Lymphedema cholestasis syndrome 1 or Aagenaes syndrome is a condition characterized by neonatal cholestasis, lymphedema, and giant cell hepatitis. The genetic background of this autosomal recessive ...disease was unknown up to now.
A total of 26 patients with Aagenaes syndrome and 17 parents were investigated with whole-genome sequencing and/or Sanger sequencing. PCR and western blot analyses were used to assess levels of mRNA and protein, respectively. CRISPR/Cas9 was used to generate the variant in HEK293T cells. Light microscopy, transmission electron microscopy and immunohistochemistry for biliary transport proteins were performed in liver biopsies.
One specific variant (c.-98G>T) in the 5’-untranslated region of Unc-45 myosin chaperone A (UNC45A) was identified in all tested patients with Aagenaes syndrome. Nineteen were homozygous for the c.-98G>T variant and seven were compound heterozygous for the variant in the 5’-untranslated region and an exonic loss-of-function variant in UNC45A. Patients with Aagenaes syndrome exhibited lower expression of UNC45A mRNA and protein than controls, and this was reproduced in a CRISPR/Cas9-created cell model. Liver biopsies from the neonatal period demonstrated cholestasis, paucity of bile ducts and pronounced formation of multinucleated giant cells. Immunohistochemistry revealed mislocalization of the hepatobiliary transport proteins BSEP (bile salt export pump) and MRP2 (multidrug resistance-associated protein 2).
c.-98G>T in the 5’-untranslated region of UNC45A is the causative genetic variant in Aagenaes syndrome.
The genetic background of Aagenaes syndrome, a disease presenting with cholestasis and lymphedema in childhood, was unknown until now. A variant in the 5’-untranslated region of the Unc-45 myosin chaperone A (UNC45A) was identified in all tested patients with Aagenaes syndrome, providing evidence of the genetic background of the disease. Identification of the genetic background provides a tool for diagnosis of patients with Aagenaes syndrome before lymphedema is evident.
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•Aagenaes syndrome/lymphedema cholestasis syndrome 1 is caused by a variant in the 5’-untranslated region of UNC45A.•Patients with Aagenaes syndrome have reduced expression of mRNA and lower UNC45A protein levels than controls.•A CRISPR/Cas9-created variant in the 5’untranslated region reproduced the reduction in mRNA and protein levels of UNC45A.•Liver biopsies demonstrated paucity of bile ducts, cholestasis, giant cell transformation and variable inflammation.•Aberrant localization of BSEP and MRP2 was observed in liver biopsies.
Growth differentiation factor 15 (GDF-15) is strongly associated with cardiovascular disease (CVD). The aim of our study was to evaluate plasma and urinary levels of GDF-15 after pediatric renal ...transplantation (Rtx) and in children with chronic kidney disease (CKD) and its associations to cardiovascular risk factors. In this cross-sectional study, GDF-15 was measured in plasma and urine from 53 children with a renal transplant and 83 children with CKD and related to cardiovascular risk factors (hypertension, obesity, and cholesterol) and kidney function. Forty healthy children served as a control group. Plasma levels of GDF-15 (median and range) for a Tx (transplantation) cohort, CKD cohort, and healthy controls were, respectively, 865 ng/L (463-3039 ng/L), 508 ng/L (183-3279 ng/L), and 390 ng/L (306-657 ng/L). The CKD and Tx cohorts both had significantly higher GDF-15 levels than the control group (p<0.001). Univariate associations between GDF-15 and hyperuricemia (p<0.001), elevated triglycerides (p=0.028), low HDL (p=0.038), and obesity (p=0.028) were found. However, mGFR (p<0.001) and hemoglobin (p<0.001) were the only significant predictors of GDF-15 in an adjusted analysis. Urinary GDF-15/creatinine ratios were 448 ng/mmol (74–5013 ng/mmol) and 540 ng/mmol (5–14960 ng/mmol) in the Tx cohort and CKD cohort, respectively. In the CKD cohort, it was weakly correlated to mGFR (r=−0.343, p=0.002). Plasma levels of GDF-15 are elevated in children with CKD and after Rtx. The levels were not associated with traditional cardiovascular risk factors but strongly associated with renal function.
To study the effect of extracellular acidosis on apoptosis and necrosis during ischemia and reoxygenation, we exposed human post‐mitotic NT2‐N neurones to oxygen and glucose deprivation (OGD) ...followed by reoxygenation. In some experiments, pH of the cell medium was lowered to 5.9 during either OGD or reoxygenation or both. Staurosporine, used as a positive control for apoptosis, caused Poly(ADP‐ribose)‐polymerase (PARP) cleavage and nuclear fragmentation, but no PARP cleavage and little fragmentation were seen after OGD. Low molecular weight DNA fragments were found after staurosporine treatment, but not after OGD. No protective effect of caspase inhibitors was seen after 3 h of OGD and 21 h of reoxygenation, but after 45 h of reoxygenation caspase inhibition induced a modest improvement in 3‐(4,5‐dimethylthiazol‐2‐yl)2,5‐diphenyltetrazolium bromide (MTT) cleavage. While acidosis during OGD accompanied by neutral medium during reoxygenation protected the neurones (MTT: 228 ± 117% of neutral medium, p < 0.001), acidosis during reoxygenation only was detrimental (MTT: 38 ± 25%, p < 0.01). We conclude that apoptotic mechanisms play a minor role after OGD in NT2‐N neurones. The effect of acidosis on neuronal survival depends on the timing of acidosis, as acidosis was protective during OGD and detrimental during reoxygenation.
Acetylsalicylic acid is used in liver-transplanted children to prevent thrombosis of the hepatic artery. We evaluated whether acetylsalicylic acid and other risk factors were associated with bleeding ...after percutaneous liver biopsy.
Medical charts, laboratory results, imaging studies, and anesthesia charts of 275 ultrasound-guided liver biopsy procedures in 190 children were reviewed. A total of 178 biopsies were performed on native livers and 97 on transplanted livers.
Three major and 28 minor bleeding incidents were found. The mortality rate was 0%. Acetylsalicylic acid had been given the last 5 days before 55 of the biopsy procedures and no increased risk of bleeding was found (odds ratio 0.96 0.37-2.26; P = 1.00). Low-molecular-weight heparin and biopsies from focal lesions were risk factors for bleeding complications. Acute liver failure was associated with increased risk for major complications (odds ratio 26.1 3.3-205; P = 0.01) and was a risk factor for major bleeding. Postbiopsy ultrasound the day after the procedure (n = 266 96% of 275 biopsies) revealed minor bleeding after 7.1% of the biopsies and after 2.6% of the ultrasounds revealed unsuspected bleeding, but none of these required intervention.
Ultrasound-guided liver biopsy in children is a procedure with a low rate of major complications and a high rate of minor bleeding not requiring intervention. Treatment with low-dose acetylsalicylic acid did not increase bleeding incidence or total complication rate. Low-molecular-weight heparin and biopsies from focal lesions were risk factors for bleeding complications. Routine ultrasound the day after the procedure did not change handling of the patients.
Although reduced cognitive function has been demonstrated after liver transplantation in children, few data are available concerning motor competence.
Thirty-five children ages 4 to 12 years were ...tested using Movement Assessment Battery for Children (M-ABC) test at a median of 5.1 (3.9-6.9) years after liver transplantation and compared with reference material of healthy children.
Children with transplantation had worse M-ABC score 8.0 (interquartile range 5.0-11.5), compared with healthy children 3.5 (1.0-6.0) (P < 0.0001). All of the subscores (manual dexterity P < 0.0001, ball skills P = 0.0037, and balance P = 0.0032) were significantly worse in the children with liver transplantation compared with the healthy reference group. Twenty-nine percent of the children with liver transplantation had impaired motor competence, compared with 9% of a healthy reference group. Seventeen of the patients with transplantation were retested 1 year later, and 11 were tested 4 years later with no changes in total M-ABC score. Ball skill competence was worse 4 years after first assessment (P = 0.013). For children with transplantation and cholestatic liver disease (n =26), renal function was a significant predictor for total M-ABC score (P = 0.018).
Children with liver transplantation had impaired motor competence compared with healthy children. Ball skills developed adversely several years after liver transplantation, and motor competence did not improve with time after transplantation. Renal function was a significant predictor for motor competence in children with liver transplantation and cholestatic liver disease.
EBV after pediatric LT is a risk factor for PTLD. We wanted to evaluate the effect of intravenous ganciclovir on EBV viremia and to identify risk factors for chronic EBV viremia. All pediatric ...patients who underwent LT in Norway from 2002 until 2015 were reviewed. Twenty‐two of 38 patients with viremia were treated with intravenous ganciclovir for a median of 22 (21‐38) days. Treated and untreated patients were not different with respect to EBV seroconversion prior to transplantation or age at transplantation, but treated patients had significantly earlier viremia after transplantation (P=.005). There was no difference in the proportion of patients with reduction in virus load in patients treated with ganciclovir compared to untreated patients at 8 weeks. After 1 year, five of 19 patients treated with ganciclovir and six of 14 untreated patients had reduced virus load compared to start of viremia (P=.27). In conclusion, treatment with intravenous ganciclovir did not change the proportion of patients with reduction in EBV load at 8 weeks and 1 year after viremia. Younger age at transplantation, short time from transplantation to viremia, and lack of EBV seroconversion prior to transplantation were significant predictors of chronic EBV viremia.
Abstract The mechanisms of neurotoxicity induced by unconjugated bilirubin (UCB) in newborns are incompletely understood. UCB may cause both necrotic and apoptotic neuronal death. We explored UCB ...toxicity and release of cytokines in human NT2-N neurons and the effect of dexamethasone on these processes. Cultured NT2-N neurons were exposed to UCB, and neuronal damage was evaluated by LDH release and MTT cleavage. After 96 hours, 2 μM UCB significantly increased release of IL-8 and MCP-1, but not IL-13, IP-10, PDGF, or VEGF. Dexamethasone significantly lowered the UCB-induced increase in MCP-1 release, and attenuated UCB-induced neuronal damage assessed with MTT cleavage and LDH release. For comparison, the effects of hydrogen peroxide on cytokine formation and neuronal damage were tested. Hydrogen peroxide increased MCP-1, IP-10, and VEGF, but not IL-8, IL-13, or PDGF. Dexamethasone inhibited the hydrogen peroxide-induced increase in MCP-1 and IP-10. We conclude that UCB causes release of IL-8 and MCP-1 in cultured human NT2-N neurons. Dexamethasone reduces UCB-induced cytokine release and protects against UCB-induced toxicity.
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