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Drosophila shows complex behaviours but performs these with relatively few neurons. Genetic tools for mapping brain circuits and perturbing neurons, together with the ability to record neural activity, offer the potential to link behaviours with brain circuits and dynamics. Neurobehavioural dynamics are complex and short experiments do not capture dynamics occurring over multiple days. To overcome this limitation, I developed a chronic imaging protocol which allows the recording of neurobehavioural dynamics of a fly living for days on a spherical treadmill under a two-photon microscope. To validate this approach, I followed the Ca2+ dynamics in a genetic line (23E10-GAL4) labelling the switch neurons controlling sleep and wake. Sleep was measured as locomotor inactivity lasting for 5 or more minutes. I explored the relationship between the dendritic Ca2+ dynamics in the switch neurons and sleep. I found that some of the features describing the dendritic Ca2+ dynamics correlate with “future sleep” and the time of day. To overcome another limitation of current experiments investigating sleep homeostasis, I developed a protocol to sleep deprive flies during imaging experiments. I proved that the protocol is effective in causing sleep rebound after sleep deprivation and does not affect the lifespan of the experimental fly. Our study paves the way for new experiments investigating sleep homeostasis and the detrimental effects of sleep deprivation over extended periods of time. It is also a validation of the virtues of Drosophila as a model for systems neuroscience.

Bio-stimulation is an injective therapy aimed to boost the anabolic functions of dermal fibroblasts to obtain skin improvement. It can be achieved with multiple intradermal injections (0.05–0.1 ml each) of a solution of 400 mg (3 ml) of injectable glucosamine sulphate, plus 5.623 mg (3 ml) of polideoxyribonucleotide, 1 ml of lidocaine and 0.5–1 ml of sodium bicarbonate, to repeat every 7, 14, 21, and 28 days. The administration of glucosamine sulphate to skin fibroblasts is believed to lead to its incorporation in glycosaminoglycans, and thereby to the stimulation of extracellular matrix synthesis, whereas polideoxyribonucleotide possesses anti-inflammatory and regenerative capability. This study aims to elucidate the in-vitro effects of this treatment by studying what happens to several genes related to connective tissue integrity. Human dermal fibroblasts were seeded in a culture medium enriched with either two drugs alone or combined: glucosamine sulphate and/or polideoxyribonucleotide. After the end of the exposure time of 24 h, 48 h, and 72 h, the cells were trypsinized and lysed for RNA extraction. Reverse transcription to cDNA was performed directly from cultured cell lysate. Finally, the cDNA was amplified by real-time PCR and a panel of genes involved in dermal integrity was tested. Gene expression of Hyaluronan synthase 1 (HAS1), Elastine (ELN), Insulin like growth factor 1 (IGF1), Growth differentiation factor 6 (GDF6) and of a series of catabolic enzymes, such as Metalloproteases (MMP) 2, 3 and 13, the neutrophyl expressed Elastase (ELANE) and the Hyaluronidase 1 (HYAL1) were tested after 24, 48 and 72 hours of exposure to glucosamine sulphate and polideoxyribonucleotide alone or combined. All the tested genes but one were up-regulated. A negative synergism on several enzymes (particularly appreciable for Insulin-like growth factor 1 and metalloprotease 13) was observed when the two drugs were delivered together. Glucosamine sulphate acts not only as building block in the biosynthesis of glycosaminoglycan chains, but also as a booster of hyaluronan synthase 1. The association of glucosamine sulphate and polideoxyribonucleotide, used in bio-stimulation therapy protocol, has a negative synergism on catabolic genes in dermal fibroblast cultures. The present observations produce further insight into the effects of glucosamine sulphate in the biosynthesis of glycosaminoglycan chains.

The platelets used in oral, maxillofacial and plastic surgery are generally grouped as concentrated platelet rich plasma. The general principle of production consists of a centrifugation, making it possible to eliminate red blood cells, then acellular plasma, to preserve only the concentrated platelets. Aim to evaluate the efficacy of a three session of injections of autologous platelet-derived growth factors for full-face rejuvenation including the perioral and periorbital regions; 16 patients female between 27 and 60 years of age, were admitted to facial rejuvenation procedure. All these patients showed mainly class II rhytides. Six patients had rejuvenation of only the perioral region (five patients had class II rhytides and the other patient had class III rhytides). Another five patients had rejuvenation of only the periorbital region (two patients with class I rhytides, the other three had class II rhytides). Approximately 0,2 cc of autologous platelet-derived growth factors was produced per tube; this activated suspension was then injected intradermally or subdermally below the perioral and periorbital regions through 30 and 27 gauge needles, respectively, as needed to achieve the most optimal correction of the rhytides with mainly a linear threading technique. All patients demonstrated some improvement. 92.88% of participants were satisfied or very satisfied with their treatment result. The investigator evaluation rates of acceptable effect (score of 2 or 3) were 76.65%. The results lasted approximately 6 months with a gradual decline to baseline. Autologous platelet-derived growth factors in one treatment session for three months can achieve effective full-face skin rejuvenation, with effects on both the epidermis and dermis.

In the cosmetic medical area facial rejuvenation with permanent or absorbable fillers requires not only biochemical and biological informations about the molecules used, but also insights into the ageing anatomy and physiology: This article reviews the results of 46 cases undergone to lip augmentation and perioral rejuvenation in the last 1 year. A total of 46 consecutive female patients, between 27 and 60 years of age, with type I and type II lips by Fulton classification were recruited to the study. In all patients both lips were treated. The product was a cross-linked hyaluronic acid (ALIAZIN(R) FL-LIP S IBSA-Lodi, Italy) injected in a single session. The aesthetic evaluation was performed at 3, 6, 9, and 12 months after injection. As to the injection technique the filler has been implanted withdrawing the needle while injecting along the lips margins. After 3 months, 87.88% of participants were satisfied or very satisfied with their treatment result. After 6, 9 and 12 months, 52.73%, 40.67% and 10.67% of participants, respectively, were satisfied or very satisfied. The investigator evaluation rates of acceptable effect (score of 2 or 3) were 96.65%, 40.43%, 28, 39 and 16.79% at 3, 6, 9 and 12 months, respectively. Fining results lasted approximately 5 months with a gradual decline to baseline. The procedure was very well tolerated with only few, mild adverse reactions that resolved spontaneously. In conclusion during first years of clinical use, crosslinked Hyaluronic Acid was proven to be reliable filler preferred by cosmetic doctors an effective and safe facial soft tissue expander and predictable treatment for lip augmentation.

Laser welding appears to be the most suitable technology for joining metallic foams. Spatially limited energy input, together with high energy density, determine a small molten material volume and a rapid solidification, which prevents the collapse of the cellular structure and the drainage phenomena. It has indeed already been shown that laser welding is a useful technique for joining metal foam panels with conventional or foamable filler. The welding techniques were also widely applied and studied for the Aluminum Foam Sandwich (AFS). Composite materials obtained by filling with metallic foam hollow profiles are nowadays available. Reliable welding processes have to be carried out in order to realize complex lightweight structures. The present paper investigates the butt-joint laser welding of a composite material made of steel tubes filled with aluminum foam core. Two pieces of a metal foam filled steel tube were prepared by chamfering the edge and foamable filler was then inserted between them. The process was divided in two parts. As a first phase of the method, the joining of the tubes was obtained by tuning the laser parameters in order to weld the whole tube thickness without damaging the metal foam. Finally, the laser beam was used in order to obtain both a sort of heat treatment in the tubes material and a temperature gradient inside the aluminum foam core. In this second stage of the laser processing, the beam parameters were tuned in order to achieve the foaming temperature inside the tubes. The hollow steel profiles were filled up with aluminum powder precursors, which were produced by cold uniaxial compaction according to the powder metallurgical route. They were then foamed in a furnace. A Laser machining center with 3 kW CW FAF CO2 laser source and five CNC axes was used. This study is a preliminary step in order to verify the reliability of the method, and further investigations will be necessary with the aim of optimizing the process. The present investigation was limited to laser beam power and process speed. At the present stage the results are very encouraging and a prototype has therefore been constructed.

Regulation of epithelial chloride flux, which is defective in patients with cystic fibrosis, may be mediated by phosphorylation of the cystic fibrosis transmembrane conductance regulator (CFTR) by cyclic AMP-dependent protein kinase (PKA) or protein kinase C (PKC). Part of the R-domain of CFTR (termed CF-2) was expressed in and purified from Escherichia coli. CF-2 was phosphorylated on seryl residues by PKA, PKC, cyclic GMP-dependent protein kinase (PKG), and calcium/calmodulin-dependent protein kinase I (CaM kinase I). Direct amino acid sequencing and peptide mapping of CF-2 revealed that serines 660, 700, 737, and 813 as well as serine 768, serine 795, or both were phosphorylated by PKA and PKG, and serines 686 and 790 were phosphorylated by PKC. CFTR was phosphorylated in vitro by PKA, PKC, or PKG on the same sites that were phosphorylated in CF-2. Kinetic analysis of phosphorylation of CF-2 and of synthetic peptides confirmed that these sites were excellent substrates for PKA, PKC, or PKG. CFTR was immunoprecipitated from T84 cells labeled with 32Pi. Its phosphorylation was stimulated in response to agents that activated either PKA or PKC. Peptide mapping confirmed that CFTR was phosphorylated at several sites identified in vitro. Thus, regulation of CFTR is likely to occur through direct phosphorylation of the R-domain by protein kinases stimulated by different second messenger pathways.

A dehydrogenative functionalization of styryl carbon-carbon double bonds with triflamide is described via dual visible-light photoredox/cobaloxime catalysis. A range of allylic triflamides (20 examples) were isolated in moderate to good yields (up to 88%) under stoichiometric acceptorless conditions. Dedicated labelling, as well as spectroscopic experiments, enabled to shed light on the concatenated photo- and chemo-catalytic cycles. (Figure presented.).

The first enantioselective [12 + 2] cycloaddition has been developed for the construction of a chiral cycl[3.2.2]azine core, a tricyclic moiety with a central ring-junction nitrogen atom, by an operationally simple one-step organocatalytic process. The reaction concept builds upon aminocatalytically generated 12π-components derived from 5H-benzo[a]pyrrolizine-3-carbaldehydes reacting with different electron-deficient 2π-components and affording the complex scaffold of benzo[a]cycl[3.2.2]azine (indolizino[3,4,5-ab]isoindole) with excellent enantio- A nd diastereoselectivity in good yields. The developed reaction is robust toward diverse substituent patterns and has been extended to different classes of electron-deficient 2π-components by minor variations in reaction setup. The obtained [12 + 2] cycloadducts are electron-deficient in nature, and their reaction with nucleophiles have been demonstrated. The enantioselective [12 + 2] cycloaddition with α,β-unsaturated aldehydes as the electron-deficient 2π-components relies upon an unconventional, simple aminocatalyst. In order to understand the high stereoselectivity of the [12 + 2] cycloaddition for this simple catalyst, combined experimental and computational investigations were performed. The investigations point to activation of both the 5H-benzo[a]pyrrolizine-3-carbaldehyde and the α,β-unsaturated aldehyde by the aminocatalyst and that the reaction proceeds by a stepwise cycloaddition, where especially the ring-closure is crucial for the stereochemical outcome. For other electron-deficient 2π-components, such as α,β-unsaturated ketoesters and nitroolefins, a more sterically demanding aminocatalyst has been applied and the corresponding [12 + 2] cycloadducts are obtained with excellent stereoselectivity. The [12 + 2] cycloaddition with vinyl sulfones afforded fully unsaturated systems, which display photoluminescence properties and for which quantum yields have been evaluated.

We present an environmentally benign methodology for the covalent functionalization (arylation) of reduced graphene oxide (rGO) nanosheets with arylazo sulfones. A variety of tagged aryl units were conveniently accommodated at the rGO surface via visible light irradiation of suspensions of carbon nanostructured materials in aqueous media. Mild reaction conditions, absence of photosensitizers, functional group tolerance and high atomic fractions (XPS analysis) represent some of the salient features characterizing the present methodology. Control experiments for the mechanistic elucidation (Raman analysis) and chemical nanomanipulation of the tagged rGO surfaces are also reported.

A carboxylative Ni-catalyzed tandem C-C σ-bond activation of cyclobutanones followed by CO2-electrophilic trapping is documented as a direct route to synthetically valuable 3-indanone-1-acetic acids. The protocol shows an adequate functional group tolerance and useful chemical outcomes (yield up to 76%) when AlCl3 is adopted as an additive. Manipulations of the targeted cyclic scaffolds and a mechanistic proposal based on experimental evidence complete the investigation.

The first stereoselective synthesis of enantioenriched axially chiral indole–quinoline systems is presented. The strategy takes advantage of an organocatalytic enantioselective Povarov cycloaddition of 3-alkenylindoles and N-arylimines, followed by an oxidative central-to-axial chirality conversion process, allowing for access to previously unreported axially chiral indole–quinoline biaryls. The methodology is also implemented for the design and the preparation of challenging compounds exhibiting two stereogenic axes. DFT calculations shed light on the stereoselectivity of the central-to-axial chirality conversion, showing unconventional behavior.

The development of the first chemo-, regio-, and stereoselective hetero-[6+4] and -[6+2] cycloadditions of heteroaromatic compounds via amino aza- and diazafulvenes is presented. Pyrroles, imidazoles, and pyrazoles substituted with a formyl group react with an aminocatalyst to generate an electron-rich hetero-6π-component that reacts in a chemo-, regio-, and stereoselective manner with electron-deficient dienes and olefins. For the hetero-[6+4] cycloaddition of the pyrrole system with dienes, a wide variation of both reaction partners is possible, providing attractive pyrrolo-azepine products in high yields and excellent enantioselectivities (99% ee). The hetero-[6+4] cycloaddition reaction concept is extended to include imidazoles and pyrazoles, giving imidazolo- and pyrazolo-azepines. The same activation concept is successfully employed to include hetero-[6+2] cycloadditions of the pyrrole system with nitroolefins, giving important pyrrolizidine-alkaloid scaffolds. Experimental NMR and mechanistic studies allowed for the identification of two different types of intermediates in the reaction. The first intermediate is the result of a rapid formation of an iminium ion, which generates a hetero-6π aminofulvene intermediate as a mixture of two isomers. Density functional theory calculations were used to determine the mechanism and sources of asymmetric induction in the hetero-[6+4] and -[6+2] cycloadditions. After formation of the reactive hetero-6π-components, a stepwise addition occurs with the diene or olefin, leading to a zwitterionic intermediate that undergoes cyclization to afford the cycloadduct, followed by eliminative catalyst release. The stereoselectivity is controlled by the second step, and computations elaborate on the various substrate and catalyst effects that alter the experimentally observed enantioselectivities. The computational studies provided a basis for improving the enantioselectivity of the hetero-[6+2] cycloaddition.

A highly stereoselective 1,3-dipolar [6+4] cycloaddition towards bridged azabicyclo[4.3.1]decane scaffolds has been developed, reacting aldehydes, 2-aminomalonates and tropone under mild conditions in the presence of a chiral phosphoric acid catalyst. The scope is demonstrated for a series of aldehydes and 2-aminomalonates, and the reaction proceeds in high yields, >95:5 d.r. and up to 99 % ee. A series of transformations, as well as a mechanistic proposal, are presented.

In this work, a new chemodivergent domino approach for the preparation of various saturated heterocycles, based on phase-transfer catalysis (PTC), is presented. The versatile nature of doubly electrophilic substrates, showing both a Michael acceptor and a ketone, tethered by a heteroatom, enables three different domino reaction pathways. The nucleophile dictates the chemoselectivity of the reaction. Sulfa-Michael/aldol, cyanide addition/oxa-Michael and Michael/H-shift/aldol processes, along with the variation of the tethering heteroatom, results in the formation of six different classes of saturated heterocycles. DFT calculations account for the observed chemo- and diastereoselectivity of the two most productive processes. Moreover, an extensive investigation on the sulfa-Michael/aldol pathway was carried out, ultimately leading to the development of a new enantioselective domino approach to multi-substituted piperidines based on PTC. (Figure presented.).

In this paper a straightforward synthesis of a novel pyrazole derivative is reported. Prominent feature of this synthetic process is a 1,3-Dipolar Cycloaddition of a suitable nitrile imine with an activated α,β-unsaturated lactam to afford directly and regioselectively the corresponding ring-fused pyrazole. Having obtained the central core of the synthetic target, a double stepwise functionalization with a “side chain” characterized by a terminal cyclic aliphatic amine was carried out. This molecular structure was designed to interact strongly with typical biological residues, and indeed it showed potent anticancer capability: in vitro cytotoxicity test on five different cancer cell lines showed interesting IC50 values in the range of 15–60 μM for exposure time of 24–72 h, thus resulting comparable with commercially available and nowadays therapeutically exploited anticancer compounds, such as 5-FU and NVP-BEZ235.

Reactive oxygen species (ROS) are chemically reactive molecules with impaired electrons that make them unstable and able to react easily with a great variety of molecules. The main targets of ROS are DNA, proteins, and membrane phospholipids. In the skin, ROS are able to affect the production of collagen and elastin, the main components of the extracellular matrix (ECM). This action contributes to the skin's aging. N-Acetylcysteine (NAC) is an acetylated cysteine residue with excellent anti-oxidant activity that boosts glutathione (GSH) levels. This study evaluates the effect of a solution of NAC and amino acids, which is used in aesthetic medicine as an intra-dermal injective treatment, on fibroblast behavior. To this aim, the expression levels of some ECM-related genes (HAS1, HYAL1 ELN, ELANE, MMP2, MMP3, MMP13, COL1A1, COL3A1) were analyzed on cultured dermal fibroblasts using real-time reverse transcription polymerase chain reaction (RT-PCR). All but two collagen genes were up-regulated after 24 hr of treatment.

La condizione dei diritti della povertà appare oggi come una nuova emergenza. Uno dei compiti necessari di ogni politica di inclusione, che non voglia porsi al tempo stesso come una semplice omologazione all'ordine, dovrebbe essere la salvaguardia - anche all'interno di quella che è pur sempre un'imposizione del diritto - di ogni eventuale spazio che la povertà e la sua differenza siano ancora in grado di rivendicare, rispetto alla norma che ne ha 'cura'. Questo spazio della differenza potrebbe forse interrogarci nuovamente anche sulla verità dell'ordine economico che, ostinatamente, nonostante le sue crisi, la 'legge' continua a replicare e a difendere.