Bile Pigment Excretion Research Articles

A Controlled, Randomized, Double-Blind Trial of Prophylaxis Against Jaundice Among Breastfed Newborns

Neonatal jaundice is a greater problem for infants fed breast milk, compared with formula. This study tested the hypotheses that feeding breastfed newborns beta-glucuronidase inhibitors during the first week after birth would increase fecal bilirubin excretion and would reduce jaundice without affecting breastfeeding deleteriously. Sixty-four breastfed newborns were randomized to 4 groups, ie, control or receiving 6 doses per day (5 mL per dose) of L-aspartic acid, enzymatically hydrolyzed casein (EHC), or whey/casein (W/C) for the first week. L-aspartic acid and EHC inhibit beta-glucuronidase. Transcutaneous bilirubin levels (primary outcome) were measured daily (Jaundice Meter [Minolta/Air Shields, Hatboro, PA] and Bilicheck [Respironics, Pittsburgh, PA]). All stools were collected, and fecal bile pigments, including bilirubin diglucuronide, bilirubin monoglucuronides, and bilirubin, were analyzed with high-performance liquid chromatography. Follow-up assessments included day 7 body weight, day 6/7 prebreastfeeding/postbreastfeeding weights, maternal ratings, and ages at formula introduction and breastfeeding cessation. The groups were comparable at entry. Overall, the L-aspartic acid, EHC, and W/C groups had significantly lower transcutaneous bilirubin levels than did the control group (75.8%, 69.6%, and 69.2%, respectively, of the control mean, 8.53 mg/dL, at the bilirubin peak on day 4). The L-aspartic acid, EHC, and W/C groups had significantly lower transcutaneous bilirubin levels on days 3 to 7. Fecal bile pigment excretion was greatest in the L-aspartic acid group, significantly greater than control values. There were no significant differences in dosages, follow-up measurements, and maternal ratings. Use of minimal aliquots of L-aspartic acid and EHC for beta-glucuronidase inhibition results in increased fecal bilirubin excretion and less jaundice, without disruption of the breastfeeding experience. Decreased jaundice in the W/C group, which lacked a beta-glucuronidase inhibitor, suggests a different mechanism.

Intestinal excretion of unconjugated bilirubin in man and rats with inherited unconjugated hyperbilirubinemia.

Patients with Crigler-Najjar syndrome and Gunn rats cannot form bilirubin glucuronides owing to a lack of bilirubin UDP-glucuronosyltransferase activity. Because increased serum and tissue bilirubin levels remain constant, an alternative excretory route has to substitute for this deficiency. Gunn rats excrete in bile only 2-13% of the bilirubins eliminated in Wistar rats. In contrast, the biliary excretion rate of urobilinogen in Gunn and Wistar rats is comparable. The sum of bilirubins and urobilinogen excreted in the bile of Gunn rats amounts to 10-30% of pigments excreted in Wistar rats. Despite this low biliary excretion, the intestinal content and fecal excretion of bile pigments in Gunn and Wistar rats were similar. These data support an extrabiliary entrance of unconjugated bilirubin into the intestine. Additional proof for this was found in that the intestinal lumen of Gunn rats still contains a high amount of bilirubins and urobilinogen after 3 d of external biliary drainage. A similar procedure in Wistar rats resulted in the complete disappearance of bile pigments from the intestine. The direct transmural transport of bilirubin from blood to all parts of the intestinal lumen was demonstrated by injecting 14C-bilirubin i.v. into Gunn rats with isolated parts of small and large intestine. In Crigler-Najjar and Gilbert's syndrome patients, the biliary excretion of bile pigments has previously been shown to be strongly reduced. Their stools, however, contained approximately the same amount of bile pigments as in normal subjects. Although only traces of unconjugated bilirubin were detected in the stool of normal persons (4 +/- 3% of total bile pigments), higher amounts were found in patients with Crigler-Najjar disease (20 +/- 12&). These results suggest a direct intestinal permeation of unconjugated bilirubin in severe unconjugated hyperbilirubinemia both in man and rats.

Hepatocyte cotransport of taurocholate and bilirubin glucuronides: role of microtubules.

Modulation of bile pigment excretion by bile salts has been attributed to modification of canalicular membrane transport or a physical interaction in bile. Based on the observation that a microtubule-dependent pathway is involved in the hepatocellular transport of bile salts, we investigated the possibility that bilirubin glucuronides are associated with bile salts during intracellular transport. Experiments were conducted in intact rats (basal) or after overnight biliary diversion and intravenous reinfusion of taurocholate (depleted/reinfused). All rats were pretreated with intravenous low-dose colchicine or its inactive isomer lumicolchicine. Biliary excretion of radiolabeled bilirubin glucuronides derived from tracer [14C]bilirubin-[3H]bilirubin monoglucuronide (co-injected iv) was unchanged in basal rats but was consistently delayed in depleted/reinfused rats. This was accompanied by a significant shift toward bilirubin diglucuronide formation from both substrates. In basal Gunn rats, with deficient bilirubin glucuronidation, biliary excretion of intravenous [14C]bilirubin monoglucuronide-[3H] bilirubin diglucuronide was unaffected by colchicine but was retarded in depleted/reinfused Gunn rats. Colchicine had no effect on the rate of bilirubin glucuronidation in vitro in rat liver microsomes. We conclude that a portion of the bilirubin glucuronides generated endogenously in hepatocytes or taken up directly from plasma may be cotransported with bile salts to the bile canalicular membrane via a microtubule-dependent (vesicular?) mechanism.

Regulation of bilirubin glucuronide synthesis in primate (Macaca fascicularis) liver

Abstract Hepatic bilirubin uridine diphosphate glucuronyltransferase (UDP-glucuronyltransferase) catalyzes the formation of bilirubin monoglucuronides (BMG, C-8 and C-12 isomers) and bilirubin diglucuronide (BDG) from bilirubin and the cosubstrate, UDP-glucuronic acid. Distinctive patterns of bile pigment excretion occur in different species and in pathologic disorders (e.g., Gilbert's syndrome). In normal human and monkey ( Macaca fascicularis ) bile, the proportion of BDG exceeds that of BMG and the C-8C-12 BMG isomer ratio approaches unity. To investigate the mechanisms responsible for the patterns of BDG and BMG isomers in bile, we used a radiochemical assay to analyze the kinetics and regulation of bilirubin UDP-glucuronyltransferase in microsomes prepared from monkey liver. The synthesis of BMG from bilirubin was a higher capacity, lower affinity step ( V max = 295 pmol / mg protein · min, K m = 24 μM ) than BDG synthesis from endogenously formed BMG ( V max = 170 pmol / mg protein · min, K m = 14 μM ). This observation was confirmed when biosynthetically prepared BMG was used as substrate. The rate of formation of BDG relative to BMG was modulated by both bilirubin and UDP-glucuronic acid concentration, whereas the C-8C-12 BMG isomer ratio was influenced exclusively by UDP-glucuronic acid concentration. The data obtained with increasing UDP-glucuronic acid concentration did not obey conventional single-site kinetics, suggesting the presence of more than one binding site on the enzyme or a membrane transporter for this nucleotide sugar. These findings demonstrate that microsomal bilirubin UDP-glucuronyltransferase has a greater capacity for BMG than BDG synthesis, and thus support the concept that the decreased BDGBMG ratio in the bile of patients with reduced hepatic enzyme activity (i.e., Gilbert's syndrome and type II Crigler-Najjar disease) reflects the diminished capacity of the enzyme to synthesize BDG from BMG. Bilirubin and UDP-glucuronic acid concentrations both appear to be pivotal in regulating the synthesis of individual bilirubin glucuronides and hence the patterns of bilirubin conjugates excreted in bile.

Regulation of bilirubin glucuronide synthesis in primate (Macaca fascicularis) liver: Kinetic analysis of microsomal bilirubin uridine diphosphate glucuronyltransferase

Hepatic bilirubin uridine diphosphate glucuronyltransferase (UDP-glucuronyltransferase) catalyzes the formation of bilirubin monoglucuronides (BMG, C-8 and C-12 isomers) and bilirubin diglucuronide (BDG) from bilirubin and the cosubstrate, UDP-glucuronic acid. Distinctive patterns of bile pigment excretion occur in different species and in pathologic disorders (e.g., Gilbert's syndrome). In normal human and monkey (Macaca fascicularis) bile, the proportion of BDG exceeds that of BMG and the C-8/C-12 BMG isomer ratio approaches unity. To investigate the mechanisms responsible for the patterns of BDG and BMG isomers in bile, we used a radiochemical assay to analyze the kinetics and regulation of bilirubin UDP-glucuronyltransferase in microsomes prepared from monkey liver. The synthesis of BMG from bilirubin was a higher capacity, lower affinity step (Vmax = 295 pmol/mg protein · min, Km = 24 μM) than BDG synthesis from endogenously formed BMG (Vmax = 170 pmol/mg protein · min, Km = 14 μM). This observation was confirmed when biosynthetically prepared BMG was used as substrate. The rate of formation of BDG relative to BMG was modulated by both bilirubin and UDP-glucuronic acid concentration, whereas the C-8/C-12 BMG isomer ratio was influenced exclusively by UDP-glucuronic acid concentration. The data obtained with increasing UDP-glucuronic acid concentration did not obey conventional single-site kinetics, suggesting the presence of more than one binding site on the enzyme or a membrane transporter for this nucleotide sugar. These findings demonstrate that microsomal bilirubin UDP-glucuronyltransferase has a greater capacity for BMG than BDG synthesis, and thus support the concept that the decreased BDG/BMG ratio in the bile of patients with reduced hepatic enzyme activity (i.e., Gilbert's syndrome and type II Crigler-Najjar disease) reflects the diminished capacity of the enzyme to synthesize BDG from BMG. Bilirubin and UDP-glucuronic acid concentrations both appear to be pivotal in regulating the synthesis of individual bilirubin glucuronides and hence the patterns of bilirubin conjugates excreted in bile.

Influence of antibiotics on the faecal excretion of bile pigments in healthy subjects.

We have evaluated the effects of 10 antibiotics, given orally for 6 days to healthy subjects, on faecal excretion of urobilinogen. Intake of bacitracin, vancomycin, clindamycin, erythromycin, and ampicillin resulted in a pronounced suppression of the faecal excretion of urobilinogen (p less than 0.05). Intake of doxycycline, metronidazole, nalidixic acid, ofloxacin, and trimethoprim/sulphamethoxazole had no significant effect. The effects of three antibiotics-ampicillin, clindamycin, and metronidazole--on faecal excretion of conjugated bilirubin were similarly evaluated. Intake of clindamycin led to a marked increase of conjugated bilirubin (p less than 0.05) in the faeces, and the pattern of separated azopigment derivatives of the bilirubin conjugates became altered. Intake of ampicillin and metronidazole resulted in far less alterations in faecal conjugated bilirubin, although a significant change was observed in the subjects receiving metronidazole (p less than 0.05). The differences between the antibiotics with regard to altered intestinal bile pigment metabolism may be due to differences in antimicrobial spectra and/or intestinal concentrations of the drugs. Our findings indicate that orally taken antibiotics may cause a suppression of the microbial deconjugation of conjugated bilirubin and urobilinogen formation, respectively. This may reflect a pronounced disturbance of the intestinal microflora.

Effects of anesthetic agents on bile pigment excretion in the rat.

Anesthesia-induced alterations in bilirubin conjugation were studied. Rats were fitted with bile duct and jugular vein catheters while anesthetized with diethyl ether, ketamine or pentobarbital. As anesthesia abated, bile was collected for the next 5 hr and analyzed for flow rate, total bilirubin excretion and bilirubin glucuronide composition. The high-performance liquid chromatography method used allowed direct analysis of bile without derivatization or extraction. Ether anesthesia was associated with a reversible suppression of diglucuronide formation and total bilirubin excretion, with reciprocal monoglucuronide changes. Bile flow and pigment excretion were variable with ketamine. Pentobarbital provided the most uniform excretion data, although the ratio of C-8:C-12 monoglucuronide varied with all drugs. These data are consistent with recently reported drug-induced alterations in hepatic uridine diphosphoglucuronic acid concentration and support the hypothesis that alterations in this substrate concentration are capable of influencing rates of hepatic glucuronide formation.

Biliverdin accumulation in the caudal intestinal segment of juvenile adult lampreys, Petromyzon marinus L.

The possibility of bile pigment excretion by the caudal intestinal region in lampreys was investigated using spectrophotometry, routine electron microscopy, and an exogenous protein tracer. The green pigment present in the caudal intestines of immediately postmetamorphic and juvenile adult lampreys was found to be biliverdin. Cytoplasmic inclusions, which resembled biliary inclusion bodies and which were not formed as a result of exocytosis of materials at the apical surface, were found in the caudal intestine in absorptive, caveolated, and mucous cells concomitant with the appearance of the biliverdin. Evidence therefore indicates that these inclusions probably contain large quantities of the bile pigment biliverdin and other substances with which it may be complexed. It is suggested that the caudal segment of the adult lamprey intestine serves as a site for the elimination of bile pigment in the form of biliverdin. This method of elimination of bile pigment may be an essential function of the intestine owing to the absence of a bile duct in this animal.

Effect of biliverdin and bilirubin on the hepatic excretion of bile pigments in the rabbit

1. 1. A study has been carried out on the effect of i.v. infusion of biliverdin and bilirubin at four different dose strengths in sodium pentobarbital-anaesthetized rabbits. 2. 2. Both pigments show similar values for the maximum excretion rate in bile. 3. 3. At doses of 0.45 and 0.60 mg/kg/min, the infusion of biliverdin does not affect bile flow in the resting state, contrary to the negative effect on flow induced by bilirubin. 4. 4. This negative effect induced by bilirubin on bile flow is explained on the basis of its inhibitory action on the bile salt independent fraction (BSIF).

Hepatic bile pigment excretion and erythrocyte turnover in various species.

Endogenous bile pigment excretory rates have now been recorded in fifteen species, and the following review briefly compares the excretion per unit of body weight in animals of varying sizes, attempts to determine if any correlation exists between bile flow and endogenous pigment excretion and discusses bile pigment excretion in relation to mean erythrocyte survival time and metabolic rate. Graphs and one table illustrating pigment metabolism and erythrocyte replacement are presented. Differences between animals excreting primarily biliverdin as opposed to bilirubin are discussed, with special attention being given throughout the article to mammals and their individual differences. Accepted theory concludes that the rate of endogenous bile pigment excretion is primarily dependent upon hemoglobin catabolism. However, further studies will be necessary to clarify basic physiological mechanisms in order to substantiate any theory concerning the correlation of erythrocyte turnover with metabolic rates.

A new animal model with hyperbilirubinemia

An unconjugated hyperbilirubinemia has been observed in all species of normal indigo snakes. The plasma clearance of large organic anions such as sulfobromophthalein and unconjugated bilirubin was markedly delayed when compared to other snake species. Endogenous bile flow and biliverdin and bilirubin excretory rates and the excretion of bile pigments after a bilirubin load were measured in various snakes. The indigo snake represents a new animal model in which to study mechanisms important to hepatic anion uptake and biliary transport.

Hereditary cholestasis combined with peripheral pulmonary stenosis and other anomalies.

A syndrome consisting of chronic intrahepatic cholestasis with retention of bile acids but with normalization of bile pigment excretion ahd blood lipids, peripheral pulmonary stenosis, vertebral anomalies and a characteristic facies is described in six patients, including a father and his daughter. An autosomal dominant mode of inheritance is suggested.

Metabolism of biliverdin: Biliary excretion of bile pigments after intravenous injection of biliverdin isomers

14C-labelled biliverdins IX α, β, γ and δ have been prepared in vitro from haemoglobin obtained from duck erythrocytes incubated with 5-amino[4- 14C]-laevulinic acid. When injected intravenously into rats with biliary fistulae, about 60% of the label was recovered in the bile in 24 h after the α isomer was given, while approximately 10% was recovered with injection of the β isomer. The γ and δ isomers gave intermediate values. In each experiment, most of the recovered isotope was found in association with conjugated bile pigment. Thus, the metabolic pathway for bile pigment excretion in the rat handles the IX α isomer preferentially but is not specific for it.

A physiological approach to jaundice

An understanding of the production, transport, hepatic uptake, biotransformation and excretion of bile pigments is necessary for the rational investigation of a patient presenting with jaundice. These aspects are reviewed, and at each step of bilirubin metabolism the clinical abnormalities giving rise to jaundice are indicated.

Proceedings: The excretion of bile pigments in rats after intravenous injection of biliverdin isomers.

Proceedings: The excretion of bile pigments in rats after intravenous injection of biliverdin isomers.

THE BINDING OF BILIRUBIN TO AGAR

In animal experiments, variation was found between agar lots in their effect on fecal excretion of bile pigments and on lowering of serum bilirubin levels in the Gunn strain of rat. The binding of bilirubin to agar was studied in vitro to account for the variation. Suspensions of agar powder in bilirubin solutions (isotonic, 37°, pH 7.9) were employed to derive adsorption isotherms. From these, the association constants (Ka) for the binding reaction was about 2×105 for all products. The capacity for binding (n) varied between 25 and 56 moles of bilirubin/50,000 g. of agar. The variation in (n) was related to the calcium and sulfate content of the agar products. The calcium and sulfate content of the agar lots each varied from less than 1 to 11 moles/50,000 g. of agar. Calcium-depleted agar lost its bilirubin binding capacity and regained it upon readdition of calcium. Salicylate, sulfonamide, BSP and HABA dyes, and conjugated bilirubin were not bound by agar. Cellulose, phosphocellulose and DEAE-cellulose did not bind bilirubin as well as agar. Binding of bilirubin to agar decreased as pH, ionic strength, or urea concentration were independently increased. There was an increase in apparent Ka with a rise in temperature. Free energy change was about 2 kcal/mole. A model is proposed in which strands of carbohydrate are cross-linked with ionic calcium disulfate bridges to form relatively apolar interstices between chains and between bridges in which bilirubin is bound possibly by hydrophobic interaction or by hydrogen bonds or both.

Bilirubin and biliverdin excretion by the chicken.

ARTICLESBilirubin and biliverdin excretion by the chickenGL Lin, JA Himes, and CE CorneliusGL Lin, JA Himes, and CE CorneliusPublished Online:01 Apr 1974https://doi.org/10.1152/ajplegacy.1974.226.4.881MoreSectionsPDF (1 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByExpanding the eggshell colour gamut: uroerythrin and bilirubin from tinamou (Tinamidae) eggshells9 July 2020 | Scientific Reports, Vol. 10, No. 1Multiple origins of green coloration in frogs mediated by a novel biliverdin-binding serpin13 July 2020 | Proceedings of the National Academy of Sciences, Vol. 117, No. 31Chronic cadmium exposure in Japanese quails perturbs serum biochemical parameters and enzyme activity21 May 2019 | Drug and Chemical Toxicology, Vol. 43, No. 1Infection of Human Tracheal Epithelial Cells by H5 Avian Influenza Virus Is Regulated by the Acid Stability of Hemagglutinin and the pH of Target Cell Endosomes9 January 2020 | Viruses, Vol. 12, No. 1Systems metabolic engineering of Corynebacterium glutamicum for the bioproduction of biliverdin via protoporphyrin independent pathway29 March 2019 | Journal of Biological Engineering, Vol. 13, No. 1A multiplatform metabolomic approach to characterize fecal signatures of negative postnatal events in chicks: a pilot study9 April 2019 | Journal of Animal Science and Biotechnology, Vol. 10, No. 1Avian Liver: The Forgotten Organ15 February 2019 | Animals, Vol. 9, No. 2Tracking Modified Vaccinia Virus Ankara in the Chicken Embryo: In Vivo Tropism and Pathogenesis of Egg Infections24 August 2018 | Viruses, Vol. 10, No. 9Comparative transcriptome analysis provides clues to molecular mechanisms underlying blue-green eggshell color in the Jinding duck (Anas platyrhynchos)12 September 2017 | BMC Genomics, Vol. 18, No. 1Unique Infectious Strategy of H5N1 Avian Influenza Virus Is Governed by the Acid-Destabilized Property of HemagglutininViral Immunology, Vol. 30, No. 6Gastrointestinal Anatomy and PhysiologyTemporal transcriptome analysis of the chicken embryo yolk sac19 August 2014 | BMC Genomics, Vol. 15, No. 1Avian Clinical BiochemistryUnconjugated bilirubin induces apoptosis in colon cancer cells by triggering mitochondrial depolarization1 January 2004 | International Journal of Cancer, Vol. 112, No. 3Gastrointestinal Anatomy and PhysiologyAvian Clinical BiochemistryThe blue-green blood plasma of marine fishComparative Biochemistry and Physiology Part B: Comparative Biochemistry, Vol. 97, No. 1Biliverdin reductase activity in cattle, sheep, rabbits and ratsInternational Journal of Biochemistry, Vol. 21, No. 5Blood chemistry for the diagnosis of hepatobiliary disease in birds. 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Influence of phenobarbital on serum bilirubin and biliary excretion of bile pigments in gunn rats

Influence of phenobarbital on serum bilirubin and biliary excretion of bile pigments in gunn rats

Studies on bile secretion (III) the uptake, conjugation and excretion of bile pigments by the liver under the obstructive jaundice

Studies on bile secretion (III) the uptake, conjugation and excretion of bile pigments by the liver under the obstructive jaundice

Bile pigment excretion: a comparison of the biliary excretion of bilirubin and bilirubin derivatives.

Bile pigment excretion: a comparison of the biliary excretion of bilirubin and bilirubin derivatives.