Appearance of spontaneously Ig secreting B cells in human peripheral blood after surgery.

Lymphoblastoid B cells, spontaneously secreting specific antibodies of IgG and IgA classes, are constantly detected after in-vivo immunization and represent a sensitive marker of a recent antigenic exposure. In this study we demonstrate that surgical trauma is followed, at a well-defined time after surgery, by the appearance of circulating lymphoblastoid B cells spontaneously secreting IgG and IgA. The kinetics and the functional behaviour of this B cell subset are identical to those of lymphoblastoid B cells observed after in-vivo immunization. Our data indicate that surgical trauma activates a humoral immune response. Antigens released by traumatized tissues or encountered through breaches in skin or mucous membranes might initiate the reaction.

Effects of iron overload on bile secretion and hepatic porphyrin metabolism in ethinyl estradiol-treated rats.

We investigated the effects of ethinyl estradiol (5 mg/kg body wt daily for 5 days, orally) and/or iron sorbitol (50 mg/kg body wt daily for 5 days, i.m.) on bile flow, bile salt independent fraction (BSIF), hepatic delta-aminolevulinate synthase (ALA-S) and uroporphyrinogen decarboxylase (URO-D) in female rats. Ethinyl estradiol administration was associated with a significant decrease of bile flow and BSIF and an increase in URO-D activity in comparison to control values. Iron alone did not modify biliary parameters, but significantly increased the activity of ALA-S. Combined treatment with ethinyl estradiol plus iron partially corrected the reduction of BSIF and restored the activity of ALA-S and URO-D to control levels. Thus iron appears to exert a partially protective effect against ethinyl estradiol-induced cholestasis. No porphyrinogenic effect was observed.

Double-blind placebo-controlled clinical trial of microporous cholestyramine in the treatment of intra- and extra-hepatic cholestasis: relationship between itching and serum bile acids.

In order to investigate the effectiveness of microporous cholestyramine in the pharmacological management of intra- and extrahepatic cholestasis, a double-blind placebo-controlled study was carried out in 10 patients. Microporous cholestyramine or placebo 3 g t.i.d. were given orally over a four-week period. The active drug resulted statistically superior to the placebo in reducing itching intensity (p less than 0.01) and serum bile acids (p less than 0.01). A positive linear relationship between itching and serum bile acids (p less than 0.01) was also demonstrated.

A comparative trial of desferrioxamine and hydroxychloroquine for treatment of porphyria cutanea tarda in alcoholic patients.

Forty male alcoholic patients with porphyria cutanea tarda (PCT) were randomly assigned to 2 groups of 20. The 1st group received desferrioxamine (30 mg/kg body weight/day for 1 week every 3 months) whereas the latter was given hydroxychloroquine (200 mg twice/wk orally). Alcohol abstinence was advised for all patients. Improvement of cutaneous signs was evident after 6 months in hydroxychloroquine-treated subjects and after 12 months in desferrioxamine-treated subjects. At the end of the 1-year clinical trial, significant decreases of serum iron and ferritin were found in all patients, irrespective of the therapy. Urinary total porphyrins were reduced significantly in both groups, but the drop was significantly more evident in hydroxychloroquine- than in desferrioxamine-treated subjects. After 1 year of therapy, 4 desferrioxamine-treated patients vs 16 hydroxychloroquine-treated subjects acquired a normal urinary porphyrin pattern. These results indicate that hydroxychloroquine is more effective than desferrioxamine in inducing clinical and biochemical remission of PCT. Accordingly, hydroxychloroquine should be the preferred alternative to phlebotomy, if the latter is contraindicated.

The treatment of retained biliary stones with monooctanoin: report of 16 patients.

The aim of this study was to estimate efficacy, safety, and tolerance of monooctanoin in 16 patients with retained radiolucent biliary stones and indwelling biliary drainage. Monooctanoin was infused continuously at a rate of 3-4 ml/h. Monitoring of pressure with a manometer broken off at 20 cm prevented the development of excessive pressure in the common bile duct. The mean volume of the compound instilled (+/- SD) was 848 +/- 393 ml (range 80-1450) and the mean duration of treatment was 13 +/- 6 days (range 2-23). Monooctanoin induced disappearance of stones in 11 of 16 patients giving a success rate of 69%. Three patients exhibited a partial dissolution of stones which were then successfully removed through the postoperative T-tube. The two failures can be attributed to the type of stones mainly composed of bile pigments. Side effects from the infusion of monooctanoin were only minor and easily controlled by reducing the infusion rate of the solution. Laboratory tests, including hepatic and pancreatic enzymes, remained stable. In one subject endoscopic and histological evidence of mild duodenitis was found when pre- and posttreatment features were compared. On the basis of these data, we recommend monooctanoin to treat biliary-retained cholesterol stones.

Decreased blood levels of ethanol and acetaldehyde by S-adenosyl-L-methionine in humans.

In view of the protective effects of SAM on alcohol-induced fatty liver degeneration, an investigation has been carried out to see if this compound accelerates the clearance of ethanol and acetaldehyde in humans. Both parameters were significantly lower after SAM, indicating the capability of exogenous SAM to favour the inactivation of ethanol without increasing blood levels of acetaldehyde.

Bile secretion and liver microsomal mixed function oxidase system in mice with griseofulvin-induced hepatic protoporphyria.

Administration of 2.5% griseofulvin in the diet to male CD1 mice produced protoporphyria and cholestasis. Protoporphyria became evident as early as after 10 days of treatment, whereas cholestasis, expressed in terms of total bile flow reduction, developed only after 45 days of griseofulvin. Bile flow impairment was due both to the length of treatment and to the severity of liver protoporphyria. Griseofulvin administration was also associated with a significant modification of the relative amounts of hepatic microsomal cytochromes P-450 and b5, a loss in concentration/mg of protein of cytochrome P-450 and a concomitant increase of b5. Despite these changes, the activity of aniline hydroxylase expressed per mg of microsomal protein, assessed in vitro, was not modified.

Hydroxychloroquine versus phlebotomy in the treatment of porphyria cutanea tarda.

Hydroxychloroquine and phlebotomy were compared in the treatment of porphyria cutanea tarda (PCT). Thirty patients received hydroxychloroquine (200 mg twice weekly) for 1 year and thirty-one underwent twice-monthly phlebotomies of 400 ml whole blood each, also for 1 year. Clinical signs of disease improved equally in both groups. At the end of the year, urinary porphyrin excretion had significantly improved in twenty-two out of thirty hydroxychloroquine-treated subjects, but in only eight out of the thirty-one patients who received phlebotomy. Liver histology showed significant regression of steatosis and siderosis in both groups compared with the pretrial biopsy, but the activity of liver disease, as judged by the extent of necrosis, inflammation and fibrosis, worsened in twelve hydroxychloroquine and in seven phlebotomy-treated patients. It is concluded that hydroxychloroquine is more effective than phlebotomy in decreasing porphyrin production. However, further work is needed to assess whether long-term hydroxychloroquine treatment favours the progression of the chronic liver disease associated with PCT.

Effects of phlebotomy on urinary porphyrin pattern and liver histology in patients with porphyria cutanea tarda.

Urinary porphyrin profiles and liver histology have been investigated in a group of adult alcoholics with porphyria cutanea tarda (PCT) before and after one year phlebotomy. Both parameters were evaluated during the same period in a group of patients who did not undergo specific therapy for PCT. All patients were advised to abstain from alcohol. At the end of the one year observation period there was a significant fall of urinary total porphyrins and in the uro/coproporphyrin ratio in treated patients compared to basal values whereas no changes were found in controls. Liver biopsy findings revealed a significant reduction of hepatic fatty degeneration and siderosis with no changes in inflammatory infiltrates and fibrosis in treated patients, so the progression of liver disease was similar to controls. These results show that clinical and biochemical remission of PCT can occur independently of the evolution of the concomitant liver disease.

3-Hydroxy-3-methylglutaric acid (HMGA) reduces dietary cholesterol induction of saturated bile in hamster.

3-Hydroxy-3-methylglutaric acid (HMGA) is a competitive inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCoAR) and strongly reduces cholesterol biosynthesis both in vitro and in vivo. Since the effects of this compound on biliary lipid composition are hitherto unknown, we have investigated whether it prevents dietary cholesterol induction of saturated bile in the hamster. Female Golden Syrian hamsters have been divided into four groups and treated for 10 weeks as follows: I) Standard diet, containing 0.8 mg cholesterol/g food; II) Standard diet plus HMGA (100 mg/kg b.w./day per os); III) Lithogenic diet containing 2.4 mg cholesterol/g food; IV) Lithogenic diet plus HMGA as above. The results indicate that HMGA is effective in reducing both bile cholesterol supersaturation and hypercholesterolemia. Inhibition of hepatic cholesterogenesis at the level of mevalonate synthesis by HMGCoAR and reduction of intestinal cholesterol absorption may be responsible for these effects.

Effect of a concomitant treatment with ethynylestradiol and ascorbic acid on bile secretion in female hamsters.

The administration of large doses (5 mg/kg b.wt./day) of ethynylestradiol to adult female hamsters did not induce cholestasis or modifications of bile lipid composition. These findings are in contrast with the data of other authors who in different experimental conditions described the sensitivity of hamsters to the estrogen-induced hepatobiliary toxicity. Ascorbic acid alone or added to ethynylestradiol did not impair bile secretion. However, it significantly increased the plasma levels of radioactivity tested 24 hours after the oral administration of a tracer dose of radiolabelled ethynylestradiol. These results confirm previous data showing in humans the capability of ascorbic acid to favour the rise of plasma concentrations of ethynylestradiol.

Effect of ethanol on biliary unconjugated bilirubin and its implication in pigment gallstone pathogenesis in humans.

Though some epidemiological investigations support the association between pigment gallstone formation and chronic alcoholism with cirrhosis, little attention has been paid to the influence of alcohol itself on biliary bilirubin secretion, so that the pathogenesis of pigment cholelithiasis in alcoholics is hitherto unknown. On different days we intravenously administered ethanol (0.7 g/kg body weight), diluted with 500 ml of saline, or saline alone to 6 non-obese patients with an indwelling T tube and reestablished enterohepatic bile circulation. At the time of the investigation bile cultures were negative for aerobic and anaerobic bacteria. Ethanol significantly increased biliary unconjugated bilirubin in respect to control values. The phenomenon reached a maximum 2 h after alcohol infusion when the value of unconjugated bilirubin averaged 2.37 +/- 0.30% of total bilirubin in contrast to 0.65 +/- 0.14% in control conditions (p less than 0.01), and subsided 6 h after the end of ethanol infusion. Since increased amounts of biliary unconjugated bilirubin predispose to pigment stone formation, it can be speculated that alcohol contributes to pigment cholelithiasis pathogenesis by enhancing the biliary concentrations of this form of pigment.

Acute ethanol administration increases biliary concentrations of total and unconjugated bilirubin in rabbits.

Epidemiological investigations have revealed that alcoholic cirrhosis is associated with a high frequency of pigment gallstones, but only scanty information is available on the effects of ethanol on biliary secretion of bilirubin. We have injected intravenously 1.0 and 1.5 g/kg body wt of ethanol into six cholecystectomized rabbits and a common bile duct fistula. Experiments were performed ten days after surgery and a stream-splitting apparatus was interposed in the circuit in order to withdraw continuously biliary samples without interruption of enterohepatic bile circulation. Analysis of hourly data showed that both ethanol doses significantly increase the biliary concentration of total bilirubin, without affecting bile flow and lipid composition. Alcohol also promoted the efflux of unconjugated bilirubin into bile. The maximum effect occurred within the first 5 hr following alcohol administration. Thereafter the bile returned to normal. Since excessive concentrations of biliary unconjugated bilirubin favor pigment gallstone development, it can be speculated that alcohol acts as a risk factor for pigment lithiasis by enhancing the biliary levels of this form of pigment.

Bile flow impairment of ventro-medial hypothalamus lesioned obese rats.

Cholesterol supersaturation of bile has been reported in human obesity. Since electrolytic lesions placed in the ventro-medial hypothalamus induce hyperphagia and obesity in the rat, bile flow and lipid composition have been studied two months after the induction of such stereotaxic lesions in a group of ten animals and in their sham-operated controls. Bile flow was significantly lower in obese rats than in controls. The bile flow reduction was attributed to a decrease of the bile acid independent fraction, since no variation in bile acid excretion rate and in bile to plasma ratio of [14C] erythritol was seen between the two groups. Whereas plasma cholesterol and triglycerides were higher in ventro-medial hypothalamus lesioned rats, biliary cholesterol and phospholipid excretion rates were similar in the two groups. These data indicate that stereotaxic lesions of ventro-medial hypothalamus in the rat significantly reduce bile flow, suggesting a correlation between active sodium transport at canalicular level and neuroendocrine hypothalamic function, but fail to induce qualitative alterations of bile lipid composition.

Ethynylestradiol-induced impairment of bile secretion in the rat: protective effects of S-adenosyl-L-methionine and its implication in estrogen metabolism.

Previous investigations demonstrated that S-adenosyl-L-methionine suppresses the cholestatic effect of ethynylestradiol and reduces the irreversible binding of the estrogen to rat liver microsomes, probably favoring ethynylestradiol transformation into its methyl-derivatives. Since it is not known whether SAMe also prevents the changes of bile lipid composition induced by EE and how methylation interferes with estrogen metabolism and biliary excretion, female rats have been treated with ethynylestradiol (5 mg/kg body wt, orally for 3 days) or with ethynylestradiol plus S-adenosyl-L-methionine (25 mg/kg body wt, i.m., t.i.d.). After bile duct cannulation, 5 muCi of [6,7-3H]ethynylestradiol were injected i.v. and an 8-h bile collection was started. Bile flow in ethynylestradiol + S-adenosyl-L-methionine treated rats was similar to controls, but significantly higher than in ethynylestradiol-treated animals (1.44 +/- 0.11; 1.66 +/- 0.30; 0.97 +/- 0.17 microliter/min/g liver, respectively as mean +/- SE). Cholesterol molar percentage was significnatly higher in ethynylestradiol (3.46 +/- 0.25) than in ethynylestradiol + S-adenosyl-L-methionine treated (2.16 +/- 0.41) or control rats (1.90 +/- 0.30). Total radioactivity excretion was similar in all groups, ranging from 74% to 76% of the administered dose, but radiogaschromatography showed a significant increase of methylated ethynylestradiol metabolites in S-adenosyl-L-methionine treated rats. These data indicate that in ethynylestradiol treated rats S-adenosyl-L-methionine can: (a) reverse both cholestasis and related abnormalities of biliary lipids; (b) modify the pattern but not the total amount of biliary ethynylestradiol excretion.

Effects of methyltestosterone on bile secretion in female and male rats.

No adverse effect on bile secretion was noted in methyltestosterone-treated female rats. In male rats, methyltestosterone administration exerted a cholestatic effect, which is not only due to the decrease of bile salt excretion rate, but also to a possible interference of the steroid with bile salt micelle size.