Assessing protein and albumin recovery rates in different ascites filtration membrane washing methods for cell-free and concentrated ascites reinfusion therapy.

Objectives: In cell-free and concentrated ascites reinfusion therapy (CART), the protein recovery rate decreases when the filtration membrane gets clogged. Employing a device with a filtration membrane washing feature prevents clogging, but it leads to the loss of ascites within the filter, resulting in reduced protein recovery. This study employed a device with a membrane washing function to investigate the relationship between protein recovery rate and the quantity of washing solution used, depending on the selected washing method. Methods: We analyzed cases of CART conducted at Fujita Health University Hospital between May 2021 and November 2022. The cases were divided and compared between two groups: one using flush and rinse as the washing method (flush+rinse group) and another using only flushing (flush group). Results: We identified nine cases and 16 sessions. In the flush+rinse group, the median amount of washing solution used per membrane washing was 259 mL per cycle, whereas it was 54 mL per cycle in the flush group. This difference was statistically significant (p<0.0001). The median total protein recovery rate was 53.8% for the flush+rinse group and 78.8% for the flush group, with the latter showing a significantly higher value (p=0.0199). Conclusions: In CART using a membrane washing function, adopting a washing method that reduces the amount of washing solution leads to an increase in the total protein recovery rate.

Effects of N-acetyl-L-tryptophan on desorption of the protein-bound uremic toxin indoxyl sulfate and effects on uremic sarcopenia.

INTRODUCTION: Indoxyl sulfate (IS) is a protein-bound uremic toxin that causes uremic sarcopenia. IS has poor dialysis clearance; however, the addition of a binding competitor improves its removal efficiency. METHODS: Dialysis experiments were performed using N-acetyl-l-tryptophan (L-NAT) instead of l-tryptophan (Trp) using pooled sera obtained from dialysis patients. The molecular structures of L-NAT and Trp were similar to that of IS. Therefore, we examined whether Trp and L-NAT were involved in muscle atrophy in the same manner as IS by performing culture experiments using a human myotube cell line. RESULTS: The removal efficiency of L-NAT was the same as that of Trp. However, L-NAT concentrations in the pooled sera increased at the end of the experiment. Trp (1 mM) decreased the area of human myocytes, similar to IS, whereas L-NAT did not. CONCLUSION: L-NAT is a binding competitor with the ability to remove protein-bound IS while preventing sarcopenia.

Dysbiosis of the gut microbiota in children with severe motor and intellectual disabilities receiving enteral nutrition: A pilot study.

BACKGROUND: Children with severe motor and intellectual disabilities (SMIDs) frequently and continuously receive enteral nutrition and medications and lack adequate exercise, which may lead to dysbiosis, an imbalance in the composition of the gut microbiota. However, studies on the composition of gut microbiota in children with SMIDs are limited. Therefore, we aimed to examine the characteristics of the gut microbiota in children with SMIDs. METHODS: 16S rRNA gene sequencing was performed using fecal samples of 10 children with SMIDs, who received enteral nutrition through a gastric fistula or gastric tube (SMID group: median age, 10.0 years), and 19 healthy children (healthy control [HC] group: median age, 9.0 years). Microbial diversity, microbial composition, and abundance of butyric acid-producing bacteria were compared between the groups. Daily dietary fiber intake in the SMID group was evaluated using questionnaires. RESULTS: The Shannon and Simpson indices (alpha diversity indices) were significantly lower in the SMID group than those in the HC group. Beta diversity analysis identified different clusters. Compared with the HC group, Clostridiales and butyric acid-producing bacteria were less abundant and Bacteroidales were more abundant in the SMID group. Dietary fiber intake in the SMID group was approximately two-thirds of the estimated average requirement for healthy Japanese children. CONCLUSION: Children with SMIDs showed dysbiosis with alteration in the microbial diversity, which could partly be attributed to their low dietary fiber intake. Further studies, with the intervention of prebiotics, probiotics, and synbiotics, are warranted to improve dysbiosis in children with SMIDs.

Chitin powder enhances growth factor production and therapeutic effects of mesenchymal stem cells in a chronic kidney disease rat model.

Previously, we fabricated a device with polylactic acid nonwoven filters and mesenchymal stem cells (MSCs), which effectively reduced urinary protein levels in a rat model of chronic kidney disease (CKD) but could not suppress CKD progression. Therefore, to improve the therapeutic effects of MSCs, in this study, we analyzed the ability of rat adipose tissue-derived MSCs (ADSCs) in contact with chitin nonwoven filters or chitin powder to produce growth factors and examined their therapeutic effect in an adriamycin (ADR)-induced CKD rat model. Hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) production was significantly enhanced by ADSCs cultured in a medium containing chitin powder (C-ADSCs) compared with that by ADSCs cultured in a standard medium without chitin (N-ADSCs). However, the production of HGF and VEGF by ADSCs on chitin nonwoven filters was not significantly enhanced compared with that by the control. Intravenous C-ADSC injection significantly increased podocin expression and improved proteinuria compared with those in saline-treated CKD rats; however, no such improvements were observed in the N-ADSC-treated group. These results showed that ADSCs cultured in a medium supplemented with chitin powder suppressed proteinuria via enhanced HGF and VEGF production in ADR-induced CKD rats to mitigate podocyte damage, offering a new strategy to reduce the dose of MSC therapy for safe and effective treatment of kidney disease.

Accuracy of diagnosing acute kidney injury by assessing urine output within the first week of life in extremely preterm infants.

BACKGROUND: Neonatal acute kidney injury (AKI) is associated with increased mortality and is often assessed with the neonatal modified Kidney Disease: improving Global Outcomes (KDIGO) classification, which uses changes in serum creatinine levels. However, because this classification has many drawbacks, a novel method, the neonatal Risk, Injury, Failure, Loss, and End-Stage Kidney Disease (nRIFLE) classification for diagnosing neonatal AKI according to urine output (UO), was recently proposed. To date, no data on the incidence of AKI according to nRIFLE are available for extremely preterm infants (born at gestational age less than 28 weeks). This study was conducted to clarify the association between incidence of AKI and in-hospital mortality in extremely preterm infants. METHODS: Of 171 extremely preterm infants hospitalized from 2006 to 2020, 84 in whom indwelling bladder catheters were placed for UO measurements within 24 h of life were included. The incidence of AKI was assessed using the nRIFLE classification. In-hospital mortality was compared between patients with AKI and those without it. RESULTS: The incidence of AKI during the first week of life was 56% and that of in-hospital mortality was significantly higher in patients with AKI (25.5%) than in those without it (2.8%). The odds ratio was 12.3 with 95% confidence interval ranging from 1.5 to 100.0. CONCLUSION: The incidence of AKI according to nRIFLE was higher than reported in most previous studies using the neonatal modified KDIGO classification, suggesting that assessment by nRIFLE criteria using UO may improve diagnostic accuracy of AKI in extremely preterm infants.

First-in-human study of blood amyloid ß removal from early Alzheimer's disease patients with normal kidney function.

INTRODUCTION: Amyloid ß (Aß) is a brain protein that causes Alzheimer's disease (AD). This study aimed to verify whether hemadsorption using a hexadecyl-alkylated cellulose bead (HexDC) column removes blood Aß and brain Aß accumulation in mild cognitive impairment/mild AD cases with normal kidney function. METHODS: Two patients with positive Aß on brain imaging underwent HexDC hemadsorption weekly for 6 months. RESULTS: The Aß removal efficiency of HexDC was 87-99%. Aß1-40 /Aß1-42 influx into the blood in one session was 596/56 and 489/48 ng for Case A and Case B, respectively. Although brain Aß accumulation did not clearly change after 6 months of hemadsorption, cognitive functions measured by the two tests were maintained or slightly improved. CONCLUSION: Blood Aß removal was performed in two early AD patients with normal kidney function without adverse events, and it slightly improved or maintained cognitive function.

Evaluation of the performance, operability, and safety of Plasauto µ, a new type of machine for cell-free and concentrated ascites reinfusion therapy, in a postmarketing clinical study.

Cell-free and concentrated ascites reinfusion therapy (CART) is performed by collecting the ascites from the patient, followed by filtration and concentration. Thereafter, concentrated cell-free ascites is reinfused into the patient intravenously. The new type of machine, Plasauto µ, for managing the process of CART was launched onto the market. We have evaluated the machine through postmarketing clinical study in 17 patients with malignant ascites. The amounts of original and concentrated ascites were 3673 ± 1920 g and 439 ± 228 g, respectively. Recovery rates were acceptable regarding values of total protein, albumin, and IgG that were 55.6% ± 17.3%, 60.2% ± 20.8%, and 58.2% ± 20.5%, respectively. Recovery rates were positively associated with amounts of original ascites and negatively associated with total protein concentration. No adverse events related to the machine were observed. The new type of machine showed preferable performance in processing malignant ascites.

Suppression of inflammation during cell-free concentrated ascites reinfusion therapy using a blood purification device.

In recent years, cell-free concentrated ascites reinfusion therapy has been used to treat patients with malignant ascites. However, concentrated ascites reinfusion therapy involves enrichment and reinfusion of useful proteins and inflammatory cytokines. Therefore, fever is a primary side effect and significant problem for patients with ascites. We removed IL-6, an inflammatory cytokine, by mixing malignant ascites and the hexadecyl group adsorbent from a ß2 -microglobulin-adsorbing column (Lixelle S-15). As a result, the hexadecyl group adsorbent did not adsorb the albumin of malignant ascites but adsorbed 43% of IL-6. To investigate the effect of the hexadecyl group adsorbent on hepatocytes, the adsorbed ascites was added to a human hepatoma cell line (HepG2), and the gene expression levels of albumin and serum amyloid A protein were examined. After absorption, ascites showed significantly suppressed serum amyloid A protein expression and significantly increased albumin gene expression compared to before adsorption. Our results suggest that incorporation of Lixelle to filter and concentrate malignant ascites can suppress inflammatory responses and reduce the inhibition of albumin synthesis in the liver after reinfusion.

Comparison Between the Internal and External Pressure Filtration Method of Cell-Free and Concentrated Ascites Reinfusion Therapy Using the Same Cancerous Ascites.

Cell-free and concentrated ascites reinfusion therapy (CART) by internal filtration pressure method (internal method) and external filtration pressure method (external method) using the same cancerous ascites was performed. The rate of rise in circuit pressure and recovered components were compared between the two methods. The factors related to circuit pressure rise were also researched. In both methods, circuit pressure rose in 50% of cases. The recovery rates of IgG, IgA, IgM, and haptoglobin were significantly higher for the internal method than for the external method, whereas the recovery rate of α1 -antitrypsin was significantly lower in the internal method than in the external method. The levels of IL-6, haptoglobin, α1 -antitrypsin, and fibrinogen/fibrindegradation products (FDP) in the original ascites were significantly higher in the group wherein circuit pressure rose than in that without circuit pressure rise. These proteins might be related to the rise in circuit pressure.

A Method for Stabilizing the Proportion of the Reduced Form of Albumin During Cell-Free and Concentrated Ascites Reinfusion Therapy in Patients with Malignant Ascites.

Cell-free and concentrated ascites reinfusion therapy (CART) is used to treat malignant ascites. However, the qualities of albumin in malignant ascites, such as antioxidative activity, may decrease owing to oxidative stress caused by cancer cells and inflammatory reactions. We investigated the fraction percentages of mercaptalbumin (HMA%, reduced form) and non-mercaptalbumin (HNA%, oxidized form) in malignant ascites from 21 patients who received CART and compared the HMA% in the malignant ascites and human serum albumin (HSA) preparations. HMA% of albumin in malignant ascites (22.5%) was significantly lower than that in HSA preparation (42.2%). To ensure a high HMA%, we added L-cysteine to the paracentesis-treated ascites followed by dialysis 1 h later. As a result, the HMA% of albumin in malignant ascites was increased to 59.1%. Our results suggest that using this method in CART will improve patient's albumin quality.

Natural course of isolated mild congenital hydronephrosis: A 2-year prospective study at a single center in Japan.

OBJECTIVES: To determine a follow-up plan for mild congenital hydronephrosis in Japanese individuals. METHODS: Neonates at Kansai Medical University Hospital (Hirakata, Osaka, Japan) who were diagnosed with mild congenital hydronephrosis - defined as a Society for Fetal Urology grade 1 or 2 - at 1-month old by abdominal ultrasonography between 2014 and 2016 were enrolled. These patients were encouraged to undergo repeated abdominal ultrasonography for 2 years every 3 months to investigate the course of congenital hydronephrosis. RESULTS: Among 1009 neonates, congenital hydronephrosis was detected in 118 affected renal units of 100 (9.9%) patients. According to the definition of the Society for Fetal Urology, 118 affected renal units were graded as grade 1 in 87 (74%), grade 2 in 30 (25%), grade 3 in one (1%) and grade 4 in 0 units. Among them, 117 affected renal units of mild congenital hydronephrosis comprising grades 1 and 2 were subjected to ultrasonographic evaluation to monitor the natural course. The rates of resolution at 7, 10, 13, 16, 19, 22 and 25 months after birth in Society for Fetal Urology grades 1 and 2 cases were 60% and 8%, 77% and 19%, 90% and 32%, 92% and 40%, 95% and 52%, 96% and 56%, and 99% and 60%, respectively. CONCLUSIONS: Grade 1 congenital hydronephrosis does not need to be followed up, because it mostly shows spontaneous resolution by 2 years of follow up without any complications. However, ultrasonographic examinations at 1-year intervals for grade 2 congenital hydronephrosis are recommended to determine the subsequent follow-up plan of patients.

Effect of Delivery Mode and Nutrition on Gut Microbiota in Neonates.

BACKGROUND/AIMS: The mode of delivery (vaginal or cesarean section) and feeding type (breastfeeding or formula feeding) of neonates are considered the most influential factors in the development of gut microbiota. OBJECTIVES: This study investigated the effect of prebiotic-rich breast milk on overcoming gut microbiota dysbiosis. METHOD: Stool samples from 36 healthy Japanese neonates were obtained at 4 days and 1 month of age, and divided into 4 groups based on mode of delivery and feeding type. The gut microbiota composition and bacterial diversity were assessed using 16S rRNA sequencing. RESULTS: At 4 days old, vaginally delivered neonates had a significantly higher diversity of bacteria than those born by cesarean section. Bacteroidales and Enterobacteriales were overrepresented in vaginally delivered neonates (p = 0.0031 and p = 0.011), while Bacillales and Lactobacillales were overrepresented in caesarean section delivered neonates (p = 0.012 and p = 0.0016). However, there was little difference in bacterial diversity and bacterial relative abundance at 1 month of age between groups. CONCLUSIONS: Cesarean section delivery appeared to reduce the diversity of neonate gut microbiota, resulting in dysbiosis, but this improved to the equivalent level seen in vaginally delivered infants by 1 month of age. Breastfeeding, even for short periods, may therefore improve neonate gut dysbiosis.

The Diagnostic Significance of Comorbidities of Congenital Heart Diseases, Low-Set Ears, and Intrauterine Growth Restriction in Neonates With Trisomies 13 and 18.

BACKGROUND: Trisomies 13 and 18 (T13/18) are autosomal trisomy syndromes with dismal prognoses. Deciding whether to perform a chromosomal analysis for the definitive diagnosis is often difficult (even for experienced pediatricians) because representative clinical signs may not be found in all T13/18 neonates. OBJECTIVES: This study aimed to investigate any clinical signs that could be useful for screening for T13/18 in participants without the representative clinical signs traditionally found in odd-looking neonates with malformation syndromes. PATIENTS AND METHODS: We retrospectively analyzed 15 T13/18 patients, 33 trisomy 21 patients, and 48 controls with other malformation syndromes, for apparent clinical signs during the neonatal period. All participants had been admitted to the neonatal intensive care unit of Kansai Medical University over a nine-year period. RESULTS: The three leading clinical signs in patients with T13/18 were congenital heart diseases (CHD; 100%), low-set ears (LSE; 80%), and intrauterine growth restriction (IUGR; 73.3%). A comorbidity of these two leading non-specific clinical signs was CHD with LSE, which showed the highest diagnostic accuracy between T13/18 and controls with a sensitivity of 80.0% and a negative predictive value of 92.5%. The chi-square test among these three groups (P < 0.01) and multiple comparison tests of proportional differences showed that the comorbidity of CHD with LSE was specific for autosomal trisomy syndromes. A comorbidity of CHD with IUGR also revealed a similar diagnostic accuracy with a sensitivity of 73.3% and a negative predictive value of 90.9% as well as a specificity for T13/18. CONCLUSIONS: The comorbidities of either CHD with LSE or CHD with IUGR should be suspected in neonates with autosomal trisomy syndromes, particularly T13/18 without the expected representative clinical signs.

Prediction of urine volume soon after birth using serum cystatin C.

BACKGROUND: Urine volume is an important clinical finding particularly during the early neonatal period. Oliguria is not a sign of impaired renal function but also a predictive factor for various complications and prognoses. It has been postulated that serum cystatin C (S-CysC) is a more sensitive biomarker for renal function than serum creatinine (S-Cr) in both adults and children. The objective of the current study was to investigate whether urine volume during 24 h after birth can be predicted using S-CysC. METHODS: The subjects were 87 infants. The average gestational age was 34.7 ± 2.9 weeks and the average birth weight was 2135 ± 614 g. Blood samples were obtained from either the umbilical cord or the peripheral veins or artery of the newborn at birth. Data regarding the amount of urine volume and fluid intake during the first 24 h of life, maternal S-Cr and S-CysC levels within 48 h before delivery, and neonatal S-Cr and S-CysC levels at birth were collected from the medical records. RESULTS: A significantly positive correlation was observed between maternal and neonatal S-Cr levels (r = 0.84, p < 0.0001) but not between maternal S-Cr levels and neonatal S-CysC levels (r = -0.069, p = 0.52). A significant negative correlation was seen between neonatal S-CysC levels and urine volume (r = -0.47, p < 0.0001). CONCLUSION: The present study findings indicate that it may be possible to use S-CysC levels at birth to predict urine volume during the first 24 h of life.

Appropriate nonwoven filters effectively capture human peripheral blood cells and mesenchymal stem cells, which show enhanced production of growth factors.

Scaffolds, growth factors, and cells are three essential components in regenerative medicine. Nonwoven filters, which capture cells, provide a scaffold that localizes and concentrates cells near injured tissues. Further, the cells captured on the filters are expected to serve as a local supply of growth factors. In this study, we investigated the growth factors produced by cells captured on nonwoven filters. Nonwoven filters made of polyethylene terephthalate (PET), biodegradable polylactic acid (PLA), or chitin (1.2-22 µm fiber diameter) were cut out as 13 mm disks and placed into cell-capturing devices. Human mesenchymal stem cells derived from adipose tissues (h-ASCs) and peripheral blood cells (h-PBCs) were captured on the filter and cultured to evaluate growth factor production. The cell-capture rates strongly depended on the fiber diameter and the number of filter disks. Nonwoven filter disks were composed of PET or PLA fibers with fiber diameters of 1.2-1.8 µm captured over 70% of leukocytes or 90% of h-ASCs added. The production of vascular endothelial growth factor (VEGF), transforming growth factor ß1, and platelet-derived growth factor AB were significantly enhanced by the h-PBCs captured on PET or PLA filters. h-ASCs on PLA filters showed significantly enhanced production of VEGF. These enhancements varied with the combination of the nonwoven filter and cells. Because of the enhanced growth factor production, the proliferation of human fibroblasts increased in conditioned medium from h-PBCs on PET filters. This device consisting of nonwoven filters and cells should be investigated further for possible use in the regeneration of impaired tissues.

Elective Cesarean section at 37 weeks is associated with the higher risk of neonatal complications.

Elective Cesarean section performed before 39 weeks of gestation may be associated with increased risk of neonatal complications. We retrospectively investigated differences in the neonatal complication rate between 684 newborns delivered by elective Cesarean section at 37 weeks of gestation (n = 390) and those delivered by the same procedure at 38 weeks (n = 294) between 2006 and 2012 at our hospital in order to ascertain whether adverse outcomes differ between the groups. Newborns delivered at 37 weeks had a significantly higher incidence of neonatal intensive care unit admission (p = 0.03), adverse respiratory complications (p < 0.01), low birth weight (p < 0.001), and hypoglycemia (p < 0.005) than those delivered at 38 weeks. Compared with normal weight neonates, low birth weight neonates were more likely to have hypoglycemia (p < 0.001). Multivariate logistic regression analysis revealed that an adverse respiratory outcome was independently associated with gestational age (p < 0.01; odds ratio [OR], 3.26; 95% confidence interval [CI], 1.36-7.81), while hypoglycemia was independently associated with birth weight (p < 0.01; OR, 16.34; 95% CI, 7.72-34.56). Respiratory disorders were significantly associated with gestational age even in normal birth weight newborns without any other complications such as hyperbirilubinemia, hypoglycemia or bacterial infections. In conclusion, the incidence of neonatal complications was higher in newborns delivered at 37 weeks of gestation than in those delivered at 38 weeks via elective Cesarean section. Thus, the procedure should be scheduled at 38 weeks to improve neonatal outcomes.

Postmortem computed tomography imaging in the investigation of nontraumatic death in infants and children.

OBJECTIVE: To determine the accuracy of postmortem computed tomography (PMCT) for the assessment of causes in nontraumatic deaths in children. STUDY DESIGN: We enrolled cases of nontraumatic deaths of infants and children who underwent PMCT at a single center. The presumed cause of death determined by PMCT was prospectively compared with the clinical and pathological diagnoses of deaths. RESULTS: Thirty-eight cases were enrolled for analysis. Among them, seven cases also underwent conventional medical autopsy. PMCT revealed an identifiable cause of death in accordance with the clinical diagnosis of death in 16 cases of the 38 cases (the concordance rate was 42%) and in accordance with the autopsy cause of death in four of the seven autopsy cases (the concordance rate was 57%). Among eight cases with unknown cause of death by clinical diagnosis, four cases (50%) were identified with cardiac tamponade as a cause of death (one case) and intracranial hemorrhage suggesting abuse (3 cases). CONCLUSIONS: PMCT seems to be a promising technique that might serve as a substitute for conventional medical autopsy and give us the complementary information to clinical diagnoses particularly in cases of child abuse. Larger multicenter trials are worthwhile to validate the general feasibility of PMCT.

Toll-like receptor 4 deficiency accelerates the development of insulin-deficient diabetes in non-obese diabetic mice.

BACKGROUND/OBJECTIVE: Toll-like receptors (TLR) mediate the recognition of microbial constituents and stress-induced endogenous ligands by the immune system. They may also be involved in the maintenance or break down of tolerance against autologous antigens. The aim of our investigation was to study the consequence of TLR4 deficiency on the development of insulin-deficient diabetes in the NOD mouse. METHODS: The TLR4 defect of the C57BL/10ScN mouse was backcrossed onto the NOD background and the effect of TLR4 deficiency on diabetes development was analysed by in vivo and in vitro studies. RESULTS: Compared to animals with wildtype TLR4 expression (TLR4(+/+)), female NOD mice carrying a homozygous TLR4 defect (TLR4(-/-)), showed significant acceleration of diabetes development, with a younger age at diabetes onset (TLR4 (+/+) 177±22 d, TLR(-/-): 118±21 d; p<0.01). Pancreata of 120 d old TLR4(-/-) NOD mice revealed increased proportions of islets with advanced stages of immune cell infiltration compared to TLR4(+/+) mice (p<0.05). TLR4 deficiency did not affect the susceptibility of islet cells to the beta cell damaging mediators nitric oxide or the inflammatory cytokines tumor necrosis factor alpha, interleukin-1 beta and interferon gamma. The lack of TLR4 further had no effect on the frequency of regulatory T-cells but reduced their capacity to inhibit T-cell proliferation. CONCLUSIONS: Our findings demonstrate that TLR4 deficiency results in an acceleration of diabetes development and immune cell infiltration of islets in NOD mice. We conclude that TLR4 is involved in the progression of the insulitis process thereby controlling the development of insulin-deficient diabetes in NOD mice.

B-type natriuretic peptide for assessment of haemodynamically significant patent ductus arteriosus in premature infants.

AIM: Haemodynamically significant patent ductus arteriosus (hsPDA) is frequently observed in premature infants. This study was conducted to explore whether the blood BNP can be a valuable biomarker to assess the necessity of treatment for hsPDA in premature infants. METHODS: Serial measurements of the blood BNP were performed during the first 5 days of life in premature infants with hsPDA (Group I) and those without hsPDA (Group N). The definition of the hsPDA was the PDA requiring treatment, such as indomethacin administration and/or surgical ligation. RESULTS: Forty-six subjects were enrolled. Compared with Group N, Group I showed significantly higher level of blood BNP at postnatal 24-96 h and demonstrated the peak value at postnatal 24-48 h. With the ROC curve using the data at postnatal 24-48 h in Group I, we deduced the predictive value of 250 pg/mL of blood BNP for indomethacin treatment. Similarly, with the ROC curve using the maximal value of blood BNP within the first 5 days of life, the predictive value of 2000 pg/mL for surgical ligation was deduced. CONCLUSIONS: Blood BNP during early postnatal period can be a useful biomarker to assess the necessity of treatment for hsPDA in premature infants.