Analysis of sequence variability and transcriptional profile of cannabinoid synthase genes in cannabis sativa l. Chemotypes with a focus on cannabichromenic acid synthase

Cannabis sativa L. has been long cultivated for its narcotic potential due to the accumulation of tetrahydrocannabinolic acid (THCA) in female inflorescences, but nowadays its production for fiber, seeds, edible oil and bioactive compounds has spread throughout the world. However, some hemp varieties still accumulate traces of residual THCA close to the 0.20% limit set by European Union, despite the functional gene encoding for THCA synthase (THCAS) is lacking. Even if some hypotheses have been produced, studies are often in disagreement especially on the role of the cannabichromenic acid synthase (CBCAS). In this work a set of European Cannabis genotypes, representative of all chemotypes, were investigated from a chemical and molecular point of view. Highly specific primer pairs were developed to allow an accurate distinction of different cannabinoid synthases genes. In addition to their use as markers to detect the presence of CBCAS at genomic level, they allowed the analysis of transcriptional profiles in hemp or marijuana plants. While the high level of transcription of THCAS and cannabidiolic acid synthase (CBDAS) clearly reflects the chemical phenotype of the plants, the low but stable transcriptional level of CBCAS in all genotypes suggests that these genes are active and might contribute to the final amount of cannabinoids

Different tool training induces specific effects on body metric representation

Morphology and functional aspects of the tool have been proposed to be critical factors modulating tool use-induced plasticity. However, how these aspects contribute to changing body representation has been underinvestigated. In the arm bisection task, participants have to estimate the length of their own arm by indicating its midpoint, a paradigm used to investigate the representation of the metric properties of the body. We employed this paradigm to investigate the impact of different actions onto tool embodiment. Our findings suggest that a training requiring actions mostly with proximal (shoulder) or distal (wrist) parts induces a different shift in the perceived arm midpoint. This effect is independent of, but enhanced by, the use of the tool during the training and in part influenced by specific demands of the task. These results suggest that specific motor patterns required by the training can induce different changes of body representation, calling for rethinking the concept of tool embodiment, which would be characterized not simply by the morphology of the tools, but also by the actions required for their specific use

Spatial limits of visuotactile interactions in the presence and absence of tactile stimulation

The presence of a light flash near to the body not only increases the ability to detect a weak touch but also increases reports of feeling a weak touch that did not occur. The somatic signal detection task (SSDT) provides a behavioural marker by which to clarify the spatial extent of such visuotactile interactions in peripersonal space. Whilst previous evidence suggests a limit to the spatial extent over which visual input can distort the perception of tactile stimulation during the rubber hand illusion, the spatial boundaries of light-induced tactile sensations are not known. In a repeated measures design, 41 participants completed the SSDT with the light positioned 1 cm (near), 17.5 cm (mid) or 40 cm (far) from the tactile stimulation. In the far condition, the light did not affect hit, or false alarm rates during the SSDT. In the near and mid conditions, the light significantly increased hit rates and led to a more liberal response criterion, that is, participants reported feeling the touch more often regardless of whether or not it actually occurred. Our results demonstrate a spatial boundary over which visual input influences veridical and non-veridical touch perception during the SSDT, and provide further behavioural evidence to show that the boundaries of the receptive fields of visuotactile neurons may be limited to reach space

Thermal-Perception-Driven Adaptive Design for Wellbeing in Outdoor Public Spaces: Case Studies in Naples

The spread of digital technologies, aiming at improving the effectiveness of the technological and environmental project proposals, has transformed the modus operandi for architects and designers who approach environmental impact assessment, especially about public space designs. Research activities aim at collecting guidelines for the sustainable regeneration of public spaces, focusing on the effectiveness of the performance of individual actions proposed by gradually checking and fixing the convenient benchmark design required by norms and sometimes by technology and building best-practices widely consolidated, even on a scientific basis. Early design optimization process relies on the combined use of appropriate IT tools for environmental control and on the interoperability of these systems with the traditional modelling tools for outdoor and indoor spaces. According to data-design-oriented logic, the core of the research methodology is applied to three case studies concerning public “complex” open spaces within the Neapolitan urban context (Italy)

Transmembrane helix structure, dynamics, and interactions: multi-nanosecond molecular dynamics simulations.

To probe the fundamentals of membrane/protein interactions, all-atom multi-nanosecond molecular dynamics simulations were conducted on a single transmembrane poly(32)alanine helix in a fully solvated dimyristoyphosphatidylcholine (DMPC) bilayer. The central 12 residues, which interact only with the lipid hydrocarbon chains, maintained a very stable helical structure. Helical regions extended beyond these central 12 residues, but interactions with the lipid fatty-acyl ester linkages, the lipid headgroups, and water molecules made the helix less stable in this region. The C and N termini, exposed largely to water, existed as random coils. As a whole, the helix tilted substantially, from perpendicular to the bilayer plane (0 degree) to a 30 degrees tilt. The helix experienced a bend at its middle, and the two halves of the helix at times assumed substantially different tilts. Frequent hydrogen bonding, of up to 0.7 ns in duration, occurred between peptide and lipid molecules. This resulted in correlated translational diffusion between the helix and a few lipid molecules. Because of the large variation in lipid conformation, the lipid environment of the peptide was not well defined in terms of "annular" lipids and on average consisted of 18 lipid molecules. When compared with a "neat" bilayer without peptide, no significant difference was seen in the bilayer thickness, lipid conformations or diffusion, or headgroup orientation. However, the lipid hydrocarbon chain order parameters showed a significant decrease in order, especially in those methylene groups closest to the headgroup

Use-dependent hemispheric balance.

In the human brain, homologous regions of the primary motor cortices (M1s) are connected through transcallosal fibers. Interhemispheric communication between the two M1s plays a major role in the control of unimanual hand movements, and the strength of this connection seems to be dependent on arm activity. For instance, a lesion in the M1 can induce an increase in the excitability of the intact M1 and an abnormal high inhibitory influence onto the damaged M1. This can be attributable to either the disuse of the affected limb or the overuse of the unaffected one. Here, to directly investigate cortical modifications induced by an abnormal asymmetric use of the two limbs, we studied both the excitability of the two M1s and transcallosal interaction between them in healthy subjects whose right hand was immobilized for 10 h. The left "not-immobilized" arm was completely free to move in one group of participants (G1) and limited in the other one (G2). We found that the non-use reduced the excitability of the left M1 and decreased the inhibitory influence onto the right hemisphere in the two groups. However, an increase in the excitability of right M1 and a deeper inhibitory interaction onto the left hemisphere were evident only in G1. Thus, modifications in the right M1 were not directly produced by the non-use but would depend on the overuse of the "not-immobilized" arm. Our findings suggest that the balance between the two M1s is strongly use dependent

When Cannabis sativa L. Turns Purple: Biosynthesis and Accumulation of Anthocyanins

Environmental cues elicit anthocyanin synthesis in plant vegetative and reproductive tissues. Their accumulation in different organs accounts for their diverse biological functions, mainly related to their antioxidant properties, and it depends on a temporally and spatially regulated mechanism controlled by the action of a well-known multi-transcription factor complex. Despite the highly recognizable value of Cannabis sativa L. as a natural biorefinery of phytochemicals, very little information is known on anthocyanin pigmentation in this species. In this work, a targeted quantification of anthocyanins via HPLC-MS/MS, combined with the transcriptional profile via RT-qPCR of genes encoding for structural and decorating enzymes and regulatory transcription factors in different C. sativa tissues, help gain insights into the anthocyanin pathway in this species. To the best of our knowledge, this is the first report on the identification of cyanidin-3-rutinoside (keracyanin) as the major anthocyanin in C. sativa vegetative and floral tissues. Keracyanin amounts were higher than in small berries, suggesting that Cannabis biomass is a valuable source of colored antioxidants to be exploited in diverse applications. Furthermore, a gene putatively encoding for an anthocyanin DTX35 type transporter and CsTTG1 were identified in silico and their transcriptional levels were assessed via RT-qPCR. The results allow us to provide the first model of anthocyanin regulation in C. sativa, opening a new research scenario in this species for both breeding purposes and phytochemical exploitation

Utilizzo del sistema CRISPR/Cas9 per ottenere partenocarpia in pomodoro

Con genome editing s\u2019intende l\u2019introduzione di una modifica in un punto preciso del genoma della cellula attraverso l\u2019utilizzo di nucleasi artificiali che operano sul gene bersaglio una rottura, la cui riparazione pu\uf2 generare mutazioni ed annullare la funzione del gene. Il sistema pi\uf9 diffuso \ue8 quello CRISPR/Cas9 di Streptococcus pyogenes, e prevede che una DNA nucleasi (Cas9) venga guidata ad una sequenza di DNA da modificare, grazie alla sua associazione con una piccola molecola di RNA (RNA guida) in grado di appaiarsi alla sequenza bersaglio. L\u2019allegagione, l\u2019inizio della crescita dell\u2019ovario a seguito della fecondazione, \ue8 un processo coordinato da segnali endogeni, soprattutto ormonali, ma che risente di fattori ambientali, che se sfavorevoli, possono avere effetti negativi sulla fecondazione e sulla produttivit\ue0. Numerosi geni che controllano l\u2019allegagione sono stati individuati. La partenocarpia \ue8 un processo alternativo in cui lo sviluppo del frutto avviene in assenza di fecondazione ed indipendentemente dalle condizioni ambientali. La manipolazione genetica del metabolismo e della trasduzione del segnale dell\u2019auxina e delle gibberelline ha permesso di identificare i geni che controllano la partenocarpia. Nel presente studio, il sistema CRISPR \ue8 stato usato per indurre mutazioni nel gene SlARF7, noto per essere un regolatore dello sviluppo partenocarpico del frutto di pomodoro. Tre sequenze nucleotidiche corrispondenti a 3 RNA guida aventi come bersaglio 3 regioni diverse del gene SlARF7 sono state clonate nel T-DNA di un vettore binario pFGC-pcoCAS9 (https://www.addgene.org; Li et al., 2013). L\u2019efficacia dei tre RNA del costrutto nell\u2019indurre il taglio nel gene ARF7- RNA guida mediato, \ue8 stata testata in vitro attraverso l\u2019impiego del kit commerciale \u201cGuide-it sgRNA in vitro Transcription and Screening Systems\u201d (Clontech). Il vettore binario \ue8 stato poi inserito in Agrobacterium tumefaciens (ceppo GV2260) ed impiegato per indurre mutazioni nella cv. UC8