Using manipulation of density-dependent fecundity to recover an endangered species: the bearded vulture Gypaetus barbatus as an example

1. Endangered species subjected to reintroduction programmes often occur as small and isolated populations with local high density and depressed fecundity. Variation in territory quality may lead to this low fecundity owing to increasing occupation of suboptimal territories as population density grows, known as the habitat heterogeneity hypothesis (HHH). In this context, food supplementation in poor territories may be used to produce extra young which could be allocated to reintroduction programmes. 2. We analyse the density-dependent fecundity pattern and the underlying mechanism in a small population of bearded vultures Gypaetus barbatus in Aragón (northeast Spain). We then use population simulations to examine the viability of a hypothetical reintroduction programme using extra young produced by supplementary feeding on poor-quality territories and the effect on the donor population. We also compare the economic cost of such a reintroduction programme in relation to the cost of a traditional captive breeding programme. 3. The wild population showed clear negative, density-dependent fecundity regulation driven by the HHH mechanism. Simulations showed that extractions for translocations had no relevant long-term effects on the donor population viability, but a marked population reduction during the extraction period. However, the implementation of supplementary feeding to produce extra young for translocation lessened significantly this expected initial population reduction. 4. Analyses showed that the annual budget of a captive breeding programme for this species could be seven times more expensive than the translocation of extra young produced by food supplementation. 5. Synthesis and applications. Reintroduction programmes based on translocation of wild-reared individuals, after a supplementary feeding programme oriented to poor-quality territories, provide a source of young at least seven times cheaper than those from captive breeding programmes. The use of this approach would decrease initial effects on donor population avoiding public criticism. Increasing the number of young released during the first years of the reintroduction decreases total financial cost and increases the final population size in the new area

Tissue engineering for total meniscal substitution : Animal study in sheep model

Objective: The aim of the study was to investigate the use of a novel hyaluronic acid/polycaprolactone material for meniscal tissue engineering and to evaluate the tissue regeneration after the augmentation of the implant with expanded autologous chondrocytes. Two different surgical implantation techniques in a sheep model were evaluated. Methods: Twenty-four skeletally mature sheep were treated with total medial meniscus replacements, while two meniscectomies served as empty controls. The animals were divided into two groups: cell-free scaffold and scaffold seeded with autologous chondrocytes. Two different surgical techniques were compared: in 12 animals, the implant was sutured to the capsule and to the meniscal ligament; in the other 12 animals, also a transtibial fixation of the horns was used. The animals were euthanized after 4 months. The specimens were assessed by gross inspection and histology. Results: All implants showed excellent capsular ingrowth at the periphery. Macroscopically, no difference was observed between cell-seeded and cell-free groups. Better implant appearance and integrity was observed in the group without transosseous horns fixation. Using the latter implantation technique, lower joint degeneration was observed in the cell-seeded group with respect to cell-free implants. The histological analysis indicated cellular infiltration and vascularization throughout the implanted constructs. Cartilaginous tissue formation was significantly more frequent in the cell-seeded constructs. Conclusion: The current study supports the potential of a novel HYAFF/polycaprolactone scaffold for total meniscal substitution. Seeding of the scaffolds with autologous chondrocytes provides some benefit in the extent of fibrocartilaginous tissue repair

Lights and shadows concerning platelet products for musculoskeletal regeneration.

Various types of platelet (PLT) products, such as Platelet Rich Plasma (PRP) and Platelet Gel (PG), derived from autologous peripheral blood, have been used for tissue repair. The good clinical outcomes, due mainly to their safety and Growth Factor (GF) content, have led to a wide use of PLT products in many fields of medicine. However, until now the existing literature adds controversies to the use of PLT concentrates. When talking about PLTs and their products, a great number of variables have to be considered. These variables are mainly related to PRP preparation methods, the type of activators, intra- and inter-species variability, types of pathology to be treated, the ways and times of administration and the association of PRP or PG with other treatments. This review considers and discusses these causes of variability with particular attention to orthopaedic implications. The possibility of improving the knowledge on variables affecting therapeutic efficacy will surely help in addressing the best combination of factors implied in the different steps of PLT concentrate preparation and use

Where do Chip and Dale come from? Origins of invasive populations of the Siberian chipmunk in Europe

Among invasive squirrels in Europe, the Siberian chipmunk Eutamias sibiricus, native to North-East Asia, shows the highest number of free-ranging populations in European countries, due to the intense pet trade it underwent between the 1960s and the 1980s. We describe 628-bp cytochrome b sequences from the Netherlands (N = 3), Belgium (N = 4) and Switzerland (N = 1), and through phylogenetic analysis show that they belong to the Korean subspecies. This confirms previous findings that the Korean subspecies (Eutamias sibiricus barberi) has been the most commonly introduced to Europe. Another subspecies (the Northern subspecies, Eutamias sibiricus sibiricus) has only been reported previously in an Italian free-living population. If the splitting of the Korean taxon as a proper species (Eutamias barberi) will be confirmed by analysis of nuclear markers, a revision of the European Regulation 1143/2014 should be conducted to explicitly ban the trade of newly identified taxa.Support to MM for this research (grant LCF/BQ/DR20/11790020) was provided by ‘La Caixa’ Foundation (ID 100010434)

Ionized jet deposition of silver nanostructured coatings: Assessment of chemico-physical and biological behavior for application in orthopedics

Infection is one of the main issues connected to implantation of biomedical devices and represents a very difficult issue to tackle, for clinicians and for patients. This study aimed at tackling infection through antibacterial nanostructured silver coatings manufactured by Ionized Jet Deposition (IJD) for application as new and advanced coating systems for medical devices. Films composition and morphology depending on deposition parameters were investigated and their performances evaluated by correlating these properties with the antibacterial and antibiofilm efficacy of the coatings, against Escherichia coli and Staphylococcus aureus strains and with their cytotoxicity towards human cell line fibroblasts. The biocompatibility of the coatings, the nanotoxicity, and the safety of the proposed approach were evaluated, for the first time, in vitro and in vivo by rat subcutaneous implant models. Different deposition times, corresponding to different thicknesses, were selected and compared. All silver coatings exhibited a highly homogeneous surface composed of nanosized spherical aggregates. All coatings having a thickness of 50 nm and above showed high antibacterial efficacy, while none of the tested options caused cytotoxicity when tested in vitro. Indeed, silver films impacted on bacterial strains viability and capability to adhere to the substrate, in a thickness-dependent manner. The nanostructure obtained by IJD permitted to mitigate the toxicity of silver, conferring strong antibacterial and anti-adhesive features, without affecting the coatings biocompatibility. At the explant, the coatings were still present although they showed signs of progressive dissolution, compatible with the release of silver, but no cracking, delamination or in vivo toxicity was observed

Differential activation of nuclear inositide-dependent signalling pathways during erythropoiesis and myelopoiesis induced by lenalidomide and azacitidine in myelodysplastic syndromes (MDS)

Inositide-dependent signalling pathways regulated by phosphoinositide-specific phospholi- pase C (PI-PLC) beta1 have been demonstrated to play important roles in MDS pathogenesis and in cell differentiation (1). Moreover, the MDS therapy aims at inducing myeloid and/or erythroid differentiation of MDS stem cells. Indeed, azacitidine is a demethylating agent that can induce myeloid differentiation. On the other hand, lenalidomide may restore a normal erythropoiesis. The exact molecular mechanisms underlying the effect of azacitidine and lenalidomide in MDS cells are still unclear, although it is clear that these therapies regulate stem cell proliferation, differentiation and apoptosis (2). The combination of azacitidine and lenalidomide in MDS therapy is now under considera- tion, given the capability of both drugs to balance proliferation and differentiation processes (3). In this study we analyzed the molecular effect of this combination therapy on PI-PLC isoenzymes, not only studying PI-PLCbeta1, but also PI-PLCgamma1, that can be associated with erythropoiesis. We analyzed 44 patients diagnosed with high-risk MDS who were given azacitidine and lenalidomide. Given the limited number of cells, we quantified the expression of these molecules by Real-Time PCR analyses and immunocytochemical experiments. Moreover, we carried out cell cycle analyses and studied both PI-PLCbeta1 methylation status and the expression of Globin genes. In our case series, 28/44 patients were evaluable, with an overall response rate of 78.6% (22/28 cases). At a molecular level, a significant increase of PI-PLCbeta1 and/or PI-PLCgamma1 expression was associated with a favourable clinical response to the combination therapy. Responder cases also showed an increase of Beta-globin expression, hinting at a specific contri- bution of lenalidomide on erythroid activation, whilst the frequent demethylation of PI-PLCbeta1 promoter could be specifically linked to azacitidine. Taken together, our results show that the combination of azacitidine and lenalidomide can be important for activating PI-PLC isoenzymes, therefore regulating myeloid and erythroid dif- ferentiation in MDS cells