Pyruvate Kinase M2 and Lactate Dehydrogenase A Are Overexpressed in Pancreatic Cancer and Correlate with Poor Outcome.

Pancreatic cancer has a 5-year survival rate of less than 4%. Despite advances in diagnostic technology, pancreatic cancer continues to be diagnosed at a late and incurable stage. Accurate biomarkers for early diagnosis and to predict treatment response are urgently needed. Since alteration of glucose metabolism is one of the hallmarks of cancer cells, we proposed that pyruvate kinase type M2 (M2PK) and lactate dehydrogenase A (LDHA) enzymes could represent novel diagnostic markers and potential therapeutic targets in pancreatic cancer. In 266 tissue sections from normal pancreas, pancreatic cystic neoplasms, pancreatic intraepithelial neoplasia (PanIN) and cancer, we evaluated the expression of PKM2, LDHA, Ki-67 and CD8+ by immunohistochemistry and correlated these markers with clinicopathological characteristics and patient survival. PKM2 and LDHA expression was also assessed by Western blot in 10 human pancreatic cancer cell lines. PKM2 expression increased progressively from cyst through PanIN to cancer, whereas LDHA was overexpressed throughout the carcinogenic process. All but one cell line showed high expression of both proteins. Patients with strong PKM2 and LDHA expression had significantly worse survival than those with weak PKM2 and/or LDHA expression (7.0 months vs. 27.9 months, respectively, p = 0.003, log rank test). The expression of both PKM2 and LDHA correlated directly with Ki-67 expression, and inversely with intratumoral CD8+ cell count. PKM2 was significantly overexpressed in poorly differentiated tumours and both PKM2 and LDHA were overexpressed in larger tumours. Multivariable analysis showed that combined expression of PKM2 and LDHA was an independent poor prognostic marker for survival. In conclusion, our results demonstrate a high expression pattern of two major glycolytic enzymes during pancreatic carcinogenesis, with increased expression in aggressive tumours and a significant adverse effect on survival

Ammonia produces pathological changes in human hepatic stellate cells and is a target for therapy of portal hypertension

BACKGROUND AND AIMS: Hepatic stellate cells (HSCs) are vital to hepatocellular function and the liver response to injury. They share a phenotypic homology with astrocytes that are central in the pathogenesis of hepatic encephalopathy, a condition in which hyperammonemia plays a pathogenic role. This study tested the hypothesis that ammonia modulates human HSC activation in vitro and in vivo, and evaluated whether ammonia lowering, by using l-ornithine phenylacetate (OP), modifies HSC activation in vivo and reduces portal pressure in a bile duct ligation (BDL) model. METHODS: Primary human HSCs were isolated and cultured. Proliferation (BrdU), metabolic activity (MTS), morphology (transmission electron, light and immunofluorescence microscopy), HSC activation markers, ability to contract, changes in oxidative status (ROS) and endoplasmic reticulum (ER) were evaluated to identify effects of ammonia challenge (50 μM, 100 μM, 300 μM) over 24–72 h. Changes in plasma ammonia levels, markers of HSC activation, portal pressure and hepatic eNOS activity were quantified in hyperammonemic BDL animals, and after OP treatment. RESULTS: Pathophysiological ammonia concentrations caused significant and reversible changes in cell proliferation, metabolic activity and activation markers of hHSC in vitro. Ammonia also induced significant alterations in cellular morphology, characterised by cytoplasmic vacuolisation, ER enlargement, ROS production, hHSC contraction and changes in pro-inflammatory gene expression together with HSC-related activation markers such as α-SMA, myosin IIa, IIb, and PDGF-Rβ. Treatment with OP significantly reduced plasma ammonia (BDL 199.1 μmol/L ± 43.65 vs. BDL + OP 149.27 μmol/L ± 51.1, p <0.05) and portal pressure (BDL 14 ± 0.6 vs. BDL + OP 11 ± 0.3 mmHg, p <0.01), which was associated with increased eNOS activity and abrogation of HSC activation markers. CONCLUSIONS: The results show for the first time that ammonia produces deleterious morphological and functional effects on HSCs in vitro. Targeting ammonia with the ammonia lowering drug OP reduces portal pressure and deactivates hHSC in vivo, highlighting the opportunity for evaluating ammonia lowering as a potential therapy in cirrhotic patients with portal hypertension

Transcriptome analysis of Paspalum notatum and Paspalum vaginatum under water deficit condition.

Drought is one of the abiotic stresses that most affect plant growth and productivity. Grasses of the genus Paspalum are successfully used as turf and forage in Australia, Argentina, Brazil and United States. Paspalum notatum has good forage quality, and P. vaginatum, high tolerance to salinity. In addition, their potential to tolerate drought has been described previously, making them interesting for transcriptome studies under water deficit. The objective of this work was to analyze the gene expression profiles of both species in response to drought.Genética 2019

The hillslope length impact on SWAT streamflow prediction in large basins

The objective of this study was to assess the impact of hillslope length on Soil and Water Assessment Tool (SWAT) streamflow predictions in large basins using three methods for hillslope length calculation (the SWAT method, L1; a 3D analysis method, L2; a constant value, L3) combined with two DEMs (pixel size of 25 and 100 m), for a total of six DEML configurations that were tested in the Upper Danube (132000 km(2)). The delineation of subbasins and HRUs were kept unchanged in all configurations, thus isolating the DEM impact on streamflow from that of subbasins delineation. The configurations were independently calibrated in 98 gauged stations located in headwater subbasins (period 1995 similar to 2006), and validated in 150 gauged stations (period 1995 similar to 2009). The analysis of streamflow prediction was extended to its components (surface runoff, lateral flow and baseflow) using performance criteria and residual analysis, and the comparison of different components of water yield was pursued. Calibration and validation showed that all configurations simulated monthly streamflow acceptably (PBIAS < 25% for more than 70% of 150 gauged stations). DEM pixel size had negligible effect of streamflow and its components. The default hillslope length (L1) resulted in large overestimations of lateral flow. L2 resulted in the best performance as well as L3 method. Given that L2 method takes into account the topographic convergence of flow, the configuration of DEM100 and L2 is recommended for SWAT application in large basins in order to obtain reliable streamflow predictions