Amoxycillin levels in human serum, bile, gallbladder, lung, and liver tissue

Amoxycillin levels were measured in the serum, bile, lung, gallbladder, and liver tissue in 19 cases of lung and 22 cases of cholecyst operations following intramuscular administration of 1 g amoxycillin. After 60-160 min, 4.4-5.6 mcg/g intact lung tissue and 1.5-3.9 mcg/g tumorous lung tissue concentrations were found, representing 41-48% and 15-32%, respectively, of the corresponding serum levels. Between 70 and 160 min when the ducts cysticus was open, the cystic bile contained 5.2-8.8 mcg/ml, the bile taken from the biliary ducts showed 10.9-13.2 mcg/ml, whereas the wall of the gallbladder and the liver tissue displayed 4.4-5.1 and 1.7-2.8 mcg/g amoxycillin levels. These levels represented 50-92, 118-136, 50-52 and 17-28% of the actual serum levels. As in the serum levels, the bile and tissue levels were about twice as high as those following ampicillin administration. The amoxycillin levels measured in the serum, bile, and other tissue tissues in most cases exceeded the minimal inhibitory concentrations for most of the bacteria usually considered. Therefore, amoxycillin can be applied successfully to treat respiratory and biliary infections.     

Serum and lung tissue levels of cephradine in thoracic surgery

Serum and lung tissue levels in fifteen patients who underwent thoracic surgery were determined by the agar-diffusion plate method after i.m. administration of cephradine (500 mg). The mean value of the serum level 30 to 120 min after administration was 6.5 mug/ml, the mean lung tissue level was 2.6 mug/g. The lung tissue levels reached 40% of the simultaneous serum level. Four patients received cephradine for the treatment of post-operative chest infections. This antibiotic has an important therapeutic role in cases of thoracic-surgical infections.     

Assessment of biliary excretion of piperacillin-tazobactam in humans

Piperacillin-tazobactam concentrations in serum and bile were measured intraoperatively in 10 patients undergoing cholecystectomy (group 1) and 5 cholecystectomized patients provided with external bile duct drainage (group 2). Each patient received a single intravenous dose of piperacillin at 4 g plus tazobactam at 0.5 g over 30 min. Drug concentrations in both serum and bile were measured by high-performance liquid chromatography. In group 1 patients, serum and bile specimens and gallbladder wall fragments were collected at mean times of 70 and 83 min postinfusion, respectively. The mean concentrations of piperacillin and tazobactam were, respectively, 69.1 +/- 41.5 (standard deviation) and 9.9 +/- 5.1 microg/ml in serum, 630.4 microg/ml (range, 24.8 to 1,194 microg/ml) and 11.8 microg/ml (range, 3.6 to 22 microg/ml) in choledochal bile, 342.3 microg/ml (range, 1.1 to 1,149 microg/ml) and 7.7 microg/ml, (range, 0.2 to 23.1 microg/ml) in gallbladder bile, and 49.3 microg/g (range, 9.7 to 223 microg/g) and 2.9 microg/g (range, 0.1 to 5.9 microg/g) in the gallbladder wall. In group 2 patients, the amounts of drugs recovered in bile drainage obtained over 12 h were 28.4 +/- 18.0 and 1.0 +/- 0.5 mg for piperacillin and tazobactam, respectively. Peak piperacillin and tazobactam concentrations in bile reached 358 +/- 242 and 10.8 +/- 4.2 microg/ml, respectively. Comparison of drug levels in serum and bile suggests an underlying active secretion process for piperacillin elimination into the bile, unlike that of tazobactam. From a therapeutic viewpoint, given the concentrations of tazobactam recorded in bile fluid and tissue, the addition of this beta-lactamase inhibitor to piperacillin therapy might be of interest in the management of biliary tract infections, mostly in patients at risk of mixed aerobic-anaerobic infections due to beta-lactamase-producing organisms.     

Residues of chlorinated hydrocarbons in livers of newborns, infants and children

In the liver of 4 newborns, 2 infants and 2 children the levels of residues of chlorinated hydrocarbons were determined. The highest levels of sigma HCH (817 mcg/kg of fat, 17.1 mcg/kg of tissue) were found in a 2-day-old newborn, the lowest ones (43 mcg/kg of fat, 1.8 mcg/kg of tissue) were in an infant aged 12 months. The content of sigma DDT was highest (3044 mcg/kg of fat, 93.8 mcg/kg of tissue) in the liver of an infant aged 13 days, the lowest values (322 mcg/kg of fat, 5.6 mcg/kg of tissue) in the liver of a child aged 12 years. The results are comparable to those obtained in this country in the liver of adults and in human placenta.     

Late-onset Wilsońs disease]

A 58-year old male with a past history of psychiatric disorders was studied for a persistent serum transaminase increase. Low serum ceruloplasmin level (lower than 3 mg/dL), increased urinary copper excretion, and increased liver tissue copper concentration (1050 mcg/g dry weight) confirmed the diagnosis of Wilsońs disease. Slit lamp examination did not show Kayser-Fleischer rings. D-penicilamin therapy was followed by serum transaminase normalization. Similar late-onset cases of Wilsońs disease are exceptional, but confirm the clinical heterogeneity of the disease.     

Serum hyaluronic acid as a reliable parameter of allograft viability in porcine liver transplantation

BACKGROUND/AIMS: Hyaluronic acid is an endogenous glycosaminoglycan which is selectively degraded by hepatic sinusoidal endothelial cells. We evaluated the significance of serum hyaluronic acid clearance as an early indicator of allograft viability in porcine liver transplantation. MATERIALS AND METHODS: According to the survival period, animals were divided into two groups: Group I (n = 8) for survival equal or over four days and Group II (n = 5) for survival less than four days. Serial serum hyaluronic acid concentrations were measured before and after reperfusion in the recipient. RESULTS: In both groups, serum hyaluronic acid levels during the anhepatic period increased rapidly 9-fold from the preoperative value due to the absence of clearance by hepatic endothelial cells. In Group I, serum hyaluronic acid peaked at 15 min postreperfusion and decreased thereafter. In contrast, Group II failed to show clearance of hyaluronic acid after reperfusion. The serum hyaluronic acid value 120 min after reperfusion was 1,029 +/- 357 micrograms/L in Group I, and 1,856 +/- 263 micrograms/L in Group II (p < 0.01). Conventional parameters of liver function such as aspartate transaminase, lactic dehydrogenase, ammonia, lactate, and total bile acids were comparable between the two groups. CONCLUSIONS: The clearance of the serum hyaluronic acid reflects hepatic sinusoidal endothelial cell function and is a reliable and early marker of hepatic allograft viability.     

Telomere length, telomerase activity and telomerase RNA expression during mouse mammary tumor progression

The effects of intraportal administration of prostaglandin E1 (PGE1) on portal venous flow, hepatic arterial flow, peripheral tissue blood flow, and systemic arterial flow before and after 60 min total liver ischemia followed by 70% partial hepatectomy in rats were investigated. Total liver ischemia was induced by occluding the hepatoduodenal ligament for 60 min. PGE1 at a dose of 0.5 microg/kg/min was infused intraportally for 15 min before inducing hepatic ischemia (preischemic period) and for 60 min after ischemia (postischemic reperfusion period) in the treatment group. Normal saline was infused in the control group. Seventy percent partial hepatectomy was performed during ischemia. Serum biochemical analysis and liver tissue histology were carried out 1, 3, and 24 h, and 1 and 24 h after reperfusion respectively. One-week survival of the PGE1 group was improved to 70% compared to that of the control group of 30%. Postischemia reperfusion values of portal and peripheral tissue blood flows in the PGE1 group were 6.33 +/- 0.600 ml/min and 27.2 +/- 23.5 (arbitrary), and were significantly different from those of the control group of 4.34 +/- 0.400 ml/min and 23.5 +/- 5.54 (arbitrary), respectively. There was no significant difference in hepatic arterial flow between the two groups. Serum alkaline phosphatase decreased significantly in the prostaglandin group. Histological examination revealed a significant portal venous congestion in the control group 1 and 24 h after reperfusion. The extent of the sinusoidal congestion was also severe in the control group 24 h after reperfusion. It was concluded that PGE1 has a protective effect against liver damage when the liver was injured by warm ischemia and reperfusion followed by partial resection.     

Diagnostic value of biliary acid serum levels in Gilbert syndrome

OBJECTIVE: The aim of the present work was to study the diagnostic value of the measurement of total serum bile acids in patients with Gilbert's syndrome as compared to the fasting test. PATIENTS AND METHODS: We have studied 17 patients, 12 males and 5 females, between the ages of 7 and 15 years. All patients showed a slight unconjugated hyperbilirubinemia (1.4 +/- 0.5 mg/dl) in the presence of repeated normal liver function test, including the total serum bile acids (7.47 +/- 2.3 mumol/l), and showed no evidence of hemolysis. A modified fasting test was performed in all of the patients. Serum bile acid levels (Merckotest, Merck Labs) and the biochemical parameters of liver function were determined before and after the fasting test. RESULTS: Our results show that to confirm the diagnosis of Gilbert's syndrome, the fasting test could be avoided in patients with increased unconjugated bilirubin levels if the basal levels of total serum bile acids are normal.     

Study of in vivo tumor state and response to therapy by analysis of DNA distribution

Comparative studies of cefamandole and cephalothin were carried out in 32 cancer patients. After rapid intravenous injection of 1 gm cefamandole or cephalothin, the peak mean serum concentrations in 11 patients achieved at 0.25 hr were 103.4 mcg/ml and 56.7 mcg/ml, respectively. Except at 6 hr, the serum concentration of cefamandole was higher (p less than 0.05) at all times. The terminal half-lives (t 1/2) were similar, being 1.2 hr for cefamandole and 1.0 hr for cephalothin. Cefamandole, 1 gm intramuscularly, induced a peak mean serum concentration of 26.6 mcg/ml at 1 hr, with a slow decay. Intermittent cefamandole (2 gm intravenously every 6 hr) induced very high mean serum concentrations (7 patients), but at 4 hr the concentrations were similar to those after 1 gm intravenously. Per cent of urinary excretion was similar for both drugs regardless of dose and mode of administration. Continuous-infusion cefamandole or cephalothin (2 gm loading followed by 2 gm every 6 hr) in 14 patients showed consistently higher serum concentrations for cefamandole (p less than 0.05) over a 5-day period. There was no evidence of drug accumulation in the multiple-dose studies. Both the single- and multiple-dose schedules were well tolerated.     

Radioimmunoassay of serum bile acid levels in biopsy-proved cirrhosis

Serum bile acid levels were determined by a radioimmunoassay for conjugates of cholic acid in 18 patients with biopsy-proved cirrhosis and compared to results of conventional liver function tests. Serum bile acid levels were abnormal in all 18 patients whereas results of the other tests were abnormal in only 50% to 72% of patients. In this group of patients with a well established diagnosis of cirrhosis, serum bile acid levels were a more sensitive indicator of liver dysfunction than standard liver function tests.     

The fundamental and clinical studies on PT-122M (doxycycline) in the otorhinolaryngological field (author's transl)

As the results of laboratory and clinical investigation with a new tetracycline derivate, PT-122M (doxycycline) was performed with the results which may lead to the following conclusions. 1) In vitro antibacterial activity: The minimal inhibitory concentration of doxycycline (DOTC) was measured by an agar plate dilution method. The MIC of DOTC against 60 strains of coagulase-positive Staphylococcus aureus isolated from otorrhoea was distributed over a range of 0.2 approximately 1.56 mcg/ml and 12.5 approximately 50 mcg/ml. Furthermore, Escherichia coli, Proteus mirabilis and Klebsiella pneumoniae was 3.13 to 100 mcg/ml. 2) Concentration in blood: The blood level of DOTC in healthy adults who were given 100 mg of DOTC intravenous reached maximum of 1.97 mcg/ml on the average 15 minutes after injection. Even at 12 hours after intravenous injection clinically effective serum DOTC concentration of 0.47 mcg/ml was still demonstrable. 3) Clinical results: PT-122M (DOCT) was intravenously injected into 30 cases with typical infection in the field of otorhinolaryngology. It was excellent in 15 cases, good in 10 cases, fair in 3 cases, and poor in 2 cases. When the cases in which it was excellent and good were considered together, good results were obtained in 25 cases, a ratio of effectiveness being 83 per cent. 4) Side effect: The comparative examination of hepatic function, electrolyte and auditory acuity before and after injection showed no significant disturbance. No side effect was shown with the intravenous injection.     

Enhancement of endogenous cyclic AMP signal: a new approach to allow for cold preservation of rat livers from non-heart-beating donors?

BACKGROUND: The organ donor shortage has led to a reconsideration of the use of non-heart-beating donors (NHBDs). However, graft injury due to warm ischemia in NHBD livers strongly affects posttransplant outcome. The present study was aimed at investigating the role of the cellular cyclic (c)AMP second messenger signal with regard to hepatic viability after cold preservation of NHBD livers. METHODS: Cardiac arrest was induced in Wistar rats by frenotomy of the anesthetized nonheparinized animal. After 30 min, the livers were excised and flushed with 20 ml of heparinized saline solution, rinsed with 10 ml of University of Wisconsin (UW) solution, and stored submerged in UW solution at 4 degrees C for 24 hr. In half of the experiments, UW solution was supplemented with glucagon (0.5 microg/ml) to increase the cAMP signal in the liver. Reperfusion was carried out in vitro after all livers were incubated at 25 degrees C in saline solution to replicate the period of slow rewarming during surgical implantation in vivo. RESULTS: Hepatic levels of cAMP (nmol/g dry weight) declined from 1.21+/-0.05 to 0.53+/-0.03 (P<0.01) at 30 min after cardiac arrest. Subsequent storage in UW solution resulted in a further decline to 0.35+/-0.04 after 24 hr in group A, whereas glucagon treatment enhanced cellular cAMP signal to 0.64+/-0.06 (P<0.01). Upon reperfusion, liver integrity was significantly improved after glucagon administration, with 66% reduction in alanine aminotransferase release and a threefold increase in hepatic bile production as compared with untreated livers. Moreover, liver ATP tissue levels were restored to only 2.19+/-0.51 micromol/g in the untreated group but reached 4.97+/-0.41 micromol/g (P<0.05) after treatment with glucagon. CONCLUSIONS: Posthoc conditioning of predamaged livers by glucagon enhances cAMP tissue levels during ischemic preservation and improves hepatic integrity upon reperfusion. This may represent a promising approach for the use of livers from non-heart-beating donors in clinical transplantation.     

Predictive value of antepartum ultrasound examination for anomalies in twin gestations

RATIONALE AND OBJECTIVES: We investigated the potential of manganese (III) mesoporphyrin (Mn-mesoporphyrin) as a hepatobiliary contrast agent for magnetic resonance (MR) imaging in rabbits given VX-2 carcinoma liver implants. METHODS: Rabbits given VX-2 carcinoma liver implants (n = 8) were imaged before and after the intravenous (i.v.) administration of 0.04 mmol/kg Mn-mesoporphyrin. MR images were correlated with gross-specimen cross-sections. The distribution of Mn in various tissues following i.v. administration of 0.04 mmol/kg Mn-mesoporphyrin was determined using atomic absorption analysis. A standard panel of serum chemistries was followed over 7 days in six rabbits following this same dose of Mn-mesoporphyrin and compared with chemistries from two control rabbits. RESULTS: I.v. administration of 0.04 mmol/kg (25 mg/kg) Mn-mesoporphyrin resulted in improvement of tumor-to-liver contrast, with enhancement of normal liver (99.7 +/- 14.7%) and the gallbladder (442 +/- 116%), but not VX-2 tumor tissue (14.8 +/- 13.9%), (n = 8, p = .05). Analysis of tissue Mn levels 100 min after i.v. Mn-mesoporphyrin injection demonstrated preferential distribution of Mn to normal liver tissue (57.8 +/- 15.3 micrograms Mn/g) compared with VX-2 tumor (4.28 +/- 1.48 micrograms Mn/g). No significant change was found in the serum chemistries of six normal rabbits over a 7-day period after the i.v. administration of 0.04 mmol/kg Mn-mesoporphyrin. CONCLUSION: I.v. Mn-mesoporphyrin improved lesion-to-liver contrast because of preferential distribution of Mn-mesoporphyrin to normal liver parenchyma and bile.     

The molecular mechanism of temperature enhancement of proton magnetic relaxation rates in methaemoprotein solutions. III. The effect of sixth ligand in high-spin derivatives

Levels of cefazolin were determined in plasma, urine, bile, and cerebrospinal fluid in humans after a bolus intravenous injection and during a controlled, continuous intravenous infusion. All the patients were studied in a steady-state and crossover fashion. In plasma, the mean peak level after bolus injection (1.5 g) studied in 12 patients was 206.5 mug/ml; during continuous infusion (6 g daily), the mean level remained stable at 52.6 mug/ml. With bolus injection and continuous infusion, respectively, 89.7 and 86.3% of the administered dose of cefazolin were excreted in the urine of nine patients over the 6-h period considered. The levels of cefazolin in common bile duct bile were studied in six cholecystectomized patients. In bile collected during the two 3-h periods of the experiment, the mean concentration of the drug in the bile after bolus injection was 66.9 and 22.0 mug/ml, respectively; during continuous infusion, the corresponding biliary levels were 50.7 and 51.3 mug/ml, respectively. In four neurosurgical patients with an intraventricular catheter, neither bolus injection nor continuous infusion resulted in a demonstrable concentration of cefazolin in the cerebrospinal fluid. The continuous intravenous administration of cefazolin might have some advantage over the intravenous bolus intermittent injections. In plasma, the area under the curve is greater with continuous infusion than with bolus injection. In bile, the levels of cefazolin are more sustained with continuous infusion than with bolus injection. This approach to intravenous administration of cefazolin deserves more pharmacological and clinical trials.     

Serum bile acids and ursodeoxycholic acid treatment in cystic fibrosis-related liver disease

Increased circulating levels of hepatotoxic bile acids may contribute to the cholestasis characteristic of cystic fibrosis-related liver disease. The aims of this study were to compare serum bile acid profiles in patients with cystic fibrosis with and without liver disease, and to evaluate the effect of treatment with ursodeoxycholic acid, a non-hepatotoxic bile acid, on liver biochemistry and serum bile acids in patients with cystic fibrosis-related liver disease. Fasting and postprandial serum bile acid levels were analysed in 15 patients (nine males; median age 18 years) with cystic fibrosis-related liver disease and compared with serum bile acid levels in 18 cystic fibrosis patients (12 males; median age 22 years) without liver disease and 10 control subjects. Fasting and postprandial serum levels of primary and secondary serum bile acids were analysed using high-performance liquid chromatography. Liver biochemistry and serum bile acids were measured in six cystic fibrosis patients with liver disease before and 6 months after treatment with ursodeoxycholic acid 20 mg/kg/day and compared with six control patients with cystic fibrosis-related liver disease. Total fasting and postprandial serum bile acid levels were significantly (P < 0.01) elevated in patients with liver disease compared to those without liver disease and controls. The fasting glycine conjugates of cholic acid, chenodeoxycholic acid and deoxycholic acid, and the fasting and postprandial taurine conjugates of cholic acid and chenodeoxycholic acid were significantly (P < 0.05) elevated in liver disease patients compared to patients without liver disease and controls. After 6 months' treatment with ursodeoxycholic acid, although the serum was significantly saturated with ursodeoxycholic acid and significant improvements in liver biochemistry were observed in the treatment group, there was no significant reduction in the levels of individual serum bile acids. Although circulating levels of potentially hepatotoxic serum bile acids are elevated in patients with cystic fibrosis-related liver disease, improvements in liver biochemistry associated with ursodeoxycholic acid treatment cannot be attributed solely to alterations in levels of endogenous bile acids.     

Hemodialysis stimulates hepatocyte growth factor release

Studies were performed in 26 patients on regular dialysis treatment with cuprophane (CU), polymethylmetacrilate (PMMA) or cuprammonium (CAM) dialyzers. Controls were six patients with chronic renal failure but not on regular dialysis treatment (CRF) and six healthy subjects (N). Blood was collected at the start (T0), and at 15 (T15) and 240 (T240) minutes of dialysis to measure the serum hepatocyte growth factor (HGF) concentration and to study HGF production by peripheral blood mononuclear cells (PBMC) in vitro. The form of HGF (that is, inactive/monomeric, active/dimeric) present in the serum was analyzed by immunoblotting. In addition, the ability of serum to stimulate proliferation of tubular cells (HK-2) and HGF release by PBMC and fibroblasts (MRC-5) was investigated. At T0, serum HGF levels were identical to that of the controls. In patients treated with CU, serum HGF rose from 0.24 ng/ml at T0 to 7.44 ng/ml at T15, and remained high at T240. PBMC collected at T15 and T240 released significantly more HGF in vitro than those collected at T0. Serum at T15 stimulated proliferation of HK-2 cells and the release of HGF by PBMC and MRC-5 cells. The PMMA and CAM dialyzers had similar effects as the CU. These results indicate that dialysis induces a striking rise in serum HGF and a prompt circulation of factor(s) stimulating HGF release. Dialysis-activated PBMC release HGF.     

Urinary bile acids in population screening for inapparent liver disease

BACKGROUND/AIMS: We performed this study in order to evaluate the diagnostic potential of bile acids in random samples of urine for detection of latent liver disease and to compare a radioimmunoassay for urine bile acids with an enzymatic method that detects bile acid sulfates. This was a prospective cohort study carried out at the VA Medical Center involving 151 adults who attended a Community Health Fair at the hospital and wanted to know if they had liver disease. METHODOLOGY: Urinary bile acids in random specimens of 5-10 ml urine were measured. Radioimmunoassay for primary bile acids and an enzymatic assay with or without sulfated bile acids, all corrected with creatinine for urine flow were performed. Serum primary bile acids were determined by radioimmunoassay. In addition, routine liver profile and clinical examination were carried out. RESULTS: In 78 of 151 subjects there was at least one recent liver profile to match with the urine bile acids. Of these 78, 52 subjects with normal urine bile acids had a normal liver profile. In 11 subjects abnormal urine bile acids were associated with an abnormal liver profile. Nine of these 11 subjects were anti HCV positive, one was HIV positive. Urine bile acids correctly predicted the outcome of routine liver tests in 89% of 78 subjects. In nine cases there was a discordance between urinary bile acids and the liver profile. Failure to correctly predict the liver profile using urine, was reduced from nine subjects to three when urine bile acids were obtained twice at separate intervals. Urine bile acids predicted the outcome of anti HCV testing in 37 subjects with similar accuracy as serum ALT or AST. Urine bile acids correlated with serum bile acids at r=0.96, 0.88 and 0.76 for the radioimmunoassay, enzymatic assay that included sulfated bile acids and enzymatic assay without the sulfates, respectively. CONCLUSION: Bile acids in a random sample of urine are useful for population screening for latent liver disease. Prediction of sub-clinical hepatitis C is comparable to that of serum ALT or AST. Inclusion of bile acid sulfates mildly increases the predictive value of urine. Urine bile acids highly correlate with serum bile acids, indicating their surrogate diagnostic value.     

Intravenous injection of Candida-derived mannan results in elevated tumor necrosis factor alpha levels in serum

Intravenous injection of Candida albicans into mice produced elevated serum tumor necrosis factor alpha (TNF-alpha) levels. We hypothesized that immunostimulants released in vivo from C. albicans during fungal sepsis might contribute to the elevated levels of TNF-alpha in serum. We tested this hypothesis in mice with C. albicans mannan (CAM). Increased serum TNF-alpha levels were observed following intravenous and intraperitoneal injections of CAM. Injection of CAM into mice resulted in increased serum TNF-alpha concentrations that reached 1,200 pg/ml of blood, compared with 2,400 microg/ml of blood following injection of 10 microg of endotoxin. The response to CAM was concentration dependent, requiring a minimum dose of 20 microg of CAM per g of body weight. Sera from mice were tested 30, 60, 90, and 120 min after intravenous injections with CAM. TNF-alpha concentrations were minimal 30 and 120 min after intravenous injection and maximal 60 and 90 min after CAM injection. The relative distribution of CAM in vivo in decreasing order was determined to be as follows: blood > liver > lung > spleen, 90 min following injection of a single 5-mg dose of CAM. CAM was confirmed as the stimulating substance by utilizing anti-CAM antibodies in vivo to block the response. Rabbit anti-mannan antibodies administered by intraperitoneal injection 24 h before CAM injection significantly suppressed (P < 0.05) the accumulation of TNF-alpha in the sera. Dexamethasone administered to mice before intravenous injection of mannan significantly reduced (40 to 90% reduction; P < 0.05) the concentrations of TNF-alpha in the sera of treated mice. Thus, when in vivo CAM clearance mechanisms are exceeded, sufficient CAM may become available to stimulate TNF-alpha production, making CAM an important part of pathogenesis in Candida sepsis.     

Carrier-mediated active transport of the glucuronide and sulfate of 6-hydroxy-5,7-dimethyl-2-methylamino-4-(3-pyridylmethyl) benzothiazole (E3040) into rat liver: quantitative comparison of permeability in isolated hepatocytes, perfused liver and liver in vivo

The hepatic uptake of glucuronic acid and sulfate conjugates of 6-hydroxy-5,7-dimethyl-2-methylamino-4-(3-pyridylmethyl) benzothiazole (E3040), a dual inhibitor of 5-lipoxygenase and thromboxane A2 synthetase, was investigated in rats. The biliary excretion clearance values for the glucuronide and the sulfate, obtained after i.v. administration of E3040, were similar and corresponded to approximately 30% of the hepatic blood flow rate. The influx clearance values of E3040 conjugates in the presence of 3% bovine serum albumin, measured by a multiple indicator dilution method in the perfused liver, were 1.20 ml/min/g liver for the glucuronide and 0.74 ml/min/g liver for the sulfate, which were twice and equal to the normal hepatic plasma flow rate, respectively, which suggests the presence of an efficient transport system(s). The uptake of E3040 conjugates into the isolated hepatocytes is mediated by Na(+)-independent active transport system(s), which is inhibited by dibromosulfophthalein and bile acids. The uptake for the sulfate had high-affinity and high-capacity transport activity (Km = 25 microM; Vmax = 7.8 nmol/min/10(6) cells) compared with that for the glucuronide (Km = 59 microM; Vmax = 2.2 nmol/min/10(6) cells). The uptakes of E3040 conjugates (glucuronide, sulfate) exhibited a mutual competitive inhibition. It is suggested that both conjugates share a multispecific organic anion transporter located on the sinusoidal membrane.     

Role of bile salt hydrophobicity in hepatic microtubule-dependent bile salt secretion

Under basal conditions, bile salt secretion by the liver is not affected by microtubule disruption. However, when a bile salt load is imposed on the liver, a microtubule-dependent secretion mechanism is recruited (J. Lipid Res. 1988. 29: 144-156). We tested the hypothesis that recruitment of this microtubule-dependent mechanism is influenced by the relative hydrophobicity of the bile salts being secreted. Intact male rats were depleted of bile salts by overnight biliary diversion, pretreated with colchicine (a microtubule inhibitor) or its inactive isomer, lumicolchicine (control), and reinfused intravenously with bile salts of increasing hydrophobicity (taurodehydrocholate < tauroursodeoxycholate < taurocholate) at 200 nmol/min.100 g. After 45 min, when steady-state bile salt secretion was achieved, tracer [3H]taurocholate was administered intravenously. The colchicine-insensitive component of bulk bile salt secretion was constant at approximately 130 nmol/min.100 g, and the colchicine-sensitive component increased from approximately 0 to 35 and 60 nmol/min.100 g, respectively, with reinfusion of the more hydrophobic bile salts. Retained bile salts accumulated in the liver and serum and were detectable in urine. Peak biliary secretion of [3H]taurocholate in control animals increased linearly from 15.3 to 18.0% administered dose/min with increasing hydrophobicity of the secreted bile salts (P < 0.002). In colchicine-pretreated animals, peak secretion rates decreased linearly from 13.8 to 9.2%/min (P < 0.001), with maximal inhibition in taurocholate-reinfused animals (P < 0.01). Utilization of a microtubule-dependent secretion mechanism increases with increasing bile salt hydrophobicity. This mechanism permits more efficient hepatic secretion of bile salts, but increases the susceptibility of bile salt secretion to microtubule disruption. We postulate that microtubule-dependent insertion of bile salt transporters into the canalicular membrane underlies the enhanced bile salt secretion observed when a bile salt load is imposed upon the liver.