Nuclear Magnetic Resonance metabolomics reveals an excretory metabolic signature of renal cell carcinoma

RCC usually develops and progresses asymptomatically and, when detected, it is frequently at advanced stages and metastatic, entailing a dismal prognosis. Therefore, there is an obvious demand for new strategies enabling an earlier diagnosis. The importance of metabolic rearrangements for carcinogenesis unlocked a new approach for cancer research, catalyzing the increased use of metabolomics. The present study aimed the NMR metabolic profiling of RCC in urine samples from a cohort of RCC patients (n = 42) and controls (n = 49). The methodology entailed variable selection of the spectra in tandem with multivariate analysis and validation procedures. The retrieval of a disease signature was preceded by a systematic evaluation of the impacts of subject age, gender, BMI, and smoking habits. The impact of confounders on the urine metabolomics profile of this population is residual compared to that of RCC. A 32-metabolite/resonance signature descriptive of RCC was unveiled, successfully distinguishing RCC patients from controls in principal component analysis. This work demonstrates the value of a systematic metabolomics workflow for the identification of robust urinary metabolic biomarkers of RCC. Future studies should entail the validation of the 32-metabolite/resonance signature found for RCC in independent cohorts, as well as biological validation of the putative hypotheses advanced

Oxidized low-density lipoproteins enhance expression and activity of CD39 and CD73 in the human aortic valve endothelium.

Extracellular nucleotides regulate thrombosis, inflammation and immune response. Ectonucleoside triphosphate diphosphohydrolase 1 (CD39) and ecto-5'-nucleotidase (CD73) convert extracellular nucleotides in a sequential order: ATP to ADP, AMP and then to adenosine. In this study, we aimed to test an effect of oxidized LDL (ox-LDL) on CD39 and CD73 in endothelial cells. Human aortic valve endothelial cells were exposed to oxidized low-density lipoprotein, for 24-48 h. Next, the activity, protein expression and mRNA transcripts level of CD39 and CD73 were characterised by: an incubation with ATP or AMP followed by HPLC analysis of media as well as western blots and qPCR. Results are presented as mean ± SEM. CD73 activity in human valve endothelial cells was increased in presence of ox-LDL (4.04±0.32 nmol/mg prot./min) as compared to control (2.75±0.21 nmol/mg prot/min). There was almost no effect of ox-LDL on CD39 activity. A similar effect was observed for mRNA and protein expression. In conclusion, we found that ox-LDL modulated CD39 and CD73 activity in the endothelium, which may contribute to relevant pathologies and featured treatments

AMP deaminase 1 gene polymorphism and heart disease-A genetic association that highlights new treatment

Nucleotide metabolism and signalling is directly linked to myocardial function. Therefore analysis how diversity of genes coding nucleotide metabolism related proteins affects clinical progress of heart disease could provide valuable information for development of new treatments. Several studies identified that polymorphism of AMP deaminase 1 gene (AMPD1), in particular the common C34T variant of this gene was found to benefit patients with heart failure and ischemic heart disease. However, these findings were inconsistent in subsequent studies. This prompted our detailed analysis of heart transplant recipients that revealed diverse effect: improved early postoperative cardiac function associated with C34T mutation in donors, but worse 1-year survival. Our other studies on the metabolic impact of AMPD1 C34T mutation revealed decrease in AMPD activity, increased production of adenosine and de-inhibition of AMP regulated protein kinase. Thus, genetic, clinical and biochemical studies revealed that while long term attenuation of AMPD activity could be deleterious, transient inhibition of AMPD activity before acute cardiac injury is protective. We suggest therefore that pharmacological inhibition of AMP deaminase before transient ischemic event such as during ischemic heart disease or cardiac surgery could provide therapeutic benefit

Reduction of hyperacute rejection and protection of metabolism and function in hearts of human decay accelerating factor (hDAF)-expressing pigs.

Objective: The use of pig hearts can solve the problem of shortage of donor hearts for transplantation. However, targeting rejection by single genetic modification was proven to be ineffective, highlighting the requirement for complex genetic modifications and more effective methods for transgenic animal production. We evaluated here whether hearts of hDAF transgenic pigs generated using our technique spermmediated gene transfer (SMGT) will be protected from structural damage, metabolic changes, and mechanical dysfunction during perfusion with human blood. Methods: Hearts from control (C, n=6) or transgenic (T, n=5) pigs were perfused ex vivo for 4 h with fresh human blood using the ex vivo working mode system allowing monitoring of the function, metabolism, and structure. Results: Cardiac output (mean±SEM) was maintained in T constant throughout the experiment, at 3.58±0.36 and 3.83±0.14 l/min after 30 min and 4 h, respectively, while cardiac output decreased to 1.95±0.35 l/min in C after 30 min of perfusion (pb0.01 vs. T). The maximum increase in coronary perfusion pressure was reduced in T to 154±16% as compared to C (237±10%, pb0.001). Myocardial ATP after 4 h was 21.1±1.1 nmol/mg dry wt (similar to initial) in T, while it decreased in C to 17.2±1.4 (pb0.05). Deposition of complement factors C3 and C5b9 was present in C but not in T after perfusion

Reduction of hyperacute rejection and protection of metabolism and function in hearts of human decay accelerating factor (hDAF)-expressing pigs.

Objective: The use of pig hearts can solve the problem of shortage of donor hearts for transplantation. However, targeting rejection by single genetic modification was proven to be ineffective, highlighting the requirement for complex genetic modifications and more effective methods for transgenic animal production. We evaluated here whether hearts of hDAF transgenic pigs generated using our technique spermmediated gene transfer (SMGT) will be protected from structural damage, metabolic changes, and mechanical dysfunction during perfusion with human blood. Methods: Hearts from control (C, n=6) or transgenic (T, n=5) pigs were perfused ex vivo for 4 h with fresh human blood using the ex vivo working mode system allowing monitoring of the function, metabolism, and structure. Results: Cardiac output (mean\ub1SEM) was maintained in T constant throughout the experiment, at 3.58\ub10.36 and 3.83\ub10.14 l/min after 30 min and 4 h, respectively, while cardiac output decreased to 1.95\ub10.35 l/min in C after 30 min of perfusion (pb0.01 vs. T). The maximum increase in coronary perfusion pressure was reduced in T to 154\ub116% as compared to C (237\ub110%, pb0.001). Myocardial ATP after 4 h was 21.1\ub11.1 nmol/mg dry wt (similar to initial) in T, while it decreased in C to 17.2\ub11.4 (pb0.05). Deposition of complement factors C3 and C5b9 was present in C but not in T after perfusion