PQ segment and QRS complex within the heart of FTO deficient mice. The sympathetic nervous system is known to play an important role in arrhythmogenesis. Catecholamines can increase automaticity and induce triggered activity, thereby increasing arrhythmic risk. Importantly, in this study we provide evidence of increased vulnerability to stress-induced tachyarrhythmias in mice lacking the FTO gene. Of note, arrhythmogenesis was almost completely absent in wild-type mice. It is interesting that arrhythmia vulnerability in FTO deficient mice was induced by stress exposure, as arrhythmic events were only sporadically noted during baseline recordings, and clearly more pronounced in response to the restraint than the injection stress. Taken together these findings point to a close link between FTO deficiency and arrhythmia vulnerability, particularly in conditions of sustained stress exposure. Investigation of potential electrophysiological changes relevant to arrhythmogenesis in the heart of FTO deficient mice revealed that no changes occurred in ventricular excitability and refractoriness, suggesting that arrhythmia vulnerability may not be linked to cellular electrophysiological abnormalities. Noteworthy, these measures were obtained in anesthetized mice, and therefore we cannot exclude that sympathetic hyperactivity in FTO deficient mice may have affected cardiac excitability and/or refractoriness in the awake state. In addition, our data indicate that arrhythmogenesis was not correlated to accumulation of fibrotic tissue in the left ventricular myocardium. Our hypothesis is that exaggerated sympathetic stress response triggered abnormal automaticity in non-pacemaker tissue. We found in FTO deficient mice signs of cardiac hypertrophy, affecting both the right and the left ventricles. The specific morphological changes were not investigated here, but might reflect structural changes in the hypertrophied myocardium altering the ion channels operating during the early repolarization phase. This hypothesis was based on the observation that the duration of QTc interval was longer in FTO deficient mice. Therefore, we hypothesize a role of ventricular hypertrophy in altering ventricular repolarization to explain QTc lengthening in these mice. The QTc interval is also influenced by the autonomic nervous system: abnormal sympathetic modulation or vagal withdrawal directly induce altered ventricular repolarization, thus leading to prolongation of the QTc interval. Therefore, exaggerated cardiac sympathetic predominance in mice lacking the FTO gene could contribute directly to both ventricular hypertrophy and abnormal ventricular repolarization independent from blood pressure, conditions that might serve as a substrate for arrhythmias. Further studies are needed in order to elucidate the biophysical mechanisms and the cellular and subcellular bases of the reported arrhythmogenesis.

DNA concentrations were converted to copy number/mL using the formula described by Fronhoffs et al.. Viral and cellular DNA copy numbers were converted to absolute weight for mutual comparison. The introduction of a new generation of sequencing technologies with high-throughput capacities has immensely impacted the field of genomics. Previous publications have provided a snapshot of the possible applications in the field of HCMV genomics and transcriptomics. We believe that the amplification, sequencing and analysis workflow that we present in this study can help to maximize the efficiency of sequencing HCMV strains in high-throughput. Given the large genetic background of HCMV, it could be interesting to routinely elucidate the complete sequence of strains that are used in mutational studies. This should no longer be considered as extremely laborious or costly. Additionally, the analysis of clinical HCMV isolates could assist in the refinement of the HCMV genetic map. It will provide a better knowledge of viral mutants and in which patient populations they are circulating. Finally, it could prove to be of value in the ongoing quest for genetic determinants of viral pathogenicity that has eluded scientists for more than a decade. To better manage health risks and costs, modern vaccines are no longer made from whole pathogens. Rather, they contain antigenic subunits that mainly provide the immune target to elicit memory responses against a broad spectrum of the pathogen’s strains and clades. For efficacy, most subunit vaccines require additional factors, so-called adjuvants; among them are pathogenassociated molecular patterns. Vaccine adjuvants can compensate for the lack of danger signals in subunit-based formulations, thereby improving the activation of innate and adaptive immune responses. Cyclic di-nucleotides are one such group of promising candidate adjuvants. They are signaling molecules which are involved in critical processes such as attachment and biofilm formation in prokaryotes and they control cell motility and proliferation states in the protozoon dictyostelium. Recently, an additional cyclic di-nucleotide, cyclic, was reported to activate the stimulator of interferon genes and to be synthesized by the mammalian enzyme cyclic GMP-AMP synthase upon stimulation with foreign DNA. Cyclic di-nucleotides such as bis–cyclic dimeric adenosine monophosphate, biscyclic dimeric guanosine monophosphate, and bis–cyclic dimeric inosine monophosphate proved to have immune modulatory activity in mice and humans. We and others have previously shown that c-di-AMP, cdi-GMP, and c-di-IMP act as potent adjuvants in immunization experiments with mice. We demonstrated that c-diAMP promotes humoral as well as cellular immune responses to model and vaccine antigens in mice immunized via the mucosal route. Immune modulation by c-di-AMP was observed to contribute to a balanced TH1/TH2/TH17 response.

However, comprehensive understanding of the biological activity of flavanols including their mechanisms of action in humans is still enigmatic. Over the past years, in vitro and in vivo studies have shown that polyphenols modulate the activity of cell signalling proteins, transcription factors and consequently the expression of both, mRNA and miRNA. For example, in animal models, supplementation of a diet with quercetin, naringenin or curcumin at nutritionally relevant doses altered the expression of hundreds of genes in lung, aorta or liver. In vitro, polyphenols impact the expression of genes in various cell types. It has been demonstrated that flavanone metabolites at physiological concentrations can modulate the expression of genes involved in inflammation in tumor necrosis factor -astimulated human umbilical vein endothelial cells. Exposure of human monocytes to green tea polyphenols affected the expression of genes related to atherosclerosis development, such as CD36, LXR-a, MYC or LDL-R. In human hepatocytes polyphenols from red grape juice were able to counteract the LDL-induced changes in gene expression of the LDL receptor, hydroxymethylglutaryl-CoA reductase and the transcription factor sterol regulatory element-binding protein -1, a key regulator of lipid homeostasis. Also genes such as ICAM1, MCP1, IL6 or IL1b of which expression is controlled by nuclear factor kappa B and of which transcriptional activation is critical in a number of pathologies, including cardiovascular diseases, are differentially expressed in the presence of polyphenols. However, only a few studies have been published up to now describing the genomic impact of polyphenols and polyphenol-rich foods in humans. It has been observed that the daily intake of 150 mg quercetin for 2 weeks significantly affected the expression of 788 genes in CD14-positive monocytes, genes related to immune system, apoptosis or cell signalling pathways. Consumption of orange juice rich in hesperidin or hesperidin alone for 4 weeks by healthy, middleaged, moderately overweight men significantly affected over 3000 and 1800 genes respectively. These genes were involved in the regulation of chemotaxis, cell adhesion, lipid transport and their expression profile could be considered as anti-inflammatory and anti-atherogenic. More recently, in a double-bind, randomized cross-over trial in menopausal women isoflavones affected in peripheral blood mononuclear cells the expression of 357 genes which are involved in inflammation, oxidative phosphorylation and cell cycle. In addition to the impact of polyphenols on the expression of genes, recent data suggest that polyphenols could also modulate DNA methylation at multiple levels. Furthermore, different lifestyle factors such as diet and smoking have been associated with DNA methylation changes in cancer and chronic inflammation related diseases such as metabolic syndrome.

To our knowledge, it is the first study directly comparing the effects of DA on AMPA/KA and NMDA receptors-mediated components of the ECSCs showing that DA exerted a stronger inhibition of NMDA than AMPA/KA receptor-mediated EPSCs. The latter finding supports the idea that there is direct D1-NMDA receptors interaction at cellular level. In addition, a recent study suggests a dopamine triggered heterosynaptic plasticity mechanism, likely mediated presynaptically located D1 recetors and expressed by presynaptic inhibition of GABA release. Most studies that have examined the effects of DA on glutamatergic neurotransmission in the nAcb have found a D1like receptors-mediated depression of excitatory responses in agreement with our results. In vitro experiments have shown that this inhibition was mediated by presynaptic dopaminergic receptors located on glutamatergic terminals and are consistent with the demonstration that DA inhibited glutamate release in nAcb. DA receptors are distributed on both presynaptic glutamatergic terminals and postsynaptic membrane of MS neurons in the nAcb. Therefore, DA may have exerted its inhibitory action on EPSCs by acting at pre- and/or postsynaptic mechanisms. We examined the changes in PPR produced during DA application as an indication of pre- or postsynaptic mechanism. We found that bath application of DA greatly reduced the EPSCs but the inhibition was less on EPSC2 than on EPSC1 resulting in an increase in PPR, suggesting that DA acted presynaptically to reduce the probability of glutamate release from presynaptic terminals. We have also found that DA did not change membrane conductance nor that it affected the extrinsic glutamate-induced currents also arguing in favor of presynaptic mechanisms. This conclusion is also consistent with that of previous investigators studying DA inhibition on EPSCs in the nAcb as well as in the nucleus of the solitary tract, the subiculum, the supraoptic nucleus and the parabrachial nucleus. In other studies, DA has also been shown to produce postsynaptic effects in MS neurons but we and others failed to observe postsynaptic effects. In the present study, we added QX314 to the recording pipette solution in order to prevent voltage-sensitive Na + channels from generating action potentials. QX314 as well as other local anaesthetics have been shown to interfere with G protein-coupled receptors as well as with other second messenger systems and it is likely to have occluded the expression of dopaminergic postsynaptic effects. QX314 is also known to interfere with cholinergic muscarinic receptors. In a previous study we found that postsynaptic action mediated by muscarinic receptor was observed only when QX314 was omitted from the recording pipette. In addition, QX-314 is also known to inhibit Gprotein-gated K + conductances and this may have also occluded dopaminergic postsynaptic effects on K + conductance reported by others.

Nanobubbles offer several advantages in targeted gene transfection. For example, nanobubbles are characterized by strong penetrating power and stable performance, which allow them to enter tumor tissues through the tumor vasculature. Therefore, in this study, we combined RNAi technology with nanotechnology by preparing nanobubbles carrying AR small interfering RNA. The physical properties of these bubbles were then assayed. Furthermore, the prepared nanobubbles were used for the siRNA transfection of AIPC cells together with ultrasonic irradiation treatment to induce the release of the siRNA transcripts. The in vitro transfection efficiency was systematically evaluated. Additionally, the anti-tumor efficacy of the nanobubbles was evaluated using imaging studies and a tumor growth inhibition assay in a mouse xenograft prostate cancer model. The results presented here provide experimental support for the use of nanobubbles carrying AR siRNA in combination with ultrasonic irradiation as a potential effective therapy for AIPC. To date, various gene therapy treatments utilizing viral vectors have been proposed. However, the use of viral vectors is a major concern because this vector system is associated with numerous safety concerns, including toxicity, immunogenicity, and tumorigenicity. Thus, the clinical application of viral vectors has been greatly restricted. The construction of an efficient, safe, and controllable in vivo gene delivery system has been a highly debated and challenging endeavor in the gene therapy community. The present study shows that ultrasound-mediated microbubble destruction may have the potential to become a new approach for targeted gene transfection. Other studies have also shown that ultrasound-destructible microbubbles not only enhanced gene transfection efficiency but were also potentially useful as a novel in vivo gene transfection vector for the delivery of any antisense oligonucleotide or DNA fragment. Thus, ultrasounddestructible microbubbles show great promise for clinical applications in the future. Despite this promise, some challenges remain in the application of conventional ultrasound-destructible microbubbles for gene or drug delivery to treat cancer. Most notably, the diameter of conventional ultrasound-destructible microbubbles, which ranges from 1-10 mm, likely prevents the bubbles from passing through the tumor vascular wall into the tumor tissues because the maximum pore size of the tumor vascular wall is 380–780 nm. This size discrepancy can affect the release and transfection of carried genes, ultimately decreasing the anti-tumor efficacy of these micro-scaled, ultrasound-destructible microbubbles. To address this size problem, researchers have proposed that nanobubbles may be a better alternative to conventional microbubbles as a gene or drug carrier for the treatment of tumors. Nanobubbles have a size of less than 1000 nm.