Differential Actions of Orexin Receptors in Brainstem Cholinergic and Monoaminergic Neurons Revealed by Receptor Knockouts: Implications for Orexinergic Signaling in Arousal and Narcolepsy

Orexin neuropeptides influence multiple homeostatic functions and play an essential role in the expression of normal sleep-wake behavior. While their two known receptors (OX1 and OX2) are targets for novel pharmacotherapeutics, the actions mediated by each receptor remain largely unexplored. Using brain slices from mice constitutively lacking either receptor, we used whole-cell and Ca(2+) imaging methods to delineate the cellular actions of each receptor within cholinergic [laterodorsal tegmental nucleus (LDT)] and monoaminergic [dorsal raphe (DR) and locus coeruleus (LC)] brainstem nuclei-where orexins promote arousal and suppress REM sleep. In slices from OX(-/-) 2 mice, orexin-A (300 nM) elicited wild-type responses in LDT, DR, and LC neurons consisting of a depolarizing current and augmented voltage-dependent Ca(2+) transients. In slices from OX(-/-) 1 mice, the depolarizing current was absent in LDT and LC neurons and was attenuated in DR neurons, although Ca(2+)-transients were still augmented. Since orexin-A produced neither of these actions in slices lacking both receptors, our findings suggest that orexin-mediated depolarization is mediated by both receptors in DR, but is exclusively mediated by OX1 in LDT and LC neurons, even though OX2 is present and OX2 mRNA appears elevated in brainstems from OX(-/-) 1 mice. Considering published behavioral data, these findings support a model in which orexin-mediated excitation of mesopontine cholinergic and monoaminergic neurons contributes little to stabilizing spontaneous waking and sleep bouts, but functions in context-dependent arousal and helps restrict muscle atonia to REM sleep. The augmented Ca(2+) transients produced by both receptors appeared mediated by influx via L-type Ca(2+) channels, which is often linked to transcriptional signaling. This could provide an adaptive signal to compensate for receptor loss or prolonged antagonism and may contribute to the reduced severity of narcolepsy in single receptor knockout mice

Mesopore-modified mordenites as catalysts for catalytic pyrolysis of biomass and cracking of vacuum gasoil processes

[EN] Mesopore-modified mordenite zeolitic materials with different Si/Al ratios have been repared and tested in the biomass pyrolysis and catalytic cracking of vacuum gasoil. Alkaline treatment was carried out to generate mesoporosity. Severity of alkaline treatment was found to be of paramount importance to tune the generated mesoporosity, while it significantly affected the crystallinity of treated mordenites. It was moreover observed that the alkaline treatment selectively extracted Si decreasing the Si/Al ratio of treated samples. Catalytic activity of parent and alkaline treated mordenites was studied in the pyrolysis of biomass. All zeolitic based materials produced less amounts of bio-oil but of better quality (lowering the oxygen content from ∼40% to as much as 21%) as compared to the non-catalytic pyrolysis experiments. On the other hand, it was found that the combination of mesopore formation and high surface area after alkaline treatment of the mordenite with a high Si/Al ratio resulted in the enhancement of its catalytic activity, despite the reduction of its acidity. The increment of the decarboxylation and dehydration reactions, combined with a reduction of carbon deposition on the catalyst, resulted in a remarkable decrease in the oxygen content in the organic fraction and therefore, resulted in a superior quality liquid product. Alkaline treated mordenites were additionally acid treated targeting dealumination and removal of the extra framework debris, thus generating mesopore-modified mordenite samples with stronger acid sites and higher total acidity, as candidate catalysts for catalytic cracking of vacuum gasoil. Desilicated and especially desilicated and dealuminated mordenites exhibited the highest activity and selectivity towards LCO with the best olefinicity in gases and higher bottoms conversion. Therefore, an optimized desilicated dealuminated mordenite additive could be an interesting candidate as a component of the FCC catalyst for a high LCO yield.The financial support of this work by the ACENET COMMON INITIATIVE HECABIO: "HEterogeneous CAtalysis for the Conversion of Solid BIOmass into Renewable Fuels and Chemicals" Project ACE.07.026 is gratefully acknowledged.Stefanidis, S.; Kalogiannis, K.; Iliopoulou, EF.; Lappas, AA.; Martínez Triguero, LJ.; Navarro Ruiz, MT.; Chica, A.... (2013). Mesopore-modified mordenites as catalysts for catalytic pyrolysis of biomass and cracking of vacuum gasoil processes. Green Chemistry. 15(6):1647-1658. doi:10.1039/c3gc40161hS1647165815

Incident Handling for Healthcare Organizations and Supply-Chains

Healthcare ecosystems form a critical type of infrastructures that provide valuable services in today societies. However, the underlying sensitive information is also of interest of malicious entities around the globe, with the attack volume being continuously increasing. Safeguarding this complex computerized setting constitutes a major challenge for the involved organizations. This paper presents an incident handling system for healthcare organizations and their supply-chain. The proposed approach utilizes swarm intelligence in order to assess the current security posture in a continuous basis and respond to attacks in real-time. The overall solution is based on the related NIST 800.61 standard and implements the operations of i) preparation, ii) detection and analysis, iii) containment, eradication, and recovery, and iv) post-incident activity. The system is developed under the EU funded project AI4HEALTHSEC and is applied in the relevant healthcare pilots

Variation in protein metabolism biomarkers during the transition period and associations with health, colostrum quality, reproduction, and milk production traits in Holstein cows

The aims of this study were to assess a) the variation of protein metabolism biomarkers and of factors affecting them during the transition period, b) the association of each biomarker with skeletal muscle reserves and their changes, and c) the association of these biomarkers with postpartum health, colostrum quality, reproduction, and milk production. For this purpose, 238 multiparous Holstein cows from 6 herds were used in a prospective cohort study. Plasma concentrations of 3-methylhistidine (3-MH) and 1-methylhistidine (1-MH) and serum concentrations of total protein (TP), albumin (ALB), urea nitrogen (BUN) and creatinine (SCR) were determined for each cow at -21d; -7d; 7d; 21d and 28d relative to calving. Clinical diseases were recorded during the first 28 d post-calving, and presence of subclinical ketosis (scKET) was investigated at 7d and 21d. Colostrum quality was estimated by Brix refractometry. Reproduction data by 150 d-in-milk (DIM) and milk production records were also available. Linear mixed models including the fixed effects of time point, herd, parity, body condition score (-21d), duration of dry period and post-parturient diseases were fitted to assess the variation in each biomarker's concentration. The association between the biomarkers' concentration during the pre-partum period with the odds for each post-parturient disease and for a combined trait (CD_1-28), defined as the presence of at least one clinical condition during the first 28 d after calving, were assessed with separate binary logistic models for time points -21d and -7d. The relationship of each biomarker's concentration with longissimus dorsi thickness (LDT) and the changes in LDT (ΔLDT) was assessed with pairwise correlations. Separate general linear models were used to assess the association of each biomarker with colostrum Brix values and milk production traits. Finally, the associated hazard for 1st artificial insemination (AI) and for pregnancy by 150 DIM (PREG_150DIM) was assessed with Cox proportional hazard models, while odds for pregnancy to the 1st AI (PREG_1stAI) were assessed with binary logistic models. 3-methylhistidine was affected mainly by herd, time points and their interaction. Higher 3-MH was associated with increased odds for metritis and CD_1-28, increased hazard for PREG_150 DIM and with increased milk production. 1-methylhistidine was affected mainly by herd, scKET and occurrence of displaced abomasum. Higher 1-MH was associated with better colostrum quality, increased odds for scKET, increased hazard for 1st AI by 150 DIM and with decreased milk production. Both 3-MH and 1-MH were weakly to moderately negatively correlated with LDT and moderately to strongly negatively correlated to ΔLDT at the corresponding time-periods. Additionally, higher TP was associated with increased odds for metritis and CD_1-28 and increased milk production, while higher ALB was associated with increased odds for scKET and increased milk production. Moreover, higher BUN was associated with decreased odds for scKET, increased odds for PREG_1stAI and increased milk production. Higher SCR was associated with decreased odds for retained fetal membranes, metritis, and CD_1-28. Peri-parturient protein metabolism is significantly associated with postpartum health, colostrum quality, reproduction, and milk production; mechanisms involved require further investigation

Exploring the Biochemical Methane Potential of Wholesale Market Waste from Jordan and Tunisia for a Future Scale-Up of Anaerobic Digestion in Amman and Sfax

3 Figuras.-- 2 TablasLocal open markets, trading fruits and vegetables, are widespread in Mediterranean countries, such as Tunisia and Jordan, producing large amounts of organic waste. Applying an anaerobic digestion process on this substrate makes it crucial to evaluate the waste mixture composition and seasonal variability properly. In this study, after defining an average composition of the fruit and vegetable waste (FVW) mixture produced in Sfax (Tunisia) and Amman (Jordan) in three seasonal intervals (autumn–winter, spring, and summer), the biochemical methane potential (BMP) of an artificially created FVW mixture was individually determined by three European institutions located in Spain, Italy, and Greece. The average BMP from all three seasons and laboratories was 286 ± 52 NmL CH4 g CODadded−1, close to the theoretical maximum yield of 350 NmL CH4 g CODadded−1, indicating a high biodegradability of the waste. Τhe biochemical methane yields of the spring mixtures were not statistically different across the three labs. The most significant differences among the BMP results were obtained for the autumn/winter and the summer mixtures used in Spain, likely due to the variety or ripeness of fruits and vegetables collected in the local markets. In the other two labs in Italy and Greece, no statistical difference was observed for the BMPs of the three season mixtures within the same lab. Therefore, not a critical difference in the biodegradability of such FVW is expected along the different seasons, indicating that the operation of a full-scale digester over a whole year would constantly benefit from the supplementation of a high biochemical methane potential feedstock. Graphical Abstract: [Figure not available: see fulltext.]This work was funded by the project entitled “Employing circular economy approach for OFMSW management within the Mediterranean countries – CEOMED” number A_B.4.2_0058, funded under the ENI CBC MED 2014–2020 programmePeer reviewe

Cholinergic Modulation of Narcoleptic Attacks in Double Orexin Receptor Knockout Mice

To investigate how cholinergic systems regulate aspects of the sleep disorder narcolepsy, we video-monitored mice lacking both orexin (hypocretin) receptors (double knockout; DKO mice) while pharmacologically altering cholinergic transmission. Spontaneous behavioral arrests in DKO mice were highly similar to those reported in orexin-deficient mice and were never observed in wild-type (WT) mice. A survival analysis revealed that arrest lifetimes were exponentially distributed indicating that random, Markovian processes determine arrest lifetime. Low doses (0.01, 0.03 mg/kg, IP), but not a high dose (0.08 mg/kg, IP) of the cholinesterase inhibitor physostigmine increased the number of arrests but did not alter arrest lifetimes. The muscarinic antagonist atropine (0.5 mg/kg, IP) decreased the number of arrests, also without altering arrest lifetimes. To determine if muscarinic transmission in pontine areas linked to REM sleep control also influences behavioral arrests, we microinjected neostigmine (50 nl, 62.5 µM) or neostigmine + atropine (62.5 µM and 111 µM respectively) into the nucleus pontis oralis and caudalis. Neostigmine increased the number of arrests in DKO mice without altering arrest lifetimes but did not provoke arrests in WT mice. Co-injection of atropine abolished this effect. Collectively, our findings establish that behavioral arrests in DKO mice are similar to those in orexin deficient mice and that arrests have exponentially distributed lifetimes. We also show, for the first time in a rodent narcolepsy model, that cholinergic systems can regulate arrest dynamics. Since perturbations of muscarinic transmission altered arrest frequency but not lifetime, our findings suggest cholinergic systems influence arrest initiation without influencing circuits that determine arrest duration