Pulmonary uptake of bupivacaine in isolated perfused rat lung

The ability of rat lung to remove the local anaesthetic drug bupivacaine from the blood was studied in isolated organs which were perfused either in an open (single-pass mode) or in a closed system (recirculating medium).Isolated perfused rat lungs exhibited a very low capacity to metabolize bupivacaine within 3 h during which the drug circulated continuously through the organ. The clearance values differed only by 0.2 ml/min from the control parameters in sham perfusions. The calculated extraction ratio was 0.2% and the elimination half-life was about 210 min. The volume of distribution of bupivacaine was 133 ml which remarkably surmounted the reference values obtained for sham perfusions.The distribution of bupivacaine into the pulmonary tissue was investigated applying the multiple indicator dilution technique to isolated lungs perfused in the single-pass mode. The mean elimination time of model compounds for distribution into the intravascular space, ¹⁴C-inulin, and the total water space, ³H-water, were 68 and 75 s at a flow rate of 6 ml/min. The volume of distribution was 5.9 ml for inulin and 6.5 ml for water. The mean transit time for concomitantly injected bupivacaine was 221 s and the volume of distribution was 14.4 ml. The respective parameters of sham perfusions performed without an isolated organ were substantially lower, i.e. mean elimination time 50, 50 and 61 s and distribution volume 4.9, 5.0 and 6.1 ml for inulin, water and bupivacaine.The volume of distribution during single-pass contact of bupivacaine to lung was not substantially influenced by an increase of the flow rate from 6 to 9 and 12 ml/min whereas the mean transit time dropped from 221 to 121 and 108 s, respectively. These results support the assumption that bupivacaine is extensively retained by the pulmonary tissue and that elimination of bupivacaine by metabolism can be neglegted for lung.The hemodynamic parameters of bronchiolar perfusion in the artificially perfused lung were determined using two fluorochrome-labeled macromolecular proteins, i.e. fluorescein-isothiocyanate (FITC)- and lissamine-rhodamine-B 200 (RB 200)-labeled globulin. After 10 min of perfusion at a flow rate of 12 ml/min in the closed system an area of 10.8070 of the peribronchiolar tissue area contained the dye-label FITC. A very similar index (10.1%) of dye-coloured capillaries was obtained when the lungs of anaesthetized rats were examined 10 min after intravenous injection of the fluorochrome into the pulmonary artery in vivo. In isolated perfused rat lungs receiving both FITC and RB 200 59.5% of FITC-labeled capillaries were reached by the second fluorochrome within 2 s. This fraction accounted for 93.3% after 10 s of circulation time. This proves that isolated rat lungs were well perfused in vitro.     

Spinal hyperbaric prilocaine vs. mepivacaine in perianal outpatient surgery

Background

The aim of this randomised, clinical trial was to compare safety and efficiency of hyperbaric prilocaine and mepivacaine at a dosage of 0.5 ml each for perianal outpatient surgery in terms of transient neurologic symptoms (TNS) and postoperative recovery.

Methods

160 patients aged 18–80 years were randomized to receive a spinal anaesthesia (SPA) with 0.5ml of mepivacaine or prilocaine. We measured the expansion of the block, evaluated postoperative recovery times and determined the incidence of TNS one week after surgery.

Results

160 patients (93 male / 67 female) were available for analysis. Prilocaine led to shorter times from SPA to micturition (prilocaine: 178 (110–254) min vs. mepivacaine: 195 (130–305) min, p=0.0008) and discharge (prilocaine: 192 (126–267) min vs. mepivacaine: 220 (140–320) min, p<0.0001). 152 / 160 patients were available for the telephone follow-up. Six patients (9%) receiving mepivacaine compared to zero patients of the prilocaine group announced typical symptoms of TNS (p=0.0284).

Conclusion

Both, hyperbaric mepivacaine 40 mg/ml and hyperbaric prilocaine 20 mg/ml can be used at a dosage of 0.5 ml each for SPA in perianal outpatient surgery. Due to the faster recovery profile and a lower incidence of TNS, we recommend the use of 10mg hyperbaric prilocaine 20 mg/ml for this indication.     

Changes in the incidence and duration of ventricular fibrillation in dependence on the extraneuronal accumulation of isoprenaline in the perfused rat heart

Summary The relationship between the accumulation of isoprenaline and the incidence and duration of ventricular fibrillation was investigated in the perfused rat heart. Isolated rat hearts were perfused with ³H-isoprenaline (1 mol/l) for 30 min at a constant flow rate of 6.5 ml/min at a temperature between 40 and 41° C. Electrocardiograms were recorded during the perfusion period and the isoprenaline content of the tissue was measured after the perfusion. The accumulation of isoprenaline was significantly increased and the duration of ventricular fibrillation was significantly prolonged by the presence of tropolone (100 mol/l). When extraneuronal uptake inhibitors such as normetanephrine (100 mol/l), 3-O-methylisoprenaline (100 mol/l) or phenoxybenzamine (1 mol/l) were added to the perfusion fluid containing ³H-isoprenaline (1 mol/l) and tropolone (100 mol/l), the accumulation of isoprenaline was sifnificantly decreased, the incidence of ventricular fibrillation was significantly reduced and the duration of ventricular fibrillation was significantly shortened. There was a significant correlation for dependence of duration of ventricular fibrillation on the isoprenaline content of rat hearts perfused with various extraneuronal uptake inhibitors in the presence of tropolone (correlation coefficient [r]=0.62, P<0.001).These results indicate that the accumulation of isoprenaline in perfused rat hearts relates to the occurrence and duration of ventricular fibrillation.This study was supported in part by a Grant-in-Aid for Scientific Research (59570980) from the Ministry of Education, Science and Culture, Japan     

Ocular and systemic bioavailability of ophthalmic flurbiprofen

Flurbiprofen, a nonsteroidal antiinflammatory agent which is not ocularly metabolized, was employed as a probe compound to investigate the drug kinetic relationship between systemic and ocular humoral circulation. The ocular and systemic bioavailabilities of topically applied flurbiprofen were also quantitated. Anesthetized albino female rabbits received flurbiprofen doses intracamerally, topically, and intravenously at 2 to 4 week intervals. Aqueous humor and plasma were used as the sampling compartments. Plasma clearance values of flurbiprofen were 6.77 and 7.87 ml/min, after 6-mg and 208-g intravenous doses, respectively. These values were not significantly different and indicated no dose-dependent disposition kinetics over a 30-fold dose range. Both ocular and systemic flurbiprofen dispositions followed a biexponential pattern with a rapid distribution phase. The systemic and ocular distribution half-lives of flurbiprofen were 12 min and 15 min, respectively. The plasma elimination half-life was 74 min and the aqueous humor elimination half-life was 93 min. The latter approximated the turnover rate of aqueous humor and suggested that aqueous humor drainage was the major process of flurbiprofen elimination from the globe. About 99% of flurbiprofen is bound to plasma protein. At distribution equilibrium, the plasma and aqueous humor concentrations of fluobiprofen differed by a hundredfold, suggesting that only free drug entered the aqueous humor after the administration of a systemic dose. In the ophthalmic studies, right eyes were instilled with 50 l of 0.3% flurbiprofen in saline (dose = 150 g), and left eyes were instilled with 50 l of 0.15% flurbiprofen in saline (dose=75 g). When the area of the aqueous humor concentration-versus-time curve values was normalized by the administered dose, the 75-g dose was 30% more available to ocular tissues than was the 150-g dose. This demonstrated a disproportionate relationship between the administered dose and the fraction absorbed. The intracameral dose was considered to be completely bioavailable for intraocular effects. The ocular bioavailability of the ophthalmic dose was defined by using intracameral administration as a standard measurement. The ocular bioavailabilities of the 75-g and 150-g topical flurbiprofen doses were 10% and 7%, respectively. Systemic bioavailability after topical administration of 225 g of flurbiprofen was 74%.     

Intrathecal substance P depresses the tail-flick response — Antagonism by naloxone

Summary Substance P injected into the lumbar subarachnoid space of rats depressed the tail-flick response to radiant heat in a dose-dependent way. The effective doses ranged from 0.1 g to 100 g per rat (ED 50: 1.5 g/rat). The maximum of the effect was reached 20 min after intrathecal injection and the effect lasted for about 30 min. An antinociceptive effect was also observed after intrathecal injection of substance P 1 g to spinal rats. The depression of the tail-flick response produced by intrathecal administration of substance P was abolished by intrathecal (5 g/rat) or i.p. (0.5 mg/kg) injections of naloxone.Supported by the Sonderforschungsbereich 38 Membranforschung     

Dose-dependent effects of berberine on cell cycle pause and apoptosis in Balb/c 3T3 cells

In determining the morphological appearance of Balb/c 3T3 cells from berberine-treated (100 and 200 g/ml) cultures by light microscopy demonstrated that the high berberine concentration (200 g/ml) treatment was associated with the accumulation of numerous apoptotic cells, as identified by condensed nuclei and decrease in cell size. On the other hand, accumulation of cells in G₂/M phase instead of induction of apoptosis was observed after 48–72 h of 100 g/ml berberine treatment. Berberine was found mainly in cytoplasm during berberine-induced (100 g/ml) cell cycle G₂/M arrest, while it was highly concentrated in nuclei in the induction of apoptosis under high dose of berberine (200 g/ml) treatment. Further addition of berberine (100–200 g/ml) had little effect on the induction of apoptosis in the cells that had already been exposed to 100 g/ml of berberine for 48 h. Our results suggest that there may exist in Balb/c 3T3 cells an important threshold for regulation of cell cycle pause and induction of apoptosis, that is dose-dependent.     

Decreasing the oxidant stress from paraquat in isolated perfused rat lung using captopril and niacin

The abilities of captopril and niacin to protect against the lung toxicity of paraquat (PQ) were studied. The anti-oxidative action of captopril, an angiotensin-converting enzyme inhibitor, appears to be attributable to the sulphahydryl group (SH) in the compound, which gives captopril the ability to scavenge reactive oxygen species. Niacin replenishes the NAD and ATP depletion caused by reactive oxygen species. PQ causes lung damage in man and in several species of laboratory animals. The damage is initially manifested by hemorrhage and edema, and later by consolidation of the lung and fibrosis development. In this study, the lungs of male Wistar rats (250–300 g in weight) were perfused by Krebs-Ringer buffer alone (control), niacin (150 M), captopril (10 M) and PQ (600 M) in perfusion fluid, and the biochemical changes that occurred in isolated rat lung were examined within 1 h and compared to PQ alone. The results show that captopril significantly decreases the lung weight/body weight ratio when used as a pretreatment and a post-treatment to captopril (p<0.0001). The results also show that captopril (10 M) and niacin (150 M) significantly decreases PQ-induced lung toxicity. Lactate dehydrogenase (LDH) activity significantly decreased in treatment groups as compared to the PQ group (p<0.0001). This study suggests that paraquat causes increased lipid peroxidation and LDH activity and decreased glutathione (GSH) and total protein in isolated perfused rat lung. These effects are reduced under these experimental conditions by captopril and niacin.     

Hepatic Disposition of Fexofenadine: Influence of the Transport Inhibitors Erythromycin and Dibromosulphothalein

Purpose. To examine the disposition of fexofenadine in the isolated perfused rat liver and the influence of erythromycin and dibromosulphthalein (DBSP) on the hepatic uptake and biliary excretion of fexofenadine. Methods. Livers from four groups of rats were perfused in a recirculatory manner with fexofenadine HCl added as a bolus (125, 250, 500, or 1000 g) to perfusate. Livers from another three groups of rats were perfused with 250 g of fexofenadine HCl. With one group as control, erythromycin (4.0 g/ml) or DBSP (136 g/ml) was added to the perfusate of the other groups. In all experiments, perfusate and bile were collected for 60 min; in addition, livers from the second experiment were retained for assay. Fexofenadine was determined in perfusate, bile, and homogenized liver by HPLC. Results. The area under the curve (AUC) of fexofenadine was linearly related to concentration. It was unchanged from control (12,800 ± 200 ng·h/ml) by erythromycin (14,400 ± 2000 ng·h/ml), but was increased 95% by DBSP (25,000 ± 2600 ng·h/ml, P <0.001). The ratios of the concentrations of fexofenadine in liver/perfusate were decreased significantly by DBSP; those for bile/liver were increased by erythromycin. Conclusions. Erythromycin reduced the canalicular transport of fexofenadine into bile, whereas DBSP reduced uptake across the sinusoidal membrane.     

Stereoselective analysis of the disposition of tosufloxacin enantiomers in man

Summary The pharmacokinetics of tosufloxacin enantiomers after oral administration of racemic tosufloxacin were examined in healthy volunteers. Only small differences were observed in time to peak concentration (2.6±0.3 [mean ± SEM] h for (+)-tosufloxacin vs 2.4±0.2 h for (–)-tosufloxacin), elimination half-life (3.61±0.24 h vs 3.49±0.23 h), and area under the curve (2.78±0.19 h·g/ml vs 2.87±0.19 h·g/ml); however, peak concentration (0.40±0.03 g/ml vs 0.44±0.03 g/ml), renal clearance (226±10 ml/min vs 202±10 ml/min), and urinary recovery (35.4±2.2% vs 32.4±1.9%) differed significantly between enantiomers.     

Über den Hydroxytyramin- und Noradrenalingehalt der Lunge

Summary 1. Dopamine represents more than 90% of the catechol amines found in lung tissue. This has been demonstrated by paperchromatography, colorimetric, fluorimetric and biological methods. The mean dopamine content of lungs is: sheep 24 g/g, ox 1,1 g/g and dog 0,3 g/g. Apart from dopamine sheep lungs contained about 0,45%, ox lungs 1,4% and dog lungs 4,8–6,5% of noradrenaline.2. No adrenaline or isopropyl-noradrenaline could be detected.3. Manometric experiments showed that dihydroxyphenylalanine (dopa) will not be decarboxylated by extracts of lung tissue although the coenzyme pyridoxal-5-phosphate had been added. Therefore the dopamine of the lung can not be formed by this tissue itself.4. Without addition of pyridoxal-5-phosphate the decarboxylation of dopa by ox liver was week compared with the activity of sheep liver. This agrees with the different dopamine content of ox and sheep lung respectively and suggests that dopamine is formed in the liver, secreted into the blood, and taken up by the lungs.

---

---

Mit 4 Textabbildungen

---

Ausgeführt mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Recovery and conversion of kinins in exsanguinated rat preparations

Summary A rat preparation in which the perfusion route bypassed the lungs, which were substituted by an artificial oxygenator, was exsanguinated and perfused with oxygenated 4% dextran (M.W. 70,000) in Tyrode's fluid; a peristaltic pump propelled the perfusing fluid at 20–25 ml/min through the aortic arch and the perfusion medium returned to the right ventricle. Known amounts of bradykinin (BK), kallidin (lysyl-bradykinin, LBK) and methionyllysylbradykinin (MLBK) were administered as single injections and samples of the perfusion fluid removed between 1.5 to 6 min following injection. Average total kinin activity remaining in the circulating fluid was calculated from assays on the isolated guinea pig ileum against respective kinin injected and found to be after 3 and 6 min respectively: 20 and 5% for BK, 54 and 21% for LBK, 60 and 30% for MLBK. When the lungs were introduced in the perfusion circuit BK recoveries decreased to 0.4% at 4 min and 0% at 6 min. In 2 experiments 1 mg MLBK and, in another two, 1 mg of LBK were recirculated for 3 to 3.5 min in the rat preparation with lung bypass; enzymic reactions were interrupted in the perfusates after removal by lowering the pH to 4.7 and placing them in a boiling water bath for 5 min. Following proper dilution, kinin activity left in the perfusates was separated on carboxymethyl-cellulose columns; in 3 experiments about 50% of the activity was identified as BK from its elution position and resistance to trypsin digestion.The average BK-inactivating potency of the perfusate obtained from the rat with lung bypass was 0.3 g BK/minxml compared to 16 g BK/minxml of rat plasma. The arylamidase activity on arginylnaphthylamide of the perfusate was 2 n moles NA/minxml and it was about 25-fold lower than that of rat plasma.Rat liver was exsanguinated and perfused in situ through the portal vein and inferior cava vein using the same conditions as for the whole animal. The perfusion rate was 12 ml/min. The recovery of injected BK in this preparation was 40% after 2 min of recirculation, declining progressively in the following minutes. When MLBK was perfused in this preparation for 3 min or glycylarginyllysylbradykinin (GALBK) for 3 and 5 min, significant amounts of BK were found in the perfusates.We conclude that LBK, MLBK and GALBK may be converted at a high rate into BK by tissue aminopeptidases found in the rat preparations used. BK inactivation in the whole rat is a fast reaction, even when the pulmonary tissue is not involved in the inactivation.Abbreviations BK Bradykinin - LBK Kallidin or lysylbradykinin - MLBK Methionyllysylbradykinin - GALBK Glycylarginyllysylbradykinin - AAMLBK Aspartylarginymethionyllysylbradikinin - NA Naphthlyamine This work was supported by grants from Convenio BIOQ/FAPESP (São Paulo) and FINEP (Rio)With a Fellowship from FAPESP     

The effect of thyroxine treatment on the dynamics of insulin release from the isolated perfused rat pancreas

Summary The isolated perfused pancreas obtained from rats treated with thyroxine (600 g/kg/day, for five days) showed a modified insulin secretory pattern. Glucose (300 mg/100 ml) induced a higher first peak of insulin secretion which appeared slightly faster than in controls. The delayed second peak, as induced by glucose in the normal rat pancreas was absent after thyroxine treatment. Tolbutamide (20 mg/100 ml) yielded a slightly higher first peak followed by a moderately increased basal insulin secretion in the treated group.     

Analysis of Nonlinear Hepatic Clearance of a Cyclopentapeptide,BQ-123, with the Multiple Indicator Dilution Method Using the Dispersion Model

Purpose. To bridge in vitro, in situ and in vivo kinetic analyses of the hepatic clearance of a cyclopentapeptide, BQ-123, by using dispersion models that assume nonlinear pharmacokinetics. Methods. Rat livers were perfused by the multiple indicator dilution method with doses of BQ-123 ranging from 1-1000 g. The outflow dilution curves were fitted to a two-compartment dispersion model that was solved numerically by the finite difference method. Further, in vivo plasma concentrations of BQ-123 after bolus injection were analyzed with a hybrid physiological model that incorporates the hepatic dispersion model. Results. The calculated Michaelis-Menten constants (K_m = 12.0 M, V_max = 321 pmol/min/10⁶ cells, P_dif = 1.2 l/min/10⁶ cells) were comparable to those obtained previously from the in vitro isolated hepatocyte experiment (K_m = 9.5 M, V_max = 517 pmol/min/10⁶ cells, P_dif =1.1 l/min/10⁶ cells). The plasma concentrations of BQ-123 at doses of 1-25 mg/kg were explained well by the hybrid physiological model. Conclusions. These results suggest that carrier-mediated transport on the sinusoidal membrane was responsible for the in vivo hepatic elimination of BQ-123.     

Dopamine and adrenaline,but not isoprenaline,are substrates for uptake and metabolism in isolated perfused lungs of rats

Summary The uptake and subsequent metabolism by catechol-O-methyltransferase (COMT) and monoamine oxidase (MAO) of dopamine, adrenaline, isoprenaline and noradrenaline in isolated perfused lungs of rats has been examined. In lung preparations in which COMT and MAO were inhibited, the uptake of ³H-labelled dopamine, (–)-adrenaline and (–)-noradrenaline, but not (±)-isoprenaline, was reduced by cocaine (10 or 100 mol/l) The rank order of the Km values of the amines that were substrates for uptake in the lungs were: dopamine (0.246 mol/l) < noradrenaline (0.967 mol/l) < adrenaline (3.32 mol/l). These results are consistent with transport of catecholamines in rat lungs by Uptake₁.In lung preparations with COMT and MAO intact, dopamine and noradrenaline were removed from the circulation (50% and 32%, respectively) and mainly metabolized. There was very little (3.0%) removal of isoprenaline by the lungs and adrenaline was not included in this part of the study. In lung preparations in which only MAO was inhibited, the rank order of COMT activity for O-methylation of the amines was dopamine noradrenaline adrenaline (k_COMT values: 4.98 min^–1, 0.357 min^–1, and 0.234 min^–1, respectively).If dopamine or adrenaline are perfused through the pulmonary circulation in isolated lungs of the rat, they are taken up and then metabolized by COMT and MAO, as also occurs for noradrenaline. Isoprenaline is not a substrate for uptake in the lungs. There was less uptake of adrenaline than noradrenaline, indicating that uptake and metabolism in the lungs may not be a significant removal process for adrenaline in the circulation of rats in vivo. The more marked uptake of dopamine (than of noradrenaline) indicates that uptake and metabolism by the lungs, at least in the rat, may play an important role in the removal of dopamine from the circulation in vivo.Abbreviations COMT catechol-O-methyltransferase - DOMA 3,4-dihydroxymandelic acid - DOPAC 3,4-dihydroxyphenylacetic acid - DOPEG 3,4-dihydroxyphenylglycol - DOPET 3,4-dihydroxyphenyl ethanol - MAO monoamine oxidase - MN metanephrine - MTA 3-methoxytyramine - NMN normetanephrine - OMDA O-methylated deaminated metabolites - OMI 3-O-methylisoprenaline - U-0521 3,4-dihydroxy-2-methylpropiophenone Some of the results of this study were presented to the Australasian Society of Clinical and Experimental Pharmacologists (Bryan and O'Donnell 1987, 1988; Bryan et al. 1989; Bryan-Lluka 1990) Send offprint requests to L.J. Bryan-Lluka at the above address     

Effect of urodilatin on platelet-activating factor-induced bronchoconstriction, vasoconstriction and edema formation in isolated rat lung

In the isolated perfused rat lung, perfusion with platelet-activating factor causes bronchoconstriction, vasoconstriction and edema formation. The bronchoconstriction and vasoconstriction are largely mediated by thromboxane, whereas the edema formation is due to enhanced vascular permeability unrelated to eicosanoids. Since natriuretic peptides are known to relax smooth muscle and were suggested to attenuate enhanced vascular permeability, we investigated the effect of urodilatin on the PAF-induced alterations in lung function. Pretreatment with urodilatin (0.25 M or 0.75 M) reduced the PAF-induced increase in airway and vascular resistance by approximately 50%. Urodilatin pretreatment, however, was completely ineffective against the PAF-induced increase in weight gain and in vascular permeability, as assessed by the vascular filtration coefficient. Furthermore, urodilatin failed to affect the release of thromboxane into the perfusate in PAF-exposed lungs. Thus, urodilatin relaxes airway and vascular smooth muscle, but fails to reduce edema formation in PAF-perfused rat lungs.     

Effects on atrio-ventricular conduction of calcium-antagonistic coronary vasodilators,local anaesthetics and quinidine injected into the posterior and the anterior septal artery of the atrio-ventricular node preparation of the dog

Summary The effects on atrio-ventricular (A-V) conduction and blood flow of calcium-antagonists (verapamil, nifedipine and diltiazem), local anaesthetics (procaine and lidocaine) and quinidine were investigated in the isolated, cross-circulated A-V node preparation of the dog. The drugs were injected individually into the posterior septal artery (PSA) through which the upper part of the A-V node is mainly perfused or into the anterior septal artery (ASA) through which the lower part of the node and the more distal conduction system are perfused. Single injections into the PSA of nifedipine (0.3–10 g), verapamil (1–30 g), diltiazem (1–30 g), quinidine (30–300 g), lidocaine (100 g–1 mg) and procaine (300 g–3 mg) produced a dose-related increase in the A-V conduction time and with higher doses of these drugs a second or third degree block of A-V conduction occurred. Nifedipine (0.3–30 g) and verapamil (1–100 g) injected into the ASA scarcely affected A-V conduction. Quinidine (30 g–1 mg) and lidocaine (100 g–3 mg) injected into the ASA prolonged the A-V conduction time in a dose-related manner, although the effects were less prominent than those produced upon injection into the PSA. High doses of quinidine (3 mg) and lidocaine (3–10 mg) injected into the ASA altered the shape of ventricular bipolar electrograms and prolonged the time interval between an electrogram of the right bundle branch and that of the ventricle. The results are consistent with the hypothesis that in excitation of A-V nodal cells a slow calcium current rather than a fast sodium current plays an important role and that in the His-Purkinje-ventricular system the fast sodium current is predominant. Single injections of the 6 drugs into the PSA produced a doserelated increase in blood flow through the PSA. All drugs but nifedipine increased the blood flow in almost the same dose range that caused impairment of A-V conduction. Nifedipine was 10 times more potent in increasing the blood flow than in impairing A-V conduction.     

Effect of nicotine or cotinine on metabolism of 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) in isolated rat lung and liver

The scope of the present study was to investigate whether nicotine or cotinine will affect the metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in isolated perfused rat lungs and livers and to study the effect of starvation on pulmonary metabolism of NNK. NNK metabolism was investigated in isolated perfused liver and lung of male F344 rats perfused with 35 nM [5-³H]NNK in presence of a 1400-fold excess of the main tobacco alkaloid nicotine and its metabolite cotinine. In perfused rat livers, nicotine and cotinine inhibited NNK elimination and metabolism and led to a substantial increase of elimination half-life from 14.6 min in controls to 25.5 min after nicotine and 36.6 min after cotinine co-administration, respectively. In parallel, the pattern of NNK metabolites was changed by nicotine and cotinine. The pathway of α-hydroxylation representing the metabolic activation of NNK was decreased to 77% and 85% of control values, whereas N-oxidation of NNK and glucuronidation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) was increased 2.6- and 1.2-fold in presence of nicotine and cotinine, respectively. When isolated rat lungs were perfused with 35 nM NNK for 3 h neither the elimination nor the pattern of metabolites were substantially affected due to co-administration of 50 μM nicotine or cotinine. Cytochrome P450 2E1 is known to participate in the activation of NNK and can be induced by starvation. However, isolated rat lungs from male Sprague Dawley rats perfused with [1-¹⁴C]NNK at about 2 μM for 3 h, revealed only small differences in pulmonary elimination and pattern of NNK metabolites between fed and starved animals. These results suggest that nicotine and its main metabolite cotinine inhibit the metabolic activation of NNK predominantly in the liver whereas activation in lung, a main target organ of NNK induced carcinogenesis, remained almost unaffected. Received: 13 March 1997 / Accepted: 21 November 1997     

Population Pharmacokinetics and Pharmacodynamics of the Anti-CD11a Antibody hu1124 in Human Subjects with Psoriasis

The pharmacokinetics of hu1124, a human anti-CD11a antibody, were investigated in human subjects with psoriasis. CD11a is a subunit of LFA-1, a cell surface molecule involved in T cell mediated immune responses. Subjects received a single dose of 0.03, 0.1, 0.3, 0.6, 1, 2, 3, or 10 mg/kg of hu1124 intravenously over 1–3 hr. Blood samples were collected at selected times from 60 min to 72 days after administration. Plasma samples were assayed for hu1124 by ELISA, and pharmacokinetic analyses were performed on the drug plasma concentrations. As the dose of hu1124 was increased, the clearance decreased from 322 ml/day per kg at 0.1 mg/kg to 6.6 ml/day per kg at 10 mg/kg of hu1124. The plasma hu1124 concentration–time profile suggested that the clearance of hu1124 was saturable above 10 g/ml. In addition, treatment with hu1124 caused a rapid reduction in the level of CD11a expression on CD3-positive lymphocytes (T cells) to about 25% of pretreatment levels. Regardless of the hu1124 dose administered, cell surface CD11a remained at this reduced level as long as hu1124 was detectable (>0.025 g/ml) in the plasma. When hu1124 levels fell below 3 g/ml, the drug was rapidly cleared from the circulation and expression of CD11a returned to normal within 7–10 days thereafter. In vitro, half-maximal binding of hu1124 to lymphocytes was achieved at about 0.1 g/ml and saturation required more than 10 g/ml. One of the receptor-mediated pharmacokinetic/pharmacodynamic models which was developed describes the dynamic interaction of hu1124 binding to CD11a, resulting in the removal of hu1124 from the circulation and reduction of cell surface CD11a. The model accounts for the continually changing number of CD11a molecules available for removing hu1124 from the circulation based on prior exposure of cells expressing CD11a to hu1124. In addition, the model also accounts for saturation of CD11a molecules by hu1124 at drug concentrations of approximately 10 g/ml, thereby reducing the clearance rate of hu1124 with increasing dose.     

Reduction of hexavalent chromium by ascorbic acid in rat lung lavage fluid

The reduction of hexavalent chromium [chromium(VI)] in lung lavage fluids, microsomal (S–9) fractions of lung and liver tissues, erythrocyte lysates and plasma prepared from adult rats was examined at pH 7.4 (37° C). Specific reducing capacity, which was defined as the amount of chromium(VI) reduced per mg of protein in the test sample, was highest in the lavage fluids. The concomitant trivalent species [chromium(III)] was detected as complexes with some of the lavage components and probably as colloidal hydroxides. By gel filtration analysis and UV spectrometry, ascorbic acid (AsA) was identified as an important reducing factor in the lavage fluids. AsA levels in the lavage fluids were about 38 g/g tissue, corresponding to 12% of total AsA in the intact lungs. The molar ratios of oxidized AsA and reduced chromium(VI) in the lavage samples were about 32.3 on an average. On the basis of this molar ratio, the AsA levels in the lavage fluids are equivalent to a reducing capacity of 8.4 g chromium(VI)/g tissue. These results suggest that the lining layers (surfactant layers) of rat lungs provide an AsA-related capacity for protection of the cells against the toxic effects of chromates and probably other oxidants.     

Lead and cadmium in breast milk higher levels in urban vs rural mothers during the first 3 months of lactation

Breast milk from 10 women each from the city of Hamburg and from a rural area was analyzed by atomic absorption spectrometry for contamination with lead and cadmium. Samples were examined at regular intervals for 3 months after birth. On day 5 a diurnal profile was analyzed; on the other days milk was taken before and after the morning feed.Daily permissable intake (DPI) for lead is 5 g/kg/day for children; the DPI for cadmium has as yet been determined only for adults as 400–500 g/week, equivalent to about 1 g/kg/day (WHO 1972). For breast milk as the main source of nutrition in infants, this study shows values of 9.1±2.5 (SD) g/l for lead in the rural population, with a tendency to decrease towards the end of lactation. Urban mothers had 13.3±5.5 (SD) g/l, with a tendency to increase. This difference was significant only on day 45. Mean cadmium content in rural mothers was 17.3±4.9 g/l, with much higher values in the colostrum and a decrease after 15 days. Urban mothers had 24.6±7.3 g/l, again with high colostrum values and a subsequent decrease. These latter values are not significantly different.Calculated daily intake according to these values is presented, based on 840 ml breast milk for a 5.5 kg infant per day. Rural infants ingested 0.9–1.3 g/kg/day of lead, and in the city 1.5–2.3 g/kg/day. Cadmium intake in rural infants amounted from 1.2–1.8 g/kg/day; in Hamburg it was 1.6–2.2 g/kg/day. Thus the daily ingestion of lead was just below the DPI, cadmium ingestion was higher than the DPI for adults. The rural population had lower values in breast milk for both heavy metals than the urban population, although not statistically significant. Compared to earlier reports there was a slight increase in lead concentration and a more significant increase for cadmium. This study shows that the increase in cadmium may have taken place during very recent years, possibly due to the increase in the pH of soil. Therefore, it is suggested that both levels be monitored in a continuous program to prevent any nutritional hazard.