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.

This study was performed to explore whether lactate, pyruvate, glucose, and glycerol levels sampled via microdialysis catheters in the transplanted liver could be used to detect ischemia and/or rejection. The metabolites were measured at the bedside every 1 to 2 hours after the operation for a median of 10 days. Twelve grafts with biopsy-proven rejection and 9 grafts with ischemia were compared to a reference group of 39 grafts with uneventful courses. The median lactate level was significantly higher in both the ischemia group [5.8 mM (interquartile range ¼ 4.0-11.1 mM)] and the rejection group [2.1 mM (interquartile range ¼ 1.9-2.4 mM)] versus the reference group [1.5 mM (interquartile range ¼ 1.1-1.9 mM), P < 0.001 for both]. The median pyruvate level was significantly increased only in the rejection group [185 lM (interquartile range ¼ 155-206 lM)] versus the reference group [124 lM (interquartile range ¼ 102-150 lM), P < 0.001], whereas the median lactate/ pyruvate ratio and the median glycerol level were increased only in the ischemia group [66.1 (interquartile range ¼ 23.9-156.7) and 138 lM (interquartile range ¼ 26-260 lM)] versus the reference group [11.8 (interquartile range ¼ 10.6-13.6), P < 0.001, and 9 lM (interquartile range ¼ 9-24 lM), P ¼ 0.002]. Ischemia was detected with 100% sensitivity and greater than 90% specificity when a positive test was repeated after 1 hour. In 3 cases of hepatic artery thrombosis, ischemia was detected despite normal blood lactate levels. Consecutive pathological measurements for 6 hours were used to diagnose rejection with greater than 80% sensitivity and specificity at a median of 4 days before the activity of alanine aminotransferase, the concentration of bilirubin in serum, or both increased. In conclusion, bedside measurements of intrahepatic lactate and pyruvate levels were used to detect ischemia and rejection earlier than current standard methods could. Discrimination from an uneventful patient course was achieved. Consequently, intrahepatic graft monitoring with microdialysis may lead to the earlier initiation of graft-saving treatment. Liver Transpl 18: 839-849, 2012. V C 2012 AASLD. Received December 9, 2011 accepted February 29, 2012.As many as 20% of transplanted liver grafts are lost within the first year despite considerable therapeutic improvements in treatment during the last decades.1 Most grafts are lost within the first week or weeks. As a result, attempts to improve overall graft survival rates should include the early detection and adequate treatment of complications such as acute rejection 2 and vascular occlusion with subsequent ischemia.3 The latter is an especially major contributor to the loss of grafts. Biliary complications and infections are other organ-threatening complications. [4][5][6]

Barbiturates differ markedly in their neuroprotective effects against combined oxygen and glucose deprivation in human NT2-N neurons. The variation in neuroprotective effects could only partly be explained by differences in antioxidant action.

Hepatocyte-like cells (HLCs) generated in vitro from human pluripotent stem cells (hPSCs) provide an invaluable resource for basic research, regenerative medicine, drug screening, toxicology, and modeling of liver disease and development. This unit describes a small-molecule-driven protocol for in vitro differentiation of hPSCs into HLCs without the use of growth factors. hPSCs are coaxed through a developmentally relevant route via the primitive streak to definitive endoderm (DE) using the small molecule CHIR99021 (a Wnt agonist), replacing the conventional growth factors Wnt3A and activin A. The small-molecule-derived DE is then differentiated to hepatoblast-like cells in the presence of dimethyl sulfoxide. The resulting hepatoblasts are then differentiated to HLCs with N-hexanoic-Tyr, Ile-6 aminohexanoic amide (Dihexa, a hepatocyte growth factor agonist) and dexamethasone. The protocol provides an efficient and reproducible procedure for differentiation of hPSCs into HLCs utilizing small molecules. © 2016 by John Wiley & Sons, Inc.

Human NTera2 teratocarcinoma cells were differentiated into postmitotic NT2-N neurons and exposed to hypoxia for 6 h. The cultures were evaluated microscopically, and percent lactate dehydrogenase (LDH) release after 24 and 46 h was used as an assay for cell death. After 48 h LDH release was 24.3 ±5.6% versus 13.8 ±3.7% in controls (p < 0.001). Cell death was greatly diminished by MK-801 pretreatment (15.4 ± 5.1%, p < 0.001). If glutamine was omitted from the medium, glutamate levels after 6 h of hypoxia were reduced from 101 ±63 to 2.3 ±0.3~tM,and cell death at 48 h was also markedly reduced (15.4 ± 4.5%, p < 0.001). The a-amino-3-hydroxy-5-methylisoxazole-4-propionate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (18.7 ± 5.1%, p < 0.001) and mild hypothermia (33.5-34°C) during hypoxia (19.5 ±2.7%, p < 0.05) were moderately protective. Basic fibroblast growth factor (24.1 ±3.2%), the nitric oxide synthase inhibitor N°-nitro-L-arginine methyl ester (22.8 ±8.1%), the antioxidant N-tert-butyl-o-phenylnitrone (18.9 ±5.9%), and the 21-aminosteroid U74389G (24.0 ±3.4%) did not protect the cells. N-Acetyl-L-cysteine even tended to increase cell death (30.1 ±2.5%, p = 0.06). Treatment with MK-801 at the end of hypoxia did not reduce cell death (23.3 ±2.3%). In separate experiments, a 15-mm exposure to 1 mM glutamate without hypoxia did not result in significant cell death (14.7 ±2.4 vs. 12.2 ±2.1%, p = 0.07). We conclude that, although somewhat resistant to glutamate toxicity when normoxic, NT2-N neurons die via an ionotropic glutamate receptor-mediated mechanism when exposed to hypoxia in the presence of glutamate. As far as we know, this isthe first reported analysis ofthe mechanism of hypoxic cell death in cultured human neuronlike cells.