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PubMedDate not supplied

Sn-protoporphyrin lowers serum bilirubin levels, decreases biliary bilirubin output, enhances biliary heme excretion and potently inhibits hepatic heme oxygenase activity in normal human subjects.

Sn-protoporphyrin, a potent competitive inhibitor of heme oxygenase, the rate-limiting enzyme in the degradation of heme to bile pigment, was administered to 10 normal volunteers: 8 males and 2 females. A significant decrease in the levels of serum (mean decrease; 38%) and biliary bilirubin (mean decrease: 47%) was demonstrated in all 10 subjects. The decrease in these parameters lasted for a minimum of 4 days after administration of the metalloporphyrin. Sn-protoporphyrin also facilitated the excretion of endogenous heme in bile during the 24- to 48-hr period following administration of the compound. The metalloporphyrin was rapidly cleared from plasma with a half-life of 3.4 hr. A small amount (3 to 6%) of Sn-protoporphyrin was excreted in both urine and bile. The activity of microsomal heme oxygenase was measured in five human liver samples freshly obtained at the time of cholecystectomy and varied from approximately 2 to 4 nmoles bilirubin formed per mg protein per hour. The addition of Sn-protoporphyrin to microsomal heme oxygenase isolated from these liver samples resulted in a dose-dependent decrease in the activity of this enzyme at concentrations of the metalloporphyrin in the range of 10(-7) to 10(-9) M. The findings reported here provide strong evidence that the suppression of biliary and serum bilirubin levels and the enhancement of heme excretion in the bile of the normal volunteers examined in this study were due to inhibition of physiological rates of heme oxidation activity by the synthetic heme analog, Sn-protoporphyrin.

Adult↗
PubMed1999

Experimental study on the bilirubin metabolism after major hepatectomy: alterations in the proportions of bile bilirubin subfractions.

Bilirubin metabolism after major hepatectomy was investigated experimentally using rats. After a 70% and an 80% hepatectomy, proportions of bilirubin diglucuronide (BDG) decreased, and reversely, those of bilirubin monoglucuronide (BMG) increased. These changes were even more remarkable after an 80% hepatectomy. Parallel to the decrease in the proportions of BDG, the concentrations of uridine diphosphoglucuronic acid (UDP-GA) in the remnant liver decreased, and there was a significant correlation between the changes in BDG and UDP-GA. Although UDP-glucuronyltransferase (UDP-GT) activity and energy charge of the remnant liver also decreased after surgery, these decreases were mild and returned to the control level earlier than BDG. And there was no significant correlation between the changes in BDG and those in UDP-GT activity and energy charge. In this study, the decrease of the proportions of BDG in the bile juice was long term after partial hepatectomy and the period of the decrease became longer according to the augmentation of the volume of the hepatectomized liver. We clarified that the process of the UDP-GA production was inhibited after hepatectomy and the decrease of the proportions of BDG was derived from a deficiency of substrate of the glucuronidation, UDP-GA.

Animals↗
PubMed1981

Bilirubin binding capacity and free bilirubin concentration: fluorescence quenching compared with peroxidase oxidation and sephadex column elution techniques.

Fluorescence quenching was compared with the techniques of peroxidase oxidation and Sephadex column elution for determining free bilirubin concentrations and the capacity of albumin to bind bilirubin and assessed as to its suitability for use as a routine method in a clinical laboratory. The poor reproducibility of the fluorescence quenching method made it unacceptable and peroxidase oxidation was found to be the most satisfactory technique. The Sephadex column elution technique did not measure free bilirubin concentration but gave a good estimate of binding capacity. This method is, however, limited by the sample size required for performing the determinations.

Bilirubin↗
PubMed1968

Determination of bilirubin glucuronide and assay of glucuronyltransferase with bilirubin as acceptor.

1. Conjugated bilirubin is conveniently determined by coupling with the diazonium salt of ethyl anthranilate. 2. This method has been used in the development of assays for UDP-glucuronyltransferase (EC 2.4.1.17), with bilirubin as substrate, in rat liver homogenates, microsomal preparations and partly purified fractions. 3. Chromatographic analysis suggests that bilirubin monoglucuronide is the product of the enzyme systems studied.

Animals↗
PubMed1984

Pulmonary excretion of carbon monoxide in the human infant as an index of bilirubin production. IIc. Evidence for the possible association of cord blood erythropoietin levels and postnatal bilirubin production in infants of mothers with abnormalities of gestational glucose metabolism.

A total of 20 infants who had levels of erythropoietin (Ep), the major hormone regulating erythropoiesis, measured in their cord blood also had determinations of the pulmonary excretion rate of CO (VECO) performed, as an index of total bilirubin production. They were either infants of normal mothers or those of mothers with diabetes, gestational diabetes, and missed abnormalities of gestational glucose metabolism. The mean VECO (13.0 +/- 3.5 mu 1/kg/hr) and the mean Ep (20.0 +/- 9.7 SD mU/ml) of the infants with normal mothers (n = 9) were not different from the means previously established by our laboratories (13.9 +/- 3.5 SD mu 1/kg/hr, n = 20; and 23.7 +/- 12.8 SD mU/ml, n = 30, respectively); they were significantly lower than those of the infants of the abnormal mothers in this study. The 5 infants who had a cord blood Ep level greater than 50 mU/ml had a higher mean VECO, 27.8 +/- 7.1 mu 1/kg/hr, compared with 17.2 +/- 4.9 SD mu 1/kg/hr, of the six infants with cord blood Ep levels that were within 2 SD of the previously established normal mean cord blood Ep level (p less than .025). These data suggest that increased cord blood Ep levels and postnatal bilirubin production in infants whose mothers had abnormalities of gestational glucose metabolism are associated phenomena. Since polycythemia did not occur in these infants, ineffective erythropoiesis or mild, compensated hemolysis remains a likely cause of the increased total bilirubin production. In some cases, perinatal hypoxic stress may have affected the Ep response.(ABSTRACT TRUNCATED AT 250 WORDS)

Bilirubin↗
PubMed1988

Efficacy of using total bilirubin values as a guide for screening direct bilirubin requests.

Two divergent laboratory approaches to the determination of direct bilirubin were assessed. The first approach, assaying direct bilirubin (DB) on all total bilirubin (TB) requests regardless of TB values, resulted in a true negative rate (normal TB level less than 1.0 mg/dL [less than 17 mumol/L], and DB level, less than 0.4 mg/dL [less than 6.8 mumol/L]) of 98.7% (984/997 specimens) and a low false negative (normal TB level, abnormal DB level) rate of 1.3% (13 of 997). The second approach, assaying DB on physician request and only if the TB level was greater than or equal to 1.0 mg/dL (1.7 mumol/L), resulted in a true negative rate (normal TB level, no liver disease) of 87.7% (150%) of 171 patients) and a relatively low false negative rate (normal TB level, liver disease) of 12.3% (21 of 171) patients. Medical chart review revealed that, with either approach, none of the patients with false negative results with hepatobiliary disease would have been missed clinically, even if the DB assay had not been done. It is possible to screen DB requests using a TB of 1.0 mg/dL (17 mumol/L) as a means to determine whether DB should be assayed.

Biliary Tract Diseases↗
PubMed2001

Influence of bilirubin uridine diphosphate-glucuronosyltransferase 1A promoter polymorphisms on serum bilirubin levels and cholelithiasis in children with sickle cell anemia.

PURPOSE: Genetic mutations in the uridine diphosphate (UDP)-glucuronosyltransferase 1A (UGT1A) enzyme promoter have been associated with unconjugated hyperbilirubinemia and Gilbert syndrome. The effects of UGT1A promoter polymorphisms on serum bilirubin levels and symptomatic gallstone formation were studied in a cohort of children with sickle cell anemia (SCA). METHODS: The UGT1A promoter genotype was deterrmined for 115 consecutive children with SCA. Steady-state laboratory parameters and previous cholecystectomy for symptomatic gallstones were recorded retrospectively, then analyzed according to UGT1A genotype. RESULTS: Children with SCA had a lower frequency of the normal (TA)6 UGT1A promoter allele (0.413) than the abnormal (TA)7 allele (0.461). A previously described shorter (TA)5 allele (frequency 0.074) and longer (TA)8 allele (frequency 0.052) were also observed. Children with the 7/7 UGT1A genotype had a significantly higher mean bilirubin level (5.8 +/- 3.1 mg/dL) than those with the 6/6 (2.4 +/- 0.8 mg/dL) or 6/7 genotype (3.0 +/- 1.1 mg/dL; P < 0.001 by analysis of variance). Patients with the 7/7 genotype were more likely to have previous cholecystectomy (87.5%) than those with the 6/6 (35.7%) or the 6/7 genotype (36.1%; P = 0.002 by chi2). CONCLUSIONS: Genetic variation in the UGT1A promoter significantly influences serum bilirubin levels and the development of symptomatic cholelithiasis in children with SCA. The UGT1A promoter polymorphisms represent an important nonglobin genetic modifier of clinical disease expression in SCA.

Adolescent↗
PubMed1993

The increase of yellow skin colour beyond that of serum bilirubin: a proposed indicator of risk for bilirubin encephalopathy in the newborn.

Forty-seven newborn infants with 1 min Apgar score < 7 were studied. On the third postnatal day the following measurements were made: yellow skin colour, serum bilirubin concentration, reserve albumin concentration and plasma pH. Given the bilirubin concentration and the regression curve between the yellow skin colour and the bilirubin concentration, delta-TcB was calculated as the difference between measured yellow skin colour and the expected yellow skin colour. There was a negative correlation between delta-TcB and Apgar score (P = 0.003), pH (P = 0.026) and reserve albumin concentration (P = 0.045). Fourteen of the included newborns had central nervous system symptoms in the days just following birth. A tendency towards higher delta-TcB was noted in this group (P = 0.08). The results suggest that further study of delta-TcB determination as a tool in the assessment of the icteric newborn infant is justified.

Apgar Score↗
PubMed1994

Cells, bilirubin and light: formation of bilirubin photoproducts and cellular damage at defined wavelengths.

Cultured cells from one human and one murine cell line were treated with bilirubin and irradiated with visible light of different wavelengths, either from phototherapy lamps or from a Xenon/Mercury lamp equipped with a monochromator. Bilirubin bound to human serum albumin was also irradiated with light. After irradiation, the bilirubin and its photoisomers were extracted and analysed with High Pressure Liquid Chromatography. The formation of single strand breaks in the DNA of treated cells was studied using a fluorescence marker. Cytotoxicity in the mouse skin cell line was measured by loss of the ability to form visible colonies in vitro. Green light exposure favours the production of lumirubin, while blue light causes more DNA damage and cytotoxicity. Green light may be more efficient and safer than shorter wavelength exposure when treating jaundiced newborns with phototherapy.

Animals↗
PubMed2004

Transcutaneous bilirubin measurements and serum total bilirubin levels in indigenous African infants.

OBJECTIVE: The objective of this study was to determine whether transcutaneous bilirubin (TcB) measurements correlate with serum total bilirubin (STB) levels in indigenous, darkly pigmented African newborns with varying degrees of skin pigmentation, some of which had developed kernicterus. METHODS: Jaundiced infants who were < or =2 weeks of age and admitted to Baptist Medical Center-Eku (Eku; n = 29) and Jos University Teaching Hospital (Jos; n = 98) in Nigeria were studied. TcB measurements using the BiliChek were made simultaneously with blood sampling for STB measurements by spectrophotometry before phototherapy. RESULTS: Using linear regression analysis, we found that measurements of TcB correlated well with those of STB with r values of.90 and.88 for Eku and Jos, respectively. Mean bias and imprecision of TcB measurements as compared with STB measurements for the total population was 0.5 +/- 7.6 mg/dL using the method of Bland and Altman. At STB > or 12 mg/dL, correlation (r =.84) and bias and imprecision (-1.2 +/- 8.6 mg/dL) of measurements were only slightly poorer. Furthermore, when infants were grouped by degree of skin pigmentation, correlations of TcB and STB measurements remained strong. CONCLUSIONS: From these results, we can conclude that TcB measurements are a useful and reliable index for estimating STB levels in pigmented neonates, including those with hyperbilirubinemia and kernicterus. In the absence of reliable STB measurements, the relatively simple and noninvasive TcB measurements can be an important adjunct in directing phototherapy and exchange transfusions, thereby preventing bilirubin-induced morbidity and mortality in low-technology clinical environments.

Bias↗
PubMed1975

[Serum bilirubin determination in newborn infants. A new micromethod for the determination of serum of plasma bilirubin in newborn infants].

A micro-method for determination of bilirubin concentration (c) of plasma or serum from newborns was developed. Photometer-readings of the extinction (E) of diluted serum (S) at the bilirubin absorption-band were done against diluted serum as blank, made free from bilirubin by the oxydative power of Chloramin T. Reagents: 1/15 M Na2(HPO4)/K(H2PO4) buffer pH 7,4 (B) and 2 g/dl Chloramin T in B(C). Blank: 0,6 ml C + 0,02 ml S; sample: 0,6 ml B + 0,02 ml S. Reaction time: 4--8 min; filter Hg 436 nm; 1 cm light-path. c (mg/dl) = E X 43,5. Accuracy was determined by a calibration curve (x: mg/dl c of diazo-method; ): E of micro-method); b = 1,008. Reproducibility in series at 14,2 mg/dl resp. 47,9 mg/dl was 1,5% resp. 1,6%. Linearity between E and c is good up to 48 mg/dl c. Specifity was only reduced by serum hemoglobin (h). Sodium heparinate (50 E./ml S) and carotin (952 microgramm/1) had no effect upon E; plasma anticoagulated with heparinate therefore can be used instead of serum. Concentration of h is without importance for clinical purposes; 100 mg/dl h causes about 0,2 mg/dl wrong depression of c.

Bilirubin↗
PubMed2002

[Using purified conjugated bilirubin as a calibrator for bilirubin measurement].

OBJECTIVES: To improve the accuracy and precision of the determination of bilirubin, especially direct bilirubin (DB), and the standardization of that as well. METHODS: Purified conjugated bilirubin (Bc) and ditaurobilirubin(DTB) and their diazo products were subjected to absorption spectrum analysis. The diazo reaction characters of their calibration solutions were compared by the method of Doumas J-G(TB & DB). RESULTS: Bc, DTB and their azopigments were found to have the similar absorption spectra with the same lambda max. Their TB standard curves almost superposed together all over. Although the slopes of their DB standard curves were not markedly different ((YBc = 0.00366X + 0.00933, rBc2 = 0.9977, P < 0.01; YDTB = 0.00391X + 0.00023, rDTB2 = 0.9987, P < 0.01; Pb1-b2 > 0.05, n1 = n2 = 5), the DB value measured for Bc differed from that for DTB(n = 5, P < 0.05). In addition, the calibrators made from Bc based different matrices, such as HSA, BSA and human serum, were significantly different in DB/Bc, but no difference was seen among the concentrations. Furthermore, the DB values determined for DTB or Bc increased linearly with the corresponding concentrations, respectively, with no difference between the slopes (YBc = 0.8300XBc + 1.9463, rBc2 = 0.9977, P < 0.01; YDTB = 0.8853XDTB-0.0251, rDTB2 = 0.9986, P < 0.01; n1 = n2 = 5, Pb1-b2 > 0.05). CONCLUSIONS: The results demonstrate that the diazo reaction characters of Bc are identified with those of DTB. However, under the condition of DB, Bc reacts differently from DTB. This study also indicates that as a calibrator of DB based human serum, Bc has the similar constant effect of HCl as serum samples do, so it is a more reliable calibrator to eliminate the matrix effects.

Bilirubin↗
PubMed1990

Light has a greater effect on direct bilirubin measured by the bilirubin oxidase method than by the diazo method.

We compared the effect of light on direct-reacting bilirubin (DBIL) measurement by the bilirubin oxidase (EC 1.3.3.5; BOX) method and by the Jendrassik-Gróf diazo method. DBIL concentrations determined by the BOX method in the sera of hyperbilirubinemic infants treated with phototherapy yielded falsely higher values than those by the direct diazo method. A similar tendency was noted when DBIL concentrations in infants' sera irradiated with light in vitro were determined by both methods, although by HPLC none of these sera had detectable DBIL (i.e., conjugated plus delta bilirubin). In general, DBIL concentrations after photoirradiation remained unchanged when measured by the diazo method, but significantly increased when the BOX method was used. Indeed, photoirradiation gave rise to material that acted like a photobilirubin product, which was oxidized at pH 3.7 and therefore was measured as DBIL. Such false increases in DBIL values generated by the BOX method may have clinical diagnostic implications in monitoring jaundiced neonates and in differentiating between physiological jaundice and incipient pathological jaundice.

Azo Compounds↗
PubMed1988

Understanding bilirubin conformation and binding. Circular dichroism of human serum albumin complexes with bilirubin and its esters.

Human serum albumin binds tightly and noncovalently to a wide variety of hydrophobic bilirubins, including (4Z,15Z)-bilirubin-IX alpha, its dimethyl ester and mono methyl esters, its mono 2-butyl esters and amides, the dimethyl ester of (4Z,15Z)-mesobilirubin-IV alpha, and even (4Z,15Z)-etiobilirubin-IV gamma. The heteroassociation complexes formed from these highly water-insoluble pigments and the protein can be prepared in pH 7.4 aqueous by using a small quantity of dimethyl sulfoxide as amphiphilic carrier. In those solutions the protein acts as a water-soluble chiral complexation agent to induce an asymmetric transformation of the bound pigment. This is recognized by positive chirality, bisignate induced circular dichroism (CD) Cotton effects that fall in the region of the bichromophoric pigment's long wavelength UV-visible absorption band and are characteristic of intramolecular exciton coupling of the bilirubin component pyrromethenone chromophores. The same-signed CD spectra shared by all the pigments of this work indicate selection at the protein binding site for a positive chirality conformer and suggest a common binding site. The CD intensities, which are greatest ([delta epsilon[ congruent to 50) for pigments with one or two free carboxyl groups, are consistent with a binding model where one salt linkage plays a major role in the enantioselectivity of the right-handed folded conformation stabilized by inter- and intramolecular hydrogen bonds.

Bilirubin↗
PubMed1979

[Study of bilirubin- albumin binding. I.-In vitro study by partition of bilirubin between an aqueous solution and an immiscible organic solvent (author's transl)].

The partition of bilirubin in vitro between water and isobutyl-methylketone/n-heptane mictures is investigated at different pH, in the absence and the presence of albumin. Results are used to compute the concentrations of unbound ionized and unionized bilirubin and the bilirubin-albumin characteristics (association constant K and nomber n of primary binding sites per mole of albumin). The effect of various factors upon these characteristics is discussed.

Animals↗
PubMed1977

Toxicity to bilirubin in neonates: infant development during first year in relation to maximum neonatal serum bilirubin concentration.

Neurologic and developmental performance during the first year of life was correlated with maximum neonatal serum bilirubin levels for 27,000 infants in the Collaborative Perinatal Project. The infants were grouped by race and by five birth weight/gestational age categories to control for the effect of these factors on hyperbilirubinemia and developmental outcome. Low mean eight-month motor scores and delayed one-year motor development were associated with serum bilirubin levels in the range of 10 to 14 mg/dl and above. This relationship was strongest for low-birth-weight/short-gestational-period infants. A persistent association of developmental outcome with hyperbilirubinemia was found over and above the variation of maturity within the birth weight/gestational age categories.

Bilirubin↗
PubMed1998

Bilirubin as an anti precipitant against copper mediated denaturation of bovine serum albumin: formation of copper-bilirubin complex.

BR forms a beautiful 1:1 greenish complex with Cu2+, having a characteristic absorption maximum at 343 nm. To our knowledge, this is the first metal complex of bilirubin in aqueous solution so far to be reported. It has also been shown that bilirubin (BR) at a concentration more than protein binding capacity has a definite role as an anti precipitant of bovine serum albumin by excess copper. At a concentration of 1 x 10(-3) moll-1 of the Cu-BR complex, the colour becomes greenish black. The rate of formation of this Cu-BR complex when BR extracts copper from copper-albumin complex as obtained in our experiment is 34.981 mol-1 s-1. The Cu-BR complex is stable at a pH ranging from 3.5 to 13.2, and also can scavenge radicals like CCl3OO., eaq- and OH.. Addition of excess copper sulfate to the solution of this complex causes a greenish black precipitate which can be re dissolved in HCl but insoluble in 99% ethanol. The complex does not give positive test in Gmelin reaction for bile pigments. The fluorescence spectrum of the complex in solution exhibits a peak at 450 nm when excited at 343 nm. The precipitated complex is insoluble in a number of solvents like ether, xylene, benzene, acetone, chloroform-methanol mixture and DMSO. These results show that BR may protect mammals from copper poisoning.

Animals↗