONS: New insights are given into pathophysiology of asthma exacerbations: Although at baseline T-cell activation is Th2-biased, a mixed Th1/Th2 activation occurs during exacerbations. The Treg cell deficiency found at baseline in SRA increases during exacerbations.

T-cell activation in occupational asthma and rhinitis.

BACKGROUND: Allergic asthma and rhinitis are described as associated with a Th2 activation. However, recent works indicate that a Th1 activation can also be associated with these diseases, concomitantly to a defect in regulatory T (Treg) cell activation. Occupational asthma (OA) and occupational rhinitis (OR) are peculiar cases of these diseases in which the T-cell activation profile is largely unknown. OBJECTIVE: To characterize T-cell activation induced after a specific inhalation test (SIT) in OA and OR. MATERIAL AND METHODS: A total of 21 subjects with OA, 10 subjects with OR, 10 exposed nonallergic (ENA) subjects, and 14 healthy volunteers were included. The SIT with the incriminated substance was performed in patients and ENA subjects. Blood and induced sputum were obtained before and after SIT. T cells were analysed for CD69, CD25, IL-13, and IFN-gamma expression by flow cytometry. IL-4 and IFN-gamma were assayed by enzyme-linked immunosorbent assay (ELISA) in cell culture supernatants. Treg cells were identified as CD4(+)CD25(+high)CD45RO(+)CD69(-) T cells in peripheral blood. RESULTS: Baseline IFN-gamma production was decreased in OA and OR compared with controls. The SIT induced an increase in both Th1 and Th2 cells in blood and sputum from OA. In this group, the proportion of peripheral Treg cells decreased after SIT. Similar results were found in the CD8(+) population. ELISA assays were concordant with flow cytometry. In OR, an attenuated activation profile was found, with an increase in the proportion of IL-13-producing T cells after SIT. By contrast, in ENA subjects, SIT induced Th2 activation, with an increase in Treg cells and a decrease in Th1 cells. CONCLUSIONS: Our results demonstrate a gradient of T-cell activation from a tolerating profile in ENA subjects to an inflammatory profile in OA, with an intermediate stage in OR.

Lack of toxicity of hydroethanolic extract from Mitragyna inermis (Willd.) O. Kuntze by gavage in the rat.

In traditional medicine in Mali, extracts derived from Mitragyna inermis (Willd.) O. Kuntze (Family: Rubiaceae) are commonly used to treat malaria. The antimalarial activity and the lack of genotoxicity in vitro and in vivo have been demonstrated in previous studies. Acute and chronic evaluation of the toxicity of the hydroethanolic extract of Mitragyna inermis leaves was performed in this study, according to the recommendations (cahier de l'Agence no. 3) of the French Drug Office. Two dosages (300 mg/kg and 3 g/kg) were given in one single administration by gavage to male and female rats. No animal died and no behavioral signs of acute toxicity were observed. Chronic toxicity studies over 28 days showed no changes in body weight and no macroscopic abnormality in the 14 organs examined after the animals were sacrificed. With the 3 g/kg/d drug dosage (100-fold higher than those proposed in man), only slight histological abnormalities were observed. Statistically significant differences, compared to control animals, in the weight of some organs and the values of some haematological or biochemical parameters were observed. However, these values always remained in the range given by the breeder for naive animals of the same strain. These investigations thus seemed to indicate the safety of repeated oral administration (up to 3 g/kg/d) of the hydroethanolic extract of Mitragyna inermis leaves, which can therefore be continuously used with safety by the African population in traditional treatment of malaria.

Assessment of T lymphocyte cytokine production in induced sputum from asthmatics: a flow cytometry study.

BACKGROUND: Asthma results from a bronchial inflammation in which Th2 lymphocytes play a pivotal role, as shown in invasive bronchial biopsies and broncho-alveolar lavages. Induced sputum (IS) is a non-invasive method of recovery of bronchial cells, which can be repeated in the same patients. However, lymphocyte activation has not been studied in IS to date, because of the low number of T cells recovered. Herein we took advantage of flow cytometry, a method suitable for the study of small cell populations, to assess T cell cytokine production in IS. OBJECTIVES: (1) To assess induced sputum T cell cytokine production by flow cytometry in asthmatic subjects and controls. (2) To compare the T cell cytokine production between symptomatic and non-symptomatic asthmatics. METHODS: Thirteen asthmatics and 19 controls were included. Sputum was induced by a hypertonic saline. Sputum cells were stimulated and intracellular IL-13 and IFN-gamma were detected in T cells by flow cytometry. RESULTS: Stimulation induced an increase of IL-13 and IFN-gamma production by T cells. This increase was higher in asthmatics. IL-13-producing T cells were increased in asthmatics after stimulation. In symptomatic asthma, IFN-gamma-producing T cells were in higher proportion than in controlled asthma. CONCLUSION: IS T cell cytokine production indicates a basic Th2 bias in asthma, accompanied during symptoms by a Th1-like activation. These results open the field for longitudinal studies of the variation of T cell activation in asthma.

[Induced sputum as a new approach of the assessment of inflammation in asthma]. / L'analyse de l'expectoration induite dans l'asthme permet une nouvelle approche de l'inflammation.

Asthma is an inflammatory condition. Traditionally bronchoalveolar lavage and bronchial biopsies obtained by bronchoscopy have been used to demonstrate inflammation. Induced-sputum is a non-invasive, reliable, reproducible and safer technique for monitoring inflammatory activity in patients with asthma. Studies have shown that induced-sputum measures aspects of inflammation distinct to that measured by bronchoalveolar lavage or bronchial biopsies. Numerous studies have suggested that induced-sputum is a potentially useful tool for early diagnosis of exacerbation, monitoring of therapy, identification of the lowest effective dose and assessing compliance in asthmatics. In this respect, we suggest that this test can be routinely used in the management of difficult asthmatics.

Dietary soybean phosphatidylcholines lower lipidemia: mechanisms at the levels of intestine, endothelial cell, and hepato-biliary axis.

The beneficial metabolic effects of dietary soybean lecithin on lipid metabolism are now more clearly established. The intestinal absorption of cholesterol is decreased by soybean phosphatidylcholine-enriched diet and results in a cholesterol-lowering effect. There is an enhancement of the cholesterol efflux by endothelial cells incubated with soybean phosphatidylcholines, and a stimulation of the reverse cholesterol transport by high density lipoprotein-phosphatidylcholines. As a result of all these processes, phosphatidylcholines provided by the soybean lecithin metabolism appear to be key molecules controlling the biodynamic exchanges of lipids. They regulate homeostasis of cholesterol and fatty acids by decreasing their synthesis and promoting cholesterol oxidation into bile salts. Finally, the outcome is the increase in bile secretion of these lipids and/or their metabolite forms. Such findings constitute promising goals in the field of nutritional effects of soybean lecithin in the treatment or prevention of hyperlipidemia and related atherosclerosis.

Determination of apolipoprotein B-48 in plasma by a competitive ELISA.

BACKGROUND: Apolipoprotein B-48 (apoB-48) is produced by the small intestine, as part of chylomicrons, and appears to be a suitable marker for clinical studies of postprandial lipoproteins and related cardiovascular risk. Our aim was to develop, for routine analysis, an assay to quantify apoB-48 in plasma samples. METHODS: A microtiter plate was coated with a C-terminal apoB-48-specific heptapeptide. Plasma samples were incubated with appropriate detergent to allow competition between immobilized antigen and plasma apoB-48. Appropriate calibration curves were obtained in the ELISA, using calibrated lymph and chylomicrons. RESULTS: Treatment of plasma samples with the mild detergent Triton X-100 allowed an efficient competition between immobilized antigen and plasma apoB-48. No cross-reactivity was found with apoB-100, as checked by ELISA and Western blot analysis. Intra- and interassay CVs were 5.4% and 5. 5%, respectively. In healthy subjects, apoB-48 concentrations markedly increased in the postprandial state, in parallel with triglycerides. CONCLUSIONS: This new ELISA allows determination of the concentration of apoB-48 in normolipidemic plasma.

Excretion of lidocaine and bupivacaine in breast milk following epidural anesthesia for cesarean delivery.

BACKGROUND: There is a lack of information and knowledge about the practical importance of even low concentrations of the excretion of local anesthetics into breast milk, particularly concerning bupivacaine. The present work aims to confirm, under practical clinical conditions of admission of parturients, the passage of local anesthetics (lidocaine and bupivacaine) into breast milk after an epidural anesthesia. METHODS: Twenty-seven pregnant women admitted for cesarean delivery received epidural anesthesia with 0.5% bupivacaine and 2% lidocaine. Blood and milk samples were simultaneously collected at 2, 6 and 12 h after the beginning of the epidural infusion. Lidocaine, bupivacaine and its main metabolite, pipecolylxylidide (PPX), were determined in serum and milk by a gas-liquid chromatographic technique. APGAR scores were systematically performed at delivery and a clinical examination was done 24 h after delivery. RESULTS: Our data indicate that lidocaine and bupivacaine as well as PPX are excreted into breast milk. The milk/serum ratio based upon area under the curve values were 1.07 +/- 0.82, 0.34 +/- 0.24 and 1.37 +/- 0.61 mean +/- SD for lidocaine, bupivacaine and PPX, respectively. Most of the newborns had a maximal APGAR score. Our study does not reveal any adverse reactions related to the excretion of local anesthetics into breast milk. CONCLUSION: This study documents the magnitude of excreted lidocaine, bupivacaine and PPX in breast milk, and indicates that the use of both lidocaine and bupivacaine for epidural anaesthesia is safe with regard to breast-feeding.

Seasonal variations in the kinetics of bupivacaine in the mouse.

Very few studies have been devoted to the influence of the time of the year, i.e. seasonal variations, on drug kinetics. This study aims to evaluate whether or not the kinetics of bupivacaine varied according to the time of the year in mice. Eight groups of 30 animals each received bupivacaine (20 mg/kg i.p.) at a specific month of the year, i.e. February (second week), March (first week), May (fourth week), July (first week), September (fourth week), November (second and third weeks) and December (first week), at the same time of the day (10.00 h). Total bupivacaine serum concentrations were determined by using a specific gas liquid chromatographic method on blood samples collected by decapitation at 0.25, 0.5, 1, 2, 4 and 6 h after drug administration. Pharmacokinetic variables were determined, according to a non linear fitting method (two compartment open model). All data were compared by ANOVA and the influence of the month of the year was estimated by Cosinor analysis. According to the month of the year, the kinetic parameters were significantly different except for Vd: the maximal Cmax occurred in July, the highest Tmax occurred in May, the maximal T1/2 beta and the maximal AUC were observed in February. The mechanisms underlying these variations may depend on seasonal variations of resorption, distribution, metabolism and elimination.

[Use of percutaneous anesthesia in cardiac pacemaker implantation]. / Emploi de l'anesthésie transcutanée dans les procédures d'implantation de stimulation cardiaque.

OBJECTIVES: Assess the efficacy of an anesthesic cream for pacemaker implantations. METHODS: Percutaneous anesthesia was studied in a series of permanent pacemaker transvenous implantations. The anesthesic cream composed of a mixture of lidocaine and prilocaine was applied precisely over operative areas after marking the skin. Percutaneous anesthesia should be applied 2 hours before entering the operating room. RESULTS: This percutaneous local anesthesia was perfectly effective for simple replacement procedures. At first implantations, it was used alone in 4 out of 10 cases while intradermal injections were needed to anesthetize the deep layers in the other patients. Serum concentrations indicate very low levels which are tolerated very well. CONCLUSION: Alone or combined with lidocaine infiltration, the use of an anesthesic cream is safe and effective in transvenous pacemaker surgery.

Kinetics of bupivacaine after clonidine pretreatment in mice.

Previous studies have reported that clonidine pretreatment causes an increase in the local anaesthetic activity of bupivacaine. This study was designed to document possible changes in the pharmacokinetic behaviour of bupivacaine and its main metabolite, desbutylbupivacaine, PPX, in mice after a single, 0.1 mg.kg-1, injection of clonidine. Kinetic variables of bupivacaine were determined after a single 20 mg.kg-1 ip dose of bupivacaine in controls (Group I) and in clonidine (0.1 mg.kg-1 ip) pretreated mice (Group 2). The maximal concentration in serum (Cmax, 2.553 +/- 0.862 micrograms.ml-1 versus 0.962 +/- 0.141 microgram.ml01 for Groups 2 and 1, respectively, P = 0.01) and the area under the concentration curve (AUC, 3.530 +/- 0.330 micrograms.ml-1.hr-1 versus 1.755 +/- 0.252 micrograms.ml-1.hr-1 for Groups 2 and 1, respectively, P < 0.01) of bupivacaine were higher in clonidine pretreated mice while the Clearance (Cl) was decreased in clonidine pretreated animals (0.603 +/- 0.054 micrograms.ml-1 versus 1.264 +/- 0.447 micrograms.ml-1 for Groups 2 and 1, respectively, P < 0.01). The ratio of AUC PPX/AUC bupivacaine (which may partially indicate the rate of metabolism) was lower in presence of clonidine (0.220 +/- 0.019 against 0.425 +/- 0.033 for Groups 2 and 1, respectively, P < 0.01). Our data indicate decreased metabolism in the clonidine-treated mice which suggests altered hepatic metabolism of bupivacaine by clonidine. This may explain the previously reported enhanced anaesthetic activity of bupivacaine in the presence of clonidine.

New regimen for interpleural block in children.

A new regimen for postoperative analgesia after thoracic surgery is proposed. Eight children received an interpleural infusion using bupivacaine 0.1% in a regimen from 0.5 ml.kg-1.h-1 up to 1 ml.kg-1.h-1, for 48 h according to the pain scores. The plasma levels after 24 h and 48 h were measured as well as the pleural level and in two patients the free fraction of plasma bupivacaine and the plasma PPX (a metabolite of bupivacaine) and one patient the orosomucoid (main plasma protein involved in bupivacaine protein binding) were also measured pre and postoperatively. The results shows the safety of such a regimen, for two days of postoperative analgesia.

Rapid simultaneous determination of lidocaine, bupivacaine, and their two main metabolites using capillary gas-liquid chromatography with nitrogen phosphorus detector.

A gas-liquid chromatographic method for the simultaneous measurement in serum of bupivacaine, lidocaine, and their main metabolites, 2,6-pipecolylxylidide (PPX) and monoethylglycine xylidide (MEGX), respectively, is described. The procedure involves a one-step extraction and injection of the extract into a gas chromatograph equipped with a capillary column and nitrogen phosphorus detector under constant temperature conditions. Recovery of all components averaged 94%, with the lowest detection limit of 15 ng/ml for the four drugs. The precision within-series coefficients of variation ranged from 7.7% for bupivacaine, 8.6% for lidocaine, 10.2% for MEGX, and 15.8% for PPX. The interday coefficients ranged from 0.7 to 6.5%. Concomitant use of caffeine and carbamazepine may interfere with MEGX and bupivacaine determination, respectively. For this reason, in patients receiving one of these two drugs (or ingesting foods and beverages containing caffeine), the described method is not recommended.

Cigarette smoke increases bupivacaine metabolism in rats.

This study was designed to document possible changes in bupivacaine kinetics in rats after exposure to cigarette smoke. Rats (n = 15) were exposed to cigarette smoke (Borgwaldt type Hamburg II) for ten minutes per day during four days (C) or eight days (B); controls (A) were used simultaneously without exposure to cigarette smoke. After bupivacaine 20 mg.kg-1 ip at day 4 (C) or day 8 (B), blood was sampled (0.5 ml of blood collected by puncture at the retro-orbital sinus 0.25, 0.5, 1, 2, 4, 6 and 8 hours after administration) and bupivacaine and its main metabolite i.e., desbutylbupivacaine (PPX) were determined by gas liquid chromatography. The sensitivity of the method was 15 ng.ml-1 and the reproductibility was < 6%. Serum bupivacaine concentrations were plotted against time and the pharmacokinetic variables were determined assuming a two compartment open model: Cmax, Tmax were derived directly from individual data. The beta phase elimination half-lives (T1/2 beta), the area under the serum concentration curve (AUC0 infinity), the total plasma clearance (Cl) and the total volume of distribution (Vd) were calculated. These variables were assessed according to non-linear fitting method. Cigarette smoking exposure did not change the pharmacokinetic variables of bupivacaine.(ABSTRACT TRUNCATED AT 250 WORDS)

Bupivacaine kinetics during hyperthermia in rats.

The aim of this study was to document possible alterations of bupivacaine pharmacokinetic behaviour in rats during hyperthermia. Two groups of Wistar AF IO PS male rats (Group A = normothermic controls, Group B = hyperthermia-induced animals) received a single 20 mg.kg-1 ip dose of bupivacaine. Two other groups (Group C = normothermic controls without bupivacaine, Group D = hyperthermia-induced animals without bupivacaine) received, under the same experimental conditions, an equivalent volume of saline. Hyperthermia-induced animals (Groups B and D) were placed in a water-bath at 40 degrees C. Bupivacaine or saline were administered (Group B and D) four hours after the beginning of the experiment and blood samples were obtained by retro-orbital sinus puncture 0.25, 0.5, 1, 2, 4 and 8 hr after administration. Bupivacaine and its main metabolite, 2,6 desbutylbupivacaine (PPX) were assayed according to a gas liquid chromatographic method. The Cmax, Tmax, t1/2, Cl, Vd and AUC were determined according to a two compartment open model. Our data have demonstrated a decrease in clearance of bupivacaine (5.85 +/- 0.23 ml.hr-1 and 4.59 +/- 0.35 ml.hr-1 for groups A and B, respectively, P < 0.05, and, Tmax of PPX during hyperthermia (0.24 +/- 0.03 hr and 0.15 +/- 0.0 hr for Groups A and B, respectively, P < 0.05). In conclusion, hyperthermia induces a decrease in bupivacaine clearance in rats which may be of importance in clinical practice.

Plasma endothelin in haemodialysis patients treated with recombinant human erythropoietin.

Predialysis plasma endothelin (ET) values were followed during the first 8 weeks of rHuEpo treatment in 12 patients on routine haemodialysis. Mean plasma ET was significantly increased in uraemic patients before rHuEpo (27.9 +/- 11.4 pmol/l), as compared to 40 healthy controls (16.5 +/- 5.7 pmol/l) (P < 0.0001). Under rHuEpo treatment, predialysis values remained unchanged although diastolic blood pressure increased after 2 and 6 weeks. We found no correlation between ET and haemoglobin or blood pressure before or under rHuEpo treatment. These results confirmed the high levels of plasma ET in haemodialysis patients, but no increase was observed during rHuEpo treatment.

Bupivacaine kinetic changes induced by diltiazem in mice.

This study was designed to document possible changes in bupivacaine and its main metabolite, des-butyl-bupivacaine (PPX) kinetics in mice after a single 5 mg/kg injection of diltiazem. After a single 20 mg/kg i.p. dose of bupivacaine, kinetic parameters of bupivacaine were not statistically different but the ratio AUC PPX/AUC bupivacaine was significantly lower when bupivacaine was associated with diltiazem: thus, it may indicate an influence of diltiazem on bupivacaine metabolism i.e. an inhibition. Diltiazem also seems to increase the free fraction of bupivacaine and thus to decrease the percentage of protein binding. These effects may contribute to the enhanced toxicity previously observed.

Lack of bupivacaine kinetic changes induced by flumazenil in mice.

This study was designed to document possible changes in bupivacaine kinetics in mice after a single 1 mg/kg injection of flumazenil. After a single 20 mg/kg i.p. dose of bupivacaine, C max, Vd, Cl and AUC were not significantly modified by flumazenil; even if T max was shown to be significantly shorter when flumazenil was associated, bupivacaine bioavailability did not seem to be modified and thus may not be involved in the explanation of previously reported increasing bupivacaine-induced mortality by flumazenil.