Evaluation of core-shell Fe3O4@Au nanoparticles as radioenhancer in A549 cell lung cancer model.

In radiotherapy, metallic nanoparticles are of high interest in the fight against cancer for their radiosensitizing effects. This study aimed to evaluate the ability of core-shell Fe3O4@Au nanoparticles to potentiate the irradiation effects on redox-, pro-inflammatory markers, and cell death of A549 human pulmonary cancer cells. The hybrid Fe3O4@Au nanoparticles were synthesized using green chemistry principles by the sonochemistry method. Their characterization by transmission electron microscopy demonstrated an average size of 8 nm and a homogeneous distribution of gold. The decreased hydrodynamic size of these hybrid nanoparticles compared to magnetite (Fe3O4) nanoparticles showed that gold coating significantly reduced the aggregation of Fe3O4 particles. The internalization and accumulation of the Fe3O4@Au nanoparticles within the cells were demonstrated by Prussian Blue staining. The reactive oxygen species (ROS) levels measured by the fluorescent probe DCFH-DA were up-regulated, as well as mRNA expression of SOD, catalase, GPx antioxidant enzymes, redox-dependent transcription factor Nrf2, and ROS-producing enzymes (Nox2 and Nox4), quantified by RT-qPCR. Furthermore, irradiation coupled with Fe3O4@Au nanoparticles increased the expression of canonical pro-inflammatory cytokines and chemokines (TNF-α, IL-1ß, IL-6, CXCL8, and CCL5) assessed by RT-qPCR and ELISA. Hybrid nanoparticles did not potentiate the increased DNA damage detected by immunofluorescence following the irradiation. Nevertheless, Fe3O4@Au caused cellular damage, leading to apoptosis through activation of caspase 3/7, secondary necrosis quantified by LDH release, and cell growth arrest evaluated by clonogenic-like assay. This study demonstrated the potential of Fe3O4@Au nanoparticles to potentiate the radiosensitivity of cancerous cells.

Impact of Green Gold Nanoparticle Coating on Internalization, Trafficking, and Efficiency for Photothermal Therapy of Skin Cancer.

Skin cancer is a global health issue and mainly composed of melanoma and nonmelanoma cancers. For the first clinical proof of concept on humans, we decided to study good prognosis skin cancers, i.e., carcinoma basal cell. In UE, the first-line treatment remains surgical resection, healing most of the tumors, but presents aesthetic disadvantages with a high reoccurrence rate on exposed areas. Moreover, the therapeutic indications could extend to melanoma and metastasis, which is a different medical strategy that could combine this treatment. Indeed, patients with late-stage melanoma are in a therapeutic impasse, despite recent targeted and immunological therapies. Photothermal therapy using gold nanoparticles is the subject of many investigations due to their strong potential to treat cancers by physical, thermal destruction. We developed gold nanoparticles synthesized by green chemistry (gGNPs), using endemic plant extract from Reunion Island, which have previously showed their efficiency at a preclinical stage. Here, we demonstrate that these gGNPs are less cytotoxic than gold nanoparticles synthesized by Turkevich's method. Furthermore, our work describes the optimization of gGNP coating and stabilization, also taking into consideration the gGNP path in cells (endocytosis, intracellular trafficking, and exocytosis), their specificity toward cancerous cells, their cytotoxicity, and their in vivo efficiency. Finally, based on the metabolic switch of cancerous cells overexpressing Glut transporters in skin cancers, we demonstrated that glucose-stabilized gGNP (gGNP@G) enables a quick internalization, fourfold higher in cancerous cells in contrast to healthy cells with no side cytotoxicity, which is particularly relevant to target and treat cancer.

Polyphenol Characterization of the Aqueous Extract from Hubertia ambavilla L. (Asteraceae) by HPLC-DAD-ESI-MSn and Assessment of Its Antioxidant Activity.

In this work, phytochemical components, and the antioxidant properties of an aqueous extract obtained from a medicinal plant Hubertia ambavilla, endemic to Reunion Island, were investigated. A total of 37 compounds were detected and identified by high-performance liquid chromatography (UHPLC) using a photodiode-array detector (DAD) coupled with electrospray ionization/mass spectrometry (ESI/MSn ). From calibration curves, the quantity of secondary metabolites in the aqueous extract was calculated. The mean amounts of phenols, flavonoids, and condensed tannins found were 158.38±1.20 mg GAE/g DE, 60.41±1.65 mg AE/g DE and 23.77±1.36 mg CE/g DE, respectively. The in vitro antioxidant properties of the Hubertia ambavilla plant were measured using three methods: DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) scavenging and ferric reducing antioxidant power. The results showed that crude aqueous extract of H. ambavilla had effective radical scavenging and reducing power in comparison with standard antioxidant compounds. In conclusion, the crude extract herein presented offers a natural alternative biosource of antioxidants with potential applications in food and health industries.

Effects of sex and estrous cycle on sleep and cataplexy in narcoleptic mice.

Narcolepsy type 1 (NT1) is a rare neurology disorder caused by the loss of orexin/hypocretin neurons. NT1 is characterized by excessive daytime sleepiness, sleep and wake fragmentation, and cataplexy. These symptoms have been equally described in both women and men, although influences of gender and hormonal cycles have been poorly studied. Unfortunately, most studies with NT1 preclinical mouse models, use only male mice to limit potential variations due to the hormonal cycle. Therefore, whether gender and/or hormonal cycles impact the expression of narcoleptic symptoms remains to be determined. To address this question, we analyzed vigilance states and cataplexy in 20 female and 17 male adult orexin knock-out narcoleptic mice, with half of the females being recorded over multiple days. Mice had access to chocolate to encourage the occurrence of cataplectic episodes. A vaginal smear was performed daily in female mice to establish the state of the estrous cycle (EC) of the previous recorded night. We found that vigilance states were more fragmented in males than females, and that females had less paradoxical sleep (p = 0.0315) but more cataplexy (p = 0.0375). Interestingly, sleep and wake features were unchanged across the female EC, but the total amount of cataplexy was doubled during estrus compared to other stages of the cycle (p = 0.001), due to a large increase in the number of cataplexy episodes (p = 0.0002). Altogether these data highlight sex differences in the expression of narcolepsy symptoms in orexin knock-out mice. Notably, cataplexy occurrence was greatly influenced by estrous cycle. Whether it is due to hormonal changes would need to be further explored.

Histamine in murine narcolepsy: What do genetic and immune models tell us?

An increased number of histaminergic neurons, identified by labeling histidine-decarboxylase (HDC) its synthesis enzyme, was unexpectedly found in patients with narcolepsy type 1 (NT1). In quest for enlightenment, we evaluate whether an increase in HDC cell number and expression level would be detected in mouse models of the disease, in order to provide proof of concepts reveling possible mechanisms of compensation for the loss of orexin neurons, and/or of induced expression as a consequence of local neuroinflammation, a state that likely accompanies NT1. To further explore the compensatory hypothesis, we also study the noradrenergic wake-promoting system. Immunohistochemistry for HDC, orexin, and melanin-concentrating hormone (MCH) was used to count neurons. Quantitative-PCR of HDC, orexin, MCH, and tyrosine-hydroxylase was performed to evaluate levels of mRNA expression in the hypothalamus or the dorsal pons. Both quantifications were achieved in genetic and neuroinflammatory models of narcolepsy with major orexin impairment, namely the orexin-deficient (Orex-KO) and orexin-hemagglutinin (Orex-HA) mice respectively. The number of HDC neurons and mRNA expression level were unchanged in Orex-KO mice compared to controls. Similarly, we found no change in tyrosine-hydroxylase mRNA expression in the dorsal pons between groups. Further, despite the presence of protracted local neuroinflammation as witnessed by the presence of reactive microglia, we found no change in the number of neurons nor the expression of HDC in Orex-HA mice compared to controls. Importantly, no correlation was found in all conditions between HDC and orexin. Our findings indicate that, in mice, the expression of histamine and noradrenalin, two wake-promoting systems, are not modulated by orexin level whether the lack of orexin is constitutive or induced at adult age, showing thus no compensation. They also show no recruitment of histamine by local neuroinflammation. Further studies will be needed to further define the role of histamine in the pathophysiology of NT1.

Dataset concerning plasmonic thermal destruction of murine melanoma by gold nanoparticles obtained by green chemistry.

The data presented in this article are related to the research paper "Proof of concept of plasmonic thermal destruction of surface cancers by gold nanoparticles obtained by green chemistry" (Ben Hadadda et al, 2019). In this article, we examined plasmonic thermal destruction of murine melanoma using gold nanoparticles obtained by green chemistry. The presented data were obtained by measuring tumor volume and mice weight in different groups of mice murine melanoma B16F10 treated or not with the nanoparticles and coupled to laser irradiation. These data were compared to the clinical reference treatment: anti-PD1 monoclonal antibody. The data were analyzed in order to be able to compare the antitumor effect of our treatment (photothermal plasmonic therapy using gold nanoparticles) and the reference treatment.

Assessment of antioxidant and dermoprotective activities of gold nanoparticles as safe cosmetic ingredient.

Hubertia ambavilla, an endemic plant originating from Reunion Island in the Indian Ocean, is traditionally used as an anti-inflammatory and in healing, both for internal and external use. Polyphenolic compounds from aqueous phase extractions can reduce metal salts into nanoparticles and stabilize them in one step. Although gold nanoparticles are well described in the literature as anti-ageing ingredients, the nanoparticles presented herein are novel and are synthesized using a green process. We demonstrate their efficiency as dermoprotective, free radical scavenger and antioxidant cosmetic ingredients. Comparison with common nanoparticles obtained by the Turkevich method clearly emphasizes the necessity to carefully screen the products used for nanoparticle coatings, as they play a major role in the biological properties of the product. Hubertia ambavilla mediated gold nanoparticles are non-toxic to human dermal fibroblasts, possess free radical scavenging potential, and protect against damage to fibroblast and dermal cells caused by ultraviolet A radiation.

Proof of concept of plasmonic thermal destruction of surface cancers by gold nanoparticles obtained by green chemistry.

A greener approach for the design of surface plasmon resonant gold nanoparticles has been obtained with a hydrosoluble fraction of an endemic asteraceae medicinal plant. This medicinal plant is originated from Indian Ocean and demonstrates its bioreducing activity in the design of stable green nanomedicine in aqueous media. This article describes the preclinical assessment of the efficacy of these novel nanocandidates on murine model by intratumoral and intravenous injections. It definitely demonstrates two key points in the treatment of cancer: 1) optimization of the tumor microenvironment targeting by specific ligands for a limited damage on healthy tissue, 2) the need to screen the specific irradiation dose (time, power) taking into account the type of tumor.

Partial homologies between sleep states in lizards, mammals, and birds suggest a complex evolution of sleep states in amniotes.

It is crucial to determine whether rapid eye movement (REM) sleep and slow-wave sleep (SWS) (or non-REM sleep), identified in most mammals and birds, also exist in lizards, as they share a common ancestor with these groups. Recently, a study in the bearded dragon (P. vitticeps) reported states analogous to REM and SWS alternating in a surprisingly regular 80-s period, suggesting a common origin of the two sleep states across amniotes. We first confirmed these results in the bearded dragon with deep brain recordings and electro-oculogram (EOG) recordings. Then, to confirm a common origin and more finely characterize sleep in lizards, we developed a multiparametric approach in the tegu lizard, a species never recorded to date. We recorded EOG, electromyogram (EMG), heart rate, and local field potentials (LFPs) and included data on arousal thresholds, sleep deprivation, and pharmacological treatments with fluoxetine, a serotonin reuptake blocker that suppresses REM sleep in mammals. As in the bearded dragon, we demonstrate the existence of two sleep states in tegu lizards. However, no clear periodicity is apparent. The first sleep state (S1 sleep) showed high-amplitude isolated sharp waves, and the second sleep state (S2 sleep) displayed 15-Hz oscillations, isolated ocular movements, and a decrease in heart rate variability and muscle tone compared to S1. Fluoxetine treatment induced a significant decrease in S2 quantities and in the number of sharp waves in S1. Because S2 sleep is characterized by the presence of ocular movements and is inhibited by a serotonin reuptake inhibitor, as is REM sleep in birds and mammals, it might be analogous to this state. However, S2 displays a type of oscillation never previously reported and does not display a desynchronized electroencephalogram (EEG) as is observed in the bearded dragons, mammals, and birds. This suggests that the phenotype of sleep states and possibly their role can differ even between closely related species. Finally, our results suggest a common origin of two sleep states in amniotes. Yet, they also highlight a diversity of sleep phenotypes across lizards, demonstrating that the evolution of sleep states is more complex than previously thought.

CD8 T cell-mediated killing of orexinergic neurons induces a narcolepsy-like phenotype in mice.

Narcolepsy with cataplexy is a rare and severe sleep disorder caused by the destruction of orexinergic neurons in the lateral hypothalamus. The genetic and environmental factors associated with narcolepsy, together with serologic data, collectively point to an autoimmune origin. The current animal models of narcolepsy, based on either disruption of the orexinergic neurotransmission or neurons, do not allow study of the potential autoimmune etiology. Here, we sought to generate a mouse model that allows deciphering of the immune mechanisms leading to orexin(+) neuron loss and narcolepsy development. We generated mice expressing the hemagglutinin (HA) as a "neo-self-antigen" specifically in hypothalamic orexin(+) neurons (called Orex-HA), which were transferred with effector neo-self-antigen-specific T cells to assess whether an autoimmune process could be at play in narcolepsy. Given the tight association of narcolepsy with the human leukocyte antigen (HLA) HLA-DQB1*06:02 allele, we first tested the pathogenic contribution of CD4 Th1 cells. Although these T cells readily infiltrated the hypothalamus and triggered local inflammation, they did not elicit the loss of orexin(+) neurons or clinical manifestations of narcolepsy. In contrast, the transfer of cytotoxic CD8 T cells (CTLs) led to both T-cell infiltration and specific destruction of orexin(+) neurons. This phenotype was further aggravated upon repeated injections of CTLs. In situ, CTLs interacted directly with MHC class I-expressing orexin(+) neurons, resulting in cytolytic granule polarization toward neurons. Finally, drastic neuronal loss caused manifestations mimicking human narcolepsy, such as cataplexy and sleep attacks. This work demonstrates the potential role of CTLs as final effectors of the immunopathological process in narcolepsy.

Differential effects of amyloid-beta 1-40 and 1-42 fibrils on 5-HT1A serotonin receptors in rat brain.

Evidence accumulates suggesting a complex interplay between neurodegenerative processes and serotonergic neurotransmission. We have previously reported an overexpression of serotonin 5-HT1A receptors (5-HT(1A)R) after intrahippocampal injections of amyloid-beta 1-40 (Aß40) fibrils in rats. This serotonergic reactivity paralleled results from clinical positron emission tomography studies with [(18)F]MPPF revealing an overexpression of 5-HT(1A)R in the hippocampus of patients with mild cognitive impairment. Because Aß40 and Aß42 isoforms are found in amyloid plaques, we tested in this study the hypothesis of a peptide- and region-specific 5-HT(1A)R reactivity by injecting them, separately, into the hippocampus or striatum of rats. [(18)F]MPPF in vitro autoradiography revealed that Aß40 fibrils, but not Aß42, were triggering an overexpression of 5-HT(1A)R in the hippocampus and striatum of rat brains after 7 days. Immunohistochemical approaches targeting neuronal precursor cells, mature neurons, and astrocytes showed that Aß42 fibrils caused more pathophysiological damages than Aß40 fibrils. The mechanisms of Aß40 fibrils-induced 5-HT(1A)R expression remains unknown, but hypotheses including neurogenesis, glial expression, and axonal sprouting are discussed.

Enhanced Cocaine-Associated Contextual Learning in Female H/Rouen Mice Selectively Bred for Depressive-Like Behaviors: Molecular and Neuronal Correlates.

BACKGROUND: Major depression has multiple comorbidities, in particular drug use disorders, which often lead to more severe and difficult-to-treat illnesses. However, the mechanisms linking these comorbidities remain largely unknown. METHODS: We investigated how a depressive-like phenotype modulates cocaine-related behaviors using a genetic model of depression: the Helpless H/Rouen (H) mouse. We selected the H mouse line for its long immobility duration in the tail suspension test when compared to non-helpless (NH) and intermediate (I) mice. Since numerous studies revealed important sex differences in drug addiction and depression, we conducted behavioral experiments in both sexes. RESULTS: All mice, regardless of phenotype or sex, developed a similar behavioral sensitization after 5 daily cocaine injections (10 mg/kg). Male H and NH mice exhibited similar cocaine-induced conditioned place preference scores that were only slightly higher than in I mice, whereas female H mice strikingly accrued much higher preferences for the cocaine-associated context than those of I and NH mice. Moreover, female H mice acquired cocaine-associated context learning much faster than I and NH mice, a facilitating effect that was associated to a rapid increase in striatal and accumbal brain-derived neurotrophic factor levels (BDNF; up to 35% 24 h after cocaine conditioning). Finally, when re-exposed to the previously cocaine-associated context, female H mice displayed greater Fos activation in the cingulate cortex, nucleus accumbens, and basolateral amygdala. CONCLUSIONS: Our data indicate that neurobiological mechanisms such as alterations in associative learning, striato-accumbal BDNF expression, and limbic-cortico-striatal circuit reactivity could mediate enhanced cocaine vulnerability in female depressive-like mice.

Enhanced anxiety observed in cocaine withdrawn rats is associated with altered reactivity of the dorsomedial prefrontal cortex.

Discontinuation of drug intake in cocaine abusers commonly produces a variety of adverse withdrawal symptoms among which anxiety and depression-related behavior are prevailing during the initial period of abstinence. The aim of this study was to provide further insight into the neurobiological dysregulations that might contribute to these pathological states. Rats were treated with cocaine or saline for 14 days (20 mg/kg; i.p) and anxiety-related behavior was assessed in different paradigms (elevated plus-maze (EPM), confinement to an open arm of the EPM and shock-probe burying tests) for up to 4 weeks after withdrawal. Depression-like behavior was assessed by the forced swim test and sucrose preference test. Altogether our results demonstrated that cocaine withdrawal induced persistent heightened levels of anxiety that last for at least 28 days but did not affect depression-like behavior. We then used Fos immunohistochemistry to map neuronal activation patterns in withdrawn rats confined to one open arm of an EPM, and a double labeling procedure using Fos immunohistochemistry and in situ hybridization of glutamic acid decarboxylase or vesicular glutamate transporter mRNAs to identify the phenotype of the activated neurons. Our data showed that the exacerbated anxiety observed in cocaine withdrawn rats exposed to an elevated open arm was accompanied by an altered reactivity of the dorsal part of the medial prefrontal cortex (anterior cingulate and dorsal prelimbic cortices), the paraventricular thalamic nucleus and the lateral and anterior areas of the hypothalamus. In the medial prefrontal cortex, we evidenced a negative correlation between Fos expression in its dorsal part and open arm-induced freezing in NaCl-treated rats but not in cocaine withdrawn rats. We also found that more than 65% of activated neurons were glutamatergic projection neurons. The present study provides new insights into the neuroanatomical regions and neuronal cell types that may underlie pathological anxiety during cocaine withdrawal.

Biosensors elaborated on gold nanoparticles, a PM-IRRAS characterisation of the IgG binding efficiency.

This work is focused on studying the grafting of gold nanoparticles (Np) on a cystamine self-assembled monolayer on gold, in order to build sensitive immunosensors. The synthesis and deposition of gold nanoparticles, 13 and 55 nm sizes, were characterised by combining Polarisation Modulation Infrared Reflection-Absorption Spectroscopy (PM-IRRAS), X-ray Photoelectron Spectroscopy (XPS) Surface Enhanced Raman Scattering (SERS), and Atomic Force Microscopy (AFM) which all indicated the formation of a dispersed layer of nanoparticles. This observation is explained by the compromise between the high reactivity of amine-terminated layers towards gold, and interparticle repulsions. Nps were then functionalised with antibody probes, and the recognition by an anti-rIgG was assayed both on planar and Np gold surfaces. The important result is that nanoparticles of 55 nm are preferable for the following reasons: they enable to build a denser and well dispersed layer and they increase both the number of receptors (IgGs) and their accessibility. Beside these geometric improvements, a net enhancement of the Raman signal was observed on the 55 nm nanoparticle layer, making this new platform promising for optical detection based biosensors.

Optimized immobilization of gold nanoparticles on planar surfaces through alkyldithiols and their use to build 3D biosensors.

This paper describes a controlled way to immobilize gold nanoparticles on planar gold surfaces and the use of the resulting 3D platform to build up a 3D biosensor. The surface was first functionalized by grafting hexanedithiol, this molecule has 2 thiol end groups, which enables its chemical grafting to planar gold while retaining a free thiol group to attach nanoparticles. This step was optimized by varying experimental parameters such as solvent, temperature and immersion time. The grafting was monitored by polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) and X-ray photoelectron spectroscopy (XPS). The high resolution XPS sulfur peak made clear the existence of two contributions, S bound to gold and free S, thus led us to determine the optimal conditions to graft hexanedithiol in an extended conformation. 15 nm spherical gold nanoparticles were then immobilized on the resulting surface and their presence was evidenced by surface enhanced Raman spectroscopy (SERS) and atomic force microscopy (AFM). The resulting gold layer was used to build up a 3D biosensor by grafting protein A (PrA), rabbit immunoglobulin (rIgG), and bovine serum albumin (BSA), respectively. Each step was characterized by PM-IRRAS then compared to the results on planar gold surface. Despite the small size of particles and their rather low density on the planar surface, the amount of immobilized proteins, starting from PrA, was almost doubled. The amount of rIgG fixed on the 3D layer was also significantly increased ( approximately 4 times higher than on planar surfaces), however accompanied by a slight decrease of their accessibility, checked by assaying the recognition of a secondary IgG. This work demonstrates the feasibility and interest of building arrays of nanoparticles to immobilize molecular receptors; it also shows that controlling the conditions of elaboration of the biosensor at each step is determining for optimizing the number of molecular receptors.

Sonochemical approach to the synthesis of Fe(3)O(4)@SiO(2) core-shell nanoparticles with tunable properties.

In this study, we report a rapid sonochemical synthesis of monodisperse nonaggregated Fe(3)O(4)@SiO(2) magnetic nanoparticles (NPs). We found that coprecipitation of Fe(II) and Fe(III) in aqueous solutions under the effect of power ultrasound yields smaller Fe(3)O(4) NPs with a narrow size distribution (4-8 nm) compared to the silent reaction. Moreover, the coating of Fe(3)O(4) NPs with silica using an alkaline hydrolysis of tetraethyl orthosilicate in ethanol-water mixture is accelerated many-fold in the presence of a 20 kHz ultrasonic field. The thickness of the silica shell can be easily controlled in the range of several nanometers during sonication. Mossbauer spectra revealed that nonsuperparamagnetic behavior of obtained core-shell NPs is mostly related to the dipole-dipole interactions of magnetic cores and not to the particle size effect. Core-shell Fe(3)O(4)@SiO(2) NPs prepared with sonochemistry exhibit a higher magnetization value than that for NPs obtained under silent conditions owing to better control of the deposited silica quantities as well as to the high speed of sonochemical coating, which prevents the magnetite from oxidizing.