Improved antitumor response to isolated limb perfusion with tumor necrosis factor after upregulation of endothelial monocyte-activating polypeptide II in soft tissue sarcoma

BACKGROUND: Experiments with tumor necrosis factor alpha (TNF) in rodents have shown that a high dose can lead to hemorrhagic necrosis in tumors. Endothelial monocyte-activating polypeptide II (EMAP-II) is a novel tumor-derived cytokine, and its expression increases the TNF-1 receptor on tumor endothelium, enhances the induction of tissue factor on tumor endothelial cells, and has an antiangiogenic effect. It has recently been shown that in vivo sensitivity of tumor vasculature to TNF is determined by tumor production of EMAP-II. METHODS: We measured the level of EMAP-II in a TNF-resistant soft tissue sarcoma. We subsequently stabile-transfected this cell line with a retroviral construct containing the EMAP gene. In an extremity perfusion model in tumor-bearing rats, we measured response rates to TNF therapy. RESULTS: Functional EMAP-II production was increased after this transfection. Immunostaining of paraffin-embedded tumor tissue sections in rats showed an overexpression of human EMAP-II. Results of the TNF perfusions in rats suggest that this tumor is more sensitive to TNF therapy. CONCLUSIONS: EMAP-II is produced in various levels. One can increase the sensitivity of tumor for TNF therapy in vivo by upregulating the EMAP-II production. This result leaves an opportunity for enhanced TNF response of tumors in future settings

Tumor Motility: Cell migration analysis and effects of EMAP-II on TNF antitumour activities

RePub contains a preprint of the article 'TNF-alpha in cancer treatment: molecular insights, antitumor effects, and clinical utility', The Oncologist, 11(4), pp 397-408, see: http://dx.doi.org/10.1634/theoncologist.11-4-397The tumour itself is at least as dynamic as the fields of research aiming to understand cancer formation and optimize treatments for patients. This thesis describes experiments to study and analyse tumour motility at various levels. Cell migration was studied of tumour and endothelial cells by means of a novel migration method. Treatment of solid tumours with TNF was used to study the mechanisms of action. For EMAP-II studies from molecule to patient were performed. Mobilisation of proteins within cells and redistribution of cells within tumours was analysed

Acknowledgments The authors are grateful to Adriaan Houtsmuller, Niels

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Lactate point-of-care testing for acidosis: Cross-comparison of two devices with routine laboratory results

Objectives: Lactate is a major parameter in medical decision making. During labor, it is an indicator for fetal acidosis and immediate intervention. In the Emergency Department (ED), rapid analysis of lactate/blood gas is crucial for optimal patient care. Our objectives were to cross-compare-for the first time-two point-of-care testing (POCT) lactate devices with routine laboratory results using novel tight precision targets and evaluate different lactate cut-off concentrations to predict metabolic acidosis. Design and methods: Blood samples from the delivery room (n=66) and from the ED (n=85) were analyzed on two POCT devices, the StatStrip-Lactate (Nova Biomedical) and the iSTAT-1 (CG4+ cassettes, Abbott), and compared to the routine laboratory analyzer (ABL-735, Radiometer). Lactate concentrations were cross-compared between these analyzers. Results: The StatStrip correlated well with the ABL-735 (R=0.9737) and with the iSTAT-1 (R=0.9774) for lactate in umbilical cord blood. Lactate concentrations in ED samples measured on the iSTAT-1 and ABL-735 showed a correlation coefficient of R=0.9953. Analytical imprecision was excellent for lactate and pH, while for pO2 and pCO2 the coefficient of variation was relatively high using the iSTAT-1. Conclusion: Both POCT devices showed adequate analytical performance to measure lactate. The StatStrip can indicate metabolic acidosis in 1 μl blood and will be implemented at the delivery room. Keywords: Lactate, Point-of-care testing, Blood gas, Fetal acidosi

Differential expression of glucose-metabolizing enzymes in multiple sclerosis lesions

Introduction: Demyelinated axons in multiple sclerosis (MS) lesions have an increased energy demand in order to maintain conduction. However, oxidative stress-induced mitochondrial dysfunction likely alters glucose metabolism and consequently impairs neuronal function in MS. Imaging and pathological studies indicate that glucose metabolism is altered in MS, although the underlying mechanisms and its role in neurodegeneration remain elusive. We investigated expression patterns of key enzymes involved in glycolysis, tricarboxylic acid (TCA) cycle and lactate metabolism in well-characterized MS tissue to establish which regulators of glucose metabolism are involved in MS and to identify underlying mechanisms. Results: Expression levels of glycolytic enzymes were increased in active and inactive MS lesions, whereas expression levels of enzymes involved in the TCA cycle were upregulated in active MS lesions, but not in inactive MS lesions. We observed reduced expression and production capacity of mitochondrial a-ketoglutarate dehydrogenase (alpha KGDH) in demyelinated axons, which correlated with signs of axonal dysfunction. In inactive lesions, increased expression of lactate-producing enzymes was observed in astrocytes, whereas lactate-catabolising enzymes were mainly detected in axons. Our results demonstrate that the expression of various enzymes involved in glucose metabolism is increased in both astrocytes and axons in active MS lesions. In inactive MS lesions, we provide evidence that astrocytes undergo a glycolytic shift resulting in enhanced astrocyte-axon lactate shuttling, which may be pivotal for the survival of demyelinated axons. Conclusion: In conclusion, we show that key enzymes involved in energy metabolism are differentially expressed in active and inactive MS lesions. Our findings imply that, in addition to reduced oxidative phosphorylation activity, other bioenergetic pathways are affected as well, which may contribute to ongoing axonal degeneration in M

Additional file 1: Table S1. of Differential expression of glucose-metabolizing enzymes in multiple sclerosis lesions

Antibodies. (DOCX 12 kb

Additional file 2: Figure S1. of Differential expression of glucose-metabolizing enzymes in multiple sclerosis lesions

MS lesion classification. MS lesions are characterized by loss of myelin staining, here visualized by labelling of proteolipid protein (PLP) (A,C). In active lesions abundant MHCII positive immune cells occupy the lesion area (B). In chronic active lesions MHCII positive cells are mainly found around the rim of the lesion, but absent in the lesions center (D). (TIF 10733 kb

Additional file 4: Figure S3. of Differential expression of glucose-metabolizing enzymes in multiple sclerosis lesions

Representative image of αKGDH production capacity in an active and inactive MS lesion with (A-C) and without appropriate substrate (B-D). Synaptophysin (E, red) and αKGDH (F,green) immunostaining in inactive MS lesions. Overlay is shown in G. (TIF 13088 kb