Category: Kinase Inhibitor Library

Based on the above observations, we hypothesized that global knockout of FTO would lead to an increased sympathetic excitation of the heart. To test this hypothesis, sympathetic and parasympathetic influences on the heart were assessed during resting and stress conditions via time- and frequency-domain analysis of heart rate variability. We also evaluated whether supposed cardiac sympathetic hyperactivity in FTO knockout mice was associated with increased arrhythmia vulnerability, and investigated potential mediating mechanisms at the electrical and structural level of the heart. A previous study in a mouse model bearing a missense mutation in the FTO gene provided preliminary evidence linking FTO deficiency to increased sympathetic nervous system activity. However, to the best of our knowledge, this study is the first description of the effects of global knockout of FTO on cardiac function and its autonomic neural regulation. In resting conditions, FTO deficient mice were characterized by higher heart rate values than wild-type mice, both during the active and inactive phase of the daily cycle. Likewise, we found signs of elevated body temperature in mice lacking the FTO gene. Clearly, differences in heart rate and body temperature may have been determined by different levels of somatomotor activity, which indeed resulted significantly higher in knockout mice during the active phase of the daily cycle. However, given that heart rate was consistently higher in FTO deficient mice even when somatomotor activity levels were not greater, we believe that autonomic mechanisms concurred to determine higher heart rate in these animals. Supporting this view, HRV analysis revealed that knockout mice were characterized by a lower vagal modulation of heart rate than wild-type counterparts. In addition, the fact that FTO deficient mice showed higher LF to HF ratio is suggestive of a larger contribution of the sympathetic modulation of heart rate in mice lacking the FTO gene. Signs that link FTO deficiency to increased cardiac sympathetic drive were evident during stress conditions. Following the injection of saline and during the restraint test, stress-induced tachycardia was greater in knockout mice, despite similar low levels of vagal modulation between the two groups. This is a clear indication of a larger sympathetic modulation of heart rate in FTO deficient mice, which consequently resulted in a shift of the sympatho-vagal balance towards an exaggerated sympathetic prevalence. Given that high expression of FTO is seen in the paraventricular and dorsomedial nuclei of the hypothalamus, which represent important brain centers for the regulation of autonomic function, especially during stress response, we hypothesize a role of FTO in these brain areas in modulating sympathetic outflow to the heart. Previous studies have demonstrated that b-adrenergic agonists increase the inward sodium current in cardiomyocytes.

A better understanding of the molecular basis of adjuvanticity could also promote the rational design of successful vaccines by supplementing them with the appropriate adjuvants or combinations thereof. Here, we studied the effector functions of the candidate adjuvant c-di-AMP on different immune cells in vitro as well as in vivo. We asked which cell types undergo activation and which cellular processes and molecules are involved. We previously reported that c-di-AMP promotes antigen specific T cell proliferation. C-di-AMP does not seem to directly exert this effect on T cells, thus we concluded that the observed T cell activation is mediated by other immune cell types. This was also suggested by previous in vitro studies showing that DCs which were antigen-loaded in the presence of c-di-AMP were more efficient at promoting the activation of antigen-specific T cells from TCR transgenic mice. Accordingly, we examined the response of different types of APCs to c-di-AMP. We showed that treatment with c-di-AMP induces the up-regulation of the expression of MHC class II as well as T cell co-stimulatory molecules on the surface of both murine and human DCs and MWs in vitro. These in vitro findings were further confirmed by ex vivo studies in which murine DCs and MWs were analyzed after i. n. cdi-AMP application. 24 h after i. n. administration especially DCs and not so much MWs were activated by c-di-AMP in the lymphoid compartments. Although early DC antigen presenting activity in the draining CLN was described for i. n. immunization experiments with viral antigens, our adjuvant candidate c-di-AMP alone did not notably induce T cell stimulatory molecules on APCs of the CLN after 24 h. The more pronounced DC activation was observed in the NALT, the lymphoid tissue directly associated with the site of administration, and also in the MLN. The early DC response in the MLN is in line with several studies reporting a similar observation upon i. n. immunization with model antigens and adjuvants as well as upon i. n. infection with influenza virus. We also demonstrated that murine DCs and MWs/monocytes/granulocytes respond to in vivo administration of c-di-AMP by the upregulation of IFN-b expression. Taken together, we observed the effects of c-di-AMP on its target cells on two different aspects of immune regulation. First, we found up-regulation of CD80 and CD86 on the surface of mouse and human DC subsets treated with c-di-AMP. These molecules are known to interact with CD28 on the T cell surface to deliver co-stimulatory signals after TCR engagement with the antigen presenting MHC molecule. This costimulation represents an important regulator of T cell immune tolerance toward activation in response, for example by promoting T cell proliferation and differentiation. Nevertheless, both CD80 and CD86 are also ligands for the T cell surface molecule CTLA-4 which facilitates negative T cell activation signaling.

Pretreatment technologies are necessarily applied to lignocellulosic material to decrease recalcitrance and to improve the yield of fermentable sugars. Many pretreatment methods have been proposed and investigated, such as alkaline, steam explosion, ammonia fiber expansion, organic solvent, dilute acid, and so on. Different pretreatment methods have different mechanisms, for example, they can decrease cellulose crystallinity and/or the polymerization degree, increase accessible surface areas. However, economic and environmental requirements limit the applicability of these methods. An effective pretreatment strategy should also minimize carbohydrate degradation and the production of enzyme inhibitors and toxic products for fermenting microorganisms. One of the most promising pretreatment processes for lignocelluloses material is liquid hot water pretreatment. Some studies have been conducted on the mechanisms of LHW pretreatment. However, different biomass types have different structures and show different reaction mechanisms. In the process of ethanol production from lignocellulosic material, enzymatic hydrolysis and fermentation can be performed separately or simultaneously. In separate hydrolysis and fermentation, these two steps are separate, and SHF can coordinate the inconsistent contradiction between the temperatures for enzymatic hydrolysis and fermentation. In simultaneous saccharification and fermentation, both steps occur in a single bioreactor where the glucose formed is rapidly converted to ethanol by the yeast. However, solid loading is limited by the higher effective mixing and high viscosity of the system in the SSF process. Semi-SSF of ethanol production is an operating mode between SSF and SHF. S-SSF consists of two phases, namely, pre-hydrolysis and SSF. To increase substrate concentration, fed-batch S-SSF process was carried out. Fed-batch S-SSF for ethanol production showed that higher substrate concentration and higher ethanol yield can be obtained compared with S-SSF and SSF when a suitable pre-hydrolytic period is selected. In our previous study, LHW pretreatment was applied to corn stover to test the efficiency of enzymatic hydrolysis, and cellulose conversion rates of almost 100% were obtained. In the present work, corn stover samples were subjected to a combination of LHW pretreatment and fed-batch S-SSF to obtain higher ethanol concentration and yield. The effects of different impact factors on the fermentation digestibility of LHW-pretreated corn stover in S-SSF and fed-batch S-SSF are discussed, and the chemical structures and morphological characteristics of corn stover during LHW pretreatment were presented. A double-blind, randomized, placebo-controlled intervention study recently revealed, that the daily consumption of monomeric and oligomeric flavanols derived from seeds of grapes for 8 weeks accomplish a vascular health benefit in male smokers.

the understanding of cellular and molecular aspects of dopaminergic and glutamatergic receptor interaction, little is known about the interaction between glutamatergic and dopaminergic function in the nAcb during postnatal development. Recent studies have shown that there are important changes in glutamatergic neurotransmission from the day of birth throughout adulthood. Of particular interest is a change in the amplitude of the NMDA receptor-mediated excitatory postsynaptic current to AMPA receptor-mediated EPSC ratio which reaches its maximum toward the end of the second postnatal week and decreases after that until adulthood. In addition to changes in glutamatergic neurotransmission, it has also been found that postnatal development is accompanied by changes in the dopaminergic innervation as well as the density and expression of dopaminergic receptors. The effects of dopamine on glutamatergic neurotransmission have been previously studied. Some studies reported that the activation of D1 receptors enhanced NMDA receptor-mediated EPSCs in dorsal striatal slices, while others reported that D1 receptor agonists attenuated NMDA EPSCs in MS striatal neurons in culture. In the nAcb, some investigators reported that DA or D1 receptor agonists potentiate NMDA receptor-mediated EPSCs in slices, while others reported no significant modulatory effects of DA on NMDA receptor-mediated EPSCs but the presynaptic inhibitory effect of DA on EPSCs was only determined on AMPA/KA receptormediated EPSCs in the nAcb. A substantial effect of DA on pharmacologically isolated NMDA and AMPA/KA receptormediated EPSCs remains to be determined. In a previous study, we showed that acetylcholine presynaptically modulated AMPA/KA and NMDA receptors mediated EPSCs in a parallel fashion in the nAcb during postnatal development. In an effort to clarify how the NMDA and AMPA/KA EPSCs might be affected in the nAcb by dopaminergic innervation, we investigated the effect of DA on NMDA and AMPA/KA excitatory synaptic transmission in this region. Our results demonstrate that DA depresses the excitatory input onto MS neurons by the activation of presynaptic D1-like receptors. While DA depressed the elicited AMPA/KA receptor-mediated EPSCs in MS neurons by 40% of the control, DA almost completely abolished NMDA receptor-mediated EPSC. The effects of DA on glutamatergic EPSCs remained constant throughout the first 3 postnatal weeks. We found that DA inhibited glutamatergic EPSCs in MS neurons recorded in an in vitro slice preparation. Whereas both AMPA/KA and NMDA receptors-mediated components of the EPSCs were significantly inhibited by DA, the inhibition of the NMDA receptor-mediated component was much more pronounced than that of the AMPA/KA receptor-mediated component. Pharmacological evidence suggests that dopaminergic inhibitory effects were mediated by the activation of presynaptic D1-like receptors and that D2-like receptors were not involved.

It should be noted that according to a recently published study, there are at least 1849 human transcripts that can be translated by alternate ATG initiation codons. Its 62 amino-terminal portion bears no similarity with the previously characterized RNase k-01 isoform or other human proteins, whereas the 63–134 region is absolutely identical to the major part of RNase k-01. Based on this amino-acid sequence, we managed to produce and purify an RNase k-02 specific polyclonal antibody. By the means of our newly developed antibody we were able to demonstrate the biogenesis of the RNase k-02 protein resulting from the RNase k02 mRNA translation in human cells. This alternative protein isoform bears a cytoplasmic topology. To our knowledge, this is the first instance of a human protein isoform encoded by a D4 subtly alternatively spliced transcript. The finding that RNase k-02 protein isoform is detected only in the detergent-insoluble fraction of cell extracts after Triton X-114 phase separation is in agreement with the predicted highly hydrophobic nature of this protein. This fact, in combination with its cytoplasmic localization, supports the hypothesis that RNase k-02 could participate in the formation of macromolecular complexes in vivo or localize in membranic structures such as the endoplasmic reticulum. It is well established that prostate cancer is dependent on androgens. Therefore, androgen deprivation has been the main treatment for advanced prostate cancer. With this therapy, however, the disease rapidly progresses to androgen-independent prostate cancer in most patients. Currently, there are no effective long-term therapies for AIPC. Developing new treatment strategies for AIPC remains attractive but is a challenging problem in clinical oncology. Previous studies have shown that the growth of AIPC is dependent on the androgen receptor. Thus, blocking AR expression has great potential for the treatment of AIPC. Studies have demonstrated that suppressing AR expression with RNA interference technology is an effective way to inhibit the growth of prostate cancer cells, indicating that this method may have the potential to overcome hurdles associated with AIPC treatment. In our previous studies, AR double-stranded RNA with a high specificity for AR genes was designed to block the expression of AR in AIPC cells and to inhibit cell growth. However, this type of gene therapy approach is difficult to apply in a clinical setting due to low gene transfection efficiency. One of the key reasons for low transfection efficiency is the lack of an effective, noninvasive in vivo targeted gene delivery system. In recent years, ultrasound-destructible microbubbles have been shown to be a promising method for gene therapy. Indeed, ultrasound-mediated microbubble destruction can not only improve gene transfection efficiency but can also be used for the tissue-specific delivery of therapeutic agents.