Unconjugated Bilirubin Levels Research Articles

A high frequency of Gilbert syndrome (UGT1A1*28/*28) and associated hyperbilirubinemia but not cholelithiasis in adolescent and adult north Indian patients with transfusion-dependent β-thalassemia.

Hyperbilirubinemia and pigment gallstones are frequent complications in transfusion-dependent β-thalassemia (TDβT) patients. Bilirubin production and clearance are determined by genetic as well as environmental variables like ineffective erythropoiesis, hemolysis, infection-induced hepatic injury, and drug- or iron-related toxicities. We studied the frequency of the Gilbert syndrome (GS), a common hereditary cause of hyperbilirubinemia in 102 TDβT patients aged 13-43years (median 26years). Total and unconjugated hyperbilirubinemia were frequent (81.4% and 84.3% patients respectively). Twenty (19.6%) patients showed total bilirubin > 3.0mg/dL; 53 (51.9%) had an elevation of either alanine or aspartate aminotransferase, or alkaline phosphatase liver enzymes. Nineteen (18.6% of the 92 tested) were positive for hepatitis B or C, or HIV. The mean total and unconjugated bilirubin levels and AST, ALT, and ALP levels in patients positive for hepatitis B or C were not significantly different from negative cases. Eighteen patients (17.7%) had GS: homozygous (TA)7/7 UGT1A1 promoter motif (the *28/*28 genotype), 48 (47.1%) were heterozygous (TA)6/7. Total + unconjugated bilirubin rose significantly with the (TA)7 allele dose. Fourteen (13.7%) patients had gallstones. There was no significant difference in total/unconjugated bilirubin in patients with/without gallstones and no significant differences in frequencies of gallstones within the three UGT1A1 genotypes. This largest study in Indian TDβT patients suggests that GS should be excluded in TDβT cases where jaundice remains unexplained after treatable causes like infections, chelator toxicity, or transfusion-related hemolysis are excluded. GS was not associated with gallstones, possibly due to a lower incidence of cholelithiasis overall, a younger age cohort, or other environmental factors.

Oxidative state in equine neonates: Anti- and pro-oxidants.

In newborns, exposure to the extrauterine environment with high oxygen tension and sudden pulmonary adaptation leads to an increase in reactive oxygen species (ROS). ROS have several physiological roles, which are essential for neonatal development, however, when unbalanced, these highly unstable molecules can cause cellular destabilisation, compromising vital processes. To characterise the oxidative status in healthy equine neonates, evaluating an indicator of lipid peroxidation and both enzymatic and nonenzymatic antioxidant systems, during the first week of life. Experimental cohort. Twenty-four foals were evaluated, with blood collections performed at 5minutes, 12, 72 and 168hours after birth. The degree of lipid peroxidation was quantified using Thiobarbituric Acid Reactive Substances (TBARS). Superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzymatic activities, and total, conjugated and unconjugated serum bilirubin levels were also analysed. Comparisons were performed using ANOVA followed by a Tukey's test. Additionally, dependent variables were also evaluated with Pearson's correlation tests. Higher GPx activity was observed at 12 and 72hours when compared to 5minutes. An increase in TBARS levels was found at 5minutes after birth, followed by a decrease at 72hours and stabilisation through subsequent moments until 168hours after birth. No differences were observed in SOD activity when comparing the four time points. Bilirubin concentrations were lower at 5minutes after birth and total and unconjugated bilirubin increased at 12hours and decreased between 72 and 168hours after birth. Lipid peroxidation at birth was high, suggesting an increase in ROS levels relating to physiological events in neonatal adaptation. Antioxidant systems, involving unconjugated bilirubin and GPx, were activated and these biomolecules act concomitantly to reduce ROS levels, thus maintaining oxidative homeostasis. Overall, our results suggest a pro-oxidant balance during the first 168hours after birth in equine neonates.

Genotype-phenotype correlation in children with hereditary spherocytosis.

Hereditary spherocytosis (HS) is a common inherited haemolytic anaemia attributed to disturbances in five different red cell membrane proteins. We performed a retrospective study of 166 children with HS and describe the clinical phenotype according to the genotype. In 160/166 (97%) children with HS a disease-causing mutation was identified. Pathogenic variants in ANK1, SPTB, SLC4A1 and SPTA1 were found in 49%, 33%, 13% and 5% of patients. Children with SLC4A1-HS had the mildest phenotype, showing the highest haemoglobin (P < 0·001), lowest reticulocyte counts (P < 0·001) and lowest unconjugated bilirubin levels (P = 0·006), and none required splenectomy in childhood (P < 0·001). Conversely, children with autosomal recessive SPTA1-HS had the most severe clinical phenotype, with almost all patients undergoing splenectomy in early childhood. Patients with ANK1 and SPTB variants showed a similar clinical phenotype. Within each gene, variant type or location did not predict disease severity or likelihood of splenectomy. Among patients with a genetic diagnosis, 47 (29%) underwent splenectomy (23 partial; 24 total) while 57 (36%) underwent cholecystectomy. Total splenectomy led to greater improvements in haemoglobin (P = 0·02). Select use of genetic testing (especially in patients without a family history) may help predict clinical phenotype in childhood and guide family counselling.

Dubin-Johnson Syndrome in Pregnancy: Case Report and Literature Review

Dubin-Johnson Syndrome in Pregnancy: Case Report and Literature Review

Disease burden of Crigler-Najjar syndrome: Systematic review and future perspectives.

Crigler-Najjar syndrome (CNS) results from biallelic mutations of UGT1A1 causing partial or total loss of uridine 5'-diphosphate glucuronyltransferase activity leading to unconjugated hyperbilirubinemia and its attendant risk for irreversible neurological injury (kernicterus). CNS is exceedingly rare and has been only partially characterized through relatively small studies, each comprising between two and 57 patients. A systematic literature review was conducted to consolidate data on the patient, caregiver, and societal burden of CNS. Twenty-eight articles on clinical aspects of CNS were identified, but no published data on its humanistic or economic burden were found. In patients with complete UGT1A1 deficiency (type 1 CNS [CNS-I]), unconjugated bilirubin levels increase 3-6mg/dL/day during the newborn period and reach neurologically dangerous levels between 5 and 14days of age. Phototherapy is the mainstay of treatment but poses significant challenges to patients and their families. Despite consistent phototherapy, patients with CNS-I have worsening hyperbilirubinemia with advancing age. Liver transplantation is the only definitive therapy for CNS-I and is increasingly associated with excellent long-term survival but also incurs high costs, medical and surgical morbidities, and risks of immunosuppression. Crigler-Najjar syndrome is associated with a substantial burden, even with existing standards of care. The development of novel disease-modifying therapies has the potential to reduce disease burden and improve the lives of CNS patients and their families.

Crigler najjar syndrome: A systematic outline

Crigler-Najjar Syndrome (CNS) is a rare genetic condition characterized by non-hemolytic unconjugated hyperbilirubinemia. It is caused by mutations in the UGT1A1 gene which codes for the enzyme uridine diphosphate glucoronosyl transferase-1, required for the conjugation and further elimination of bilirubin from the body. Affected individuals are usually asymptomatic apart from the jaundice and investigations reveal isolated indirect hyperbilirubinemia. It can be conveniently diagnosed by evaluating the response to phenobarbital in terms of decrease in bilirubin levels. Genetic testing of the UGT1A1 gene for mutations is the investigative clincher. The hallmark outcome of Crigler-Najjar syndrome is a determined yellowing of the skin, mucous membranes and the sclera of eye. There are two patterns of this disorder: Crigler-Najjar syndrome type I, characterized by a nearly complete lack of enzyme activity and severe symptoms; and Crigler-Najjar syndrome type II, characterized by inadequate enzyme activity and milder symptoms. Both forms are inherited as autosomal recessive characters and are caused by faults or mutations of the UGT1A1 gene. Treatment is engaged toward dropping the level of unconjugated bilirubin in the blood. Early treatment is vibrant in Crigler-Najjar syndrome type I to prevent the development of encephalopathy during the first few months of life. Because Crigler-Najjar syndrome type II is milder and responds to phenobarbital, treatment is different. The purpose of the current review article is to emphasize on causes, types, clinical symptoms, autosomal pattern, complications, management and future direction for treatment of Crigler-Najjar syndrome. Keywords: Hyperbilirubinemia, Phenobarbitone, UGT1A1 gene, Bilirubin, Jaundice, Mutations

2223 Zieve’s Syndrome: Case Report, Review of the Literature and Electronic Patient Database

INTRODUCTION: Zieve’s syndrome (ZS) is rare condition of alcoholic hepatitis, hyperlipidemia and hemolytic anemia1. Clinicians need to be aware of ZS for several reasons: First, hemolysis increases unconjugated bilirubin levels, leading to false elevation of the Maddrey’s discriminant function (MDF), which may lead to the unnecessary prescription of steroids.2 Second, the anemia that occurs from hemolysis may be misinterpreted as a gastrointestinal bleed, leading to extra endoscopic procedures. Finally, since patients can present with fever and abdominal pain, diagnosis may be construed for other causes such as sepsis or acute cholecystitis.2-5 CASE DESCRIPTION/METHODS: A 39-year-old female with long-standing alcohol abuse had three successive admissions resulting in refractory ZS. We performed a review of the literature to assess other case reports of patients with ZS. A total of 38 patients were included, 8 in women and 30 in men. Patients’ age ranged from 28 to 65 years old (mean 45.6 years). Average total bilirubin was 9.12. Average hemoglobin was 9.15. Average total cholesterol was 486. Average LDH was 243. Most patients did not receive prednisolone and fully recovered with strict alcohol cessation. A retrospective analysis using the IBM Explorys database of over 63 million de-identified, unique patients in the United States revealed that of 17,990 patients with alcoholic hepatitis, only 80 had ZS. The odds of a patient with alcoholic hepatitis also having hemolytic anemia is 7. ZS is rare, only seen in 0.4% of patient’s with alcoholic hepatitis. DISCUSSION: Our patient was 39, younger than the average age of 45.6 years. She was also female, a gender seen in a minority of previously reported cases. She had a total bilirubin 3 times above average, and a hemoglobin 33% lower than average. Despite 12 units of packed red blood cells, prednisolone, and pursuit of inpatient/outpatient alcohol rehabilitation, she ultimately succumbed to her liver injury 6 weeks after initially presenting to the hospital. This is the first reported case of death from ZS. Clinicians should maintain a high index of suspicion for ZS in patients with alcoholic hepatitis and transfusion refractory anemia. Evaluating for hemolysis and hyperlipidemia are recommended. Alcohol abstinence is essential. There is no role for steroids. Early diagnosis is essential. Our goal is that increased case reporting will raise clinician awareness.

Neonatal hyperbilirubinaemia necessitating exchange transfusion due to maternal sickle cell crisis

Abstract Background Pregnancy in women with sickle cell disease (SCD) is associated with a number of fetal complications such as intra-uterine death, intra-uterine growth restriction (IUGR), preterm birth, low birth weight and an increased perinatal mortality and morbidity. Hyperbilirubinaemia necessitating exchange transfusion in an infant of a mother with SCD, to the best of our knowledge, has not been previously described. Case presentation An infant was delivered at 33 weeks and 5 days of gestation due to a maternal sickle cell crisis. The infant had an unconjugated bilirubin level of 153 μmol/L on admission to the neonatal intensive care unit at 30 min of age. Phototherapy was immediately commenced, intravenous immunoglobulin administered and then a double-volume exchange transfusion was performed. There was, however, no evidence of haemolysis in the infant and the infant’s haemoglobin level remained stable following the exchange. No further exchange transfusions were required. The mother had a high unconjugated bilirubin level (151 μmol/L) prior to delivery. Conclusion High neonatal unconjugated bilirubin levels necessitating exchange transfusion can occur due to haemolysis in the maternal circulation, in this case due to SCD.

Effect of Neonatal Hyperbilirubinemia on Ventricular Functions

Background: Unconjugated hyperbilirubinemia(UCH) is a common systemic disease in the neonate. Many researches on etiology, risk factors and treatment have been made and guidelines have been drawn up. Because it is a systemic disease, it affects the body, organelle, cells, organs and systems. Researchers have also done a lot of research to clarify which levels, which mechanisms, and which organs the unkonjuge bilirubin affects. We aimed to investigate the effect of high unconjugated bilirubin levels on left and right ventricular systolic and diastolic functions in newborns by echocardiography and doppler. Methods: This single-center, prospective, case-control study was conducted on 19 full-term newborn infants with severe UHB requiring phototherapy and 20 healthy infants as controls. Ventrikuler fonction assessment was performed by using M-mode ekokardiyografi and doppler. Results: There was no significant difference in right and left ventriculer fonctions between two groups. Conclusions: In our study, unkonjugated hyperbilirubinemia did not affect left and right ventricular function. However, we believe that new studies are needed with higher bilirubin values and wider case groups.

Prebiotics for the prevention of hyperbilirubinaemia in neonates.

Hyperbilirubinaemia occurs in approximately two-thirds of all newborns during the first days of life and is frequently treated with phototherapy. Although generally seen as safe, there is rising concern regarding phototherapy and its potentially damaging effects on DNA and increased side effects particularly for preterm infants. Other methods, such as enteral feeding supplementation with prebiotics, may have an effective use in the management of hyperbilirubinaemia in neonates. To determine whether administration of prebiotics reduces the incidence of hyperbilirubinaemia among term and preterm infants compared with enteral supplementation of milk with distilled water/placebo or no supplementation. We used the standard search strategy of Cochrane Neonatal to search the Cochrane Central Register of Controlled Trials (CENTRAL 2018, Issue 5), MEDLINE via PubMed (1966 to 14 June 2018), Embase (1980 to 14 June 2018), and CINAHL (1982 to 14 June 2018). We also searched clinical trials databases, conference proceedings, and the reference lists of retrieved articles for randomised controlled trials (RCTs) and quasi-randomised trials. We considered all RCTs that studied neonates comparing enteral feeding supplementation with prebiotics versus distilled water/placebo or no supplementation. Two reviewers screened papers and extracted data from selected papers. We used a fixed-effect method in combining the effects of studies that were sufficiently similar. We then used the GRADE approach to assess the quality of the evidence. Three small studies evaluating 154 infants were included in this review. One study reported a significant reduction in the risk of hyperbilirubinaemia and rate of treatment with phototherapy associated with enteral supplementation with prebiotics (risk ratio (RR) 0.75, 95% confidence interval (95% CI) 0.58 to 0.97; one study, 50 infants; low-quality evidence). Meta-analyses of two studies showed no significant difference in maximum plasma unconjugated bilirubin levels in infants with prebiotic supplementation (mean difference (MD) 0.14 mg/dL, 95% CI -0.91 to 1.20, I² = 81%, P = 0.79; two studies, 78 infants; low-quality evidence). There was no evidence of a significant difference in duration of phototherapy between the prebiotic and control groups, which was only reported by one study (MD 0.10 days, 95% CI -2.00 to 2.20; one study, 50 infants; low-quality evidence). The meta-analyses of two studies demonstrated a significant reduction in the length of hospital stay (MD -10.57 days, 95% CI -17.81 to -3.33; 2 studies, 78 infants; I² = 0%, P = 0.004; low-quality evidence). Meta-analysis of the three studies showed a significant increase in stool frequency in the prebiotic groups (MD 1.18, 95% CI 0.90 to 1.46, I² = 90%; 3 studies, 154 infants; high-quality evidence). No significant difference in mortality during hospital stay after enteral supplementation with prebiotics was reported (typical RR 0.94, 95% CI 0.14 to 6.19; I² = 6%, P = 0.95; 2 studies; 78 infants; low-quality evidence). There were no reports of the need for exchange transfusion and incidence of acute bilirubin encephalopathy, chronic bilirubin encephalopathy, and major neurodevelopmental disability in the included studies. None of the included studies reported any side effects. Current studies are unable to provide reliable evidence about the effectiveness of prebiotics on hyperbilirubinaemia. Additional large, well-designed RCTs should be undertaken in neonates that compare effects of enteral supplementation with prebiotics on neonatal hyperbilirubinaemia with supplementation of milk with any other placebo (particularly distilled water) or no supplementation.

A novel UGT1A1 gene mutation causing severe unconjugated hyperbilirubinemia: a case report

BackgroundCrigler-Najjar syndrome (CNs) presents as unconjugated hyperbilirubinemia, as a result of UGT1A1 deficiency, and can be categorized in a severe (type I) and mild (type II) phenotype. CNs type II patients usually benefit from phenobarbital treatment that induces residual UGT1A1 activity.Case presentationHere we present a CNs type II patient that is not responsive to phenobarbital treatment, which can be explained by two heterozygous mutations in the UGT1A1 gene. A 3 nucleotide insertion in the HNF-1α binding site in the proximal promoter previously reported in a Crigler-Najjar patient on one allele and a novel two nucleotide deletion in exon 1, resulting in a frameshift and a premature stop codon.ConclusionIn newly diagnosed CNs patients with unconjugated bilirubin levels consistent with CNs type II but that are unresponsive to phenobarbital treatment, disruption of the HNF-1α binding site in the proximal promoter should be considered as a probable cause. Upon confirming a mutation in the HNF-1α site, phenobarbital treatment should be stopped or at least be reconsidered because of its sedative effects and its teratogenic properties.

Spectrum of UGT1A1 variants in Pakistani children affected with inherited unconjugated hyperbilirubinemias

Inherited unconjugated hyperbilirubinemias are a group of disorders characterized by increased levels of serum unconjugated bilirubin and arise because of the imbalance between its production and elimination from the body. It includes Crigler-Najjar syndrome and Gilbert syndrome. Crigler-Najjar syndrome type 1 represents the extreme severe end of the spectrum with complete absence of hepatic bilirubin uridine diphosphoglucuronate glucuronosyltransferase (UGT1A1). Crigler-Najjar syndrome type 2 patients have intermediate levels of bilirubin owing to incomplete deficiency of UGT1A1, and Gilbert syndrome lies at the extreme mild end of the spectrum with only slightly raised bilirubin level. Here, we present spectrum of UGT1A1 genetic variants in 25 Pakistani children from 23 unrelated families affected with persistent unconjugated hyperbilirubinemias. The promoter region, coding exons and splice junctions of the UGT1A1 were PCR amplified and subjected to Sanger sequencing. Eleven sequence variants were identified underlying disease phenotype including a novel c.582delC variant. Overall, c.622_625dupCAGC was the most frequent variant followed by c.1021C>T found in Crigler-Najjar syndrome type 1 patients. The evaluation of promoter polymorphism A(TA)nTAA in the affected children and their families further supported the body of evidence that the A(TA)7TAA allele could enhance the effect of other structural variants in Crigler-Najjar syndrome patients. To our knowledge, this is the first comprehensive study on molecular genetics of persistent unconjugated hyperbilirubinemias from Pakistan. This study expands the spectrum of UGT1A1 variants and should help in improved clinical diagnosis, genetic counseling and prenatal diagnosis of the affected families.

Interference in the anti-Xa heparin activity assay due to hemolysis and icterus during pediatric extracorporeal life support.

Chromogenic anti-Xa assays for unfractionated heparin monitoring (heparin activity) are susceptible to interference from hemolysis and icterus. The purpose of this study was to better understand the effect of hemolysis and icterus on anti-Xa heparin activity and to predict the magnitude of the error. Increasing levels of hemoglobin and unconjugated bilirubin were added to pooled normal plasma or buffer containing known levels of heparin. Increased plasma hemoglobin or bilirubin produced falsely increased residual factor Xa activity as measured by the absorbance change (OD/min) in the Stago heparin activity assay. This increased absorbance change slope resulted in falsely lower estimates of heparin activity. The falsely lower heparin activity measurement occurred even when heparin was not present, indicating it was not due to heparin neutralization. In a sample containing 0.62±0.06 U/mL heparin and 228 mg/dL hemoglobin, the measured heparin activity was 0.41±0.03 U/mL, underestimating heparin activity by 0.21±0.07 U/mL. Interference occurred if plasma hemoglobin was above 70 mg/dL or bilirubin was above 16 mg/dL, which happened in 16%-26% of samples from pediatric patients on extracorporeal life support (ECLS). In conclusion, hemolysis and icterus were common in ECLS patients, leading to underestimates of unfractionated heparin activity and potentially higher doses of heparin than intended. The magnitude of the heparin activity measurement error could be predicted based on plasma hemoglobin and bilirubin levels until these levels exceeded the technical limits of the assay, ~230 mg/dL hemoglobin and 55 mg/dL bilirubin.

Unconjugated bilirubin and schizophrenia: a systematic review.

Schizophrenia is a complex syndrome of unknown etiology and difficult to manage. Unconjugated bilirubin has been researched as a potential biological marker of this syndrome. The objective of this review article was to gather the studies published to date on the relationship between this molecule and schizophrenia. Broad inclusion criteria have been used (PRISMA) to include as many relevant studies as possible. Fourteen studies were selected: 3 analyzed the effects of unconjugated hyperbilirubinemia in animal models; 6 demonstrated an increased incidence of schizophrenia in patients with increased unconjugated bilirubin; 2 reported an increased incidence of the disease in patients with decreased unconjugated bilirubin; and 3 linked an increased incidence of schizophrenia with an increased excretion of the oxidative product of bilirubin, the so-called biopyrrins. Because of apparently contradictory reported results, the hypothesis that the relationship between schizophrenia and unconjugated bilirubin was not linear and that there was an inflammatory dysfunction explaining this was considered. The 2 most accepted models for the pathophysiology of schizophrenia are described, and the possible role of the molecule in each is clarified. The bilirubin buffer system and its role in antioxidant defense was explored. The average levels of unconjugated bilirubin in patients with schizophrenia, schizoaffective disorder, and bipolar disorder were also compared, having been hypothesized that these diseases could be different points of a same pathological spectrum. Finally, it was concluded that unconjugated bilirubin is a promising molecule that could be used as a possible biological marker for schizophrenia, and the necessity of subsequent efforts for its research was considered.

Modulation of Cav2.3 channels by unconjugated bilirubin (UCB) – Candidate mechanism for UCB-induced neuromodulation and neurotoxicity

Elevated levels of unbound unconjugated bilirubin (UCB) can lead to bilirubin encephalopathy and kernicterus. In spite of a large number of studies demonstrating UCB-induced changes in central neurotransmission, it is still unclear whether these effects involve alterations in the function of specific ion channels. To assess how different UCB concentrations and UCB:albumin (U/A) molar ratios affect neuronal R-type voltage-gated Ca2+ channels, we evaluated their effects on whole-cell currents through recombinant Cav2.3 + β3 channel complexes and ex-vivo electroretinograms (ERGs) from wildtype and Cav2.3-deficient mice. Our findings show that modestly elevated levels of unbound UCB (U/A = 0.5) produce subtle but significant changes in the voltage-dependence of activation and prepulse inactivation, resulting in a stimulation of currents activated by weak depolarization and inhibition at potentials on the plateau of the activation curve. Saturation of the albumin binding capacity (U/A = 1) produced additional suppression that became significant when albumin was omitted completely and might involve a complete loss of channel function.Acutely administered UCB (U/A = 0.5) has recently been shown to affect transsynaptic signaling in the isolated vertebrate retina. The present report reveals that sustained exposure of the murine retina to UCB significantly suppresses also late responses of the inner retina (b-wave) from wildtype compared to Cav2.3-deficient mice. In addition, recovery during washout was significantly more complete and faster in retinae lacking Cav2.3 channels.Together, these findings show that UCB affects cloned and native Cav2.3 channels at clinically relevant U/A molar ratios and indicate that supersaturation of albumin is not required for modulation but associated with a loss of channel functional that could contribute to chronic neuronal dysfunction.

The effect of intestinal flora on the neural development of severe hyperbilirubinemia neonates.

To investigate the effect of intestinal flora on the neural development of severe hyperbilirubinemia neonates. The clinical data of 108 severe hyperbilirubinemia neonates admitted to the Dezhou People's Hospital from January 2015 through January 2018 were analyzed, and all newborns had a serum total bilirubin level > 342 μmol/L. Based on whether they suffered from neural development abnormalities, the neonatal patients were divided into the neural abnormality group (n=52) and the non-neural abnormality group (n=56). The unconjugated bilirubin levels in serum and cerebrospinal fluid (CSF) and the composition of intestinal flora were compared. Among 108 neonates, there were 55 cases with developmental abnormalities, in which 52 (48.13%) cases had neural developmental abnormalities, mainly epileptic patients. The serum and CSF unconjugated bilirubin levels of the neonatal patients in the neural abnormality group were (466.25±97.64) μmol/L and (9.64±2.98) μmol/L, respectively, which were higher than those in neonatal patients of the non-neural abnormality group [(357.89±72.53) μmol/L and (6.73±3.11) μmol/L], with statistically significant differences (p<0.05). The abundance of intestinal flora genus in the neonates in the neural abnormality group was lower than that in the non-neural abnormality group, and the comparisons of Fusobacterium, Catabacter, Succinivibrio, Clostridium and Bacteroides between the two groups showed statistically significant differences (p<0.05). The intestinal micro-ecological environment of newborns was vulnerable and easily affected by many factors such as methods of delivery, feeding ways and eating habits of their mothers. This study investigated the effects of intestinal flora on the neural development of neonates with severe hyperbilirubinemia. The results showed that, due to decreased intestinal flora diversity, the serum and cerebrospinal fluid bilirubin levels were elevated, and the abnormal rate of neural development was increased. Severe hyperbilirubinemia neonates with neural abnormalities have decreased diversity of intestinal flora genus and relatively high serum and CSF bilirubin levels, probably because the decrease in the diversity of intestinal flora genus leads to the change of the blood-CSF barrier permeability, leading to raised levels of bilirubin in serum and CSF, thus affecting the neural development of neonatal patients.

Hyptis verticillata attenuates dyslipidaemia, oxidative stress and hepato-renal damage in streptozotocin-induced diabetic rats

AimsChronic hyperglycaemia in diabetes mellitus (DM) increases the production of free radicals which results in oxidative stress and related disorders such as cardiovascular diseases, compromised hepatic and renal functions. Hyptis verticillata reportedly demonstrated glucose lowering activity in previous studies. The present study therefore evaluated the effect of H. verticillata on hyperglycaemia-induced dyslipidaemia, hepatorenal distortions, oxidative stress, as well as calculated indices of cardiovascular function. MethodsWistar rats employed for this study consisted of normoglycaemic and diabetic rats in nine experimental groups. The normoglycaemic and diabetic rats were either treated with metformin (500 mg/kg b.w.), quercetin (10 mg/kg b.w.), or ethanol extract of H. verticillata leaf (250 mg/kg b.w. and 500 mg/kg b.w.) administered orally for 28 days. Key findingsResults revealed that H. verticillata significantly lowered blood glucose level, attenuated dyslipidaemia, decreased atherogenic coefficient, atherogenic and coronary risk indices, and increased cardioprotective index in diabetic rats. Also, H. verticillata significantly decreased serum urea, creatinine, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and unconjugated bilirubin levels, relative to untreated diabetic rats. Further, H. verticillata increased serum superoxide dismutase, catalase and glutathione peroxidase activities and glutathione level, and decreased malondialdehyde level in diabetic rats in a manner similar to metformin and quercetin. Histopathological investigation of the liver and kidney revealed restored hepatocytes and amelioration of congested interstitial blood vessel of the Bowman's space of the kidneys upon intervention with H. verticillata. SignificanceH. verticillata in addition to its anti-hyperglycaemic activity ameliorates oxidative stress, dyslipidaemia, atherogenicity and hepatorenal lesions in DM.

Hepatoprotective Effect of Inonotus obliquus Melanins: In Vitro and In Vivo Studies

The purpose of this study is to identify hepatoprotective properties of melanins from aqueous extracts of Inonotus obliquus distinguished by microwave modes used in extraction in both in vitro and in vivo studies. In vitro tests were used in studies of the effect of Chaga mushroom melanins on the vitality of cells of a normal human hepatocyte line Chang Liver, as well as their hepatoprotective effect and influence on the cell cycle. The hepatoprotective effect was studied in the context of the influence of the toxicant d-galactosamine, at a concentration of 150 mM. The results show that the melanin of the aqueous extract of Chaga, obtained in the process of microwave-assisted extraction at 180 W, at concentrations of 10−5 and 10−3 g/l, displays a hepatoprotective effect, as it increases the vitality of cells under the toxic influence of d-galactosamine by 2–2.5 times. In vivo tests were used in studies of the hepatoprotective properties of the melanin of the aqueous extract of Chaga obtained in the process of microwave-assisted extraction at 180 W on white male Sprague Dawley rats. The melanin was administered to rats for 14 days at a dose of 100 mg/kg. Toxic damage was inflicted on the liver using carbon tetrachloride on days 5 to 12 of administering the melanin; the liver was studied and the blood biochemical parameters were determined on day 15. It was shown that melanin produces a hepatoprotective effect which is expressed in the minimization of liver injury signs such as steatosis, necrosis, fat accumulation, and normalization of the total and unconjugated bilirubin, total protein, serum cholinesterase, and gamma-glutamyl transpeptidase levels.

Circadian Clock Gene Bmal1 Regulates Bilirubin Detoxification: A Potential Mechanism of Feedback Control of Hyperbilirubinemia.

Controlling bilirubin to a low level is necessary in physiology because of its severe neurotoxicity. Therefore, it is of great interest to understand the regulatory mechanisms for bilirubin homeostasis. In this study, we uncover a critical role for circadian clock in regulation of bilirubin detoxification and homeostasis.Methods: The mRNA and protein levels of Bmal1 (a core clock gene), metabolic enzymes and transporters were measured by qPCR and Western blotting, respectively. Luciferase reporter, mobility shift and chromatin immunoprecipitation were used to investigate transcriptional gene regulation. Experimental hyperbilirubinemia was induced by injection of bilirubin or phenylhydrazine. Unconjugated bilirubin (UCB) and conjugated bilirubin were assessed by ELISA.Results: We first demonstrated diurnal variations in plasma UCB levels and in main bilirubin-detoxifying genes Ugt1a1 and Mrp2. Of note, the circadian UCB levels were antiphase to the circadian expressions of Ugt1a1 and Mrp2. Bmal1 ablation abrogated the circadian rhythms of UCB and bilirubin-induced hepatotoxicity in mice. Bmal1 ablation also decreased mRNA and protein expressions of both Ugt1a1 and Mrp2 in mouse livers, and blunted their circadian rhythms. A combination of luciferase reporter, mobility shift, and chromatin immunoprecipitation assays revealed that Bmal1 trans-activated Ugt1a1 and Mrp2 through specific binding to the E-boxes in the promoter region. Further, Bmal1 ablation caused a loss of circadian time-dependency in bilirubin clearance and sensitized mice to chemical induced-hyperbilirubinemia. Moreover, bilirubin stimulated Bmal1 expression through antagonism of Rev-erbα, constituting a feedback mechanism in bilirubin detoxification.Conclusion: These data supported a dual role for circadian clock in regulation of bilirubin detoxification, generating circadian variations in bilirubin level via direct transactivation of detoxifying genes Ugt1a1 and Mrp2, and defending the body against hyperbilirubinemia via Rev-erbα antagonism. Thereby, our study provided a potential mechanism for management of bilirubin related diseases.

Glucuronidated bilirubin: Significantly increased in hepatic encephalopathy.

Bilirubin is produced by the breakdown of hemoglobin in senescent erythrocytes by macrophages and carried by albumin from blood circulation to the liver for removal in normal physiology. Glucuronic acid modification of bilirubin by UDP-glucuronyltransferase in the liver is the key event for its subsequent elimination from human body. Conditions that accelerate the breakdown of erythrocytes may cause an elevated blood level of unconjugated bilirubin whereas the factors affect the glucuronidated bilirubin formation and subsequent elimination may cause decreased or increased blood level of glucuronidated bilirubin, the water soluble "direct bilirubin" measured by clinical blood test. Studies showed that increased total serum bilirubin has a protective effect on cardiovascular and other related diseases, but it is unknown how direct bilirubin levels were related to different diseases. By taking advantage of the data collected in the clinical laboratory of our hospital, the direct bilirubin data from 192,535 patients with 72 clinically defined diseases were compared to that of healthy controls (10,497). Based on the mean, median, and p values, we found that patients with hepatic encephalopathy had the highest serum direct bilirubin level, which resembled acute hepatic encephalopathy caused by increased serum direct bilirubin level in neonates. In contrast, patients with uremia, nephrotic syndrome, and preeclampsia had significantly lower levels of serum direct bilirubin. Taken together, our data revealed that serum direct bilirubin levels were either increased or decreased in a disease-dependent manner. The possible molecular mechanisms of increased direct bilirubin levels in patients suffering hepatic encephalopathy are discussed.