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Biomedical subjects

A Lorenzen

Publications and source records attributed to A Lorenzen.

At least 37 records · Page 2Linked to original sources
PubMed1997

Halogenated aromatic hydrocarbon-mediated porphyrin accumulation and induction of cytochrome P4501A in chicken embryo hepatocytes.

Concentration-dependent induction of cytochrome P4501A (CYP1A) and intracellular porphyrin accumulation were observed following treatment of chicken embryo hepatocyte (CEH) cultures with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 3,3',4,4'-tetrachlorobiphenyl (PCB 77, IUPAC nomenclature), 2,3',4,4',5-pentachlorobiphenyl (PCB 118), 3,3',4,4',5-pentachlorobiphenyl (PCB 126), 3,3',4,4',5,5'-hexachlorobiphenyl (PCB 169), and a commercial mixture of PCBs (Aroclor 1254). For these halogenated aromatic hydrocarbons (HAHs), or mixture, maximal CYP1A activity [measured as ethoxyresorufin-O-deethylase (EROD) activity] and immunodetectable protein were observed at concentrations just prior to, or coincident with, the concentrations at which porphyrin accumulation became evident. Both immunodetectable CYP1A protein and catalytic activity decreased at high concentrations of these compounds, but the rate and extent of decrease of immunodetectable CYP1A protein varied. Time-course studies with PCB 77 indicated a decrease in potency and an increase in maximal CYP1A induction between 24 and 48 hr of exposure which may indicate in vitro metabolism of this HAH. Intracellular accumulation of total porphyrins without CYP1A induction, was observed for 2,2',5,5'-tetrachlorobiphenyl (PCB 52), 2,2',6,6'-tetrachlorobiphenyl (PCB 54), 2,2',3,5',6-pentachlorobiphenyl (PCB 95), 2,2',4,5,5'-pentachlorobiphenyl (PCB 101), 2,2',3,3',6,6'-hexachlorobiphenyl (PCB 136), and 2,2',4,4',5,5'-hexachlorobiphenyl (PCB 153). Overall, these results are consistent with a role for CYP1A induction and/or Ah receptor activation in porphyrin accumulation mediated by HAHs with a planar configuration, whereas those that are not planar may mediate porphyrin accumulation by a mechanism not involving induction of CYP1A.

Animals↗
PubMed1997

Sensitivity of common tern (Sterna hirundo) embryo HepatocyteCultures to CYP1A induction and porphyrin accumulation by halogenated aromatic hydrocarbons and common tern egg extracts.

Experiments were conducted to compare the sensitivity ofprimary cultures of common tern (Sterna hirundo) and chicken(Gallus domesticus) embryo hepatocytes to cytochrome P4501A (CYP1A)induction and porphyrin accumulation after exposure to halogenated aromatichydrocarbons (HAHs) or tern egg extracts. The HAHs tested were2,3,7,8-tetrachlorodibenzo-p -dioxin (TCDD),2,3,7, 8-tetrachlorodibenzofuran (TCDF) 3,3',4,4'-tetrachlorobiphenyl (PCB 77;IUPAC nomenclature), 3,3',4,4',5-pentachlorobiphenyl (PCB 126),3, 3',4,4',5,5'-hexachlorobiphenyl (PCB 169) and Aroclor(R) 1254 (a commercialmixture of PCBs). Extracts were prepared from common tern eggs collected fromthree sites in the Great Lakes basin and one reference site on the east coastof Canada. CYP1A induction was assayed asethoxyresorufin-O-deethylase (EROD) activity and/or immunodetectableCYP1A protein, and total intracellular porphyrin accumulation was measuredfluorometrically. Unlike chicken embryo hepatocyte (CEH) cultures, nointracellular porphyrin accumulation was observed after treatment of ternembryo hepatocyte (TEH) cultures with HAHs or egg extracts. TEH cultures were approximately 50 to >1600 times less sensitive than CEH cultures to HAH-mediated CYP1Ainduction. In contrast, TEH cultures were either approximately equallysensitive or only 3.5-15 times less sensitive than CEH cultures to CYP1Ainduction mediated by tern egg extracts. These data suggest that common ternembryos may be more susceptible to the CYP1A inducing effects mediated bycomplex mixtures of environmental contaminants than indicated by theirresponse to individual HAHs.

Animals↗
PubMed1997

Comparative toxicity of polychlorinated biphenyls to Japanese quail (Coturnix c. japonica) and American kestrels (Falco sparverius).

Polychlorinated biphenyls (PCBs) and related halogenated hydrocarbons bioaccumulate to high concentrations in top predators, such as raptorial birds, yet little is known of PCB toxicity to such species. This study explored several aspects of both the acute and chronic response of American kestrels (Falco sparverius) to three purified PCB congeners and a commercial mixture, Aroclor 1254, and compared the response to that of the Japanese quail (Coturnix c. japonica), a more studied species known to be PCB sensitive. In one experiment, adult female birds were given single oral doses of either Aroclor 1254, 3,3',4,4'-TCB (PCB 77, IUPAC nomenclature), 3,3',4,4',5-PCB (PCB 126) or 2,2',4,4',5,5'-HCB (PCB 153) and sacrificed after 5 d. In kestrels, neither the pure compounds nor the mixture affected hepatic or renal porphyrin levels. There was slight but significant hepatic and renal ethoxyresorufin O-deethylase (EROD) induction in birds dosed with PCBs 77 and 126. A cytochrome P-4501A (CYP1A) cross-reactive protein was detected in liver and kidney of kestrels given PCBs 77 and 126, but not in Aroclor 1254-dosed birds. In quail, an acute dose of Aroclor 1254 caused significant liver weight increases, hepatic and renal EROD and aminopyrine n-demethylase (APND) induction, and dose-related hepatic and renal porphyria. Quail treated with PCB 126 developed hepatic and renal porphyria; EROD and APND were also induced. Administration of PCB 77 caused only slight induction of hepatic EROD activity. PCB 153 caused some hepatic and renal porphyria and induced EROD to the same degree as PCB 126. A hepatic CYP1A cross-reactive protein was induced about 200-fold in all individual quail that exhibited significant EROD induction and was also induced in kidney of 1 quail given Aroclor 1254. A second experiment examined chronic exposure to Aroclor 1254 by feeding adult females of both species a daily dose of 7 mg/kg/d for 4-, 8-, and 12-wk periods. There were no effects on hepatic porphyrins in kestrels. APND and aldrin epoxidase (AE) were induced; EROD was not induced, although a hepatic CYP1A-like protein was detected in 1 kestrel dosed for 12 wk. Chronic exposure of quail to Aroclor 1254 caused highly significant increases in mean hepatic porphyrin levels and in activity of EROD, APND, and 4-chlorobiphenyl hydroxylase; a CYP1A-like protein was also induced about 200-fold. In both studies, Aroclor 1254 residues accumulated in tissues of both species, but there was no significant relationship between residue levels and effects. In conclusion, adult American kestrels were relatively insensitive to the effects of PCBs, from both acute and chronic exposure, on hepatic and renal porphyrin levels. Although concentrations of a CYP1A-like protein were increased in some kestrels given PCBs, EROD activity was only marginally increased, suggesting that catalytic activity of this protein differed among the two species.

Aminopyrine N-Demethylase↗
PubMed1996

Characterization of a novel adenosine binding protein sensitive to cyclic AMP in rat brain cytosolic and particulate fractions.

A novel binding site for the adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA), which was enriched in rat forebrain, was characterized in cytosolic and particulate preparations. The site showed a pharmacological profile different from other [3H]NECA binding proteins and was named adenotin 2. [3H]NECA was bound in the presence of 100 microM 2-chloroadenosine with a Kd of 45.4 nM and a Bmax of 4711 fmol/mg in the cytosol and a Kd of 72.4 nM and a Bmax of 4844 fmol/mg in the crude membrane fraction. The presence of two different binding sites on adenotin 2 for [3H]NECA was shown in kinetic experiments. This protein showed identical pharmacological profiles in both subcellular preparations. [3H]NECA was displaced by purine analogues with a rank order of potency of NECA > 3'5' cyclic AMP (cAMP) > 5'-deoxy-5'-chloroadenosine > S-adenosylhomocysteine approximately 5'-deoxy-5'-methylthioadenosine (MeSA) > adenosine approximately adenine. cAMP inhibited [3H]NECA binding allosterically, whereas adenine and MeSA acted competitively. Inhibitors and activators of protein kinases such as N-(2-aminoethyl)-5-isoquinolinesulfonamide, Sp-adenosine cyclic monophophothioate and (8R*, 9S*, 11S*)-(-)-9-hydroxy-9-methoxy -carbonyl-8-methyl-2,3,9, 10-tetrahydro-8,11-epoxy-1H, 8H, 11H-2, 7b, 11a-triazadibenzo-(a,g)cycloocta(cde)-trinden-1-one (K 252a) interacted with [3H]NECA binding to adenotin 2 in nanomolar concentrations. Adenosine-5'-O-(3-thiotriphosphate) (100 microM) increased the affinity of [3H]NECA to a Kd of 9 nM and diminished the affinity of cAMP. The pharmacological characteristics of this novel binding site for [3H]NECA resemble those of the inhibition of phosphorylation processes by adenosine and its derivatives in heart and smooth muscle but are distinct from known adenosine receptors, adenosine binding proteins and protein kinases.

Adenosine↗
PubMed1996

Cytochrome P4501A induction in avian hepatocyte cultures: a promising approach for predicting the sensitivity of avian species to toxic effects of halogenated aromatic hydrocarbons.

Concentration-dependent effects of halogenated aromatic hydrocarbons (HAHs) on cytochrome P4501A (CYP1A) induction in primary hepatocyte cultures prepared from embryos of chickens (four breeds), pheasants, turkeys, ducks (three breeds), and herring gulls were determined. CYP1A activity was estimated by measuring ethoxyresorufin O-deethylase (EROD) activity and the concentration of immunodetectable CYP1A was estimated using mouse monoclonal antibody 1-12-3 that was prepared against scup (Stenotomus chrysops) CYP1A1. The HAHs studies were 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 3,3',4,4'-tetrachlorobiphenyl (PCB 77, IUPAC nomenclature), 3,4,4',5-tetrachlorobiphenyl (PCB 81), 3,3',4,4',5-pentachlorobiphenyl (PCB 126), 3,3',4,4',5,5'-hexachlorobiphenyl (PCB 169), 2,3,3',4,4'-pentachlorobiphenyl (PCB 105), and 2,3',4,4',5-pentachlorobiphenyl (PCB 118). Two general types of comparisons were made: (1) relative potencies of compounds within a species (expressed relative to TCDD as induction equivalency factors, IEFs) and (2) relative sensitivity of each species to EROD induction by each compound. Three methods for estimating potency were compared. These were: (1) the concentration of inducer that produced a half-maximal (EC50) EROD response, (2) the concentration producing a response equivalent to 10% of the maximal response produced by TCDD (ECTCDD 10%), and (3) a slope ratio method. For each method, the rank order in potency was TCDD > or = TCDF > PCB 126 > PCB 81 > PCB 77 > PCB 169 in chicken, pheasant, and turkey hepatocytes. The rank order was similar in duck and herring gull hepatocytes with the following exceptions: TCDF was approximately 2- to 4-fold more potent than TCDD in duck hepatocytes; PCB 169 induced EROD in gulls, but PCB 77 had no measurable effect in this species. PCB 118 was a relatively weak EROD inducer in most species/breeds, but it did not induce EROD in Pekin ducks or gulls. PCB 105 was a weak inducer in White Leghorn chicken and turkey hepatocytes, but it did not induce EROD in other species. The EC50, ECTCDD10% and slope ratio methods for estimating potencies generally gave similar IEFs for compounds that produced a maximal response that was at least 60% of the maximal response produced by TCDD. For compounds that caused a response that was 50% or lower than that produced by TCDD, EC50-based IEFs were greater (10- to 100-fold) than ECTCDD10%-based IEFs or slope-ratio-based IEFs. Among species, the rank order in sensitivity to EROD induction was chicken > pheasant > turkey > or = duck > or = herring gull. The relative sensitivity of avian hepatocyte cultures to EROD induction by PCB 77 was similar to the relative sensitivity of these species (reported elsewhere) to lethality after in ovo injection of PCB 77. Chicken hepatocyte cultures were 5-10 times more sensitive to EROD induction by TCDD than were pheasant hepatocyte cultures, which is identical to the difference in sensitivity of these species to the lethal effect of TCDD after in ovo injection. Measuring the sensitivity of hepatocyte cultures to EROD induction might be useful for estimating the sensitivity of avian species (including rare or endangered species, where it is impossible to conduct in vivo studies) to the embryotoxic effects of TCDD, non-ortho substituted PCBs, and other aryl hydrocarbon receptor agonists.

Animals↗
PubMed1996

Interaction of full and partial agonists of the A1 adenosine receptor with receptor/G protein complexes in rat brain membranes.

Full and partial agonists of the A1 adenosine receptor were characterized with respect to their influence on G protein activation and their thermodynamic parameters of receptor binding in rat brain membranes. G protein activation was determined through measurement of [35S]guanosine-5'-(gamma-thio)triphosphate ([35S]GTP[S]) binding, and receptor binding was studied under identical conditions through the displacement of [3H]-1,3-dipropyl-8-cyclopentylxanthine ([3H]DPCPX) in equilibrium binding studies. The intrinsic activity in stimulating [35S]GTP[S] binding did not correlate with the affinity of the ligands. 5'-Deoxy-5'-methylthioadenosine, 2-phenylaminoadenosine, and 2-chloro-2'-deoxyadenosine were identified as partial A1 agonists in the G protein activation assay. Depending on the temperature, these ligands showed agonistic and antagonistic properties to varying extents. EC50 values for G protein stimulation and KH and KL values of the partial agonists decreased when the incubations were performed at lower temperatures, indicating a mainly enthalpy-driven process of interaction with the receptor. Thermodynamic parameters of receptor binding of the partial agonists resembled the characteristics of the antagonist DPCPX more closely than those of the agonist 2-chloro-N6-cyclopentyladenosine. In addition, partial agonists detected fewer A1 adenosine receptors in the high affinity state than did full agonists. The lower efficacy in stimulation of the binding of [35S]GTP[S] is probably the consequence of an impaired ability of the partial agonists to release GDP from the G protein, as was shown by an impaired release of prebound [35S]GDP[S] from the membranes.

Animals↗
PubMed1993

Ethoxyresorufin-O-deethylase and porphyrin analysis in chicken embryo hepatocyte cultures with a fluorescence multiwell plate reader.

Rapid and sensitive methods for measuring ethoxyresorufin-O-deethylase (EROD) induction and total intracellular porphyrin accumulation in cultured chicken embryo hepatocytes exposed to polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are described. Cells were cultured in 48-well cell culture plates, and EROD assays were carried out directly within the wells. EROD activity and porphyrin concentrations were determined with a computer-operated fluorescence multiwell plate reader. Porphyrin and EROD assays could be carried out on the same plates or on separate plates. The assays allow one to obtain dose-response curves at rates that are approximately 100 times faster than conventional methods.

Animals↗
PubMed1993

Affinity of central adenosine A1 receptors is decreased in spontaneously hypertensive rats.

Functional defects in purinergic neurotransmission have been related to the development of arterial hypertension in spontaneously hypertensive rats. In order to elucidate the molecular basis of this perturbation, we have directly characterized adenosine A1 receptors using radioligand binding to rat brain membranes of Wistar Kyoto (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP). Saturation studies with [3H]1,3-dipropylcyclopentylxanthine ([3H]DPCPX) showed a lower affinity in both 5- and 48-week-old SHRSP in comparison with age-matched WKY. Similarly, competition experiments with [3H]DPCPX showed lower affinity of R-N6-phenylisopropyladenosine for the low-affinity binding site in 5- and 48-week-old SHRSP in comparison with WKY. In both studies, the difference in KD values was abolished by guanosine-5'-triphosphate in 5-week-old rats and mitigated in 48-week-old animals. No differences in Bmax values were observed in 5-week-old rats, whereas in 48-week-old SHRSP the number of receptors was significantly higher in comparison with age-matched WKY. Saturation experiments with the A1-selective agonist [3H]2-chloro-N6-cyclopentyladenosine ([3H]CCPA) demonstrated a higher affinity in 5-week-old SHRSP, whereas in 48-week-old hypertensive animals it was lower than in control WKY rats. No difference in receptor number was detected in comparison with age-matched WKY. In conclusion, our data demonstrated a diminished affinity of central adenosine A1 receptors for antagonists and for the low affinity state of the agonist binding site in genetically hypertensive rats. This might be due to structural changes of the receptor protein, to an altered G protein or defective receptor-G protein coupling in arterial hypertension.

Adenosine↗
PubMed1993

Characterization of membrane-bound and solubilized high-affinity binding sites for 5'-N-ethylcarboxamido[3H]adenosine from bovine cerebral cortex.

A high-affinity binding site for 5'-N-ethylcarboxamido[3H]adenosine ([3H]NECA) from bovine cerebral cortex has been characterized in its membrane-bound and solubilized state after gel filtration on Sepharose CL-6B. For detection of this site in membranes, it was necessary to remove metabolites with high affinities for this site enzymatically, e.g., adenosine by addition of adenosine deaminase and inosine by addition of nucleoside phosphorylase. The pore-forming peptide antibiotic alamethicin further enhanced binding of [3H]NECA to this site in membranes. In contrast to adenosine receptors and the adenotin-like low-affinity binding protein, this novel site was extremely sensitive against treatment with the sulfhydryl alkylating agent N-ethylmaleimide. In competition experiments, this site could be differentiated from adenosine receptors by its high affinity for adenine nucleotides and its lack of affinity for adenosine receptor antagonists. Inosine and its derivative S-(4-nitrobenzyl)-6-thioinosine were relatively potent ligands with Ki values in the high nano- and low micromolar range, respectively. We conclude that the high-affinity NECA binding site described previously in bovine striatum is not exclusively located in the striatum, but can also be detected in membrane preparations and soluble extracts of bovine brain cortex.

Adenosine↗
PubMed1993

Measurement of guanine nucleotide-binding protein activation by A1 adenosine receptor agonists in bovine brain membranes: stimulation of guanosine-5'-O-(3-[35S]thio)triphosphate binding.

Signal transduction by A1 adenosine receptors was investigated by measuring the modulation by adenosine agonists of guanosine-5'-O-(3-[35S]thio)triphosphate ([35S]GTP[S]) binding to guanine nucleotide-binding proteins (G proteins). The extent of stimulation of [35S]GTP[S] binding was dependent on the presence of high concentrations of Mg2+ (1-10 mM), GDP (10 microM), and NaCl (100 mM). Under optimal conditions, the agonist (R)-N6-phenylisopropyladenosine [(R)-PIA] stimulated binding of [35S]GTP[S] to G proteins approximately 2.3-fold. All adenosine receptor agonists tested stimulated the binding of [35S]GTP[S] with a rank order of potency typical of A1 adenosine receptors in bovine tissues, (R)-PIA > 2-chloro-N6-cyclopentyladenosine > (S)-PIA > 5'-N-ethylcarboxamidoadenosine > 2-[4-(2-carboxyethyl)phenethylamino]-5'-N-ethylcarboxamidoadenosin e. The EC50 values for G protein activation correlated with the Ki value of agonists for inhibition of radioligand binding to the high affinity state of the A1 adenosine receptor. The inclusion of 100 mM NaCl as well as increasing GDP concentrations led to a parallel increase of Ki values and EC50 values. In addition, both compounds induced a shift of A1 adenosine receptors from the high affinity state for agonists to the low affinity state.

Animals↗
PubMed1992

Reduction of postischemic leukocyte-endothelium interaction by adenosine via A2 receptor.

The adhesion of leukocytes to the endothelium of postcapillary venules hallmarks a key event in ischemia-reperfusion injury. Adenosine has been shown to protect from postischemic reperfusion injury, presumably through inhibition of postischemic leukocyte-endothelial interaction. This study was performed to investigate in vivo by which receptors the effect of adenosine on postischemic leukocyte-endothelium interaction is mediated. The hamster dorsal skinfold model and fluorescence microscopy were used for intravital investigation of red cell velocity, vessel diameter, and leukocyte-endothelium interaction in postcapillary venules of a thin striated skin muscle. Leukocytes were stained in vivo with acridine orange (0.5 mg kg-1 min-1 i.v.). Parameters were assessed prior to induction of 4 h ischemia to the muscle tissue and 0.5 h, 2 h, and 24 h after reperfusion. Adenosine, the adenosine A1-selective agonist 2-chloro-N6-cyclopentyladenosine (CCPA), the A2-selective agonist CGS 21,680, the non-selective adenosine receptor antagonist xanthine amine congener (XAC), and the adenosine uptake blocker S-(p-nitrobenzyl)-6-thioinosine (NBTI) were infused via jugular vein starting 15 min prior to release of ischemia until 0.5 h after reperfusion. Adenosine and CGS 21,680 significantly reduced postischemic leukocyte-endothelium interaction 0.5 h after reperfusion (p less than 0.01), while no inhibitory effect was observed with CCPA. Coadministration of XAC blocked the inhibitory effects of adenosine. Infusion of NBTI alone effectively decreased postischemic leukocyte-endothelium interaction. These findings indicate that adenosine reduces post-ischemic leukocyte-endothelium interaction via A2 receptor and suggest a protective role of endogenous adenosine during ischemia-reperfusion.

Adenosine↗
PubMed1992

Identification of a novel high affinity adenosine binding protein from bovine striatum.

In solubilized extracts from bovine striatal membranes three different binding sites for 5'-N-ethylcarboxamidoadenosine ([3H]NECA) were observed after separation of the extract by gel filtration on Sepharose CL-6B. The first peak was eluted in the void volume and contained the A2 adenosine receptor. In the second peak, [3H]NECA binding sites were eluted with a pharmacological profile characteristic of adenotin, a low affinity non-receptor adenosine binding protein. The third peak represented approximately 50% of the [3H]NECA binding activity. This site bound [3H]NECA in a reversible and saturable manner with KD of 17 nmol/l and a binding capacity of 11.3 pmol/mg protein. In competition experiments, adenosine, NECA, NAD, inosine, 5'-AMP and S-adenosyl-L-homocysteine were the most potent ligands. In contrast to adenosine receptors, this site did neither bind adenosine receptor antagonists nor the A2 selective agonist CGS 21,680 (2-[p-(2-carboxyethyl)phenethylamino]5'-N-ethylcarboxamidoadeno sin e). These results suggest the existence of a novel high affinity binding site for adenosine of unknown function in bovine striatum.

Adenosine↗
PubMed1991

Adenosine receptors in the central nervous system.

Two major subclasses of adenosine receptors have been distinguished in the central nervous system, termed A1 and A2. They are coupled to G-proteins and regulate the activity of adenylyl cyclase, potassium channels and several other effector systems. Autoradiographic studies have shown that A1 receptors are mainly found in the hippocampus and the cerebellum, whereas A2 receptors are almost exclusively located in the striatum and olfactory tubercle. Furthermore, a novel adenosine binding protein was identified in bovine striatum by radioligand binding with [3H]5'-N-ethylcarboxamidoadenosine ([3H]NECA). The pharmacological profile of this NECA binding protein has been determined in competition experiments with adenosine receptor ligands. It can be distinguished from that of A2 adenosine receptors and other adenosine binding proteins such as S-adenosylhomocysteine hydrolase and the adenosine transporter.

Adenosine↗
PubMed1991

Multiple-dose pharmacokinetics of epirubicin at four different dose levels: studies in patients with metastatic breast cancer.

Pharmacokinetic analysis of epirubicin and its metabolites epirubicinol and 7-deoxy-13-dihydro-epirubicinol aglycone during the first and the fourth courses of treatment was performed in 78 patients with metastatic breast cancer. The patients were treated every 3 weeks with epirubicin given as 10-min i.v. infusions at four different dose levels: 40, 60, 90 and 135 mg/m2. In most cases (76 of 78 cases), plasma concentration-time curves fitted to a three-compartmental pharmacokinetic model. The terminal half-life of epirubicin was independent of dose and duration of treatment. Large interindividual differences were demonstrated (mean t1/2 gamma, 21.6 +/- 7.9 h; range, 10.6-69 h; n = 110). In two subjects, extremely long half-lives and high serum bilirubin concentrations indicated impaired liver function. No correlation was found between the half-life and levels of liver alanine aminotransferase (ALAT) or serum creatinine. The metabolite epirubicinol appeared quickly after epirubicin administration and its half-lives were shorter than that of the parent compound (mean t1/2 gamma, 18.1 +/- 4.8 h; range, 8.2-38.4 h; n = 105). Formation of the aglycone metabolite was delayed and the half-life of this metabolite was shorter than that of epirubicin (mean t1/2 gamma, 13 +/- 4.6 h; range, 2.7-29 h; n = 104). The AUC of epirubicin and the total AUC (drug and metabolites) were linearly proportional to the dose, with the former value constituting two-thirds of the latter. A correlation was found between AUC and the plasma concentration of epirubicin at two time points (2 and 24 h after administration). The proposed model was AUC = 9.44 x c2 + 62.5 x c24 + 157.7 (r = 0.953).

Breast Neoplasms↗
PubMed1991

Detection and characterization of Ah receptor in tissue and cells from human tonsils.

Ah receptor was identified and characterized in cytosol and nuclear extracts from human tonsils obtained at surgery from children 2 to 6 years of age. Ah receptor was found in cytosol prepared from whole-tonsil homogenates as well as in cytosol and nuclear fractions prepared from tonsil lymphocytes or tonsil fibroblasts grown in primary culture. Cytosolic Ah receptor was detectable in tonsillar tissue with either halogenated (2,3,7,8-[3H]tetrachlorodibenzo-p-dioxin (TCDD)) or nonhalogenated (3-[3H]methylcholanthrene and [3H]benzo[a]pyrene) aromatic hydrocarbons and sedimented at approximately 9 S after velocity sedimentation on sucrose gradients. The apparent binding affinity (Kd) of [3H]TCDD for Ah receptor ranged from 3 to 12 nM in cytosols from seven different donors. The same analyses indicated a concentration of Ah receptor in human tonsils of approximately 100-300 fmol/mg cytosolic protein. Incubation of either tonsil lymphocytes or tonsil fibroblasts with [3H]TCDD resulted in transformation of cytosolic Ah receptor to a nuclear binding form which could be detected as a specifically labeled peak sedimenting at approximately 6 S on sucrose gradients. These data demonstrate the existence of Ah receptor in human tonsils and suggest that this immune organ may be an appropriate model for further studies on the mechanism and manifestation of aromatic hydrocarbon-induced immunotoxicity in man.

Cell Nucleus↗
PubMed1990

Detection and characterization of [3H]2,3,7,8-tetrachlorodibenzo-p-dioxin binding to Ah receptor in a rainbow trout hepatoma cell line.

Ah receptor was identified and characterized in cytosol and nuclear extracts from the rainbow trout hepatoma cell line RTH-149. The cytosolic receptor was detectable with both halogenated ([3H]2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)) and non-halogenated ([3H]3-methylcholanthrene and [3H]benzo[a]pyrene) aromatic hydrocarbons and sedimented at approximately 9 S after velocity sedimentation on sucrose gradients. The apparent binding affinity (kd) of cytosolic Ah receptor was always less than 1 nM as derived from Scatchard or Woolf plot analyses. The same analyses indicated a concentration of Ah receptor in the RTH-149 cells of approximately 20 fmol/mg cytosolic protein or approximately 4400 receptor sites per cell. Thus, this trout hepatoma cell line has a low concentration of high-affinity binding sites in comparison to Ah receptor concentrations in cytosol obtained from rodent tissues. Incubation of whole cells with the radioligand [3H]TCDD resulted in transformation of the cytosolic Ah receptor to a nuclear binding form which could be detected as a specifically labeled peak sedimenting at approximately 6 S on sucrose gradients. Aryl hydrocarbon hydroxylase was induced after exposure of RTH-149 cells to TCDD or benz[a]anthracene for 24 hr in culture. These data demonstrate the existence of the Ah receptor in a cell line derived from a nonmammalian species and provide an additional step toward understanding the mechanisms by which fish respond to specific aquatic contaminants.

Animals↗
PubMed1989

Human choriogonadotropin entrapped into liposomes: characterization, biologic effects and interaction with purified mouse Leydig cells in vitro.

Liposomes containing human choriogonadotropin (hCG) were prepared from phosphatidylserine by the ether injection method. hCG adsorbed to the outer surface of the liposomes (77% of total liposome-associated hCG) was removed by proteolytic digestion with subtilisin. hCG-containing liposomes digested and not digested with subtilisin stimulated testosterone biosynthesis by Leydig cells in a dose-dependent way; both preparations had identical biologic activities (32% of the activity of free, not liposome-associated hCG) when equal doses of liposome-associated hCG were applied. The onset of stimulation was delayed when compared to the action of free hCG. Liposomes without hCG did not stimulate testosterone biosynthesis. Association of liposomes with Leydig cells was determined by measurement of transfer of radioactive label from liposomes to Leydig cells. The association was not mediated by the hormone receptor. hCG entrapped in liposomes was incorporated by Leydig cells and translated to the cellular surface. This process was impaired by colchicine (10(-5) M). hCG translocated to the external surface of the cell membrane contained a modified alpha-subunit (Mr 16,200 instead of 20,600) which was not detected in unentrapped hCG bound to Leydig cells. We suggest that liposomally entrapped hCG is taken up by Leydig cells and re-exported to the cell membrane by a mechanism resembling retroendocytosis.

Animals↗