SLCO1B1 c.388A > G variant incidence and the severity of hyperbilirubinemia in Indonesian neonates.

OBJECTIVE: It has been established that genetic factors play a substantial role in the development of neonatal hyperbilirubinemia. The population of Indonesia and other Southeast Asian countries has similar, yet different genetic makeup compared to the rest of Asia. Aside from UGT1A1, variants of SLCO1B1 have also been known to contribute to the severity of neonatal hyperbilirubinemia in Asian populations. Since there has been no report on SLCO1B1 polymorphism in relation with hyperbilirubinemia in Indonesia, this study aims to explore incidence of SLCO1B1*1B polymorphism in Indonesia based on 3 hospitals from different provinces and population, and their association with hyperbilirubinemia severity. METHODS: Our study included 88 neonates with mild and moderate-severe hyperbilirubinemia from 3 NICU in hospitals representing homogenous and heterogenous populations: Biak General Hospital Papua, Cipto Mangunkusumo Hospital (Jakarta), and M Yunus Hospital (Bengkulu). We collected samples between November 2016 and September 2017. DNA was obtained from existing samples of the patients from previous studies and were subjected to Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP). We analyzed the *1B variant located in exon 5 of SLCO1B1 with TaqI restriction endonuclease. Clinical, demographic, and laboratory data was also collected from medical records and parents' interviews. RESULTS: The most dominant variant of SLCO1B1*1B in our population is the homozygous G/G (68.18%), followed by heterozygous A/G (26.14%), and wild type A/A (5.68%). The heterozygous A/G had an Odds Ratio (OR) of 0.73 (95% CI 0.10-5.2) and homozygous G/G with OR of 0.51 (95%CI 0.08-3.27), both were not significant. Genotypic distribution across the different centers were also similar and not significant. The significant risk factors for moderate-severe hyperbilirubinemia were the population the neonate originated from (p = < 0.001) and the delivery location (p = 0.001), while SLCO1B1*1B was not associated with the different severity of hyperbilirubinemia. CONCLUSIONS: SLCO1B1*1B is not associated with higher bilirubin levels among neonates with hyperbilirubinemia in Indonesia. Further study is needed to find other potentially important genetic polymorphisms in the development of severe hyperbilirubinemia in Indonesia.

Agreement test of transcutaneous bilirubin and bilistick with serum bilirubin in preterm infants receiving phototherapy.

BACKGROUND: This study compares the minimally invasive Bilistick and a noninvasive method with standard Total Serum Bilirubin (TSB) measurement in preterm newborns receiving phototherapy. We assess the agreement of Transcutaneous Bilirubinometer (TcB) and Bilistick bilirubin measurements with standard TSB measurement in preterm infants receiving phototherapy. METHODS: Bilirubin was measured by using TcB and Bilistick in 94 preterm infants in RSCM Jakarta Neonatal Ward from October 2016 to March 2017, with gestational ages of < 35 weeks, before phototherapy and after 24 and 48 h of phototherapy. RESULTS: There was significant correlation before, at 24 and 48 h of phototherapy between TSB and either TcB (r = 0.874; r = 0.889; r = 0.878 respectively; p < 0.0001), or Bilistick (r = 0.868; r = 0.877; r = 0.918 respectively; p < 0.0001). The mean difference and limits of agreement before, at 24 and 48 h of phototherapy between TcB and TSB were 0.81 ± 1.51 mg/dL (- 2.14 to 3.77 mg/dL); 0.43 ± 1.57 mg/dL (- 2.66 to 3.51 mg/dL); 0.41 ± 1.58 mg/dL (- 2.69 to 3.50 mg/dL), respectively. For Bilistick they were - 1.50 ± 1.47 mg/dL (- 4.38 to 1.38 mg/dL); - 1.43 ± 1.47 mg/dL (- 4.32 to 1.46 mg/dL); - 1,15 ± 1.31 mg/dL (- 3,72 to 1,42 mg/dL), respectively. CONCLUSIONS: Both methods are reliable for measuring TSB before, during, and after phototherapy in preterm infants. TcB tends to overestimate while Bilistick underestimates TSB.

Metagenomic profiles of the early life microbiome of Indonesian inpatient neonates and their influence on clinical characteristics.

Determining the initial normal neonatal gut microbiome is challenging. The debate regarding the sterile fetal environment is still ongoing. Therefore, studying and comparing normal and dysbiotic microbiomes requires the elucidation of both the fetal and infant microbiomes. Factors influencing the normal microbiome also include regional and genetic factors specific to different countries. Determining the normal microbiome population in our center and their association with the clinical conditions of infants is helpful as a tool for both the prevention and treatment of related diseases during neonatal care. Here, we employed metagenomic sequencing to characterize meconium and the subsequent early-life gut microbiome of preterm neonates in Jakarta, Indonesia. Microbiome diversity and complexity was higher in the meconium and on day 4 than on day 7. At the genus level, the most abundant genus overall was unidentified Enterobacteriaceae, with meconium samples dominated by Ureaplasma, day 4 fecal samples dominated by Staphylococcus, and day 7 samples dominated by Clostridiales, while at the phylum level the most abundant was Proteobacteria and Firmicutes. Perinatal factors of PROM and mother's diet influenced the meconium microbiome, while day 4 and day 7 microbiome was associated with bacteremia and early administration of antibiotics. One of our sample sets was derived from triplets, and they had varying diversity despite being triplets. These data are valuable for understanding the formation of a healthy microbiome specific to neonates and devising a strategy to improve both the gut health and related clinical outcomes of the neonate.

Clinical characteristics influence cultivable-bacteria composition in the meconium of Indonesian neonates.

BACKGROUND: Microbial colonization of a neonate's gastrointestinal tract has significant perinatal and lifetime health consequences. However, information regarding the profile of meconium microbiota in neonates and the influence of clinical parameters are lacking in the Indonesian population. This study aimed to preliminary investigate the profile of cultivable bacterial diversity of meconium isolated from neonates born at Cipto Mangunkusumo Hospital (CMH), Jakarta. The cultivable bacteria were isolated from meconium samples and were then processed for cultivation and molecular identification. RESULTS: Fourteen neonates were enrolled as described, i.e., seven hyperbilirubinemia (Hyp) and seven non-Hyp with ten neonates delivered by cesarean section (CS) and four others by vaginal route (VR), and with five exclusive breastfeeding (Ebf), four formula milk, and five combinations. Microbiological identification, molecular 16S rDNA PCR-Sanger sequencing, and PCA analysis of cultivable bacteria isolated from meconium showed Firmicutes' predominance (84.41%), with an abundant population of Staphylococcus, which consist of S. hominis, S. epidermidis, and S. haemolyticus. The influence of mode of delivery showed a lower diversity than the CS populates the VR, but their composition was similar. Concurrently, between feeding patterns, the genera profile did not show much difference; in the non-Ebf group, the total amount of Staphylococcus and Bacillus showed a higher amount but a less diverse. Interestingly, the non-Hyp group showed more abundant and diverse Staphylococcus than that of the Hyp group. In contrast, neonates diagnosed with NEC and proven sepsis showed the same pattern of Staphylococcus domination. CONCLUSION: Staphylococcus predominated the composition of cultivable bacteria in neonates meconium. Due to the small sample size, only the hyperbilirubinemia parameter significantly influenced the profile, i.e., Staphylococcus's proportion (p = 0.037).

Profiling of UGT1A1*6, UGT1A1*60, UGT1A1*93, and UGT1A1*28 Polymorphisms in Indonesian Neonates With Hyperbilirubinemia Using Multiplex PCR Sequencing.

Background: Single nucleotide polymorphism (SNP) variants of the uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) gene have been studied as an important factor in neonatal hyperbilirubinemia (jaundice) severity. Specific ethnicities, including Asians, have certain SNPs that appear more frequently than others. Aim: To identify the most common SNPs in Indonesian neonates and their association with the severity of neonatal hyperbilirubinemia. Methods: Eighty-eight inborn and outborn jaundiced infants from three different hospitals (Bengkulu, Jakarta, Biak Papua) across Indonesia were enrolled in this cross-sectional study and their peak total serum bilirubin (TSB) levels assessed. SNP variant analyses of the TATAA box, promoter, and exon 1 regions of UGT1A1 gene from 78 of the 88 infants were carried out using the SNaPshotR Multiplex Polymerase Chain Reaction (PCR) System followed by DNA sequencing. Results: We detected SNP variants UGT1A1 * 28, UGT1A1 * 60, UGT1A1 * 93, and UGT1A1 * 6 in our population. Mean total serum bilirubin (TSB) was 14.59 ± 5.57 mg/dL. Bivariate analyses using delivery location, gestational age, birth weight, mother's age, and ethnicity were shown to be associated with moderate-to-severe hyperbilirubinemia (p < 0.05). None of the four SNPs appeared to be associated with moderate-to-severe hyperbilirubinemia. In multivariate analysis, however, only the "other ethnic group" (e.g., Chinese, Bengkulu, Papua, Bima) category showed an association with moderate-to-severe hyperbilirubinemia, with an odds ratio of 6.49 (95% CI 1.01-41.67; p < 0.05). Conclusions: We found that the UGT1A1 * 60 is the most common SNP detected in neonates with hyperbilirubinemia in the Indonesian population. Interestingly, in Indonesia, UGT1A1 polymorphisms do not appear to be associated with differences in the severity of hyperbilirubinemia.

UGT1A1 gene and neonatal hyperbilirubinemia: a preliminary study from Bengkulu, Indonesia.

OBJECTIVE: The genetic involvement in unconjugated neonatal hyperbilirubinemia has been extensively studied. Despite the high incidence of hyperbilirubinemia in Indonesia, studies are lacking. The objective of this study is to elucidate the role of polymorphism in the UGT1A1 in Neonatal Hyperbilirubinemia in Bengkulu, Indonesia. RESULTS: There were 41 neonates enrolled in the study; 30 had a total serum bilirubin level ≥ 15 mg/dL (hyperbilirubinemia neonates) while 11 has < 15 mg/dL (control neonates). Genetic mutations in Exon 1, UGT1A1*6 (c211g > a) and one in promoter region, UGT1A1*60 (c3279t > g) were determined by polymerase chain reaction-restriction fragment length polymorphism. We found 18 (60%) mutation in exon 1 in hyperbilirubinemia group and 7 (64%) in the control group with an identical allele frequency of 0.3 in both groups. We found heterozygous UGT1A1*60 4 times (13.3%) and homozygous 26 times (86.7%) in the hyperbilirubinemia group, with an identical allele frequency of 0.935 in hyperbilirubinemia and 1 in control group. This study supports the involvement of genetic factors in the development of unconjugated hyperbilirubinemia in Bengkulu population.