The study of cellular central carbon metabolism modulations induced by viruses is an emerging field. Human cytomegalovirus (HCMV), herpes simplex virus (HSV), Kaposi's sarcoma-associated herpesvirus (KSHV), and hepatitis C virus (HCV) have been shown recently to reprogram cell metabolism to support their replication. During HCV infection the global glucidolipidic metabolism of hepatocytes is highly impacted. It was suggested that HCV might modify glucose uptake and glycolysis to increase fatty acids synthesis, but underlying mechanisms have not been completely elucidated. We thus investigated how HCV may modulate glycolysis. We observed that in infected Huh7.5 cells and in subgenomic replicon-positive Huh9.13 cells, glucose consumption as well as lactate secretion was increased. Using protein complementation assays and coimmunoprecipitation, we identified a direct interaction between the HCV NS5A protein and cellular hexokinase 2 (HK2), the first rate-limiting enzyme of glycolysis. NS5A expression was sufficient to enhance glucose consumption and lactate secretion in Huh7.5 cells. Moreover, determination of HK activity in cell homogenates revealed that addition of exogenous NS5A protein, either the full-length protein or its D2 or D3, but not D1, domain, was sufficient to increase enzyme activity. Finally, determination of recombinant HK2 catalytic parameters ( V _max and K _m ) in the presence of NS5A identified this viral protein as an activator of the enzyme. In summary, this study describes a direct interaction between HCV NS5A protein and cellular HK2 which is accompanied by an increase in HK2 activity that might contribute to an increased glycolysis rate during HCV infection. IMPORTANCE Substantial evidence indicates that viruses reprogram the central carbon metabolism of the cell to support their replication. Nevertheless, precise underlying mechanisms are poorly described. Metabolic pathways are structured as connected enzymatic cascades providing elemental biomolecular blocks necessary for cell life and viral replication. In this study, we observed an increase in glucose consumption and lactate secretion in HCV-infected cells, revealing higher glycolytic activity. We also identified an interaction between the HCV NS5A nonstructural protein and cellular hexokinase 2, the first rate-limiting enzyme of glycolysis. This interaction results in an enhancement of catalytic parameters of the enzyme, which might explain, at least in part, the aerobic glycolysis shift observed in HCV-infected cells.

Liver disease is a major cause of morbidity and mortality worldwide. As in other fields of medicine, there is a stringent need for non-invasive markers to improve patient diagnostics, monitoring and prognostic ability in liver pathology. Cell-free circulating RNA molecules have been recently acknowledged as an important source of potential medical biomarkers. However, many aspects related to the biology of these molecules remain to be elucidated. In this review, we summarize current concepts related to the origin, transportation and possible functions of cell-free RNA. We outline current development of extracellular RNA-based biomarkers in the main forms of non-inherited liver disease: chronic viral hepatitis, hepatocellular carcinoma, non-alcoholic fatty liver, hepato-toxicity, and liver transplantation. Despite recent technological advances, the lack of standardization in the assessment of these markers makes their adoption into clinical practice difficult. We thus finally review the main factors influencing quantification of circulating RNA. These factors should be considered in the reporting and interpretation of current findings, as well as in the proper planning of future studies, to improve reliability and reproducibility of results.

Hepatitis B virus (HBV) genome transcription is highly dependent on liver‐enriched, metabolic nuclear receptors (NRs). Among others, NR farnesoid X receptor a (FXRa) enhances HBV core promoter activity and pregenomic RNA synthesis. Interestingly, two food‐withdrawal‐induced FXRα modulators, peroxisome proliferator‐activated receptor‐γ coactivator 1α (PGC‐1α) and deacetylase SIRT1, have been found to be associated with HBV genomes ex vivo. Whereas PGC‐1α induction was shown to increase HBV replication, the effect of SIRT1 on HBV transcription remains unknown. Here, we showed that, in hepatocar‐cinoma‐derived Huh‐7 cells, combined activation of FXRα by GW4064 and SIRT1 by activator 3 increased HBV core promoter‐controlled luciferase expression by 25‐fold, compared with a 10‐fold increase with GW4064 alone. Using cell lines differentially expressing FXRα in overexpression and silencing experiments, we demonstrated that SIRT1 activated the core promoter in an FXRα‐ and PGC‐1 α‐dependent manner. Maximal activation (> 150‐fold) was observed in FXRα‐and PGC‐1α‐overexpressing Huh‐7 cells treated with FXRα and SIRT1 activators. Similarly, in cells transfected with full‐length HBV genomes, maximal induction (3.5‐fold) of core promoter‐controlled synthesis of 3.5‐kb RNA was observed in the same conditions of transfection and treatments. Thus, we identified a subnetwork of metabolic factors regulating HBV replication, strengthening the hypothesis that transcription of HBV and metabolic genes is similarly controlled.—Curtil, C., Enache, L. S., Radreau, P., Dron, A.‐G., Scholtès, C., Deloire, A., Roche, D., Lotteau, V., André, P., and Ramière, C. The metabolic sensors FXRα, PGC‐1 α, and SIRT1 cooperatively regulate hepatitis B virus transcription. FASEB J. 28, 1454–1463 (2014). www.fasebj.org

Foster care young people have an increased overall risk for the development of chronic health conditions.Smoking is a major risk factor for many conditions with inflammatory component. We performed a cross-sectional pilot study to assess the correlation between tobacco consumption status and inflammatory profile among adolescents living with foster parents in the central region of Romania. A number of 35 teenagers aged 10-18 were enrolled. Blood samples were collected by venipuncture for complete blood count (CBC), fibrinogen, Interleukin-6 (IL 6), C reactive protein (CRP). Exhaled CO was measured and urine samples were collected in the same session for cotinine assessment. Of the 123 teenagers initially enrolled, 35 completed the entire study protocol. Urinary cotinine accurately reflected smoker status. Smoking did not affect hemoglobin levels or other hematological parameters in foster care teenagers. The value of C-reactive protein was higher in smokers. Plasma fibrinogen correlated with the daily number of cigarettes. Interleukin-6 did not correlate with the smoker status or the daily number of cigarettes consumed. Our study highlights the existence of an inflammatory response to smoking in foster care adolescents, a social category otherwise predisposed to various addictive behaviors.

Background and objective: Analysis of inflammatory biomarkers, along with the neutrophil/lymphocyte ratio (NLR) or platelet/lymphocyte ratio (PLR), supports the connection between inflammation and carcinogenesis. Methods: We conducted a retrospective observational study at the Clinical County Hospital Mureș involving patients with lung cancer. The parameters analyzed included histopathological type (NSCLC: squamous cell carcinoma or adenocarcinoma; SCLC), molecular mutations (EGFR, ALK, PD-L1), parameters from the complete blood count, inflammatory parameters, and associated comorbidities. Results: A total of 380 patients were included: 115 patients in the cancer group and 265 patients in the control group. Among patients in the lung cancer group, 88 were diagnosed with NSCLC (44 adenocarcinomas, 44 squamous cell carcinomas) and 27 with SCLC. Both NLR and PLR were significantly higher in cancer patients than in the control group (5.30 versus 2.60, p < 0.001; 217 versus 136, p < 0.001, respectively). NLR and PLR differ between men and women (p = 0.005 and p = 0.056, respectively). C-reactive protein was not correlated with either NLR (p-value: 0.0669) or PLR (p-value: 0.6733) in lung cancer patients. Conclusions: The NLR and PLR values may serve as new predictive biomarkers for the diagnosis of disease in patients with lung cancer, especially those with NSCLC.

Background: There are a multitude of factors that influence smoking status, and minors from the social protection system are a vulnerable category in terms of smoking. Methods: The objective of this research was to assess the degree of smoking dependence and to identify potential predictors of smoking status in foster care teenagers. Smoker status was confirmed by dosing CO in the exhaled air, and the degree of dependence was assessed using the Fagerström score. We performed a multivariate logistic regression analysis. Results: From the 275 foster care minors, 22.5% were current smokers. Exhaled CO was not influenced by general demographic factors, was associated with the frequency of smoking, and was positively correlated with the estimated number of cigarettes consumed daily and with the Fagerström score. The calculated probability of being a smoker was less than 20.4% in 75% of nonsmokers, whereas 75% of actual smokers had a predicted probability higher than 30.3%. Conclusions: In addition to age, gender, social environment, previous exposure to secondhand smoking, and residential type of foster care system, the expressed opinions regarding the health effects of tobacco use were associated with smoking in foster care teenagers.