101,869 research outputs found
First results of Waterborne Geophysical surveys around the Malpasso site (Tuoro sul Trasimeno, Italy) for geological and archeological characterization
No full textWith the aims of both reconstructing the geological setting of the area around the Malpasso
site and to eventually find some localized remains of the battle we carried out several
waterborne geophysical surveys on the area. Adopted methodologies were: magnetic surveys,
seismic reflection Chirp Sonar surveys and Continuous Vertical Electric Soundings (CVES)
profiles
Novel mitochondrial protein interactors of immunoglobulin light chains causing heart amyloidosis
No full textIn immunoglobulin (Ig) light-chain (LC) (AL) amyloidosis, AL deposition translates into life-threatening cardiomyopathy. Clinical and experimental evidence indicates that soluble cardiotoxic LCs are themselves harmful for cells, by which they are internalized. Hypothesizing that interaction of soluble cardiotoxic LCs with cellular proteins contributes to damage, we characterized their interactome in cardiac cells. LCs were purified from patients with AL amyloidosis cardiomyopathy or multiple myeloma without amyloidosis (the nonamyloidogenic/noncardiotoxic LCs served as controls) and employed at concentrations in the range observed in AL patients' sera. A functional proteomic approach, based on direct and inverse coimmunoprecipitation and mass spectrometry, allowed identifying LC-protein complexes. Findings were validated by colocalization, fluorescence lifetime imaging microscopy (FLIM)-fluorescence resonance energy transfer (FRET), and ultrastructural studies, using human primary cardiac fibroblasts (hCFs) and stem cell-derived cardiomyocytes. Amyloidogenic cardiotoxic LCs interact in vitro with specific intracellular proteins involved in viability and metabolism. Imaging confirmed that, especially in hCFs, cardiotoxic LCs (not controls) colocalize with mitochondria and spatially associate with selected interactors: mitochondrial optic atrophy 1-like protein and peroxisomal acyl-coenzyme A oxidase 1 (FLIM-FRET efficiencies 11 and 6%, respectively). Cardiotoxic LC-treated hCFs display mitochondrial ultrastructural changes, supporting mitochondrial involvement. We show that cardiotoxic LCs establish nonphysiologic protein-protein contacts in human cardiac cells, offering new clues on the pathogenesis of AL cardiomyopathy.-Lavatelli, F., Imperlini, E., Orrù, S., Rognoni, P., Sarnataro, D., Palladini, G., Malpasso, G., Soriano, M. E., Di Fonzo, A., Valentini, V., Gnecchi, M., Perlini, S., Salvatore, F., Merlini, G. Novel mitochondrial protein interactors of immunoglobulin light chains causing heart amyloidosis
Tre città per una comunità: note sulle rifondazioni di Malpasso in Sicilia
No full textLe ricostruzioni della città di Malpasso a seguito dei due funesti eventi naturali succedutisi a breve distanza fra loro, offrono lo spunto per analizzare, pratiche, consuetudini e tecniche fondative in due diversi casi studio afferenti una stessa comunità e legati a una delle maggiori famiglie feudali del Regno di Sicilia nel XVII secolo
Cord blood derived endothelial progenitor cells: a superior buiding company in angiogenesis market.
No full textObjective: Endothelial progenitor cells (EPCs) are a promising
tool in regenerative medicine. We designed a comparative study of peripheral (adult, PB) versus umbilical cord blood (CB) derived EPCs in order to investigate the angiogenic properties of these two sources.
Methods: Mononuclear cells (MNCs) were isolated from PB (n=4) and CB (<24 hours, n=4) and cultured on fibronectin-coated cell culture plates in endothelial cell culture medium until the formation of cobblestone-shaped colonies. Colony-derived cells were expanded for further investigation and their endothelial
phenotype was confi rmed evaluating the uptake of Di-Iacetylated-low-density lipoprotein (Ac-LDL) and expression of
endothelial cell-surface antigens (CD34, CD31, CD146, vWF, KDR) and CD45. Proliferation rates of CB and PB derived EPCs were determined using the MTT assay. The angiogenic properties of colony-derived EPCs were assessed by in vitro capillary-like network formation assay: early-passage cells were seeded (20,000 cells per well) onto 96-wells plates coated with Matrigel. Capillary-like network formation and maintenance were evaluated with an inverted microscope after 4-18-24-48-72 hours of incubation; images were taken from 3 random fields of Matrigel wells to assess the number of branch points per field to quantify the degree of tubulogenesis. Each experiment was performed in triplicate.
Results: The median frequency of colonies obtained was higher in CB than PB (0.49/107MNCs vs 0.05/107MNCs) and CB-derived EPCs had higher proliferative potential than PB-derived EPCs. Both CB and PB colony-derived cells incorporated Ac-LDL, expressed the endothelial cell-surface antigens and were CD45-. Both CB and PB derived EPCs formed capillary-like structures in Matrigel, but CB-derived tubes formed earlier and more complex networks than PB-derived EPCs. Moreover, the number of branch points in CB-derived capillary-like networks was higher than PB-derived networks (16.3±4.3 vs 7.1±-2.6). Finally, CB-derived capillary-like networks were maintained for at least 72 hours, while PB-derived networks started to disassembly in 48 hours.
Conclusion: Our study confirms the different clonogenic and proliferative potential of EPCs derived from PB and CB and demonstrates a superior angiogenic potential of CB-derived cells. Our preliminary data indicate that CB-derived EPCs have a better angiogenic potential than PB-derived EPCs and CB is a promising source of EPCs for regenerative medicine purpose
Testing the paracrine properties of human mesenchymal stem cells using conditioned medium
No full textMesenchymal stem cells (MSC) produce and secrete a great variety of cytokines and chemokines that play beneficial paracrine actions when MSC are used for tissue repair. The conditioned medium (CM) derived from MSC can be used both in vitro and in vivo to test specific paracrine effects or to screen putative paracrine/autocrine mediators by proteomics.In this chapter, we describe a straightforward method to prepare MSC-derived CM. Furthermore, we summarize some in vitro assays useful for testing the cytoprotective, angiogenic, and regenerative activity of CM. These assays are very helpful when studying the role of MSC in cardiac repair and regeneration
Paracrine mechanisms of mesenchymal stem cells in tissue repair
No full textTissue regeneration from transplanted mesenchymal stromal cells (MSC) either through transdifferentiation or cell fusion was originally proposed as the principal mechanism underlying their therapeutic action. However, several studies have now shown that both these mechanisms are very inefficient. The low MSC engraftment rate documented in injured areas also refutes the hypothesis that MSC repair tissue damage by replacing cell loss with newly differentiated cells. Indeed, despite evidence of preferential homing of MSC to the site of myocardial ischemia, exogenously administered MSC show poor survival and do not persist in the infarcted area. Therefore, it has been proposed that the functional benefits observed after MSC transplantation in experimental models of tissue injury might be related to the secretion of soluble factors acting in a paracrine fashion. This hypothesis is supported by pre-clinical studies demonstrating equal or even improved organ function upon infusion of MSC-derived conditioned medium (MSC-CM) compared with MSC transplantation. Identifying key MSC-secreted factors and their functional role seems a reasonable approach for a rational design of nextgeneration MSC-based therapeutics. Here, we summarize the major findings regarding both different MSC-mediated paracrine actions and the identification of paracrine mediators
Protocols for in vitro differentiation of human mesenchymal stem cells into osteogenic, chondrogenic and adipogenic lineages
No full textMesenchymal stem cells (MSC) possess high plasticity and the potential to differentiate into several different cell types; this characteristic has implications for cell therapy and reparative biotechnologies. MSC have been originally isolated from the bone marrow (BM-MSC), but they have been found also in other tissues such as adipose tissue, cord blood, synovium, skeletal muscle, and lung. MSC are able to differentiate in vitro and in vivo into several cell types such as bone, osteocytes, chondrocytes, adipocytes, and skeletal myocytes, just to name a few.During the last two decades, an increasing number of studies have proven the therapeutic potential of MSC for the treatment of neurodegenerative diseases, spinal cord and brain injuries, cardiovascular diseases, diabetes mellitus, and diseases of the skeleton. Their immuno-privileged profile allows both autologous and allogeneic use. For all these reasons, the scientific appeal of MSC is constantly on the rise.The identity of MSC is currently based on three main criteria: plastic-adherence capacity, defined epitope profile, and capacity to differentiate in vitro into osteocytes, chondrocytes, and adipocytes. Here, we describe standard protocols for the differentiation of BM-MSC into the osteogenic, chondrogenic, and adipogenic lineages
IRES-based lentivirus co-espressing TGFbeta1 and FGF2 improves cell survival and angiogenesis in bone marrow-derived mesenchymal stem cells.
No full textBackground. Bone marrow mesenchymal stem cells (BM-MSC) repair infarcted hearts mainly through paracrine mechanisms. However, donor age negatively influences the production of paracrine factors from BM-MSC. In particular, we have shown that in BM-MSC from elderly patients the expression of TGFbeta1 (T) and FGF2 (F) is decreased. We hypothesized that the overexpression of these two factors involved in cytoprotection and angiogenesis may improve MSC repair capacity.
Methods. Rat BM-MSC were transduced with control virus (GFP-MSC) or TF virus (TF-MSC). To study cytoprotective paracrine properties H9c2 cells were exposed to 24h of hypoxia in the presence of control medium (CTRL-M) or media conditioned by GFP- (GFP-CM) or TF-MSC (TF-CM). Cell viability was measured by MTS assay. Apoptosis was evaluated through caspase-3 activation (luminometric assay and western blot). Angiogenesis was assessed by quantifying HUVEC tube formation on Matrigel. Transcriptional levels of known survival genes in H9c2 cells and of soluble factors other than transgenes in MSC were measured by RT-PCR. Activation of FGF2 and TGF1 pathways (Akt, ERK1/2, and SMAD2) was evaluated by western blot.
Results. Compared with CTRL-M, TF-CM increased H9c2 viability (+46,3% p<0.001), while GFP-CM had no effect. Caspase-3 activation was reduced by TF-CM of 60,3% (p<0.001) vs CTRL-M and of 44,7 % (p<0.05) vs GFP-MSC. H9c2 treated with TF-CM showed a strong activation of Akt, ERK1/2 and SMAD2 anti-apoptotic pathways, enhanced expression of Bcl-2 and Stat3 pro-survival genes, and inhibition of FasL and TNF pro-apoptotic genes. HUVEC tube formation were enhanced by TF-CM of 70% (p<0.01) vs CTRL-M and of 59% (p<0.05) vs GFP-CM. Finally, we documented that TF-MSC upregulated transcriptional levels of other soluble factors like IGF1, PDGF-β, BMP2, VEGF and IL11.
Conclusions. Simultaneous overexpression of TF transgenes improves cytoprotective and pro-angiogenic paracrine properties and enhances expression of soluble factors in BM-MSC
Impiego di cellule staminali mesenchimali in cardiologia.
No full textL'infarto miocardico acuto è la principale causa di morte prematura nei paesi occidentalizzati. I pazienti che sopravvivono alla fase acuta vanno spesso incontro ad irreversibile perdita di cellule miocardiche che provoca progressiva riduzione della funzione ventricolare fino a determinare indufficienza cardiaca severa. lo scompenso si manifesta soprattutto in quei pazienti che non vengono prontamente riperfusi e che vanno quindi incontro a fenomeni di rimodellamento del ventricolo sinistro.
Le terapie oggi disponibili possono solo parzialmente reallentare il rimodellamento ventricolare negativo senza tuttavia evitare la comparsa d'insufficienza cardiaca nei pazienti con esteso infarto del miocardio. Ad oggi il trapianto di cuore rappresenta l'unica terapia salvavita per la maggior parte di questi malati. Sarebbe quindi estremamente importante scoprire nuove terapie che permettano di prevenire o curare il rimodellamento ventricolare e quindi l'insufficienza cardiaca. recentamente molti studi preclinici hanno suggerito che l'iniezione intramiocardica di cellule staminali adulte, ed in particolare di cellule staminali mesenchimali, possieda un'enorme potenziale per la cura del danno da infarto del miocardio
Bibliographie Hilarion G. Petzold 1958 – 2009 mit Anhang als Einführung
No full textDieses Archiv enthält die Gesamtbibliographie der Werke des Autors nebst einiger Texte „Über H. G. Petzold“ im Schlussteil der Bibliographie sowie einen Anhang mit einer Einführung in die Architektur des Werkes in seinem wissenslogischen Aufbau als Ausarbeitung seines „Tree of Science Modells“ (2007).This archive contains the complete bibliography of the author and some texts about H. G. Petzold, moreover an epilogue with an introduction to the architecture of the works in its epistemological structure and composition and as an elaborations of Petzold’s „Tree of Science Modell (2007).https://www.fpi-publikation.de/polyloge/01-2009-petzold-h-g-gesamtbibliographie-h-g-petzold-1958-2009-updating-november2009/peerReviewedpublishedVersio
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