Application of Encapsulation Technology: In Vitro Screening of Two Ficus carica L. Genotypes under Different NaCl Concentrations

Salinity stress represents an increasing issue for agriculture and has a great negative impact on plant growth and crop production. The selection of genotypes able to tolerate salt stress could be a suitable solution to overcome the problem. In this context, in vitro cultures can represent a tool for identifying the NaCl tolerant genotypes and quickly producing large populations of them. The possibility of exerting selection for tolerance to NaCl by using encapsulation technology was investigated in two genotypes of fig: 'Houmairi' and 'Palazzo'. The effects of five concentrations of NaCl (0, 50, 100, 150 and 200 mM) added to the artificial endosperm were tested on the conversion of synthetic seeds and on the growth of derived shoots/plantlets. Moreover, proline (Pro) and malondialdehyde (MDA), the enzymatic activities of catalase (CAT), guaiacol peroxidase (POD), and EL (Electrolytic Leakage), as well as the chlorophyll content, flavanols, anthocyanins, and Nitrogen Balance Index (NBI) were determined on shoots/plantlet. The obtained results clearly showed that 'Houmairi' and 'Palazzo' could tolerate salt stress, although a strong difference was found depending on each specific physiological pathway. Indeed, 'Houmairi' was revealed to be more tolerant than 'Palazzo', with different response mechanisms to salt stress. The use of encapsulated vitro-derived explants proved to be a useful method to validate the selection of genotypes tolerant to salinity stress. Further investigation in the field must validate and confirm the legitimacy of the approach

A Coevolutionary Residue Network at the Site of a Functionally Important Conformational Change in a Phosphohexomutase Enzyme Family

Coevolution analyses identify residues that co-vary with each other during evolution, revealing sequence relationships unobservable from traditional multiple sequence alignments. Here we describe a coevolutionary analysis of phosphomannomutase/phosphoglucomutase (PMM/PGM), a widespread and diverse enzyme family involved in carbohydrate biosynthesis. Mutual information and graph theory were utilized to identify a network of highly connected residues with high significance. An examination of the most tightly connected regions of the coevolutionary network reveals that most of the involved residues are localized near an interdomain interface of this enzyme, known to be the site of a functionally important conformational change. The roles of four interface residues found in this network were examined via site-directed mutagenesis and kinetic characterization. For three of these residues, mutation to alanine reduces enzyme specificity to ∼10% or less of wild-type, while the other has ∼45% activity of wild-type enzyme. An additional mutant of an interface residue that is not densely connected in the coevolutionary network was also characterized, and shows no change in activity relative to wild-type enzyme. The results of these studies are interpreted in the context of structural and functional data on PMM/PGM. Together, they demonstrate that a network of coevolving residues links the highly conserved active site with the interdomain conformational change necessary for the multi-step catalytic reaction. This work adds to our understanding of the functional roles of coevolving residue networks, and has implications for the definition of catalytically important residues

Ribosomal oxygenases are structurally conserved from prokaryotes to humans

2-Oxoglutarate (2OG)-dependent oxygenases have important roles in the regulation of gene expression via demethylation of N-methylated chromatin components1,2 and in the hydroxylation of transcription factors3 and splicing factor proteins4. Recently, 2OG-dependent oxygenases that catalyse hydroxylation of transfer RNA5,6,7 and ribosomal proteins8 have been shown to be important in translation relating to cellular growth, TH17-cell differentiation and translational accuracy9,10,11,12. The finding that ribosomal oxygenases (ROXs) occur in organisms ranging from prokaryotes to humans8 raises questions as to their structural and evolutionary relationships. In Escherichia coli, YcfD catalyses arginine hydroxylation in the ribosomal protein L16; in humans, MYC-induced nuclear antigen (MINA53; also known as MINA) and nucleolar protein 66 (NO66) catalyse histidine hydroxylation in the ribosomal proteins RPL27A and RPL8, respectively. The functional assignments of ROXs open therapeutic possibilities via either ROX inhibition or targeting of differentially modified ribosomes. Despite differences in the residue and protein selectivities of prokaryotic and eukaryotic ROXs, comparison of the crystal structures of E. coli YcfD and Rhodothermus marinus YcfD with those of human MINA53 and NO66 reveals highly conserved folds and novel dimerization modes defining a new structural subfamily of 2OG-dependent oxygenases. ROX structures with and without their substrates support their functional assignments as hydroxylases but not demethylases, and reveal how the subfamily has evolved to catalyse the hydroxylation of different residue side chains of ribosomal proteins. Comparison of ROX crystal structures with those of other JmjC-domain-containing hydroxylases, including the hypoxia-inducible factor asparaginyl hydroxylase FIH and histone Nε-methyl lysine demethylases, identifies branch points in 2OG-dependent oxygenase evolution and distinguishes between JmjC-containing hydroxylases and demethylases catalysing modifications of translational and transcriptional machinery. The structures reveal that new protein hydroxylation activities can evolve by changing the coordination position from which the iron-bound substrate-oxidizing species reacts. This coordination flexibility has probably contributed to the evolution of the wide range of reactions catalysed by oxygenases

Effects of Anacetrapib in Patients with Atherosclerotic Vascular Disease

BACKGROUND: Patients with atherosclerotic vascular disease remain at high risk for cardiovascular events despite effective statin-based treatment of low-density lipoprotein (LDL) cholesterol levels. The inhibition of cholesteryl ester transfer protein (CETP) by anacetrapib reduces LDL cholesterol levels and increases high-density lipoprotein (HDL) cholesterol levels. However, trials of other CETP inhibitors have shown neutral or adverse effects on cardiovascular outcomes. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 30,449 adults with atherosclerotic vascular disease who were receiving intensive atorvastatin therapy and who had a mean LDL cholesterol level of 61 mg per deciliter (1.58 mmol per liter), a mean non-HDL cholesterol level of 92 mg per deciliter (2.38 mmol per liter), and a mean HDL cholesterol level of 40 mg per deciliter (1.03 mmol per liter). The patients were assigned to receive either 100 mg of anacetrapib once daily (15,225 patients) or matching placebo (15,224 patients). The primary outcome was the first major coronary event, a composite of coronary death, myocardial infarction, or coronary revascularization. RESULTS: During the median follow-up period of 4.1 years, the primary outcome occurred in significantly fewer patients in the anacetrapib group than in the placebo group (1640 of 15,225 patients [10.8%] vs. 1803 of 15,224 patients [11.8%]; rate ratio, 0.91; 95% confidence interval, 0.85 to 0.97; P=0.004). The relative difference in risk was similar across multiple prespecified subgroups. At the trial midpoint, the mean level of HDL cholesterol was higher by 43 mg per deciliter (1.12 mmol per liter) in the anacetrapib group than in the placebo group (a relative difference of 104%), and the mean level of non-HDL cholesterol was lower by 17 mg per deciliter (0.44 mmol per liter), a relative difference of -18%. There were no significant between-group differences in the risk of death, cancer, or other serious adverse events. CONCLUSIONS: Among patients with atherosclerotic vascular disease who were receiving intensive statin therapy, the use of anacetrapib resulted in a lower incidence of major coronary events than the use of placebo. (Funded by Merck and others; Current Controlled Trials number, ISRCTN48678192 ; ClinicalTrials.gov number, NCT01252953 ; and EudraCT number, 2010-023467-18 .)

Biochemical Insights into the Ability of Lemna minor L. Extract to Counteract Copper Toxicity in Maize

Metal trace elements (MTE) can damage crops if present in excessive amounts in the environment. This research investigated the effect of a plant extract of an aquatic species, Lemna minor L. (duckweed) (LE), on the ability of maize to cope with copper (Cu) toxicity. LE reversed the effects of Cu2+ on photosynthetic activity (Pn), evapotranspiration (E), stomatal conductance (gs), sub-stomatal CO2 concentration (Ci) and biomass which did not differ from the untreated controls. LE did not regulate the amount of copper in maize leaves, but compared to Cu-treated samples, the extract decreased the hydrogen peroxide (H2O2; −26% on average) and malondialdehyde (MDA; −47% on average) content, regardless of the dosage applied. Furthermore, the activity of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) was significantly increased by LE compared to samples treated with Cu alone. Untargeted metabolomic profiling revealed that LE activated maize secondary metabolism, eliciting the content of non-enzymatic antioxidants (flavonoids, glutathione and glutathione-related compounds, tocopherols and tocotrienols) and modulating plant stress-related hormones (brassinosteroids and ABA derivatives). The results of this study are promising and pave the way for using duckweed as a biostimulant to trigger beneficial effects in maize and increase its resistance to MTEs

Le relazioni intergenerazionali, tra risorsa e vincolo

L\u2019intergenerazionalit\ue0 identifica in generale le relazioni tra membri di generazioni diverse come passaggio fondamentale per avanzare verso una societ\ue0 nella quale tutte le et\ue0 abbiano cittadinanza, secondo una concezione age friendly. Indica quindi una capacit\ue0/volont\ue0 di assumere da parte di tutte le generazioni implicate, sia a livello degli orientamenti sia a livello dell\u2019azione, una posizione generativa nei confronti delle generazioni, in particolare le pi\uf9 giovani. In tal modo essi possono diventare una risorsa per la societ\ue0 nel suo complesso. Difatti oggigiorno le persone di una stessa famiglia condividono pi\uf9 anni insieme, hanno maggiori opportunit\ue0 di entrare in relazione, un numero maggiore di passaggi critici da affrontare, in cui possono rientrare sia rischi sia risorse. L\u2019osservazione della famiglia come realt\ue0 oggi multigenerazionale (cio\ue8 oltre i confini della famiglia nucleare con due sole generazioni) richiede esplicitamente di prendere in considerazione le relazioni tra le diverse generazioni presenti sulla scena familiare . Questa prospettiva risulta particolarmente adeguata per comprendere le dinamiche familiari presenti in Italia, Paese tradizionalmente considerato a legami familiari forti (Castiglioni, Dalla Zuanna, 2017) in cui la maggior parte delle persone considera coincidenti l\u2019interesse proprio con quello della propria famiglia, e in cui la propria felicit\ue0 \ue8 ricercata insieme a quella dei propri genitori e dei propri figli (almeno dove presenti). In questo capitolo si prover\ue0 a mostrare come la prospettiva assunta dalla mediazione intergenerazionale sia particolarmente adeguata al nostro contesto sociale in cui, pur con importanti trasformazioni, i legami familiari e di solidariet\ue0 tra le generazioni continuano a essere considerati centrali per il benessere delle persone

Le relazioni intergenerazionali, tra risorsa e vincolo

L\u2019intergenerazionalit\ue0 identifica in generale le relazioni tra membri di generazioni diverse come passaggio fondamentale per avanzare verso una societ\ue0 nella quale tutte le et\ue0 abbiano cittadinanza, secondo una concezione age friendly. Indica quindi una capacit\ue0/volont\ue0 di assumere da parte di tutte le generazioni implicate, sia a livello degli orientamenti sia a livello dell\u2019azione, una posizione generativa nei confronti delle generazioni, in particolare le pi\uf9 giovani. In tal modo essi possono diventare una risorsa per la societ\ue0 nel suo complesso. Difatti oggigiorno le persone di una stessa famiglia condividono pi\uf9 anni insieme, hanno maggiori opportunit\ue0 di entrare in relazione, un numero maggiore di passaggi critici da affrontare, in cui possono rientrare sia rischi sia risorse. L\u2019osservazione della famiglia come realt\ue0 oggi multigenerazionale (cio\ue8 oltre i confini della famiglia nucleare con due sole generazioni) richiede esplicitamente di prendere in considerazione le relazioni tra le diverse generazioni presenti sulla scena familiare . Questa prospettiva risulta particolarmente adeguata per comprendere le dinamiche familiari presenti in Italia, Paese tradizionalmente considerato a legami familiari forti (Castiglioni, Dalla Zuanna, 2017) in cui la maggior parte delle persone considera coincidenti l\u2019interesse proprio con quello della propria famiglia, e in cui la propria felicit\ue0 \ue8 ricercata insieme a quella dei propri genitori e dei propri figli (almeno dove presenti). In questo capitolo si prover\ue0 a mostrare come la prospettiva assunta dalla mediazione intergenerazionale sia particolarmente adeguata al nostro contesto sociale in cui, pur con importanti trasformazioni, i legami familiari e di solidariet\ue0 tra le generazioni continuano a essere considerati centrali per il benessere delle persone