Single cell tracing of Pomc neurons reveals recruitment of 'Ghost' subtypes with atypical identity in a mouse model of obesity.

The hypothalamus contains a remarkable diversity of neurons that orchestrate behavioural and metabolic outputs in a highly plastic manner. Neuronal diversity is key to enabling hypothalamic functions and, according to the neuroscience dogma, it is predetermined during embryonic life. Here, by combining lineage tracing of hypothalamic pro-opiomelanocortin (Pomc) neurons with single-cell profiling approaches in adult male mice, we uncovered subpopulations of 'Ghost' neurons endowed with atypical molecular and functional identity. Compared to 'classical' Pomc neurons, Ghost neurons exhibit negligible Pomc expression and are 'invisible' to available neuroanatomical approaches and promoter-based reporter mice for studying Pomc biology. Ghost neuron numbers augment in diet-induced obese mice, independent of neurogenesis or cell death, but weight loss can reverse this shift. Our work challenges the notion of fixed, developmentally programmed neuronal identities in the mature hypothalamus and highlight the ability of specialised neurons to reversibly adapt their functional identity to adult-onset obesogenic stimuli.

Neocortex saves energy by reducing coding precision during food scarcity.

Information processing is energetically expensive. In the mammalian brain, it is unclear how information coding and energy use are regulated during food scarcity. Using whole-cell recordings and two-photon imaging in layer 2/3 mouse visual cortex, we found that food restriction reduced AMPA receptor conductance, reducing synaptic ATP use by 29%. Neuronal excitability was nonetheless preserved by a compensatory increase in input resistance and a depolarized resting potential. Consequently, neurons spiked at similar rates as controls but spent less ATP on underlying excitatory currents. This energy-saving strategy had a cost because it amplified the variability of visually-evoked subthreshold responses, leading to a 32% broadening of orientation tuning and impaired fine visual discrimination. This reduction in coding precision was associated with reduced levels of the fat mass-regulated hormone leptin and was restored by exogenous leptin supplementation. Our findings reveal that metabolic state dynamically regulates the energy spent on coding precision in neocortex.

Hypothalamic bile acid-TGR5 signaling protects from obesity.

Bile acids (BAs) improve metabolism and exert anti-obesity effects through the activation of the Takeda G protein-coupled receptor 5 (TGR5) in peripheral tissues. TGR5 is also found in the brain hypothalamus, but whether hypothalamic BA signaling is implicated in body weight control and obesity pathophysiology remains unknown. Here we show that hypothalamic BA content is reduced in diet-induced obese mice. Central administration of BAs or a specific TGR5 agonist in these animals decreases body weight and fat mass by activating the sympathetic nervous system, thereby promoting negative energy balance. Conversely, genetic downregulation of hypothalamic TGR5 expression in the mediobasal hypothalamus favors the development of obesity and worsens established obesity by blunting sympathetic activity. Lastly, hypothalamic TGR5 signaling is required for the anti-obesity action of dietary BA supplementation. Together, these findings identify hypothalamic TGR5 signaling as a key mediator of a top-down neural mechanism that counteracts diet-induced obesity.

Chemometric Discrimination of the Varietal Origin of Extra Virgin Olive Oils: Usefulness of 13C Distortionless Enhancement by Polarization Transfer Pulse Sequence and 1H Nuclear Magnetic Resonance Data and Effectiveness of Fusion with Mid-Infrared Spectroscopy Data.

The label authentication of monovarietal extra virgin olives is of great relevance from a socio-economical point of view. This work aims to gain insights into the prediction of the varietal origin of extra virgin olive oil (EVOO) samples obtained from single olive cultivars, French cultivars Olivière, Salonenque, and Tanche and Portuguese cultivars Blanqueta, Carrasquenha, and Galega Vulgar, collected in 2016-2017 and 2017-2018 harvest seasons. To pursue this study, spectroscopic approaches based on one-dimensional nuclear magnetic resonance (1D NMR) spectroscopy, namely, 1H and 13C NMR distortionless enhancement by polarization transfer (DEPT) 45 pulse sequence, and Fourier transform mid-infrared spectroscopy (FT-MIR) are used in combination with partial least square discriminant analysis (PLS1-DA). The results obtained by PLS1-DA models using 1H and 13C NMR DEPT 45 data are compared to those of PLS1-DA models using MIR data. The application of a control chart method allows for the optimization of the interpretation of the PLS1-DA results, and an efficient two-step strategy is proposed to improve the discrimination of the six studied cultivars. Then, NMR and MIR data are combined by either a mid- or high-level data fusion approach to further improve the discrimination. The models are also tested on samples from other cultivars to check their ability to reject varieties that were not considered in the calibration process.

Chiroptical fingerprints to characterize lavender and lavandin essential oils.

Polarimetric HPLC detector was used to acquire chromatograms for lavender/lavandin essentials oils (EOs) on different chiral stationary phases. Amylose tris-(3,5-dichloro-phenylcarbamate) immobilized on silica allowed the best separation of the numerous chiral phytomarkers and was thus used to obtain a chiroptical fingerprint for 158 samples of French lavender/lavandin EOs. Samples from different varieties (Abrial, Fine, Grosso, Maillette, Matherone, Sumian and Super) were collected over three crop years (2012, 2013 and 2014), from four different locations in France (``Alpes-de-Haute-Provence'', ``Ardèche'', ``Drôme'' and ``Vaucluse'') to take into account seasonal and geographic origin variations. The combination of the polarimetric fingerprints and chemometrics was tested for varietal discrimination. We assessed the ability to discriminate the EOs samples and their variety by partial least squares-discriminant analysis (PLS-DA). This case study showed that liquid chromatography with polarimetric detector in tandem with chemometric analysis was efficient to differentiate the varietal origins of French lavender/lavandin EOs.

Pre-treatment of a sugarcane bagasse-based substrate prior to saccharification: Effect of coffee pulp and urea on laccase and cellulase activities of Pycnoporus sanguineus.

Production of second-generation bioethanol uses lignocellulose from agricultural by-products such as sugarcane bagasse (SCB). A lignocellulose pre-treatment is required to degrade lignin, ensuring further efficient saccharification. Two experimental designs were set up to define culture conditions of Pycnoporus sanguineus in mesocosms to increase laccase activities and thus delignification. The first experimental design tested the effect of phenolic complementation (via coffee pulp) and the use of urea as a simple nitrogen source and the second defined more precisely the percentages of coffee pulp and urea to enhance delignification. The responses measured were: lignocellulolytic activities, laccase isoform profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the chemical transformation of the substrate using solid-state NMR of 13C. Adding 10% of coffee pulp increased laccase activities and fungal biomass (32.5% and 16% respectively), enhanced two constitutive isoforms (Rf 0.23 and 0.27), induced a new isoform (Rf 0.19) and led to a decrease in total aromatics. However, higher concentrations of coffee pulp (25%) decreased laccase and cellulase activities but no decrease in aromaticity was observed, potentially due to the toxic effect of phenols from coffee pulp. Moreover, laccase production was still inhibited even for lower concentrations of urea (0-5%). Our findings revealed that an agricultural by-product like coffee pulp can enhance laccase activity -though to a threshold- and that urea limited this process, indicating that other N-sources should be tested for the biological delignification of SCB.

mTORC1-dependent increase in oxidative metabolism in POMC neurons regulates food intake and action of leptin.

OBJECTIVE: Nutrient availability modulates reactive oxygen species (ROS) production in the hypothalamus. In turn, ROS regulate hypothalamic neuronal activity and feeding behavior. The mechanistic target of rapamycin complex 1 (mTORC1) pathway is an important cellular integrator of the action of nutrients and hormones. Here we tested the hypothesis that modulation of mTORC1 activity, particularly in Proopiomelanocortin (POMC)-expressing neurons, mediates the cellular and behavioral effects of ROS. METHODS: C57BL/6J mice or controls and their knockout (KO) littermates deficient either for the mTORC1 downstream target 70-kDa ribosomal protein S6 kinase 1 (S6K1) or for the mTORC1 component Rptor specifically in POMC neurons (POMC-rptor-KO) were treated with an intracerebroventricular (icv) injection of the ROS hydrogen peroxide (H2O2) or the ROS scavenger honokiol, alone or, respectively, in combination with the mTORC1 inhibitor rapamycin or the mTORC1 activator leptin. Oxidant-related signal in POMC neurons was assessed using dihydroethidium (DHE) fluorescence. RESULTS: Icv administration of H2O2 decreased food intake, while co-administration of rapamycin, whole-body deletion of S6K1, or deletion of rptor in POMC neurons impeded the anorectic action of H2O2. H2O2 also increased oxidant levels in POMC neurons, an effect that hinged on functional mTORC1 in these neurons. Finally, scavenging ROS prevented the hypophagic action of leptin, which in turn required mTORC1 to increase oxidant levels in POMC neurons and to inhibit food intake. CONCLUSIONS: Our results demonstrate that ROS and leptin require mTORC1 pathway activity in POMC neurons to increase oxidant levels in POMC neurons and consequently decrease food intake.

A FTIR/chemometrics approach to characterize the gamma radiation effects on iodine/epoxy-paint interactions in Nuclear Power Plants.

The effects of radiation on polymeric materials are a topic of concern in a wide range of industries including the sterilization, and the nuclear power industry. While much work has concentrated on systems like polyolefins that are radiation sterilized, some work has been done on epoxy systems. The epoxy system studied is an epoxy/amine paint which is representative of the paint that covers the inner surfaces of the French nuclear reactor containment buildings. In case of a severe accident on a Nuclear Power Plant, fission products can be released from the nuclear fuel to the reactor containment building. Among them, volatile iodine (I2) can be produced and can interact with the epoxy-paint. This paint is also subjected to gamma radiation damages (due to the high dose in the containment coming from radionuclides released from the fuel). So the epoxy-paint studied was exposed to gamma radiation under air atmosphere after being loaded with I2 or not. The aim of this study is to characterize by FTIR spectroscopy the iodine-paint interactions, then to identify the radiation damages on the epoxy-paint, and to check their effects on these iodine-paint interactions. This work shows the potential of multi-block analysis method (ANOVA-PCA and COMDIM = AComDim) for such a study as it allows to identify the nature of iodine/epoxy-paint interactions and to characterize the gamma radiation damages on the epoxy-paint. AComDim method conduces to the extraction of Common Components to different tables and highlights factors of influence and their interactions.

Artemisia arborescens Essential Oil Composition, Enantiomeric Distribution, and Antimicrobial Activity from Different Wild Populations from the Mediterranean Area.

Aerial parts of Artemisia arborescens were collected from different sites of the Mediterranean area (southwestern Algeria and southern Italy) and the chemical composition of their essential oil (EO) extracted by hydrodistillation was studied by both gas chromatography (GC) equipped with an enantioselective capillary column and GC/mass spectrometry (GC/MS). The EOs obtained were tested against several Listeria monocytogenes strains. Using GC and GC/MS, 41 compounds were identified, accounting for 96.0 - 98.8% of the total EO. All EOs showed a similar terpene profile, which was rich in chamazulene, ß-thujone, and camphor. However, the concentration of such compounds varied among the EOs. A. arborescens EO inhibited up to 83.3% of the L. monocytogenes strains, but the inhibitory spectrum varied among the EOs, with those from Algeria showing a higher inhibition degree than the Italian EOs. Such effect likely depended on the ketone (ß-thujone + camphor) content of the EO. The differences in the EO composition support the hypothesis that A. arborescens has at least two different chemotypes: a ß-thujone and a chamazulene type. The EO inhibitory spectrum indicates the A. arborescens EO as a valuable option in the control of the food-borne pathogens.

Analysis of the major chiral compounds of Artemisia herba-alba essential oils (EOs) using reconstructed vibrational circular dichroism (VCD) spectra: En route to a VCD chiral signature of EOs.

An unprecedented methodology was developed to simultaneously assign the relative percentages of the major chiral compounds and their prevailing enantiomeric form in crude essential oils (EOs). In a first step the infrared (IR) and vibrational circular dichroism (VCD) spectra of the crude essential oils were recorded and in a second step they were modelized as a linear weighted combination of the IR and VCD spectra of the individual spectra of pure enantiomer of the major chiral compounds present in the EOs. The VCD spectra of enantiomer of known enantiomeric excess shall be recorded if they are not yet available in a library of VCD spectra. For IR, the spectra of pure enantiomer or racemic mixture can be used. The full spectra modelizations were performed using a well known and powerful mathematical model (least square estimation: LSE) which resulted in a weighting of each contributing compound. For VCD modelization, the absolute value of each weighting represented the percentage of the associate compound while the attached sign addressed the correctness of the enantiomeric form used to build the model. As an example, a model built with the non-prevailing enantiomer will show a negative sign of the weighting value. For IR spectra modelization, the absolute value of each weighting represented the percentage of the compounds without of course accounting for the chirality of the prevailing enantiomers. Comparison of the weighting values issuing from IR and VCD spectra modelizations is a valuable source of information: if they are identical, the EOs are composed of nearly pure enantiomers, if they are different the chiral compounds of the EOs are not in an optically pure form. The method was applied on four samples of essential oil of Artemisia herba-alba in which the three major compounds namely (-)-α-thujone, (+)-ß-thujone and (-)-camphor were found in different proportions as determined by GC-MS and chiral HPLC using polarimetric detector. In order to validate the methodology, the modelization of the VCD spectra was performed on purpose using the individual VCD spectra of (-)-α-thujone, (+)-ß-thujone and (+)-camphor instead of (-)-camphor. During this work, the absolute configurations of (-)-α-thujone and (+)-ß-thujone were confirmed by comparison of experimental and calculated VCD spectra as being (1S,4R,5R) and (1S,4S,5R) respectively.

Origin of French virgin olive oil registered designation of origins predicted by chemometric analysis of synchronous excitation-emission fluorescence spectra.

The authentication of virgin olive oil samples requires usually the use of sophisticated and very expensive analytical techniques, so there is a need for fast and inexpensive analytical techniques for use in a quality control methodology. Virgin olive oils present an intense fluorescence spectra. Synchronous excitation-emission fluorescence spectroscopy (SEEFS) was assessed for origin determination of virgin olive oil samples from five French registered designation of origins (RDOs) (Nyons, Vallée des Baux, Aix-en-Provence, Haute-Provence, and Nice). The spectra present bands between 600 and 700 nm in emission due to chlorophylls a and b and pheophytins a and b. The bands between 275 and 400 nm in emission were attributed to alpha-, beta-, and gamma-tocopherols and to phenolic compounds, which characterize the virgin olive oils compared to other edible oils. The chemometric treatment (PLS1) of synchronous excitation-emission fluorescence spectra allows one to determine the origin of the oils from five French RDOs (Baux, Aix, Haute-Provence, Nice, and Nyons). Results were quite satisfactory, despite the similarity between two denominations of origin (Baux and Aix) that are composed by some common cultivars (Aglandau and Salonenque). The interpretation of the regression coefficients shows that RDOs are correlated to chlorophylls, pheophytins, tocopherols, and phenols compounds, which are different for each origin. SEEFS is part of a global analytic methodology that associates spectroscopic and chromatographic techniques. This approach can be used for traceability and vindicates the RDOs.