Defining the perivascular niche in the early stages of breast cancer bone metastasis

Advanced breast cancer is frequently associated with skeletal metastases. During dissemination to bone, breast cancer cells locate in a putative ‘metastatic niche’. Its components are not fully elucidated, however there is evidence of at least partial overlap between the hematopoietic stem cell (HSC), endosteal, metastatic and perivascular niches in bone. Two-photon microscopy was used to detect single tumour cells and map their location within the bone microenvironment. Irrespective of ER status, route of injection and animal age, breast cancer cells preferentially located in the trabecular region of the bone adjacent to the bone surface. Animals pre-treated with AMD3100 to mobilise HSCs prior injection of tumour cells displayed a higher number of breast cancer cells homing to bone, indicating that HSCs and breast cancer cells compete for space within the niche. The microenvironment of tumour growth-promoting (6-week old) and dormancy supporting (12-week old) animal models were investigated. In mature animals, bone volume, length and number of H-vessels and expression of CD31+ and CD34+ vasculature were reduced, while TSP-1 expression was increased, compared to the young model. To investigate the effect of Zoledronic acid (ZOL) on the bone microvasculature, young and mature animal model were treated with a single dose of ZOL and changes in the vasculature visualised with immunofluorescent protocols. In young animals ZOL did not change the microvasculature, while mature animals showed shorter and more numerous vessels. Alteration of microvascular activity, decreased with cediranib and increased with Deferoxaminemesylate, was investigated. Preliminary data did not show substantial changes in either the bone or the microvasculature structure and further studies are required. Overall, my work supports that the bone metastatic niche consists of several overlapping and interconnected niches. My data suggest that new approaches for 9 treatment of bone metastasis should evaluate the effect on several components of the bone microenvironmen

Endogenous production of IL-1B by breast cancer cells drives metastasis and colonisation of the bone microenvironment

Background: Breast cancer bone metastases are incurable highlighting the need for new therapeutic targets. After colonizing bone, breast cancer cells remain dormant, until signals from the microenvironment stimulate outgrowth into overt metastases. Here we show that endogenous production of IL-1B by tumor cells drives metastasis and growth in bone. Methods: Tumor/stromal IL-B and IL-1R1 expression was assessed in patient samples and effects of the IL-1R antagonist, Anakinra or the IL-1B antibody Canakinumab on tumor growth and spontaneous metastasis were measured in a humanized mouse model of breast cancer bone metastasis. Effects of tumor cell-derived IL-1B on bone colonisation and parameters associated with metastasis were measured in MDA-MB-231, MCF7 and T47D cells transfected with IL-1B/control. Results: In tissue samples from >1300 patients with stage II/III breast cancer, IL-1B in tumor cells correlated with relapse in bone (hazard ratio 1.85; 95% CI 1.05-3.26; P=0.02) and other sites (hazard ratio 2.09; 95% CI 1.26-3.48; P=0.0016). In a humanized model of spontaneous breast cancer metastasis to bone, Anakinra or Canakinumab reduced metastasis and reduced the number of tumor cells shed into the circulation. Production of IL-1B by tumor cells promoted EMT (altered E-Cadherin, N-Cadherin and G-Catenin), invasion, migration and bone colonisation. Contact between tumor and osteoblasts or bone marrow cells increased IL-1B secretion from all three cell types. IL-1B alone did not stimulate tumor cell proliferation. Instead, IL-1B caused expansion of the bone metastatic niche leading to tumor proliferation. Conclusion: Pharmacological inhibition of IL-1B has potential as a novel treatment for breast cancer metastasis

Osakeyhtiön tasekirjapohja: Case: Kouvola Innovation Oy

Kunnan Taitoa Oy:n Kouvolan toimipisteen asiakkaan, Kouvolan kaupungin omistaman yhtiön, Kouvola Innovation Oy:n kirjanpitoa on hoitanut useampi eri henkilö, eikä tilinpäätöstä ja tasekirjaa varten ole ollut valmista pohjaa tai yhtenäistä mallia. Eri vuosien tasekirjoissa on eroja sekä ulkonäössä että sisällössä. Tämän opinnäytetyön tavoitteena oli luoda Kunnan Taitoa Oy:n Kouvolan toimipisteeseen tasekirjapohja vanhoja tasekirjoja kehittämällä. Pohja tulisi Kouvola Innovation Oy:n tilinpäätöksen laatijan apuvälineeksi. Tarkoituksena oli tehdä pohjasta sellainen, että sitä voidaan pienillä muutoksilla hyödyntää muidenkin yhtiöiden tilinpäätökseen ja että se hakee tilinpäätöslaskelmien luvut suoraan konsernikirjanpito-ohjelman tietokannasta. Työssä perehdytään tilinpäätöksen ja tasekirjan sisältöön lainsäädännön, asetuksien, yleisohjeiden ja kirjallisuuden avulla. Teoriaa on kirjoitettu samaan aikaan, kun tasekirjapohjaa on laadittu. Aikaisempia tasekirjoja tutkimalla ja teoriaa selvittämällä pyrittiin löytämään kehittämiskohtia niin ulkoasusta kuin asiasisällöstä. Työn tuloksena on toimiva tasekirjapohja, jota voidaan hyödyntää pienillä muutoksilla muidenkin yhtiöiden, kuin Kouvola Innovation Oy:n tilinpäätökseen. Pohja saatiin rakennettua niin, että se poimii laskelmiin ja liitetietoihin lukuja tietokannasta. Kehittämiskohtia ja niiden ratkaisuja löydettiin sekä niihin liittyviä tarpeellisia jatkotoimenpiteitä.Kouvola Innovation Ltd is a limited liability company owned by City of Kouvola and it is a client of Kunnan Taitoa Ltd, which main line of business is combined office administrative service activities. Different persons have been in charge of accounting at Kouvola Innovation and there has been no ready layout or common pattern for financial statement or balance sheet book. There are differences in appearance and content between annual balance sheet books of different years. The objective of this thesis was to create a new balance sheet book layout for Kunnan Taitoa Ltd’s Kouvola office by improving previous balance sheet books. The layout would be used to assist the making of the financial statement of Kouvola Innovation. The purpose was to create a layout which could be used with small adjustments in financial statements of different companies and which would retrieve numbers of financial statements from group accounting program’s database. The content of financial statement and balance sheet book are discussed in the light of legislation, regulation, guidelines and literature on this thesis. The theory was written simultaneously with the layout compilation. The improvements to appearance and content were aimed to be found by studying previous balance sheet books and theory

Additional file 1: Table S1. of Zoledronic acid alters hematopoiesis and generates breast tumor-suppressive bone marrow cells

Flow cytometry cell-surface markers used to quantify indicated hematopoietic stem and progenitor cell populations. (PDF 16 kb

Development of a liver-specific tet-on inducible system for AAV vectors and its application in the treatment of liver cancer

Recombinant adeno-associated virus (rAAV) are effective gene delivery vehicles that can mediate long-lasting transgene expression. However, tight regulation and tissue-specific transgene expression is required for certain therapeutic applications. For regulatable expression from the liver we designed a hepatospecific bidirectional and autoregulatory tetracycline (Tet)-On system (Tet bidirAlb) flanked by AAV inverted terminal repeats (ITRs). We characterized the inducible hepatospecific system in comparison with an inducible ubiquitous expression system (Tet bidirCMV) using luciferase (luc). Although the ubiquitous system led to luc expression throughout the mouse, luc expression derived from the hepatospecific system was restricted to the liver. Interestingly, the induction rate of the Tet bidir Alb was significantly higher than that of Tet bidirCMV, whereas leakage of Tet bidir Alb was significantly lower. To evaluate the therapeutic potential of this vector, an AAV-Tet bidir-Alb-expressing interleukin-12 (IL-12) was tested in a murine model for hepatic colorectal metastasis. The vector induced dose-dependent levels of IL-12 and interferon-γ (IFN-γ), showing no significant toxicity. AAV-Tet bidir-Alb-IL-12 was highly efficient in preventing establishment of metastasis in the liver and induced an efficient T-cell memory response to tumor cells. Thus, we have demonstrated persistent, and inducible in vivo expression of a gene from a liver-specific Tet-On inducible construct delivered via an AAV vector and proved to be an efficient tool for treating liver cancer. © The American Society of Gene & Cell Therapy.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

La Menzogna: Le altre facce della realtà

Il volume presenta contributi scientifici sul tema della menzogna nelle sue più diversificate ‘manifestazioni’ testuali e declinazioni ‘comunicative’, potendo essere intesa come immaginazione, invenzione, finzione, artificio, dissimulazione, deformazione ma anche come manipolazione.Il volume si articola in tre macrosezioni: letteraria, filologica-linguistica e artistica. La sezione letteraria contiene una riflessione ramificata che offre uno sguardo multiprospettico. La menzogna è analizzata dapprima come tema per poi tramutarsi in oggetto di studio della critica letteraria, giungendo, infine, in un mutevole paesaggio geoculturale. La seconda sezione, quella filologica-linguistica, segue invece un principio diacronico: si indaga la menzogna a partire dalla tradizione manoscritta fino alla comunicazione digitale. La terza e ultima sezione, che conferma la natura interdisciplinare e transmediale del volume, è dedicata alle strategie di rappresentazione e dissimulazione in rapporto alle diverse forme d’arte, quali il teatro, le serie TV, la docufiction, nonché le arti performative. Il volume si chiude con un’appendice che contiene le riproduzioni delle opere pittoriche di alcuni artisti, presentate in occasione della mostra che ha affiancato la Graduate Conference