On \u3cem\u3ek\u3c/em\u3e-minimum and \u3cem\u3em\u3c/em\u3e-minimum Edge-Magic Injections of Graphs

An edge-magic total labelling (EMTL) of a graph G with n vertices and e edges is an injection λ:V(G) ∪ E(G)→[n+e], where, for every edge uv ∈ E(G), we have wtλ(uv)=kλ, the magic sum of λ. An edge-magic injection (EMI) μ of G is an injection μ : V(G) ∪ E(G) → N with magic sum kμ and largest label mμ. For a graph G we define and study the two parameters κ(G): the smallest kμ amongst all EMI’s μ of G, and m(G): the smallest mμ amongst all EMI’s μ of G. We find κ(G) for G ∈ G for many classes of graphs G. We present algorithms which compute the parameters κ(G) and m(G). These algorithms use a G-sequence: a sequence of integers on the vertices of G whose sum on edges is distinct. We find these parameters for all G with up to 7 vertices. We introduce the concept of a double-witness: an EMI μ of G for which both kμ=κ(G) and mμ=m(G) ; and present an algorithm to find all double-witnesses for G. The deficiency of G, def(G), is m(G)−n−e. Two new graphs on 6 vertices with def(G)=1 are presented. A previously studied parameter of G is κEMTL(G), the magic strength of G: the smallest kλ amongst all EMTL’s λ of G. We relate κ(G) to κEMTL(G) for various G, and find a class of graphs B for which κEMTL(G)−κ(G) is a constant multiple of n−4 for G ∈B. We specialise to G=Kn, and find both κ(Kn) and m(Kn) for all n≤11. We relate κ(Kn) and m(Kn) to known functions of n, and give lower bounds for κ(Kn) and m(Kn)

Impact of Seed Inoculation with Trichoderma afroharzianum Strains on Plant Growth, Root Morphology, and Leaf Phenolic Content in Hemp (Cannabis sativa L.) at Early Growth Stages

Industrial hemp (Cannabis sativa L.) is receiving increasing attention for its multiple end-uses; therefore, an improvement in its production is needed to meet the increased demand. In the present study, the effect of seed inoculation with two Trichoderma afroharzianum strains, T-AA and T-22, on plant growth and root morphology of hemp plants at sixth-leaf (S6) and tenth-leaf (S10) stages was assessed for two consecutive years (2020 and 2021). In addition, the ability of the two strains to enhance the accumulation of phenolic compounds in hemp leaves was also evaluated. The results obtained revealed the ability of T-22 to improve the growth and root morphology of hemp plants both in 2020 and 2021, although with different impact, probably ascribable to the different weather conditions in the two years. In 2020, the positive effects of T-22 were detected at S10 stage with significant increases in the shoot and root length (38% and 17%, respectively) and dried biomass (35% and 30%, respectively) compared to untreated plants. The total root surface area and the number of tips, forks, and crossings also increased significantly (24–36%) at this stage. In 2021, significant increases in the shoot length and dried biomass (40% and 30%, respectively) were observed at S6 stage, whereas root length and dried biomass increased significantly at S6 (55% and 47%, respectively) and S10 stage (121% and 40%, respectively). Significant increases in the total surface area and volume, as well as in the number of tips, forks, and crossings were also observed at both S6 and S10 stage (50–63% and 105–187%, respectively). Interestingly, in both years and at both stages, the two strains induced significant increases in the leaf accumulation of phenolic compounds and the antioxidant activity, which were greater in T-22- compared to T-AA-treated plants (18–102% and 13–34%, respectively). The results are discussed in light of the potential practical applications of T-22 as a biostimulant of hemp plant growth under favorable and unfavorable environmental conditions, and of both strains as promising tools for the improvement of the leaves’ economic value as a source of health-promoting compounds

hMENA11a contributes to HER3-mediated resistance to PI3K inhibitors in HER2-overexpressing breast cancer cells.

Human Mena (hMENA), an actin regulatory protein of the ENA/VASP family, cooperates with ErbB receptor family signaling in breast cancer. It is overexpressed in high-risk preneoplastic lesions and in primary breast tumors where it correlates with HER2 overexpression and an activated status of AKT and MAPK. The concomitant overexpression of hMENA and HER2 in breast cancer patients is indicative of a worse prognosis. hMENA is expressed along with alternatively expressed isoforms, hMENA11a and hMENAΔv6 with opposite functions. A novel role for the epithelial-associated hMENA11a isoform in sustaining HER3 activation and pro-survival pathways in HER2-overexpressing breast cancer cells has been identified by reverse phase protein array and validated in vivo in a series of breast cancer tissues. As HER3 activation is crucial in mechanisms of cell resistance to PI3K inhibitors, we explored whether hMENA11a is involved in these resistance mechanisms. The specific hMENA11a depletion switched off the HER3-related pathway activated by PI3K inhibitors and impaired the nuclear accumulation of HER3 transcription factor FOXO3a induced by PI3K inhibitors, whereas PI3K inhibitors activated hMENA11a phosphorylation and affected its localization. At the functional level, we found that hMENA11a sustains cell proliferation and survival in response to PI3K inhibitor treatment, whereas hMENA11a silencing increases molecules involved in cancer cell apoptosis. As shown in three-dimensional cultures, hMENA11a contributes to resistance to PI3K inhibition because its depletion drastically reduced cell viability upon treatment with PI3K inhibitor BEZ235. Altogether, these results indicate that hMENA11a in HER2-overexpressing breast cancer cells sustains HER3/AKT axis activation and contributes to HER3-mediated resistance mechanisms to PI3K inhibitors. Thus, hMENA11a expression can be proposed as a marker of HER3 activation and resistance to PI3K inhibition therapies, to select patients who may benefit from these combined targeted treatments. hMENA11a activity could represent a new target for antiproliferative therapies in breast cancer

KAP1 targets actively transcribed genomic loci to exert pleomorphic effects on RNA polymerase II activity

KAP1 (KRAB-associated protein 1) is best known as a co-repressor responsible for inducing heterochromatin formation, notably at transposable elements. However, it has also been observed to bind the transcription start site of actively expressed genes. To address this paradox, we characterized the protein interactome of KAP1 in the human K562 erythro-leukaemia cell line. We found that the regulator can associate with a wide range of nucleic acid binding proteins, nucleosome remodellers, chromatin modifiers and other transcription modulators. We further determined that KAP1 is recruited at actively transcribed polymerase II promoters, where its depletion resulted in pleomorphic effects, whether expression of these genes was normally constitutive or inducible, consistent with the breadth of possible KAP1 interactors. This article is part of a discussion meeting issue 'Crossroads between transposons and gene regulation'

Impact of Chitosan-Based Foliar Application on the Phytochemical Content and the Antioxidant Activity in Hemp (Cannabis sativa L.) Inflorescences

In the present study, the phytochemical content and the antioxidant activity in the inflorescences of the monoecious hemp cultivar Codimono grown in southern Italy were assessed, and their elicitation was induced by foliar spray application of 50 mg/L and 250 mg/L of chitosan (CHT) at three different molecular weights (low, CHT L; medium, CHT M; high CHT H). The analysis of the phytochemical profile confirmed that cannabinoids were the most abundant class (54.2%), followed by flavonoids (40.3%), tocopherols (2.2%), phenolic acids (1.9%), and carotenoids (1.4%). Cannabinoids were represented almost exclusively by cannabidiol, whereas cannabigerol and Δ9-tetrahydrocannabinol were detected at very low levels (the latter was below the legal limit of 0.3%). The most abundant flavonoids were orientin and vitexin, whereas tocopherols were mainly represented by α-tocopherol. The antioxidant activity was found to be positively correlated with flavonoids and tocopherols. Statistical analysis revealed that the CHT treatments significantly affected the phytochemical content and the antioxidant activity of hemp inflorescences. Notably, a significant increase in the total phenolic content (from +36% to +69%), the α-tocopherol (from +45% to +75%) and β+γ-tocopherol (from +35% to +82%) contents, and the ABTS radical scavenging activity (from +12% to +28%) was induced by all the CHT treatments. In addition, treatments with CHT 50 solutions induced an increase in the total flavonoid content (from +12% to +27%), as well as in the vitexin (from +17% to +20%) and orientin (from +20% to +30%) contents. Treatment with CHT 50 L almost always resulted in the greatest increases. Overall, our findings indicated that CHT could be used as a low-cost and environmentally safe elicitor to improve the health benefits and the economic value of hemp inflorescences, thus promoting their employment in the food, pharmaceutical, nutraceutical, and cosmetic supply chains

In-parallel polar monitoring of chemiluminescence emission anisotropy at the solid-liquid interface by an optical fiber radial array

Chemiluminescence (CL) detection is widely employed in biosensors and miniaturized analytical devices since it offers high detectability and flexible device design (there are no geometry requirements for the measurement cell, except the ability to collect the largest fraction of emitted photons). Although the emission anisotropy phenomenon for an emitting dipole bound to the interface between two media with different refractive index is well known for fluorescence, it is still poorly investigated for CL reactions, in which the excited-state reaction products can diffuse in solution before the photon emission event. In this paper, we propose a simple method for the realtime evaluation of the CL emission anisotropy based on a radial array of optical fibers, embedded in a poly(methyl methacrylate) semicylinder and coupled with a Charge-Coupled Device (CCD) camera through a suitable interface. The polar-time evolutions of the CL emission have been studied for catalyzing enzymes immobilized onto a solid surface (heterogeneous configuration) or free in solution (homogeneous configuration). Evidence of the anisotropy phenomenon is observed, indicating that the lifetime of the excited-state products of the enzyme-catalyzed reactions is shorter than the time required for their diffusion in solution at a distance at which the CL can be considered isotropic. These results open new perspectives in the development of CL-based miniaturized analytical devices

Ectopic expression of the beta-cell specific transcription factor Pdx1 inhibits glucagon gene transcription

Aims/hypothesis: The transcription factor Pdx1 is required for the development and differentiation of all pancreatic cells. Beta-cell specific inactivation of Pdx1 in developing or adult mice leads to an increase in glucagon-expressing cells, suggesting that absence of Pdx1could favour glucagon gene expression by a default mechanism. Method: We investigated the inhibitory role of Pdx1 on glucagon gene expression in vitro. The glucagonoma cell line InR1G9 was transduced with a Pdx1-encoding lentiviral vector and insulin and glucagon mRNA levels were analysed by northern blot and real-time PCR. To understand the mechanism by which Pdx1 inhibits glucagon gene expression, we studied its effect on glucagon promoter activity in non-islet cells using transient transfections and gel-shift analysis. Results: In glucagonoma cells transduced with a Pdx1-encoding lentiviral vector, insulin gene expression was induced while glucagon mRNA levels were reduced by 50 to 60%. In the heterologous cell line BHK-21, Pdx1 inhibited by 60 to 80% the activation of the α-cell specific element G1 conferred by Pax-6 and/or Cdx-2/3. Although Pdx1 could bind three AT-rich motifs within G1, two of which are binding sites for Pax-6 and Cdx-2/3, the affinity of Pdx1 for G1 was much lower as compared to Pax-6. In addition, Pdx1 inhibited Pax-6 mediated activation through G3, to which Pdx1 was unable to bind. Moreover, a mutation impairing DNA binding of Pdx1 had no effect on its inhibition on Cdx-2/3. Since Pdx1 interacts directly with Pax-6 and Cdx-2/3 forming heterodimers, we suggest that Pdx1 inhibits glucagon gene transcription through protein to protein interactions with Pax-6 and Cdx-2/3. Conclusion/interpretation: Cell-specific expression of the glucagon gene can only occur when Pdx1 expression extinguishes from the early α cell precurso

A KRAB/KAP1-miRNA Cascade Regulates Erythropoiesis Through Stage-Specific Control of Mitophagy

During hematopoiesis, lineage- and stage-specific transcription factors work in concert with chromatin modifiers to direct the differentiation of all blood cells. We explored the role of KRAB-containing zinc finger proteins (KRAB-ZFPs) and their cofactor KAP1 in this process. In mice, hematopoietic-restricted deletion of Kap1 resulted in severe hypoproliferative anemia. Kap1-deleted erythroblasts failed to induce mitophagy-associated genes and retained mitochondria. This was due to persistent expression of microRNAs (miRNAs) targeting mitophagy transcripts, itself secondary to a lack of repression by stage-specific KRAB-ZFPs. The KRAB/KAP1-miRNA regulatory cascade is evolutionarily conserved, as it also controls mitophagy during human erythropoiesis. Thus, a multilayered transcription regulatory system is present, in which protein- and RNA-based repressors are superimposed in combinatorial fashion to govern the timely triggering of an important differentiation event