OLHA (Nα-oleoylhistamine) modulates activity of mouse brain histaminergic neurons.

Histaminergic (HA) neurons are located in the tuberomamillary nucleus (TMN) of the posterior hypothalamus, from where they project throughout the whole brain to control wakefulness. We examined the effects of Nα-oleoylhistamine (OLHA), a non-enzymatic condensation product of oleic acid (OLA) and histamine, on activity of mouse HA neurons in brain slices. OLHA bidirectionally modulated the firing of HA neurons. At 10 nM OLHA inhibited or had no action, whereas at 1 µM it evoked excitatory and inhibitory responses. Inhibition was not seen in presence of the histamine receptor H3 (H3R) antagonist clobenpropit and in calcium-free medium. Pre-incubation with a histamine-reuptake blocker prevented the decrease in firing by OLHA. OLHA-evoked increase in firing (EC50 ∼44 nM) was insensitive to blockers of cannabinoid 1 and 2 receptors and of the capsaicin receptor, but was significantly impaired by the peroxisome proliferator-activated receptor-alpha (PPAR-alpha) antagonist MK886, which suppressed also the rise in intracellular calcium level caused by OLHA. The OLHA-evoked excitation was mimicked by synthetic PPAR-alpha agonists (gemfibrozil and GW7647) and was abolished by the PKA inhibitor H-89. The H3R affinity (Ki) for histamine, measured in HEK293 cells with stable expression of human H3R, was higher than for OLHA (Ki: 42 vs 310 nM, respectively). Expression of PPAR-alpha was not different between TMN regions of males and females, responses to OLHA did not differ. Molecular modelling of PPAR-alpha bound to either OLHA or OEA showed similar binding energies. These findings shed light on a novel biotransformation product of histamine which may play a role in health and disease.

Deprogramming metabolism in pancreatic cancer with a bi-functional GPR55 inhibitor and biased ß2 adrenergic agonist.

Metabolic reprogramming contributes to oncogenesis, tumor growth, and treatment resistance in pancreatic ductal adenocarcinoma (PDAC). Here we report the effects of (R,S')-4'-methoxy-1-naphthylfenoterol (MNF), a GPR55 antagonist and biased ß2-adrenergic receptor (ß2-AR) agonist on cellular signaling implicated in proliferation and metabolism in PDAC cells. The relative contribution of GPR55 and ß2-AR in (R,S')-MNF signaling was explored further in PANC-1 cells. Moreover, the effect of (R,S')-MNF on tumor growth was determined in a PANC-1 mouse xenograft model. PANC-1 cells treated with (R,S')-MNF showed marked attenuation in GPR55 signal transduction and function combined with increased ß2-AR/Gαs/adenylyl cyclase/PKA signaling, both of which contributing to lower MEK/ERK, PI3K/AKT and YAP/TAZ signaling. (R,S')-MNF administration significantly reduced PANC-1 tumor growth and circulating L-lactate concentrations. Global metabolic profiling of (R,S')-MNF-treated tumor tissues revealed decreased glycolytic metabolism, with a shift towards normoxic processes, attenuated glutamate metabolism, and increased levels of ophthalmic acid and its precursor, 2-aminobutyric acid, indicative of elevated oxidative stress. Transcriptomics and immunoblot analyses indicated the downregulation of gene and protein expression of HIF-1α and c-Myc, key initiators of metabolic reprogramming in PDAC. (R,S')-MNF treatment decreased HIF-1α and c-Myc expression, attenuated glycolysis, shifted fatty acid metabolism towards ß-oxidation, and suppressed de novo pyrimidine biosynthesis in PANC-1 tumors. The results indicate a potential benefit of combined GPR55 antagonism and biased ß2-AR agonism in PDAC therapy associated with the deprogramming of altered cellular metabolism.

Analysis of the Effect of Photo and Hydrodegradation on the Surface Morphology and Mechanical Properties of Composites Based on PLA and PHI Modified with Natural Particles.

Biodegradable polymer materials are increasingly used in the packaging industry due to their good properties and low environmental impact. Therefore, the work was performed on the injection molding of the bio-based composites of polylactide (PLA) and polyhydroxyalcanates (PHI) modified with two phases: reinforcing (walnut shell flour and cellulose) and coloring (beta carotene and anthocyanin). The produced materials were subjected to wide mechanical characteristics-tensile, flexural, and fatigue tests. Additionally, the influence of photo and hydrodegradation on the change of the surface structure and mechanical properties of the composites was assessed. The addition of natural fillers contributed to the improvement of the stiffness of the tested composites. PHI composites withstood a higher number of cycles during cyclic loading, but the stress values obtained in the static tensile test were higher for PLA composites. Moreover, a clear change of color was observed after both the photo and hydrodegradation process for all tested materials; however, after the degradation processes, the filler-modified materials underwent greater discoloration. For the composites based on PHI, the type of degradation did not affect the mechanical properties. On the other hand, for PLA composites, hydrolytic degradation contributed to a higher decrease in properties-the decrease in tensile strength for unmodified PLA after photodegradation was 4%, while after hydrodegradation it was 24%.

Electron Beam-Treated Enzymatically Mineralized Gelatin Hydrogels for Bone Tissue Engineering.

Biological hydrogels are highly promising materials for bone tissue engineering (BTE) due to their high biocompatibility and biomimetic characteristics. However, for advanced and customized BTE, precise tools for material stabilization and tuning material properties are desired while optimal mineralisation must be ensured. Therefore, reagent-free crosslinking techniques such as high energy electron beam treatment promise effective material modifications without formation of cytotoxic by-products. In the case of the hydrogel gelatin, electron beam crosslinking further induces thermal stability enabling biomedical application at physiological temperatures. In the case of enzymatic mineralisation, induced by Alkaline Phosphatase (ALP) and mediated by Calcium Glycerophosphate (CaGP), it is necessary to investigate if electron beam treatment before mineralisation has an influence on the enzymatic activity and thus affects the mineralisation process. The presented study investigates electron beam-treated gelatin hydrogels with previously incorporated ALP and successive mineralisation via incubation in a medium containing CaGP. It could be shown that electron beam treatment optimally maintains enzymatic activity of ALP which allows mineralisation. Furthermore, the precise tuning of material properties such as increasing compressive modulus is possible. This study characterizes the mineralised hydrogels in terms of mineral formation and demonstrates the formation of CaP in dependence of ALP concentration and electron dose. Furthermore, investigations of uniaxial compression stability indicate increased compression moduli for mineralised electron beam-treated gelatin hydrogels. In summary, electron beam-treated mineralized gelatin hydrogels reveal good cytocompatibility for MG-63 osteoblast like cells indicating a high potential for BTE applications.

Mechanical Behavior and Morphological Study of Polytetrafluoroethylene (PTFE) Composites under Static and Cyclic Loading Condition.

The key goal of this study was to characterize polytetrafluoroethylene (PTFE) based composites with the addition of bronze particles and mineral fibers/particles. The addition of individual fillers was as follows: bronze-30-60 wt.%, glass fibers-15-25 wt.%, coke flakes-25 wt.% and graphite particles-5 wt.%. Both static and dynamic tests were performed and the obtained results were compared with the microscopic structure of the obtained fractures. The research showed that the addition of 60 wt.% bronze and other mineral fillers improved the values obtained in the static compression test and in the case of composites with 25 wt.% glass fibers the increase was about 60%. Fatigue tests have been performed for the compression-compression load up to 100,000 cycles. All tested composites show a significant increase in the modulus as compared to the values obtained in the static compression test. The highest increase in the modulus in the dynamic test was obtained for composites with 25 wt.% of glass fibers (increase by 85%) and 25 wt.% of coke flakes (increase by 77%), while the lowest result was obtained for the lowest content of bronze particles (decrease by 8%). Dynamic tests have shown that composites with "semi-spherical" particles are characterized by the longest service life and a slower fatigue crack propagation rate than in the case of the long glass fibers. In addition, studies have shown that particles with smaller sizes and more spherical shape have a higher ability to dissipate mechanical energy, which allows their use in friction nodes. On the other hand, composites with glass fiber and graphite particles can be successfully used in applications requiring high stiffness with low amplitude vibrations.

Mechanical and Hydrothermal Aging Behaviour of Polyhydroxybutyrate-Co-Valerate (PHBV) Composites Reinforced by Natural Fibres.

Biodegradable composites based on poly (3-hydroxybutyrate-co-3-hydroxyvalerate), reinforced with 7.5% or 15% by weight of wood fibers (WF) or basalt fibers (BF) were fabricated by injection molding. BF reinforced composites showed improvement in all properties, whereas WF composites showed an increase in Young's modulus values, but a drop in strength and impact properties. When compared with the unmodified polymer, composites with 15% by weight of BF showed an increase of 74% in Young's modulus and 41% in impact strength. Furthermore, the experimentally measured values of Young's modulus were compared with values obtained in various theoretical micromechanical models. The Haplin-Kardas model was found to be in near approximation to the experimental data. The morphological aspect of the biocomposites was studied using scanning electron microscopy to obtain the distribution and interfacial adhesion of the fibers. Additionally, biodegradation tests of the biocomposites were performed in saline solution at 40 °C by studying the weight loss and mechanical properties. It was observed that the presence of fibers affects the rate of water absorption and the highest rate was seen for composites with 15% by weight of WF. This is dependent on the nature of the fiber. After both the first and second weeks mechanical properties decreased slightly about 10%.

Mechanisms of N-oleoyldopamine activation of central histaminergic neurons.

Histaminergic (HA) neurons located in the tuberomamillary nucleus (TMN) of the posterior hypothalamus fire exclusively during waking and support many physiological functions. We investigated the role of the endovanilloid N-oleoyldopamine (OLDA) in TMN, where dopamine synthesis and its conjugation with oleic acid likely occur. We show that several known targets of OLDA including TRPV1 and cannabinoid receptors are expressed in HA neurons. In contrast to capsaicin, which failed to increase firing of HA neurons in TRPV1 knockout mice (TRPVI KO), OLDA was still able to induce excitation. This excitation was not sensitive to the blockade of cannabinoid receptors 1 and 2 and could result from OLDA interaction with GPR119, as the ligand of GPR119, oleoylethanolamide (OEA), also increased the firing of HA neurons. However, we ruled out this possibility as OEA- (but not OLDA-) excitation was abolished by the PPAR (peroxisome proliferator activated receptor) alpha antagonist MK886. The dopamine uptake blocker nomifensine blanked OLDA-excitation and dopamine receptor antagonists abolished the OLDA action in TRPV1 KO mice. Therefore OLDA excites HA neurons through multiple targets suggesting a central role of the histaminergic system in the behavioral stimulation seen after systemic OLDA application.

N-oleoyldopamine modulates activity of midbrain dopaminergic neurons through multiple mechanisms.

N-oleoyl-dopamine (OLDA) is an amide of dopamine and oleic acid, synthesized in catecholaminergic neurons. The present study investigates OLDA targets in midbrain dopaminergic (DA) neurons. Substantia Nigra compacta (SNc) DA neurons recorded in brain slices were excited by OLDA in wild type mice. In transient receptor potential vanilloid 1 (TRPV1) knockout (KO) mice, however, SNc DA neurons displayed sustained inhibition of firing. In the presence of the dopamine type 2 receptor (D2R) antagonist sulpiride or the dopamine transporter blocker nomifensine no such inhibition was observed. Under sulpiride OLDA slightly excited SNc DA neurons, an action abolished upon combined application of the cannabinoid1 and 2 receptor antagonists AM251 and AM630. In ventral tegmental area (VTA) DA neurons from TRPV1 KO mice a transient inhibition of firing by OLDA was observed. Thus OLDA modulates the firing of nigrostriatal DA neurons through interactions with TRPV1, cannabinoid receptors and dopamine uptake. These findings suggest further development of OLDA-like tandem molecules for the treatment of movement disorders including Parkinson's disease.

Comparison of combinatory effects of PCBs (118, 138, 153 and 180) with 17 beta-estradiol on proliferation and apoptosis in MCF-7 breast cancer cells.

We analyzed whether polychlorinated biphenyls (PCBs) interfere with the activity of 17 ß-estradiol in the proliferation and apoptosis of the MCF-7 cell line. MCF-7 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) without phenol red supplemented with 5% charcoal-treated fetal bovine serum (CD-FBS) for 3 days with 10 nM 17 ß-estradiol or 0.1 µM, 0.5 µM and 1 µM of the tested PCB congeners (118, 138, 153 and 180), or both. Cell proliferation was determined by measuring 5-bromo-2'-deoxyuridine (BrdU) incorporation, and cell apoptosis was measured by caspase-9 activity. From the PCB congeners tested, PCB138 and 153 had the highest stimulatory effects on basal cell proliferation as well as the highest inhibitory actions on basal caspase-9 activity. The proliferative and anti-apoptotic actions of PCB138 and 153 were still observed in the presence of 17 ß-estradiol, while the actions of PCB118 and 180 were reversed. In conclusion, the results of this study suggest the possibility that PCB138 and 153 contribute to the action of endogenous 17 ß-estradiol on cell proliferation and apoptosis in the breast cancer cell line MCF-7.