Sleep Loss Reduces the DNA-Binding of BMAL1, CLOCK, and NPAS2 to Specific Clock Genes in the Mouse Cerebral Cortex

We have previously demonstrated that clock genes contribute to the homeostatic aspect of sleep regulation. Indeed, mutations in some clock genes modify the markers of sleep homeostasis and an increase in homeostatic sleep drive alters clock gene expression in the forebrain. Here, we investigate a possible mechanism by which sleep deprivation (SD) could alter clock gene expression by quantifying DNA-binding of the core-clock transcription factors CLOCK, NPAS2, and BMAL1 to the cis-regulatory sequences of target clock genes in mice. Using chromatin immunoprecipitation (ChIP), we first showed that, as reported for the liver, DNA-binding of CLOCK and BMAL1 to target clock genes changes in function of time-of-day in the cerebral cortex. Tissue extracts were collected at ZT0 (light onset), −6, −12, and −18, and DNA enrichment of E-box or E'-box containing sequences was measured by qPCR. CLOCK and BMAL1 binding to Cry1, Dbp, Per1, and Per2 depended on time-of-day, with maximum values reached at around ZT6. We then observed that SD, performed between ZT0 and −6, significantly decreased DNA-binding of CLOCK and BMAL1 to Dbp, consistent with the observed decrease in Dbp mRNA levels after SD. The DNA-binding of NPAS2 and BMAL1 to Per2 was also decreased by SD, although SD is known to increase Per2 expression in the cortex. DNA-binding to Per1 and Cry1 was not affected by SD. Our results show that the sleep-wake history can affect the clock molecular machinery directly at the level of chromatin binding thereby altering the cortical expression of Dbp and Per2 and likely other targets. Although the precise dynamics of the relationship between DNA-binding and mRNA expression, especially for Per2, remains elusive, the results also suggest that part of the reported circadian changes in DNA-binding of core clock components in tissues peripheral to the suprachiasmatic nuclei could, in fact, be sleep-wake driven

Prisutnost pijavice Pontobdella muricata (Hirudinea: Piscicolidae) na hrskavičnjačama u Tirenskom moru (Srednje Sredozemlje)

This paper provides the first report of the leech, Pontobdella muricata (Linnaeus, 1758), in the Tyrrhenian Sea. The leech was found on the brown ray, Raja miraletus (Linnaeus 1758), and on the spotted ray, Raja montagui (Fowler, 1910), caught by trawling during autumn 2014. Complete sequence of 18S rRNA gene, COI mitochondrial gene and partial sequences of the mitochondrial 12S rRNA gene corroborate the determination based on morphological characteristics.U radu se iznosi prvi nalaz pijavice, Pontobdella muricata (Linnaeus, 1758), u Tirenskom moru. Pijavica je pronađena na modropjegoj raži, Raja miraletus (Linnaeus 1758) i na crnopjegoj raži, Raja montagui (Fowler, 1910), iz koćarskih lovina tijekom jeseni 2014. godine. Kompletna sekvenca 18S rRNA gena, COI mitohondrijskog gena i djelomične sekvence mitohondrijskog 12S rRNA gena potvrđuju određivanje koje se zasniva na temelju morfoloških svojstava

Altered sleep homeostasis in rev-erbα knockout mice

Study Objectives: The nuclear receptor REV-ERBα is a potent, constitutive transcriptional repressor critical for the regulation of key circadian and metabolic genes. Recently, REV-ERBα's involvement in learning, neurogenesis, mood, and dopamine turnover was demonstrated suggesting a specific role in central nervous system functioning. We have previously shown that the brain expression of several core clock genes, including Rev-erbα, is modulated by sleep loss. We here test the consequences of a loss of REV-ERBα on the homeostatic regulation of sleep.Methods: EEG/EMG signals were recorded in Rev-erbα knockout (KO) mice and their wild type (WT) littermates during baseline, sleep deprivation, and recovery. Cortical gene expression measurements after sleep deprivation were contrasted to baseline.Results: Although baseline sleep/wake duration was remarkably similar, KO mice showed an advance of the sleep/wake distribution relative to the light-dark cycle. After sleep onset in baseline and after sleep deprivation, both EEG delta power (1–4 Hz) and sleep consolidation were reduced in KO mice indicating a slower increase of homeostatic sleep need during wakefulness. This slower increase might relate to the smaller increase in theta and gamma power observed in the waking EEG prior to sleep onset under both conditions. Indeed, the increased theta activity during wakefulness predicted delta power in subsequent NREM sleep. Lack of Rev-erbα increased Bmal1, Npas2, Clock, and Fabp7 expression, confirming the direct regulation of these genes by REV-ERBα also in the brain.Conclusions: Our results add further proof to the notion that clock genes are involved in sleep homeostasis. Because accumulating evidence directly links REV-ERBα to dopamine signaling the altered homeostatic regulation of sleep reported here are discussed in that context

First Report of Root and Basal Stem Rot Caused by Phytophthora cryptogea and P. inundata on Dwarf Banana in Italy

In Sicily (southern Italy) local cultivars of dwarf banana (Musa acuminata) are cultivated for edible fruit and as ornamental plants. During the summer of 2015, in an ornamental nursery of Aci San Filippo (Catania province), eastern Sicily, ten out of forty mature plants of dwarf banana grown in the field showed leaf chlorosis, wilt and sudden collapse of the entire plant associated with root and basal stem rot. Two Phytophthora species (overall 24 and 22 isolates, respectively) were consistently recovered directly from rotted roots and stems on BNPRA-HMI selective medium (Masago et al. 1977). Pure cultures of both species were obtained by single-hypha isolations. The first species formed slight petaloid colonies on potato dextrose agar (PDA) and slightly fluffy colonies on V-8 juice agar (V8A). It grew between 2 and 30°C, with an optimum of 25°C. On V8A discs flooded with non-sterile soil extract this species produced persistent, ovoid to obpyriform, non-papillate, internally proliferating sporangia (35 ..

Ferrous iron- and ammonium-rich diffuse vents support habitat-specific communities in a shallow hydrothermal field off the Basiluzzo Islet (Aeolian Volcanic Archipelago)

Ammonium- and Fe(II)-rich fluid flows, known from deep-sea hydrothermal systems, have been extensively studied in the last decades and are considered as sites with high microbial diversity and activity. Their shallow-submarine counterparts, despite their easier accessibility, have so far been under-investigated, and as a consequence, much less is known about microbial communities inhabiting these ecosystems. A field of shallow expulsion of hydrothermal fluids has been discovered at depths of 170-400 meters off the base of the Basiluzzo Islet (Aeolian Volcanic Archipelago, Southern Tyrrhenian Sea). This area consists predominantly of both actively diffusing and inactive 1-3 meters-high structures in the form of vertical pinnacles, steeples and mounds covered by a thick orange to brown crust deposits hosting rich benthic fauna. Integrated morphological, mineralogical, and geochemical analyses revealed that, above all, these crusts are formed by ferrihydrite-type Fe3+ oxyhydroxides. Two cruises in 2013 allowed us to monitor and sampled this novel ecosystem, certainly interesting in terms of shallow-water iron-rich site. The main objective of this work was to characterize the composition of extant communities of iron microbial mats in relation to the environmental setting and the observed patterns of macrofaunal colonization. We demonstrated that iron-rich deposits contain complex and stratified microbial communities with a high proportion of prokaryotes akin to ammonium- and iron-oxidizing chemoautotrophs, belonging to Thaumarchaeota, Nitrospira, and Zetaproteobacteria. Colonizers of iron-rich mounds, while composed of the common macrobenthic grazers, predators, filter-feeders, and tube-dwellers with no representatives of vent endemic fauna, differed from the surrounding populations. Thus, it is very likely that reduced electron donors (Fe2+ and NH4+) are important energy sources in supporting primary production in microbial mats, which form a habitat-specific trophic base of the whole Basiluzzo hydrothermal ecosystem, including macrobenthic fauna

Environmental and Agro-Economic Sustainability of Olive Orchards Irrigated with Reclaimed Water under Deficit Irrigation

This study explores the effects of the adoption of reclaimed water (RW) as source of irrigation in conjunction with the application of deficit irrigation strategies in an olive orchard (different genotypes) located within the “Valle dei Margi” farmhouse (Eastern Sicily). Specifically, the RW was obtained in situ by treating the wastewater coming from the farmhouse throughout a treatment wetland system (TW). The effects of RW on crop water status (CWS) was assessed by conducting plant-based measurements (i.e., leaf water potential, Ψ, and leaves relative water content, RWC) and determining satellite-based biophysical indicators. An economical and environmental evaluation of the proposed sustainable irrigation practices was carried out by using the life cycle assessment (LCA) approach.The RW quality showed high variability due to fluctuations in the number of customers at the farmhouse during the Covid-19 pandemic period. However, high removal efficiency of the overall TW was reached even if the RW quality did not always accomplish with the limits of the Italian regulations. A strong impact in the variation of Ψ was observed among the olive orchard under the different water regimes, evidencing how CWS performances are greatly conditioned by the genotype. However, no differences in leaves RWC and in satellite-based biophysical indicators were detected, despite the severe water deficit imposed (i.e., 50% of irrigation water reduction). Finally, the results of the LCA analysis underlined that the use of RW may permit to obtain important gains both in economic and environmental terms, thus representing a valid strategy for the olive cultivation

Microbial community of the deep-sea brine Lake <em>Kryos </em>seawater-brine interface is active below the chaotropicity limit of life as revealed by recovery of mRNA

Within the complex of deep, hypersaline anoxic lakes (DHALs) of the Mediterranean Ridge we identified a new, unexplored DHAL and named it "Lake Kryos" after a nearby depression. This lake is filled with MgCl2-rich, athalassohaline brine (salinity >470 practical salinity units), presumably formed by the dissolution of Messinian bischofite. Compared to the DHAL Discovery, it contains elevated concentrations of kosmotropic sodium and sulfate ions, which are capable of reducing the net chaotropicily of MgCl2-rich solutions. The brine of Lake Kryos may therefore be biologically permissive at MgCl2 concentrations previously considered incompatible with life. We characterized the microbiology of the seawater-Kryos brine interface and managed to recover mRNA from the 2.27-3.03 M MgCl2 layer (equivalent to 0.747-0.631 water-activity) thereby expanding the established chaotropicity window-for-life. The primary bacterial taxa present there were KB1 candidate division and DHAL-specific group of organisms, distantly related to Desulfohalobium. Two euryarchaeal candidate divisions MSBL1 and HC1, detected in minority in the overlaying layers, accounted for more than 85% of the rRNA-containing archaeal clones analyzed in 2.27-3.03 M MgCl2 layer. These findings shed light on the plausibility of life in highly chaotropic environments, geochemical windows for microbial extremophiles, and have implications for habitability elsewhere in the Solar System

Fermi Large Area Telescope Third Source Catalog

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