CRISPR/Cas-assisted Nanoneedle Sensor for ATP Detection in Living Cells

The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) (CRISPR/Cas) systems have recently emerged as a powerful molecular biosensing tool based on their collateral cleavage activity due to their simplicity, sensitivity, specificity, and broad applicability. However, the direct application of collateral cleavage activity for in-situ intracellular detection is still challenging. Here, we debut a CRISPR/Cas-assisted nanoneedle sensor (nanoCRISPR) for intracellular adenosine triphosphate (ATP), which avoids the challenges associated with intracellular collateral cleavage by introducing a two-step process of intracellular target recognition followed by extracellular transduction and detection. ATP recognition occurs by first presenting in the cell cytosol an aptamer-locked Cas12a activator conjugated to nanoneedles; the recognition event unlocks the activator immobilized on the nanoneedles. The nanoneedles are then removed from the cells and exposed to the Cas12a/crRNA complex, where the activator triggers the cleavage of a ssDNA fluorophore-quencher pair, generating a detectable fluorescence signal. NanoCRISPR has an ATP detection limit of 246 nM and a dynamic range from 1.56 μM to 50 μM. Importantly, nanoCRISPR can detect intracellular ATP in 30 min in live cells without impacting cell viability. We anticipate that the nanoCRISPR approach will contribute to broaden the biomedical applications of CRISPR/Cas sensors for the detection of diverse intracellular molecules in living systems

Nanoneedles Induce Targeted siRNA Silencing of p16 in the Human Corneal Endothelium

: Nanoneedles can target nucleic acid transfection to primary cells at tissue interfaces with high efficiency and minimal perturbation. The corneal endothelium is an ideal target for nanoneedle-mediated RNA interference therapy aimed at enhancing its proliferative capacity, necessary for tissue regeneration. This work develops a strategy for siRNA nanoninjection to the human corneal endothelium. Nanoneedles can deliver p16-targeting siRNA to primary human corneal endothelial cells in vitro without toxicity. The nanoinjection of siRNA induces p16 silencing and increases cell proliferation, as monitored by ki67 expression. Furthermore, siRNA nanoinjection targeting the human corneal endothelium is nontoxic ex vivo, and silences p16 in transfected cells. These data indicate that nanoinjection can support targeted RNA interference therapy for the treatment of endothelial corneal dysfunction

Extreme Bendability of Atomically Thin MoS2 Grown by Chemical Vapor Deposition Assisted by Perylene-Based Promoter

Shaping two-dimensional (2D) materials in arbitrarily complex geometries is a key to designing their unique physical properties in a controlled fashion. This is an elegant solution, taking benefit from the extreme flexibility of the 2D layers but requiring the ability to force their spatial arrangement from flat to curved geometries in a delicate balance among free-energy contributions from strain, slip-and-shear mechanisms, and adhesion to the substrate. Here, we report on a chemical vapor deposition approach, which takes advantage of the surfactant effects of organic molecules, namely the tetrapotassium salt of perylene-3,4,9,10-tetracarboxylic acid (PTAS), to conformally grow atomically thin layers of molybdenum disulphide (MoS2) on arbitrarily nanopatterned substrates. Using atomically resolved transmission electron microscope images and density functional theory calculations, we show that the most energetically favorable condition for the MoS2 layers consists of its adaptation to the local curvature of the patterned substrate through a shear-and-slip mechanism rather than strain accumulation. This conclusion also reveals that the perylene-based molecules have a role in promoting the adhesion of the layers onto the substrate, no matter the local-scale geometry

Optical Conductivity of Two-Dimensional Silicon: Evidence of Dirac Electrodynamics

The exotic electrodynamics properties of graphene come from the linearly dispersive electronic bands that host massless Dirac electrons. A similar behavior was predicted to manifest in freestanding silicene, the silicon counterpart of graphene, thereby envisaging a new route for silicon photonics. However, the access to silicene exploitation in photonics was hindered so far by the use of optically inappropriate substrates in experimentally realized silicene. Here we report on the optical conductivity of silicon nanosheets epitaxially grown on optically transparent Al2O3(0001) from a thickness of a few tens of nanometers down to the extreme two-dimensional (2D) limit. When a 2D regime is approached, a Dirac-like electrodynamics can be deduced from the observation of a low-energy optical conductivity feature owing to a silicene-based interfacing to the substrate

Second Harmonic Generation Circular Dichroism from Self-Ordered Hybrid Plasmonic-Photonic Nanosurfaces

Hybrid structures composed by self-ordered dielectric nanospheres and partially covered by gold nanocrescents, produce efficient second harmonic generation signals due to the cooperative effect of localized plasmon excitation and 2D Bragg gratings. Asymmetries in the geometric shape of the gold nanocrescents induce a circularly polarized optical response

Ecosistemi da progettare. Esercizi progettuali per la conservazione della biodiversit\ue0, il ripristino funzionale degli ecosistemi e l\u2019accessibilit\ue0 alle risorse naturali.

In un\u2019epoca di grande pressione sugli ecosistemi naturali, l\u2019uso del territorio per la conservazione della biodiversit\ue0, il recupero funzionale degli ecosistemi pi\uf9 degradati e di sistemi produttivi che si coniughino alle esigenze di biodiversit\ue0, hanno un ruolo strategico di primaria importanza per assicurare la sostenibilit\ue0 delle risorse. La biodiversit\ue0 \ue8 tutto ci\uf2 che abbiamo. Biodiversit\ue0 \ue8 ossigeno, cibo, medicine, risorse economiche, resilienza e adattamento ai cambiamenti naturali, cos\uec come a quelli indotti dalle attivit\ue0 umane. Un mondo senza uccelli, insetti, prati, aree umide o boschi non sosterebbe la popolazione umana esistente, e certamente non renderebbe possibile la ripresa economica che tutti ci auspichiamo. Abbiamo chiesto agli studenti del corso di \u201cEcosistemi, habitat protetti e ripristini ambientali\u201d della Laurea Magistrale in \u201cProgettazione e gestione degli ecosistemi agro-territoriali, forestali e del paesaggio\u201d dell\u2019Alma Mater Studiorum, Universit\ue0 degli Studi di Bologna, di fare un esercizio teorico-pratico sulla base di questa prospettiva e cio\ue8 di elaborare progetti di ripristino ambientale, conservazione della natura, e uso sostenibile delle risorse. Gli studenti hanno risposto con grande seriet\ue0 ed entusiasmo a questa proposta formativa. Sono loro gli ideatori, interlocutori e anche i beneficiari dei progetti che vengono presentati in questo booklet. In alcuni casi hanno ritenuto opportuno coinvolgere la cittadinanza e le associazioni di volontariato locali per poter sviluppare i loro progetti sulla base delle esigenze dei fruitori di queste aree. Queste sono vere e proprie azioni di Citizen Science che integrano l\u2019uso sostenibile dell\u2019ambiente con le esigenze economiche e della societ\ue0, e forniscono una prova della professionalit\ue0 acquisita dai ragazzi in questo percorso. Nell\u2019elaborazione dei progetti i ragazzi sono stati affiancati e stimolati da esperti e professionisti di vari settori che hanno permesso di indirizzare l\u2019esercizio teorico necessario per il sostenimento dell\u2019esame alle esigenze specifiche e attuali dei territori studiati. Con questo booklet desideriamo far conoscere agli addetti ai lavori, enti di gestione, aree protette, comuni, regioni, ai proprietari delle aree scelte come modello per l\u2019esercizio progettuale e alla cittadinanza, i progetti sviluppati nei loro territori. Possono servire come spunto per la pianificazione e la gestione dei territori specifici, ma esprimono anche la necessit\ue0 dei nostri ragazzi di un ambiente fatto di natura e biodiversit\ue0

Long-term proactive management of psoriasis with calcipotriol and betamethasone dipropionate foam: an Italian consensus through a combined nominal group technique and Delphi approach

none85Background: Although long-term management of psoriasis is paramount, this approach is challenging in clinical practice. In the recent PSO-LONG trial, a fixed-dose combination of betamethasone dipropionate (BD) and calcipotriol (Cal) foam applied twice a week on non-consecutive days for 52 weeks (proactive treatment) reduced the risk of relapse. However, the role of Cal/BD foam in the long-term management of psoriasis needs further clarifications. The ProActive Management (PAM) program, a nationwide Italian project, aims at reaching a consensus on the role of proactive management of psoriasis. Methods: A steering committee generated some statements through the nominal group technique (NGT). The statements were voted by an expert panel in an adapted Delphi voting process. Results: Eighteen statements were proposed, and the majority of them (14/18) reached a consensus during the Delphi voting. The need to provide long-term proactive topical treatment to reduce the risk of relapse for the treatment of challenging diseases sites or in patients where phototherapy or systemic therapies are contraindicated/ineffective was widely recognized. A consensus was reached about the possibility to associate the proactive treatment with systemic and biological therapies, without the need for dose intensification, thus favoring a prolonged remission. Moreover, the proactive treatment was recognized as more effective than weekend therapy in increasing time free from relapses. Approaches to improve adherence, on the other hand, need further investigation. Conclusions: The inclusion in guidelines of a proactive strategy among the effective treatment options will be a fundamental step in the evolution of a mild-moderate psoriasis therapeutic approach.noneDe Simone, Clara; Dapavo, Paolo; Malagoli, Piergiorgio; Martella, Alessandro; Campanati, Anna; Campione, Elena; Errichetti, Enzo; Franchi, Chiara; Gambardella, Alessio; Megna, Matteo; Osti, Federica; Ribero, Simone; Zagni, Giovanni; Calzavara-Pinton, Piergiacomo; Fabbrocini, Gabriella; Amoruso, Giuseppe Fabrizio; Baglieri, Francesco; Biamonte, Anna Silvia; Bianchelli, Tommaso; Bigi, Laura; Bortoli, Jarno; Brunetti, Bruno; Buligan, Cinzia; Cagni, Elisabetta; Calderoni, Ombretta; Calzavara-Pinton, Piergiacomo; Campanati, Anna; Caputo, Alighiero; Carrera, Carlo Giovanni; Carugno, Andrea; Chersi, Karin; Cicchelli, Stefano; De Natale, Flora; De Simone, Clara; Dapavo, Paolo; Di Maria, Domenico; Errichetti, Enzo; Fabbrocini, Gabriella; Ferrari, Angelo Salvatore; Fogli, Emanuela; Forconi, Riccardo; Franchi, Chiara; Galeazzi, Augusto; Gambardella, Alessio; Giovannini, Andrea; Giura, Maria Teresa; Iuculano, Massimo; Lazzaretti, Giuseppe; Leporati, Claudia; Magnanini, Massimiliano; Malagoli, Piergiorgio; Marconi, Barbara; Martella, Alessandro; Maruccia, Adriana; Megna, Matteo; Miglietta, Roberta; Minuti, Anna; Mocci, Luigi; Modica, Sonia; Narcisi, Alessandra; Odorici, Giulia; Osti, Federica; Pazzaglia, Massimiliano; Peila, Rossana; Pertusi, Ginevra; Pezza, Michele; Pezzullo, Elio; Puccia, Nunzio; Raulo, Umberto; Ribero, Simone; Rossi, Mariateresa; Rusignuolo, Sergio; Sapienza, Giada; Savarese, Catello; Scalisi, Mariaelena; Strippoli, Davide; Stroppiana, Elena; Tiberio, Rossana; Trischitta, Antonino; Tucci, Maria Giovanna; Vaira, Fabrizio; Verrone, Anna; Villa, Lucia; Zagni, Fabio; Zoccali, AndreaDe Simone, Clara; Dapavo, Paolo; Malagoli, Piergiorgio; Martella, Alessandro; Campanati, Anna; Campione, Elena; Errichetti, Enzo; Franchi, Chiara; Gambardella, Alessio; Megna, Matteo; Osti, Federica; Ribero, Simone; Zagni, Giovanni; Calzavara-Pinton, Piergiacomo; Fabbrocini, Gabriella; Amoruso, Giuseppe Fabrizio; Baglieri, Francesco; Biamonte, Anna Silvia; Bianchelli, Tommaso; Bigi, Laura; Bortoli, Jarno; Brunetti, Bruno; Buligan, Cinzia; Cagni, Elisabetta; Calderoni, Ombretta; Calzavara-Pinton, Piergiacomo; Campanati, Anna; Caputo, Alighiero; Carrera, Carlo Giovanni; Carugno, Andrea; Chersi, Karin; Cicchelli, Stefano; De Natale, Flora; De Simone, Clara; Dapavo, Paolo; Di Maria, Domenico; Errichetti, Enzo; Fabbrocini, Gabriella; Ferrari, Angelo Salvatore; Fogli, Emanuela; Forconi, Riccardo; Franchi, Chiara; Galeazzi, Augusto; Gambardella, Alessio; Giovannini, Andrea; Giura, Maria Teresa; Iuculano, Massimo; Lazzaretti, Giuseppe; Leporati, Claudia; Magnanini, Massimiliano; Malagoli, Piergiorgio; Marconi, Barbara; Martella, Alessandro; Maruccia, Adriana; Megna, Matteo; Miglietta, Roberta; Minuti, Anna; Mocci, Luigi; Modica, Sonia; Narcisi, Alessandra; Odorici, Giulia; Osti, Federica; Pazzaglia, Massimiliano; Peila, Rossana; Pertusi, Ginevra; Pezza, Michele; Pezzullo, Elio; Puccia, Nunzio; Raulo, Umberto; Ribero, Simone; Rossi, Mariateresa; Rusignuolo, Sergio; Sapienza, Giada; Savarese, Catello; Scalisi, Mariaelena; Strippoli, Davide; Stroppiana, Elena; Tiberio, Rossana; Trischitta, Antonino; Tucci, Maria Giovanna; Vaira, Fabrizio; Verrone, Anna; Villa, Lucia; Zagni, Fabio; Zoccali, Andre