Crigler-Najjar Syndrome Type 1: Pathophysiology, Natural History, and Therapeutic Frontier.

Background And Aims: We describe the pathophysiology, treatment, and outcome of Crigler-Najjar type 1 syndrome (CN1) in 28 UGT1A1 c.222C>A homozygotes followed for 520 aggregate patient-years.

Approach And Results: Unbound ("free") bilirubin (B ) was measured in patient sera to characterize the binding of unconjugated bilirubin (B ) to albumin (A) and validate their molar concentration ratio (B /A) as an index of neurological risk.

Bilirubin binding in jaundiced newborns: from bench to bedside?

Background: Bilirubin-induced neurologic dysfunction (BIND) is a spectrum of preventable neurological sequelae in jaundiced newborns. Current total plasma bilirubin (B) concentration thresholds for phototherapy and/or exchange transfusion poorly predict BIND.

Methods: The unbound (free) bilirubin (B) measured at these B thresholds provides additional information about the risk for BIND.

In Vitro Study on the Effect of Maraviroc or Dolutegravir on Bilirubin to Albumin Binding.

We performed an in vitro evaluation of the effect of maraviroc or dolutegravir on bilirubin to albumin binding. At typical treatment and low albumin concentrations, maraviroc had no impact, while dolutegravir affected bilirubin to albumin binding to an equivalent extent as sulfisoxazole. However in vivo, neither is likely to significantly impact bilirubin to albumin binding because of their low concentrations relative to albumin.

The Bilirubin Binding Panel: A Henderson-Hasselbalch Approach to Neonatal Hyperbilirubinemia.

Poor plasma bilirubin binding increases the risk of bilirubin neurotoxicity in newborns with hyperbilirubinemia. New laboratory tests may soon make it possible to obtain a complete bilirubin binding panel when evaluating these babies. The 3 measured components of the panel are the plasma total bilirubin concentration (B), which is currently used to guide clinical care; the bilirubin binding capacity (BBC); and the concentration of non-albumin bound or free bilirubin (B).

Optimizing exchange transfusion for severe unconjugated hyperbilirubinemia: studies in the Gunn rat.

Background: Severe unconjugated hyperbilirubinemia carries the risk of neurotoxicity. Phototherapy (PT) and exchange transfusion (ET) are cornerstones in the treatment of unconjugated hyperbilirubinemia. Studies to improve ET efficacy have been hampered by the low application of ET in humans and by the lack of an in vivo model.

Beyond plasma bilirubin: the effects of phototherapy and albumin on brain bilirubin levels in Gunn rats.

Background & Aims: Severe unconjugated hyperbilirubinemia, as occurs in Crigler-Najjar disease and neonatal jaundice, carries the risk of neurotoxicity. This neurotoxicity is related to the increased passage of free bilirubin (UCB(free)), the fraction of bilirubin that is not bound to plasma proteins, into the brain. We hypothesized that albumin treatment would lower the UCB(free) fraction, and thus decrease bilirubin accumulation in the brain.

Bilirubin binding contributes to the increase in total bilirubin concentration in newborns with jaundice.

Objective: This study tests the hypothesis that the hourly rate of increase in plasma bilirubin concentration (DeltaBT) would increase significantly with increasing binding avidity.

Methods: The plasma total bilirubin concentration (B(T)), unbound bilirubin concentration, and albumin concentration values for healthy newborns with jaundice (or=35 weeks of gestation, and >or=2.5 kg at birth) were obtained from medical records.

Predicting bilirubin neurotoxicity in jaundiced newborns.

Purpose Of Review: The management of jaundice in the newborn infant is an area of clinical practice sorely lacking an evidence-based foundation, and neonatal bilirubin neurotoxicity (kernicterus) continues to occur worldwide.

Recent Findings: Studies suggest that measuring serum or plasma bilirubin binding, in particular the nonalbumin-bound or unbound bilirubin concentration (Bf), would improve jaundice management as it better predicts bilirubin neurotoxicity than the conventionally used total bilirubin concentration (BT). However, many misconceptions persist regarding the relationships between BT, Bf, the magnitude and distribution of the neonatal bilirubin load, and the risk of bilirubin neurotoxicity.

Influence of clinical status on the association between plasma total and unbound bilirubin and death or adverse neurodevelopmental outcomes in extremely low birth weight infants.

Objectives: To assess the influence of clinical status on the association between total plasma bilirubin and unbound bilirubin on death or adverse neurodevelopmental outcomes at 18-22 months corrected age in extremely low birth weight infants.

Method: Total plasma bilirubin and unbound bilirubin were measured in 1101 extremely low birth weight infants at 5 +/- 1 days of age. Clinical criteria were used to classify infants as clinically stable or unstable.

Intervention guidelines for neonatal hyperbilirubinemia: an evidence based quagmire.

The American Academy of Pediatrics has proposed guidelines for treating term/near term infants with hyperbilirubinemia based primarily on maintaining the total serum bilirubin concentration (TSB) below a "critical" level of 25 mg/dL (426 micromol/L). We estimated the sensitivity and specificity of this critical TSB using patient data from published reports. A TSB >25 mg/dL is recorded in about 92% of term/near term infants with kernicterus.

Unbound (free) bilirubin: improving the paradigm for evaluating neonatal jaundice.

Background: The serum or plasma total bilirubin concentration (B(T)) has long been the standard clinical laboratory test for evaluating neonatal jaundice, despite studies showing that B(T) correlates poorly with acute bilirubin encephalopathy (ABE) and its sequelae including death, classical kernicterus, or bilirubin-induced neurological dysfunction (BIND). The poor correlation between B(T) and ABE is commonly attributed to the confounding effects of comorbidities such as hemolytic diseases, prematurity, asphyxia, or infection. Mounting evidence suggests, however, that B(T) inherently performs poorly because it is the plasma non-protein-bound (unbound or free) bilirubin concentration (B(f)), rather than B(T), that is more closely associated with central nervous system bilirubin concentrations and therefore ABE and its sequelae.

Unbound bilirubin predicts abnormal automated auditory brainstem response in a diverse newborn population.

Objective: The objective of this study was to determine if plasma unbound or 'free' bilirubin concentration (B(f)) measured during the first 30 days of life is associated with subsequent abnormal hearing screening testing by automated auditory brainstem response (AABR) in a diverse population of newborns.

Study Design: An observational study of newborns receiving AABR, plasma total bilirubin concentration (TBC) and B(f) measurements and without underlying conditions known to affect hearing was conducted. Logistic regression was used to determine associations between abnormal AABR and B(f) or TBC.

Effect of storage and freezing on unbound bilirubin measurement.

Background: Unbound bilirubin (Bf) is a better predictor of bilirubin toxicity than total serum bilirubin (TSB) in the postnatal period. The Food and Drug Administration approved peroxidase test for Bf analysis is not currently available in clinical laboratories. The samples are often stored at varying temperature and for varying duration before Bf analysis.

Unbound bilirubin concentration is associated with abnormal automated auditory brainstem response for jaundiced newborns.

Objective: This study was conducted to determine whether incidental jaundice affects automated auditory brainstem response results.

Methods: We reviewed the medical charts of jaundiced newborns of > or = 34 weeks of gestation who underwent automated auditory brainstem response testing within 4 hours of plasma total bilirubin concentration and unbound bilirubin concentration measurements. We tested the hypothesis that the likelihood of abnormal automated auditory brainstem response results would increase as total bilirubin and unbound bilirubin concentrations increased.

Usefulness of the bilirubin/albumin ratio for predicting bilirubin-induced neurotoxicity in premature infants.

Unconjugated hyperbilirubinaemia occurs in almost all premature infants and is potentially neurotoxic. Treatment is based on total serum bilirubin (TSB), but treatment thresholds are not evidence based. Free bilirubin (Bf)-that is, not bound to albumin, seems a better parameter for bilirubin neurotoxicity, but measurements of Bf are not available in clinical practice.

Effects of sample dilution, peroxidase concentration, and chloride ion on the measurement of unbound bilirubin in premature newborns.

Objectives: To assess the effects of sample dilution, peroxidase concentration, and chloride ion (Cl(-)) on plasma unbound bilirubin (B(f)) measurements made using a commercial peroxidase methodology (UB Analyzer) in a study population of ill, premature newborns.

Design And Methods: B(f) was measured with a UB Analyzer in 74 samples at the standard 42-fold sample dilution and compared with B(f) measured at a 2-fold sample dilution using a FloPro Analyzer. B(f) was measured at two peroxidase concentrations to determine whether the peroxidase steady state B(f) (B(fss)) measurements were significantly less than the equilibrium B(f) (B(feq)), in which case it was necessary to calculate B(feq) from the two B(fss) measurements.

Factors affecting the binding of bilirubin to serum albumins: validation and application of the peroxidase method.

The unbound "free" bilirubin concentration (Bf), not the total bilirubin concentration, is the critical determinant of cellular uptake and toxicity of bilirubin. We compared Bf measured by a modified peroxidase method with published data obtained with ultrafiltration and examined conditions that affect the affinity (KF) of human (HSA) and bovine (BSA) serum albumin for bilirubin. The peroxidase and ultrafiltration methods yielded similar KF values that decreased with increasing HSA concentration and the presence of 50 mM chloride.

Toward understanding kernicterus: a challenge to improve the management of jaundiced newborns.

Purpose: We sought to evaluate the sensitivity and specificity of total serum bilirubin concentration (TSB) and free (unbound) bilirubin concentration (Bf) as predictors of risk for bilirubin toxicity and kernicterus and to examine consistency between these findings and proposed mechanisms of bilirubin transport and brain uptake.

Methods: A review of literature was undertaken to define basic principles of bilirubin transport and brain uptake leading to neurotoxicity. We then reviewed experimental and clinical evidence that relate TSB or Bf to risk for bilirubin toxicity and kernicterus.

Evaluation of a model for brain bilirubin uptake in jaundiced newborns.

A model for brain bilirubin uptake (BBU) predicts that BBU in jaundiced newborns typically depends on the plasma total bilirubin concentration (TBC) and the bilirubin-albumin dissociation rate constant (k1) rather than the unbound bilirubin (Bf). The model's validity was tested by 1) evaluating its requirement that k3>>>k2, where k3 and k2 are the rate constants for BBU and Bf-albumin association, respectively, and 2) determining whether the calculated BBU is View Article and Find Full Text PDF

Measurement of unbound bilirubin by the peroxidase test using Zone Fluidics.

Background: Measuring plasma unbound bilirubin concentration by the peroxidase test is useful in the management of jaundiced newborns. However, the commercially available peroxidase technology is manual, and the unbound bilirubin may be seriously underestimated at the 42-fold sample dilution and single peroxidase concentration used. We investigated improving the test by adapting it to Zone Fluidics, which is a system for automating reactant handling that requires small sample volumes and dilution.

Bilirubin-albumin binding and neonatal jaundice.

Bilirubin-albumin binding weakens the correlation between the plasma total bilirubin concentration (TBC) and bilirubin encephalopathy (kernicterus), which has led to considerable interest over the years in measuring binding as part of the evaluation of jaundiced newborns. Despite development of several bilirubin-albumin binding tests, technical, ethical, and logistical factors have prevented the prospective studies needed to validate their routine clinical use. Consequently, it has been necessary to adopt aggressive exchange transfusion criteria based on the TBC that require the unnecessary treatment of large numbers of babies to prevent the rare case of kernicterus.