A 3D‑scaffold of PLLA induces the morphological differentiation and migration of primary astrocytes and promotes the production of extracellular vesicles

The present study analyzed the ability of primary rat astrocytes to colonize a porous scaffold, mimicking the reticular structure of the brain parenchyma extracellular matrix, as well as their ability to grow, survive and differentiate on the scaffold. Scaffolds were prepared using poly-L-lactic acid (PLLA) via thermally-induced phase separation. Firstly, the present study studied the effects of scaffold morphology on the growth of astrocytes, evaluating their capability to colonize. Specifically, two different morphologies were tested, which were obtained by changing the polymer concentration in the starting solution. The structures were characterized by scanning electron microscopy, and a…

Antimicrobial peptides in the tunic of Ciona intestinalis (Tunicata)

The identification and localization of two intermediate filament proteins in the tunic of Styela plicata (Tunicata, Styelidae).

The intermediate filament (IF) proteins Styela C and Styela D from the tunicate Styela (Urochordata) are co-expressed in all epidermal cells and they are thought to behave as type I and type II keratins. These two IF proteins, Styela C and Styela D, were identified in immunoblots of proteins isolated from the tunic of Styela plicata. The occurrence and distribution of these proteins within the tunic of this ascidian was examined by means of immunofluorescence and immunoperoxidase techniques, using anti-Styela C and anti-Styela D antibodies. In addition, immuno-electron microscopy of the tunic showed that the two proteins are located in the cuticle layer and in the tunic matrix. These result…

Ultrastructural aspects of the tunic wound repair in Styela plicata and Ciona intestinalis (Tunicata)

Individuazione di citocheratine nella tunica delle ascidie

Identificazione e localizzazione di citocheratine nelle Ascidie.

Screening of subtelomeric rearrangements in autistic disorder: identification of a partial trisomy of 13q34 in a patient bearing a 13q;21p translocation.

Within the framework of a FISH screening protocol to detect cryptic subtelomeric rearrangements in autistic disorder (AD), a patient bearing three copies of the subtelomeric portion of the q arm of chromosome 13 has been identified. Beside AD, the patient also has severe mental retardation and displays several dysmorphic features. Further FISH analyses revealed that the trisomy was caused by the translocation of a 13q subtelomeric fragment to the acrocentric tip of one chromosome 21 [46,XY.ish der(21) t(13;21) (q34;p13)(D13S1825+)]. Gene dosage experiments carried out with three multiallelic polymorphisms of the subtelomeric region of chromosome 13q showed that the putative length of the tr…

Screening of subtelomeric rearrangements in autism spectrum disorder. Identification of a partial trisomy of 13q in a patient

Screening of subtelomeric rearrangements in autism spectrum disorder. Identification of a partial trisomy of 13q in a patient.

Modificazioni ultrastrutturali delle cellule della tunica di Styela plicata durante i processi di riparazione e rigenerazione.

Cellular components and tunic architecture of the solitary ascidian Styela canopus (Stolidobranchiata, Styelidae).

Abstract Cell distribution and tunic morphology in the ascidian Styela canopus were examined by electron microscopy. The observations showed that the outer covering is composed of a thin sinuous cuticle with several protrusions and a deep layer of ground substance. The fibrous component and its arrangement in the tunic were demonstrated: elementary fibrils exhibit a ‘microtubular’ structure and an elliptical cross-sectional shape. Four types of cells were described: clear vesicular tunic granulocytes, tunic microgranulocytes, unilocular tunic granulocytes, and globular tunic granulocytes. Morphofunctional aspects of the tunic tissue and certain phylogenetic relationships are discussed.

« Study of proliferating blood cells in Ciona intestinalis (Tunicata) immune response»

Adenocarcinomas and their derived lymphovascular emboli contain dual compartmental populations of tumor microvesicles which differ in size and possibly content and function.

Adenocarcinomas and their associated lymphovascular emboli metastasize as tight aggregates of tumor cells. Using a xenograft model of human inflammatory breast cancer (Mary-X), which exhibited florid lymphovascular emboli in mice and high density tumor aggregates (spheroids) in vitro, we previously demonstrated that both emboli and spheroids are mediated by an intact overexpressed E-cadherin axis which regulates homotypic tumor cell adhesion. We now report that Mary-X spheroids secrete 5-10 fold higher levels of microvesicles (MVs) than most other carcinoma cell lines. Despite the high density of tumor cells and the strong juxtaposition of tumor cells to each other in the spheroids, interce…

LA GENETICA GENERALE

Exosomes from metastatic cancer cells transfer amoeboid phenotype to non-metastatic cells and increase endothelial permeability: their emerging role in tumor heterogeneity

AbstractThe goal of this study was to understand if exosomes derived from high-metastatic cells may influence the behavior of less aggressive cancer cells and the properties of the endothelium. We found that metastatic colon cancer cells are able to transfer their amoeboid phenotype to isogenic primary cancer cells through exosomes, and that this morphological transition is associated with the acquisition of a more aggressive behavior. Moreover, exosomes from the metastatic line (SW620Exos) exhibited higher ability to cause endothelial hyperpermeability than exosomes from the non metastatic line (SW480Exos). SWATH-based quantitative proteomic analysis highlighted that SW620Exos are signific…

Genetica generale e umana

libro di testo

Citrus limon-derived nanovesicles inhibit cancer cell proliferation and suppress CML xenograft growth by inducing TRAIL-mediated cell death

// Stefania Raimondo 1 , Flores Naselli 1 , Simona Fontana 1 , Francesca Monteleone 1 , Alessia Lo Dico 1 , Laura Saieva 1 , Giovanni Zito 2 , Anna Flugy 1 , Mauro Manno 3 , Maria Antonietta Di Bella 1 , Giacomo De Leo 1 , Riccardo Alessandro 1 1 Dipartimento di Biopatologia e Biotecnologie Mediche, Universita degli Studi di Palermo, sezione di Biologia e Genetica, Palermo, Italy 2 Laboratorio di Ingegneria Tissutale – Piattaforme Innovative per l’Ingegneria Tissutale (PON01–00829), Istituto Ortopedico Rizzoli, Palermo, Italy 3 Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Palermo, Italy Correspondence to: Riccardo Alessandro, e-mail: riccardo.alessandro@unipa.it Keywords: canc…

Contribution of microscopy to the study of proliferating blood cells in Ciona intestinalis immune response

Following an inflammatory stimulus such as the injection of erythrocytes into the tunic of Ciona intestinalis, hemocytes respond to events occurring in the tunic within a short time, and infiltrate the inflamed tissue being massively involved in the acute inflammatory response and capsule formation. The proliferative response of circulating hemocytes and pharynx assume particular interest as it is significantly enhanced during these defence reactions. Microscopy clearly may contribute to extend our understanding of the phenomenon showing interesting mitotic figures and hematogenic nodules with proliferative characteristics.

Hemocyte migration during inflammatory-like reaction of Ciona intestinalis (Tunicata, ascidiacea).

Following an injuring factor such as the injection of erythrocytes into the tunic of Ciona intestinalis, an inflammatory-like response occurs and blood cells are massively involved in these mechanisms. Electron microscope observations illustrate the infiltration and migration of blood cells throughout the mantle epithelium into the tunic and show several prominent morphological changes.

GENETICA UMANA

Comparative study of T84 and T84SF human colon carcinoma cells: in vitro and in vivo ultrastructural and functional characterization of cell culture and metastasis.

The HDAC6 Inhibitor tubacin induces release of CD133+ extracellular vesicles from cancer cells

Tumor-derived extracellular vesicles (EVs) are emerging as an important mode of intercellular communication, capable of transferring biologically active molecules that facilitate the malignant growth and metastatic process. CD133 (Prominin-1), a stem cell marker implicated in tumor initiation, differentiation and resistance to anti-cancer therapy, is reportedly associated with EVs in various types of cancer. However, little is known about the factors that regulate the release of these CD133+ EVs. Here, we report that the HDAC6 inhibitor tubacin promoted the extracellular release of CD133+ EVs from human FEMX-I metastatic melanoma and Caco-2 colorectal carcinoma cells, with a concomitant dow…

Ultrastructural aspects of naturally occurring wound in the tunic of two ascidians: Ciona intestinalis and Styela plicata (Tunicata).

Efficient wound healing is essential for all animals from insects to mammals. Ciona intestinalis and Styela plicata are solitary ascidians belonging to urochordates, a subphylum that occupies a key phylogenetic position as it includes the closest relative to vertebrates. Urochordate first physical barrier against invaders is the tunic, an extracellular matrix that is constantly exposed to all kinds of insults. Thus, when damage occurs, an innate immune response is triggered to eliminate impaired tissue and potentially pathogenic microbes, and restore tissue functionality. Ultrastructural aspects of the tunic in the wound healing process of two ascidians are described. In the injured areas, …

A preliminary study on antimicrobial peptides in the naturally damaged tunic of Ciona intestinalis (Tunicata)

Tumor-Derived Small Extracellular Vesicles Induce Pro-Inflammatory Cytokine Expression and PD-L1 Regulation in M0 Macrophages via IL-6/STAT3 and TLR4 Signaling Pathways

Tumor-associated macrophages play a key role in promoting tumor progression by exerting an immunosuppressive phenotype associated with the expression of programmed cell death ligand 1 (PD-L1). It is well known that tumor-derived small extracellular vesicles (SEVs) affect the tumor microenvironment, influencing TAM behavior. The present study aimed to examine the effect of SEVs derived from colon cancer and multiple myeloma cells on macrophage functions. Non-polarized macrophages (M0) differentiated from THP-1 cells were co-cultured with SEVs derived from a colorectal cancer (CRC) cell line, SW480, and a multiple myeloma (MM) cell line, MM1.S. The expression of PD-L1, interleukin-6 (IL-6), a…

Aspects of cell production in mantle tissue of Ciona intestinalis L. (Tunicata, Ascidiacea)

Renewal of cell population is needed in the tunic of ascidians, as the tunic cells are involved in many biological functions. Tunic cells are thought to arrive by migrating across the mantle epithelium into the tunic from the blood lacunae or the mesenchymal space. Electron microscope observations show that the mantle epithelium of Ciona intestinalis shares some proliferative characteristics, releasing cells into the tunic and thus providing an increase renewal of tunical cells in restricted zones of adult animals.

3D cultures of primary astrocytes on Poly-L-lactic acid scaffolds

Tissue engineering is an emerging multidisciplinary field that aims at reproducing in vitro tissues with morphological and functional features similar to the biological tissue of the human body. Polymeric materials can be used in contact with biological systems in replacing destroyed tissue by transplantation [1]. Several biopolymers, including poly L (lactic acid) (PLLA), have been used in biomedical applications to set scaffolds with ductile proprieties and biodegradation kinetics [2]. In particular, the PLLA scaffold topography mimics the natural extracellular matrix and makes it a good candidate for neural tissue engineering. We report about of 3D system the PLLA porous scaffolds prepar…

Microfluidic Fabrication of Physically Assembled Nanogels and Micrometric Fibers by Using a Hyaluronic Acid Derivative

The employ of a hyaluronic acid (HA) derivative, bearing octadecyl (C18) and ethylenediamine (EDA) groups, for microfluidic fabrication of nanogels and microfibers is reported in this study. Two HA-EDA-C18 derivatives (125 and 320 kDa) having ionic strength sensitive properties are synthesized and characterized. The control of the rheological properties of HA-EDA-C18 aqueous dispersions by formation of inclusion complexes with hydroxypropyl-β-cyclodextrins (HPCD) is described. Reversibility of C18/HPCD complexation and physical crosslinking is detected in media with different ionic strength through oscillation frequency tests. HA-EDA-C18 125 kDa is employed for nanogel fabrication. Control …

The hydrophilie of the larval test of Ascidiae: functional role played by test cells

Ascidian swimming larvae are entirely surrounded by a hyaline extracellular coat, called as tunic or test, on which numerous test cells adhere. The functional role played by test cells in larvae of various ascidian species consists in depositing submicroscopic structures known as ornaments and/or substances of proteoglycan nature in the larval test surface. The deposition of ornaments would render the larval test hydrophilic and thus allow the larvae to swim being immersed in sea water. Ultrastructural investigations reported in literature on larvae of Cionidae and Ascididae families have not evidenced the presence of ornaments in the swimming larval test. For these Ascididae families it ha…

Le basi dell'organizzazione biologica

Comparative study of T84 and T84SF human colon carcinoma cells: in vitro and in vivo ultrastructural and functional characterization of cell culture and metastasis

To better understand the relationship between tumor heterogeneity, differentiation, and metastasis, suitable experimental models permitting in vitro and in vivo studies are necessary. A new variant cell line (T84SF) exhibiting an altered phenotype was recently selected from a colon cancer cell line (T84) by repetitive plating on TNF-alpha treated human endothelial cells and subsequent selection for adherent cells. The matched pair of cell lines provides a useful system to investigate the extravasation step of the metastatic cascade. Since analysis of morphological differences can be instructive to the understanding of metastatic potential of tumor cells, we compared the ultrastructural and …

MUTAZIONI: TIPI, ORIGINI, CONSEGUENZE

«Correlation between granulocytes and tunic cuticle of Ciona intestinalis (Tunicata, Ascidiacea) as evaluated by microscopy»

The tunic is a peculiar integumentary tissue covering the epidermis of ascidians and other tunicates. It is an extracellular matrix whose outermost surface is revealed to be a continuos layer called tunic cuticle, composed of electron dense fibrous materials. Mechanisms occurring during the growth of the animal after metamorphosis, must assure the maintenance of the thin tunic cuticle density and integrity as it represents the primary body surface barrier preventing the mechanical and microbial assaults. Investigations by optical and electron microscopy on tunic samples from young Ciona intestinalis, allowed us to study morphological relationship between the tunic cuticle architecture and g…

Encapsulation Response ofCiona intestinalis(Ascidiacea) to Intratunical Erythrocyte Injection

Abstract Electron microscopic studies on the encapsulation induced by erythrocyte injection into the tunic of the ascidian Ciona intestinalis were carried out. The observations reported in the present paper complete the description previously given of capsule architecture and contribute to the characterization of the cells involved in the inflammatory reaction. The inflamed area is surrounded by an ample and peculiar “three-layered coat” respectively composed of flattened and packed extratunical hemocytes, the monolayered epithelium, and a layer of intratunical electron-dense particles. The latter are also clustered, variously arranged, and distributed in the tunic ground substance. The epi…

Microscopic features of the tunic in young Styela canopus (Tunicata, Styelidae)

The tunic is a unique tissue in metazoans covering the epidermis of ascidians and other tunicates. The tunic is a multifunctional tissue that provides support, mechanical rigidity, and protection against infection and predation. The outermost surface of it is covered by a thin continuous layer called tunic cuticle. The tunic cuticle density and integrity must be maintained during the growth of the animal after metamorphosis, as it represents the primary body surface barrier preventing the mechanical damage. Using transmission electronic microscopy we attempted to characterize the cuticle morphology of young Styela canopus, a solitary ascidian (Styelidae), in comparison with the cuticle arch…

Overview and Update on Extracellular Vesicles: Considerations on Exosomes and their Application in Modern Medicine

In recent years, there has been a rapid growth in the knowledge of cell-secreted extracellular vesicle functions. They are membrane enclosed and loaded with proteins, nucleic acids, lipids, and other biomolecules. After being released into the extracellular environment, some of these vesicles are delivered to recipient cells; consequently, the target cell may undergo physiological or pathological changes. Thus, extracellular vesicles as biological nano-carriers, have a pivotal role in facilitating long-distance intercellular communication. Understanding the mechanisms that mediate this communication process is important not only for basic science but also in medicine. Indeed, extracellular …

LA GENETICA GENERALE ED UMANA

libro di biologia e genetica

Preliminary data on antiproliferative effects in haemocyte extracts of the ascidian Styela plicata (Stolidobranchiata, Styelidae)

Effetti antiproliferativi degli estratti di emociti dell'ascidia Styela plicata ( Tunicata)

Microscopic evidence of the primary astrocytes' morphological differentiation and migration inside porous Poly-L-lactic acid 3D‑scaffolds

Tissue engineering is an emerging multidisciplinary field that aims at reproducing in vitro and/or in vivo tissues with morphological and functional features similar to the biological tissue of the human body [1]. In the attempt to construct suitable tissue models, a critical step is the setting of 3D scaffolds that mimic the supportive structures of a natural extracellular matrix microenvironment into which cells are normally embedded. In this context, the generation of 3D cultures of brain cells is of particular interest. For instance, the poly L‐lactic acid (PLLA) polymer is wildly used because of its biocompatible and biodegradable potential; the PLLA scaffold topography simulates the n…

Isolation and characterization of Citrus limon L. derived nanovesicles: potential use as antineoplastic agent

We isolated and characterized nanovesicles from edible Citrus limon with size and composition similar to mammalian-derived exosomes. Furthermore we show an in vitro and in vivo anti-proliferative and pro-apoptotic effect of these vesicles. This study opens to the possibility of using this natural plant-derived nanovesicles as antineoplastic agents.

Caratterizzazione morfologica, ultrastrutturale e funzionale della linea cellulare umana di carcinoma del colon T84SF: un modello per lo studio dell’extravasazione tumorale

Colorectal cancer-derived small extracellular vesicles induce TGFβ1-mediated epithelial to mesenchymal transition of hepatocytes

Abstract Background Metastatic disease is the major cause of cancer-related deaths. Increasing evidence shows that primary tumor cells can promote metastasis by preparing the local microenvironment of distant organs, inducing the formation of the so-called “pre-metastatic niche”. In recent years, several studies have highlighted that among the tumor-derived molecular components active in pre-metastatic niche formation, small extracellular vesicles (sEVs) play a crucial role. Regarding liver metastasis, the ability of tumor-derived sEVs to affect the activities of non-parenchymal cells such as Kupffer cells and hepatic stellate cells is well described, while the effects on hepatocytes, the m…

Immunolocalization and involvement of antimicrobial peptides in local inflammatory-like reactions in the tunic of Ciona intestinalis (Tunicata).

Immunolocalizzazione di un peptide antimicrobico nella tunica di Ciona intestinalis (Tunicata)

Immunolocalization of an antimicrobial peptide in Ciona intestinalis (Tunicata ) tunic

Itraconazole inhibits nuclear delivery of extracellular vesicle cargo by disrupting the entry of late endosomes into the nucleoplasmic reticulum

ABSTRACT Extracellular vesicles (EVs) are mediators of intercellular communication under both healthy and pathological conditions, including the induction of pro‐metastatic traits, but it is not yet known how and where functional cargoes of EVs are delivered to their targets in host cell compartments. We have described that after endocytosis, EVs reach Rab7+ late endosomes and a fraction of these enter the nucleoplasmic reticulum and transport EV biomaterials to the host cell nucleoplasm. Their entry therein and docking to outer nuclear membrane occur through a tripartite complex formed by the proteins VAP‐A, ORP3 and Rab7 (VOR complex). Here, we report that the antifungal compound itracona…

Mutazioni: tipi, origini, conseguenze

Amphiregulin contained in NSCLC-exosomes induces osteoclast differentiation through the activation of EGFR pathway

AbstractNon-small cell lung cancer (NSCLC) remains the leading cause of cancer-related deaths worldwide. The majority of patients are diagnosed in advanced disease stage. Bone metastasis is the most frequent complication in NSCLC resulting in osteolytic lesions. The perfect balance between bone-resorbing osteoclasts and bone-forming osteoblasts activity is lost in bone metastasis, inducing osteoclastogenesis. In NSCLC, the epidermal growth factor receptor (EGFR) pathway is constitutively activated. EGFR binds Amphiregulin (AREG) that is overexpressed in several cancers such as colon, breast and lung. Its levels in plasma of NSCLC patients correlate with poor prognosis and AREG was recently …