Metformin protects against diclofenac-induced toxicity in primary rat hepatocytes by preserving mitochondrial integrity via a pathway involving EPAC.

BACKGROUND AND PURPOSE: It has been shown that the antidiabetic drug metformin protects hepatocytes against toxicity by various stressors. Chronic or excessive consumption of diclofenac (DF) - a pain-relieving drug, leads to drug-induced liver injury via a mechanism involving mitochondrial damage and ultimately apoptotic death of hepatocytes. However, whether metformin protects against DF-induced toxicity is unknown. Recently, it was also shown that cAMP elevation is protective against DF-induced apoptotic death in hepatocytes, a protective effect primarily involving the downstream cAMP effector EPAC and preservation of mitochondrial function. This study therefore aimed at investigating whether metformin protects against DF-induced toxicity via cAMP-EPACs. EXPERIMENTAL APPROACH: Primary rat hepatocytes were exposed to 400 µmol/L DF. CE3F4 or ESI-O5 were used as EPAC-1 or 2 inhibitors respectively. Apoptosis was measured by caspase-3 activity and necrosis by Sytox green staining. Seahorse X96 assay was used to determine mitochondrial function. Mitochondrial reactive oxygen species (ROS) production was measured using MitoSox, mitochondrial MnSOD expression was determined by immunostaining and mitochondrial morphology (fusion and fission ratio) by 3D refractive index imaging. KEY RESULTS: Metformin (1 mmol/L) was protective against DF-induced apoptosis in hepatocytes. This protective effect was EPAC-dependent (mainly EPAC-2). Metformin restored mitochondrial morphology in an EPAC-independent manner. DF-induced mitochondrial dysfunction which was demonstrated by decreased oxygen consumption rate, an increased ROS production and a reduced MnSOD level, were all reversed by metformin in an EPAC-dependent manner. CONCLUSION AND IMPLICATIONS: Metformin protects hepatocytes against DF-induced toxicity via cAMP-dependent EPAC-2.

Association between a comprehensive smoking ban and hospitalization for acute myocardial infarction: An observational study in the Autonomous Community of Valencia, Spain.

OBJECTIVE: To assess the association between a comprehensive smoking ban and hospitalization rates for acute myocardial infarction (AMI). METHODS: An observational study was conducted to assess changes in hospital admission rates for AMI in the Autonomous Community of Valencia, Spain (population 5 million), during the period 1995-2013. Law 28/2005 prohibited smoking in all enclosed spaces (public and private), and Law 42/2010 extended the ban to bars and restaurants as well as children's playgrounds and access areas of schools and hospitals. Data on hospital admissions were obtained from the Hospital Discharge Database (CMBD) of the Autonomous Community. Annual hospital admission rates per 100000 population for AMI (ICD-9-CM code 410) for men and women were calculated. RESULTS: Adjusted hospital admission rates per 100000 population for AMI decreased markedly from 141.1 in 2005 to 119.2 in 2007, with a further reduction to 102.9 in 2013. Reductions in hospital admission were recorded in both men and women, but the downward trends were stronger in women. CONCLUSION: The Spanish comprehensive smoking ban was associated with a marked reduction in the adjusted rate of hospital admissions due to AMI in the Autonomous Community of Valencia. This decrease in the number of persons requiring in-patient care due to AMI is important from both a health care and a societal perspective.

Label-free bioanalysis of Leishmania infantum using refractive index tomography with partially coherent illumination.

In this work, we report the use of refractive index (RI) tomography for quantitative analysis of unstained DH82 cell line infected with Leishmania infantum. The cell RI is reconstructed by using a modality of optical diffraction tomography technique that employs partially coherent illumination, thus enabling inherent compatibility with conventional wide-field microscopes. The experimental results demonstrate that the cell dry mass concentration (DMC) obtained from the RI allows for reliable detection and quantitative characterization of the infection and its temporal evolution. The RI provides important insight for studying morphological changes, particularly membrane blebbing linked to an apoptosis (cell death) process induced by the disease. Moreover, the results evidence that infected DH82 cells exhibit a higher DMC than healthy samples. These findings open up promising perspectives for clinical diagnosis of Leishmania.