Characterization of the Resistance to Powdery Mildew and Leaf Rust Carried by the Bread Wheat Cultivar Victo

Leaf rust and powdery mildew are two important foliar diseases in wheat. A recombinant inbred line (RIL) population, obtained by crossing two bread wheat cultivars ('Victo' and 'Spada'), was evaluated for resistance to the two pathogens at seedling stage. Upon developing a genetic map of 8726 SNP loci, linkage analysis identified three resistance Quantitative Trait Loci (QTLs), with 'Victo' contributing the resistant alleles to all loci. One major QTL (QPm.gb-7A) was detected in response to Blumeria graminis on chromosome 7A, which explained 90% of phenotypic variation (PV). The co-positional relationship with known powdery mildew (Pm) resistance loci suggested that a new source of resistance was identified in T. aestivum. Two QTLs were detected in response to Puccinia triticina: a major gene on chromosome 5D (QLr.gb-5D), explaining a total PV of about 59%, and a minor QTL on chromosome 2B (QLr.gb-2B). A positional relationship was observed between the QLr.gb-5D with the known Lr1 gene, but polymorphisms were found between the cloned Lr1 and the corresponding 'Victo' allele, suggesting that QLr.gb-5D could represent a new functional Lr1 allele. Lastly, upon anchoring the QTL on the T. aestivum reference genome, candidate genes were hypothesized on the basis of gene annotation and in silico gene expression analysis

Cooperazione interistituzionale nelle Pubbliche Amministrazioni: l'Ufficio intercomunale

Nell'ambito dello scenario istituzionale caratterizzato da profonde trasformazioni (si pensi ad esempio alla riforma del titolo V della Costituzione), l'innovazione nella pubblica amministrazione risulta assumere un importanza rilevante. Gli Enti Locali dovranno prepararsi ed adeguarsi a ricevere le funzioni e le responsabilita' derivanti dall'attuazione della riforma. In questo quadro di riferimento si inserisce il presente progetto che individua come idea forza la collaborazione inter-istituzionale come fattore decisivo nella valorizzazione e nell'uso strategico delle risorse turistico culturali del territorio preso in esame. L'area analizzata e' quella dell'alto Casertano, nello specifico i nove Comuni che hanno dimostrato una buona propensione alla collaborazione: Alife, Castello del Matese, Gallo Matese, Letino, Piedimonte Matese, San Gregorio Matese, San Potito Sannitico, Sant'Angelo d'Alife e Valle Agricola. L'idea progetto consiste nella stipulazione di una convenzione tra questi Comuni al fine di istituire un ufficio intercomunale per la gestione associata di diverse funzioni che risultano essere fondamentali per un maggiore coordinamento istituzionale. Il progetto e' strutturato secondo il seguente schema: analisi del cambiamento istituzionale risorse culturali come fattore di sviluppo analisi del quadro di riferimento territoriale analisi swot formulazione della strategia e degli obiettivi idea forza idea progetto aspetti tecnic

Characterization and gene expression of cancer stem cells grown as sarcospheres from human primary sarcomas

This study aimed to identify, isolate and characterize cancer stem cells from human primary sarcomas. We performed cytometric analyses for stemness and differentiation antigens including CD29, CD34, CD44, CD90, CD117 and CD133 on 21 human primary sarcomas on the day of surgery. From sarcoma biopsies, we obtained two chondrosarcoma stabilized cell lines and two osteosarcoma stabilized cell lines on which spheres formation, side population profile, stemness gene expression and in vivo and in vitro assays were performed. On a chondrosarcoma cell line, the whole genoma microarray analyses were performed (sarcospheres versus adherent cells). All samples expressed the CD133, CD44 and CD29 markers. We selected a CD133+ subpopulation from stabilized cell lines that displayed the capacity to grow as sarcospheres able to initiate and sustain tumour growth in NOD/SCID mice, to express stemness genes including OCT3/4, Nanog, Sox2 and Nestin and to differentiate into mesenchymal lineages. Microarray analyses pointed out a huge gene expression difference between sarcospheres and adherent cells. About 2000 genes, including ones related to cell cycle, stemness and epigenetic regulation, resulted to be very differentially expressed in the two population considered. With signed ratio of 2,806 genes were over-expressed and 1,029 under-expressed on sarcospheres when compared with adherent cells. The most highly overexpressed gene were thioredoxin interacting protein (fold +25) that modulates the cellular redox state, growth differentiation factor 15 (fold +10), member of the TGF–β superfamily , histone cluster 1, H2ad (fold +9) and solute carrier family 2 member 14 that facilitate glucose transport (fold +8,9). Our findings evidence the existence of cancer stem cells in human primary bone sarcomas and highlight CD133 as pivotal marker for identification of these cells. This may be of primary importance in the development of new therapeutic strategies and new prognostic procedures against these highly aggressive and metastatic tumours

Pharmacological targeting of the ephrin receptor kinase signalling by GLPG1790 in vitro and in vivo reverts oncophenotype, induces myogenic differentiation and radiosensitizes embryonal rhabdomyosarcoma cells

EPH (erythropoietin-producing hepatocellular) receptors are clinically relevant targets in several malignancies. This report describes the effects of GLPG1790, a new potent pan-EPH inhibitor, in human embryonal rhabdomyosarcoma (ERMS) cell lines

Methylation and epigenetic modification by 5’ azacytidine and valproic acid treatment increase stemness attributes in bone sarcoma cell lines

Bone sarcoma is an aggressive malignancy with high mortality rate. Despite recent advances, the prognosis is still extremely poor. Bone sarcomas contain a small cell population with stem cell like properties, referred to as cancer stem cells (CSCs) expressing CD133 (Tirino et al, 2009; 2011). The biological relevance and regulatory mechanism of CD133 expression are not yet understood. The aim of this study is to elucidate mechanisms regulating aberrant expression of CD133 and stemness phenotype. Saos-2, MG63 and BS15 cell lines were treated with 0,5 mM valproic acid (VPA) and 3μM 5’azacytidine (5-AZA) for 48 hours alone and in combination. CD133 and stemness markers expression including OCT4, Sox2 and Nanog were analyzed by flow cytometry and real-time PCR. Vimentin and osteocalcin levels were also tested. Sarcospheres formation rate was assessed as spheres number/seed single cell number. After treatment with 5-AZA or VPA, the expression level of CD133 mRNA as well as of protein was significantly increased in all three cell lines. Also OCT4, Sox2 and Nanog, stemness markers, and vimentin, mesenchymal marker resulted to be upregulated after treatment by real time-PCR. On the contrary, the expression level of osteocalcin remained similar before and after treatment. Interestingly, combined treatment with 5-AZA and VPA induced an increase of CD133 expression in a synergistic manner in all three cell lines. In addition, sarcospheres formation rate was increased after drug treatment compared to untreated cells. Also in this case, the drug combination lead to synergistic increase of formation rate of spheres. In conclusion, our results indicate that DNA methylation is an important determinant of CD133 and stemness profile in human bone sarcomas and this mechanism may be associated with histone deacetilase inhibition

Cancer stem cells in head and neck tumors: evidence for metastatic spread and treatment resistance

The major challenge in the management of patients with oral squamous cell carcinomas (OSCC) is the development of resistance to therapy leading to disseminated disease. Since cancer stem cells (CSC) have emerged as important players in OSCC metastasis, our objectives were to explore the implications of CSC in OSCC tumor progression, invasion and response to conventional therapies. Methods: A panel of well-characterized cell lines originated from the most common sites in the head and neck area was used. Cells were cultured as floating spheres or under normal adherent conditions and analyzed for CD44, ALDH, CD24, CD29, CD56 by flow cytometry, PCR arrays for genes related to stemness, metastasis and EMT . We also investigated sLeX expression, known to play a key-role in many cancers metastasis by promoting tumor cells binding to endothelial E-selectin. We analyzed the tumorigenic potential of OSCC cells by invasion assays and in vivo OSCC experimental models comparatively to CSC cells. Moreover resistance to cisplatin and radiation was assessed by annexin V/PI assay and by colony forming assay. Results: The highest levels of sLeX expression were found in cell lines originated from oral cavity (9%-47%) compared to other head and neck locations (0.1%-7%). Cells grown as spheres were 95-100% positive for sLeX compared to 10-40% of adherent counterpart. Although sLeX+ and sLeX- cells were both able to form spheres, sLeX+ spheres were predominant and larger. Flow cytometry and PCR arrays indicated that the spheres were highly enriched in CSC and metastatic markers. Consistently, the spheres showed increased invasive and tumorigenic potential, and resistance to conventional chemotherapy and radiations. Conclusion: these studies are the first to unveil a novel link between sLeX expression, stem cell formation and metastatic spread in OSCC, and provide supportive evidence for CSC resistance to treatment. Understanding the mechanisms of tumor invasion and metastasis will improve patient outcome and survival

The Rediscovery of Traditional Maize Agrobiodiversity: A Study Case from Northern Italy

Nowadays, agriculture is under the pressure of climate change and new pathogen outbreaks while farmers are requiring breeders to develop more resistant and resilient genotypes. The genetic base for breeding may be increased through appropriate conservation, description and characterization of local varieties and germplasm collections that have never been used in breeding, and which could be sources of useful alleles. In this framework, the present paper focuses on eight maize landraces of the eastern part of Emilia-Romagna, derived from the Italian maize collection sampled in 1954. Landraces are characterized by a short cycle length and different kernel types—mainly flint-like or an intermediate type of yellow or yellow–orange color—while dent landraces are less represented. Pigmented and white corns are absent even though one landrace (Va213) showed the presence of scattered blue kernels on yellow ears. Ear shape is frequently conical, a trait associated with drought-resistance and common in Italian traditional landraces. Genetic characterization was carried out on 529 individuals by using 10 SSR markers. A total of 68 different alleles, ranging from 4 for markers (phi084 and umc1401) to 11 (phi031) and from 27 (Va217) to 50 (Va211), were evidenced at the individual and population level. AMOVA analysis revealed a small amount (19%) of variability between populations, as supported also by PCoA, with the only exception of Va217, which is different from the others, as evidenced also by phylogenetic analysis. Population structure analysis resulted in the identification of three and four population levels, which are consistent with previous results

Morphological and Genetic Characterization of Local Maize Accessions from Emilia Romagna Region, Italy

Italian maize germplasm is particularly rich in local materials and each region is characterized by the presence of peculiar local varieties deriving from centuries of adaptation, selection and cultivation. While the introduction of hybrids, during the 1950s, led to the disappearing of many of these varieties, some have been maintained in cultivation by farmers, frequently in marginal areas, as a kind of family heritage. Local varieties were identified throughout field surveys carried out in recent years. The discovery of a traditional popcorn variety over the most common flint and semi-flint materials used for production of polenta was interesting. Since these varieties have never been adequately described and reported in scientific literature, this study was aimed to solve this lack of knowledge on recently discovered local maize populations. Characterization represents the first step of a process focused on the preservation and possible exploitation of important genetic resources. Traditional materials are a useful reservoir of genes for adaptation to local conditions and climate changes. Adequate breeding programs can use such germplasm for developing new and more resilient varieties. These local materials have been characterized at the morphological level highlighting plant, ear and kernel differences. Genetic characterization, carried out on 455 individuals by the use of 10 SSR markers, revealed 62 different alleles ranging from four for markers phi127, phi076 and phi084 to nine for marker p-bnlg176. The landraces are well distinguishable at genetic level since 40% of genetic variability is present among accessions. Five landraces are characterized by the presence of private alleles and heterozygosity levels are generally good. These findings support the possibility to correctly preserve local materials through in situ conservation. Phylogenetic analysis evidenced the presence of varietal clusters, the clearest one formed by three red-pigmented accessions. STRUCTURE analysis revealed that five landraces have a well-defined genetic attribution while the remaining two (EMR04-Mais Rosso di Rasora and EMR10-Mais del Principe di Scavolino) are both constituted by two different backgrounds

Human DPSCs fabricate vascularized woven bone tissue : a new tool in bone tissue engineering

Human dental pulp stem cells (hDPSCs) are mesenchymal stem cells that have been successfully used in human bone tissue engineering. To establish whether these cells can lead to a bone tissue ready to be grafted, we checked DPSCs for their osteogenic and angiogenic differentiation capabilities with the specific aim of obtaining a new tool for bone transplantation. Therefore, hDPSCs were specifically selected from the stromal-vascular dental pulp fraction, using appropriate markers, and cultured. Growth curves, expression of bone-related markers, calcification and angiogenesis as well as an in vivo transplantation assay were performed. We found that hDPSCs proliferate, differentiate into osteoblasts and express high levels of angiogenic genes, such as vascular endothelial growth factor and platelet-derived growth factor A. Human DPSCs, after 40 days of culture, give rise to a 3D structure resembling a woven fibrous bone. These woven bone (WB) samples were analysed using classic histology and synchrotron-based, X-ray phase-contrast microtomography and holotomography. WB showed histological and attractive physical qualities of bone with few areas of mineralization and neovessels. Such WB, when transplanted into rats, was remodelled into vascularized bone tissue. Taken together, our data lead to the assumption that WB samples, fabricated by DPSCs, constitute a noteworthy tool and do not need the use of scaffolds, and therefore they are ready for customized regeneration

Human DPSCs fabricate vascularized woven bone tissue: a new tool in bone tissue engineering

Human dental pulp stem cells (hDPSCs) are mesenchymal stem cells that have been successfully used in human bone tissue engineering. To establish whether these cells can lead to a bone tissue ready to be grafted, we checked DPSCs for their osteogenic and angiogenic differentiation capabilities with the specific aim of obtaining a new tool for bone transplantation. Therefore, hDPSCs were specifically selected from the stromal-vascular dental pulp fraction, using appropriate markers, and cultured. Growth curves, expression of bone-related markers, calcification and angiogenesis as well as an in vivo transplantation assay were performed. We found that hDPSCs proliferate, differentiate into osteoblasts and express high levels of angiogenic genes, such as vascular endothelial growth factor and platelet-derived growth factor A. Human DPSCs, after 40 days of culture, give rise to a 3D structure resembling a woven fibrous bone. These woven bone (WB) samples were analysed using classic histology and synchrotron-based, X-ray phase-contrast microtomography and holotomography. WB showed histological and attractive physical qualities of bone with few areas of mineralization and neovessels. Such WB, when transplanted into rats, was remodelled into vascularized bone tissue. Taken together, our data lead to the assumption that WB samples, fabricated by DPSCs, constitute a noteworthy tool and do not need the use of scaffolds, and therefore they are ready for customized regeneration