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The Italian seed bank network (RIBES) aims to improve the quality and safety of the germplasm reserves of native plant species in Italy to ensure the long-term conservation of endangered and/or endemic flora. The strategy includes traditional methods to secure seed conservation. A comprehensive priority list for seed collection is being defined, it was prepared by crossing data of various checklists (red lists, endemics) and will soon be cross-referenced with an updated list of accessions of the whole network. A safety-backup program of duplicates will quickly be implemented to secure the conservation of the most threatened species in at least two seed banks of the network. On the other hand, the RIBES strategy also includes research by applying modern techniques. In collaboration with the Millennium Seed Bank, research on the storage behaviour of seeds and spores through thermal analysis is ongoing to inform conservation. Using the Differential Scanning Calorimetry (DSC), we could evaluate seed lipid properties such as glass transition temperature, melting, crystallization, oxidation behaviour, and thermal stability. Finally, RIBES participates as a co-funder in the LIFE Nature project SEEDFORCE, coordinating 11 seed banks of the network for collecting seeds/spores of 29 threatened species of EU interest. [ABSTRACT FROM AUTHOR]

Centaurea cinerariasubsp.circaeis an endemic plant with a distribution area limited to Circeo mountain (Lazio, Italy), whose population was estimated in a very low number of individuals. The aim of this work was to investigateex situconservation strategies such as achene collection andin vitroplant propagation, which will permit to carry out restoration programmes. The test carried out on the achenes demonstrated that only 5.5% of them were morphologically healthy. Seed germination tests showed that seeds do not display dormancy and that germination does not require pre-treatments. The higher germination rate (67.5%) was observed under a photoperiod of 12/12 h (light/dark) and temperature regime +20/+10°C. Thein vitrostudies demonstrated that micropropagation, acclimatization and the transfer outdoors ofC. cinerariasubsp.circaeare not particularly difficult: 74% of shoot explants in a Murashige and Skoog (MS) medium added with 0.5 mg/l benzylaminopurine and 2 mg/l kinetin formed multiple shoots; 100% of shoots rooted in the MS medium added with 0.5 mg/l indole-3-butyric acid and over 90% survived the acclimatization phase. After been transferred outdoors, the totality ofin vitro-propagated plants bloomed and appeared morphologically indistinguishable from wild plants. Preliminary chemical analyses showed a similar profile forin vitro-propagated and wild plants. [ABSTRACT FROM AUTHOR]

A reintroduction experiment of Bidens cernua L., a species included in the Red List of Italian Flora, was carried out at Lake Posta Fibreno (Lazio, central Italy). There were no significant differences in the length of the phenological phases between the reestablished population (Pr) and the natural one (Pn). The length of the phenological cycle, from seedling emergence to the end of the fruiting phase, was 207 ± 3 days. The relative growth rate in height (RGRH) and relative growth rate in mass (RGRm) were significantly (p < 0.05) higher for Pr (0.047 ± 0.001 cm cm-1 d-1 and 0.057 ± 0.001 g g-1 d-1, respectively) than for Pn (0.045 ± 0.001 cm cm-1 d-1 and 0.055 ± 0.001 g g-1 d-1, respectively). There were significant (p < 0.05) differences for plant height (H) and total leaf area per plant (TLA) between Pr and Pn, the latter having the lower values. Total plant dry mass (PDM) was significantly higher for Pr, and the above-ground/below-ground dry mass ratio was 6.7 ± 0.4 and 4.7 ± 0.2 for Pr and Pn, respectively. On the whole, results demonstrate that the Pr of B. cernua has the potential to develop in a manner that is demographically similar to the Pn present in the Posta Fibreno Lake protected area. Abbreviations: H, plant height; LA, mean leaf area; LM, leaf dry mass; PCA, principal component analysis; PDM, total plant dry mass; Pn, natural population; Pr, reestablished population; RGRH, relative growth rate in plant height; RGRm, relative growth rate in plant dry mass; SLA, specific leaf area; TLA, total leaf area per plant [ABSTRACT FROM AUTHOR]

Conservation translocations are becoming common conservation practice, so there is an increasing need to understand the drivers of plant translocation performance through reviews of cases at global and regional levels. The establishment of the Italian Database of Plant Translocation (IDPlanT) provides the opportunity to review the techniques used in 186 plant translocation cases performed in the last 50 years in the heart of the Mediterranean Biodiversity Hotspot. We described techniques and information available in IDPlanT and used these data to identify drivers of translocation outcomes. We tested the effect of 15 variables on survival of translocated propagules as of the last monitoring date with binomial logistic mixed-effect models. Eleven variables significantly affected survival of transplants: life form, site protection, material source, number of source populations, propagation methods, propagule life stage, planting methods, habitat suitability assessment, site preparation, aftercare, and costs. The integration of vegetation studies in the selection of suitable planting sites significantly increased the success of translocation efforts. Although posttranslocation watering had a generally positive effect on translocation outcome, other aftercare techniques did not always increase transplant survival. Finally, we found that how funds were spent appeared to be more important than the actual amount spent. Plant translocations in Italy and in the Mediterranean area should account for the complexity of speciation, gene flow, and plant migrations that has led to local adaptations and has important implications for the choice and constitution of source material., (© 2024 Society for Conservation Biology.)

Condensation of RNA and proteins is central to cellular functions, and the ability to program it would be valuable in synthetic biology and synthetic cell science. Here we introduce a modular platform for engineering synthetic RNA condensates from tailor-made, branched RNA nanostructures that fold and assemble co-transcriptionally. Up to three orthogonal condensates can form simultaneously and selectively accumulate fluorophores through embedded fluorescent light-up aptamers. The RNA condensates can be expressed within synthetic cells to produce membrane-less organelles with a controlled number and relative size, and showing the ability to capture proteins using selective protein-binding aptamers. The affinity between otherwise orthogonal nanostructures can be modulated by introducing dedicated linker constructs, enabling the production of bi-phasic RNA condensates with a prescribed degree of interphase mixing and diverse morphologies. The in situ expression of programmable RNA condensates could underpin the spatial organization of functionalities in both biological and synthetic cells., (© 2024. The Author(s).)

Recent discoveries in biology have highlighted the importance of protein and RNA-based condensates as an alternative to classical membrane-bound organelles. Here, we demonstrate the design of pure RNA condensates from nanostructured, star-shaped RNA motifs. We generate condensates using two different RNA nanostar architectures: multi-stranded nanostars whose binding interactions are programmed via linear overhangs, and single-stranded nanostars whose interactions are programmed via kissing loops. Through systematic sequence design, we demonstrate that both architectures can produce orthogonal (distinct and immiscible) condensates, which can be individually tracked via fluorogenic aptamers. We also show that aptamers make it possible to recruit peptides and proteins to the condensates with high specificity. Successful co-transcriptional formation of condensates from single-stranded nanostars suggests that they may be genetically encoded and produced in living cells. We provide a library of orthogonal RNA condensates that can be modularly customized and offer a route toward creating systems of functional artificial organelles for the task of compartmentalizing molecules and biochemical reactions., (© 2024. The Author(s).)

Thanks to its biocompatibility, versatility, and programmable interactions, DNA has been proposed as a building block for functional, stimuli-responsive frameworks with applications in biosensing, tissue engineering, and drug delivery. Of particular importance for in vivo applications is the possibility of making such nanomaterials responsive to physiological stimuli. Here, we demonstrate how combining noncanonical DNA G-quadruplex (G4) structures with amphiphilic DNA constructs yields nanostructures, which we termed "Quad-Stars", capable of assembling into responsive hydrogel particles via a straightforward, enzyme-free, one-pot reaction. The embedded G4 structures allow one to trigger and control the assembly/disassembly in a reversible fashion by adding or removing K + ions. Furthermore, the hydrogel aggregates can be photo-disassembled upon near-UV irradiation in the presence of a porphyrin photosensitizer. The combined reversibility of assembly, responsiveness, and cargo-loading capabilities of the hydrophobic moieties make Quad-Stars a promising candidate for biosensors and responsive drug delivery carriers.

DNA nanotechnology enables the construction of sophisticated biomimetic nanomachines that are increasingly central to the growing efforts of creating complex cell-like entities from the bottom-up. DNA nanostructures have been proposed as both structural and functional elements of these artificial cells, and in many instances are decorated with hydrophobic moieties to enable interfacing with synthetic lipid bilayers or regulating bulk self-organisation. In this feature article we review recent efforts to design biomimetic membrane-anchored DNA nanostructures capable of imparting complex functionalities to cell-like objects, such as regulated adhesion, tissue formation, communication and transport. We then discuss the ability of hydrophobic modifications to enable the self-assembly of DNA-based nanostructured frameworks with prescribed morphology and functionality, and explore the relevance of these novel materials for artificial cell science and beyond. Finally, we comment on the yet mostly unexpressed potential of amphiphilic DNA-nanotechnology as a complete toolbox for bottom-up synthetic biology - a figurative and literal scaffold upon which the next generation of synthetic cells could be built.

Hypertrophic pachymeningitis (HP) is a rare disorder that causes thickening of the dura mater. Inflammatory lesions may be located in the cerebral or spinal dura mater or, less frequently, in both locations simultaneously. Numerous clinico-pathological entities cause thickening of the pachymeninges. Indeed, HP is a potential manifestation of many different diseases, but the diagnosis often remains uncertain. Cases in which the pachymeningitis has no known aetiology are termed "idiopathic" HP (IHP). Recently, it has been suggested that IgG4-related disease represents a subset of cases previously diagnosed as idiopathic hypertrophic pachymeningitis. Little is known regarding the pathogenic events of IHP. In a general theory, the inflammatory infiltrate, mainly consisting of B and T lymphocytes, activates fibroblasts and induces collagen deposition, leading to tissue hypertrophy and increased dural thickness. Clinical manifestations of IHP depend upon the location of the inflammatory lesions and compression of the adjacent nervous structures. Three central pathological features are lymphoplasmacytic infiltration, obliterative phlebitis, and storiform fibrosis. MRI is the examination of choice for the preliminary diagnosis of IHP. Histopathological examination of a biopsy specimen of the dura mater would finally confirm the diagnosis. The differential diagnosis for HP is broad and includes infections, autoimmune disorders, and neoplasia. Currently, there is no consensus about treatment for patients with IHP. There is a preference for glucocorticoid treatment on diagnosis followed by the addition of other immunosuppressive agents in the event of a recurrence. Rituximab is used in patients who did not respond to glucocorticoids or to conventional steroid-sparing agents.

gamma-Glutamyltranspeptidase (gamma GT) appears in serum in multiple forms; their significance and clinical utility in hepatobiliary and pancreatic diseases are still a matter of controversy. Electrophoretic separation of the multiple forms of gamma GT on agarose gel was performed in 20 alcoholic patients (six with cirrhosis and 14 with fatty liver) and the results compared with those obtained in 50 healthy volunteers, 43 patients affected with chronic hepatitis C, 36 patients with posthepatitic cirrhosis and in 52 epileptic patients on long-term anti-epileptic medication. Multiple forms of gamma GT were separated into several bands (up to 11), labelled 0a, 0b, 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4a, 4b. In the alcoholic patients nine fractions were detected, and the electrophoretic pattern observed was significantly different from that observed in healthy volunteers and in patients with chronic hepatitis C or posthepatitic cirrhosis. No differences were observed in the electrophoretic patterns in the alcohol abusers and epileptic patients. In alcoholic patients significant differences were observed in the electrophoretic patterns in relation to the degree of liver injury; the electrophoretic patterns in patients with alcohol-related cirrhosis and posthepatitic cirrhosis differed significantly. The separation of multiple forms of gamma GT has high sensitivity and good reproducibility. It may be proposed as a complementary test in the diagnosis of alcoholic liver disease.

After a large uncontrolled experience, in April 1976, a randomized clinical trial between side-to-side portacaval anastomosis and a mesentericocaval shunt with internal jugular vein interposition was initiated. Up to April 1979, 23 patients were operated upon with a mesentericocaval shunt and 26 with a portacaval anastomosis. With a mean follow-up time of 23 months, no statistically significant differences in terms of operative mortality, hepatic encephalopathy and long term survival were found between the two operations. Therefore, a mesentericocaval shunt does not appear to offer effective advantages compared with the portacaval anastomosis, and it should be performed in particular instances in which, for anatomic reasons, a portacaval anastomosis cannot be performed.