Tumor Microenvironment In Experimental Models Of Human Cancer: Morphological Investigational Approaches

Introduction. Tumor microenvironment (TME) is defined as the non-tumoral part of tumors. It is composed of different cell populations and structures (such as tumor-associated vasculature, immune-inflammatory cells, fibroblasts…) (Hanahan and Coussens, 2012). TME could either promote or antagonize tumor growth and has a great potential as target for novel therapeutic strategies. Along with several methods (i.e. molecular assays), morphological techniques allow to evaluate the components of TME in the setting of their action. The aim of this work was to set up and define valuable morphological approaches useful in the investigation of the TME.Materials and methods. Histological and immunohistochemical techniques, along with digital image analysis, were tested on experimental mouse models (both xenograft and genetically engineered mice) of four different human tumors (ovarian cancer, pancreatic ductal adenocarcinoma (PDAC), colon adenocarcinoma, thyroid carcinoma).Results. Concerning the vascular compartment, CD31 immunostaining and double-immunofluorescence with CD31 and a-SMA (pericytes marker) allowed to respectively quantify vessels and evaluate their maturation degree. Immunohistochemical detection of previously administrated Pimonidazole, revealed variable extended areas of hypoxia within tumoral masses in a consistent pattern between frozen and formalin-fixed paraffin-embedded samples.Concerning the stromal component, anti-human MHC I and specie-specific markers for Vimentin demonstrated the host-derivation of stroma in xenotumors, while Sirius Red histochemical staining allowed the quantification of desmoplasia in models of PDAC.Concerning immune-inflammatory cells, an immunohistochemical panel with CD3 (T lymphocytes), B220 (B lymphocytes), MPO (neutrophils) and Iba-1 (macrophages), showed high reliability in characterizing the tumoral infiltrate. Moreover, the application of markers specific for different macrophage subsets confirmed the higher prevalence of M2 (Arginase I positive) on M1 (iNOS positive) macrophages. YM1 demonstrated a low performance in detecting the M2 population (Fig. 1).Discussion and conclusions. Due to the microenvironmental heterogeneity which influence tumor development and biological behavior, a sole quantification is unreliable for characterizing the TME. Considering that, morphological techniques proved to be a valuable approach, allowing the evaluation of the spatial distribution and mutual interaction between the different elements. Additional studies are needed for further investigate the biological significance of spatial distribution of the components of the TME

Identification and Preliminary Validation in Mouse Models of Circulating Biomarkers of Pancreatic Cancer

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal oncological malignancies in humans. Not-specific symptoms and lack of early diagnostic strategies, frequently lead to late diagnosis which limited therapeutic possibilities (Korc, 2007). The present study aimed at identifying novel potential serum biomarkers for early detection of PDAC.In the first phase, two different mouse models of PDAC were characterized: genetically engineered mice (GEMs) (Hingorani et al., 2003) which developed PanIN (pancreatic intraepithelial neoplasia) lesions and three PDAC patient-derived xenograft.In the second phase, the two mouse models were used to evaluate the reliability of 3 circulating molecules as early diagnostic biomarkers of PDAC. The plasma levels of matrix metalloproteinase-7 (MMP-7), tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) and thrombospondin-2 (THBS-2) were tested on GEMs and PDAC-PDXs bearing mice by ELISA tests, during tumor development, and at sacrifice by immunohistochemistry performed on pancreatic tissue.The three established PDAC-PDXs were found to better reproduce the tumor of origin after intra-pancreas transplantation compared to the subcutaneous ones, and to maintain molecular and morphological features over different passages.At sacrifice, histopathological analysis demonstrated different stages of PanIN lesions in GEMs and the presence of a well-developed pancreatic tumor in all the mice orthotopically inoculated with the PDAC-PDXs.Plasma levels of MMP-7, TIMP-1 and THBS-2 were progressively upregulated, over the time, in GEMs and in PDAC-PDX bearing mice.In both animal models, immunohistochemistry revealed stromal immunoreactivity for TIMP-1 and THBS-2, while MMP-7 expression was mainly localized on epithelial cells. All the markers showed progressive increase of staining intensity along with PanIN progression.In conclusion, the investigated circulating molecules represent promising biomarkers for early diagnosis of PDAC and to monitor the response to treatment in human patients. Both tumoral cells and associated stroma play a role in the production and release of such biomarkers

Presentazione

L’estetica animale come campo di studi ha mostrato negli ultimi due decenni uno sviluppo importante all’interno dell’estetica ambientale. Mentre gli approcci sperimentali e speculativi (teoretici) hanno in qualche modo dominato l’estetica ambientale dalla sua nascita come campo specifico di studi dell’estetica, le ricerche specifiche indirizzate all’estetica animale hanno visto uno sviluppo per certi versi simile ma con un maggior apporto interdisciplinare.[1] Linee di ricerca filosofiche anche molto diverse fra loro prendono avvio spesso da osservazioni tratte dall’etologia o da branche specifiche della zoologia, come l’entomologia. Allo stesso tempo, biologi impegnati nelle aree più disparate della vita animale e vegetale arrivano a toccare, forse più marcatamente che nel passato, questioni squisitamente filosofiche, e in particolare, estetologiche. Se una tale commistione di approcci e linee di ricerca ha certamente generato un fiorire di studi sul comportamento artistico animale – studi spesso arricchiti da preziose osservazioni tratte dal lavoro “sul campo” – allo stesso tempo, il richiamo a concetti tradizionalmente adottati dal pensiero filosofico (arte, sensibilità, tecnica, istinto, etc.) ha generato un bisogno sempre più impellente di chiarezza concettuale a cui autori dalle formazioni teoriche più diverse stanno cercando di dare risposta. In questa direzione, è sembrato puntuale e necessario proporre un numero della nostra rivista incentrato sull’estetica animale e su alcuni aspetti rilevanti della problematica animale. Abbiamo deciso di raccogliere alcuni contributi dalla natura anche molto diversa tra loro e con impostazioni metodologiche diverse (dalla fenomenologia, alla critica letteraria, ad esempio), in modo che il lettore possa farsi un’idea, la più problematizzante possibile, rispetto agli attuali sviluppi dell’estetica animale. In ogni caso, non abbiamo escluso dal nostro volume approcci più storici rispetto al problema dell’animale, in modo che le tematiche affrontate siano contestualizzate anche nella tradizione del pensiero occidentale. In queste brevi righe introduttive ci limiteremo dunque a indicare alcune linee di lettura che il paziente lettore potrà (o meno) decidere di seguire, per costruire un filo conduttore che unisca tutti i contributi brevemente introdotti al termine di queste pagine, e che armonizzi voci così diverse fra loro, in un panorama tanto vario e complesso come quello degli animal studies.   [1] Si veda, A. Carlson, “Ten Steps in the Development of Western Environmental Aesthetics” in M. Drenthen, J. Keulatz (Ed.) Environmental aesthetics: crossing divides and breaking ground, New York, Fordham University Press, 2014, pp. 14-15

TYPE AND EXPERIENCE. AN INQUIRY INTO THE ROLE AND FUNCTION OF TYPE AND TYPIFYING-APPERCEPTION IN EXPERIENCE AND COGNITION

The work concentrates on the early works and production of Edmund Husserl, and especially, on the time we consider fundamental for the development of Husserl's later phenomenology. That will also cover a period normally less taken into consideration by the critic. Ideality is in our understanding linked as label-term to the Husserlian \uabeidetics\ubb, which is for its part explicitly derived in the first Book of Ideas from eidos, a term introduced in reality years earlier by Husserl, in connection with essence [Wesen]. It is in fact only around the time of the works belonging to this fundamental introduction to phenomenology that the concept of eidos is assumed as an equivalent for \uabpure essence\ubb and phenomenology is established for its part as \u201ceidetics\u201d or \u201ceidetic science\u201d. Around the time of the Logical Investigations indeed, the term essence comprehends a large variety of concepts less distinguished in comparison with to later Husserlian works, for example, for what concern the concept of eidos. Among the different definitions and functions showed by essence and idea in the 1900 work and in the course on logic and epistemology before the Ideas, we will try to stress one peculiar aspect of such overly complex thematic, which is the partial definition of essence in terms of conceptual universality. With respect to this characterization, we will therefore try to indicate the kind of definition given to it by Husserl, which partially recalls the traditional interpretation of the Universal in the sense of an ideal terms, or a common element, over against the multiplicity. This latter is therefore interpreted in the sense of a universal object. Our aim will be consequently, to analyze a series of manuscripts on the period prior to the Logical Investigations, where, according to our interpretation, all the features assigned to the same concept are present. That will be basically, show the \u201corigin\u201d of one important aspect of the future doctrine of essence, even besides the later interpretation via Lotze and Bolzano of Ideality

In situ characterization of immiscible three-phase flow at the pore scale for a water-wet carbonate rock

X-ray micro-tomography is used to image the pore-scale configurations of fluid in a rock saturated with three phases - brine, oil and gas - mimicking a subsurface reservoir, at high pressure and temperature. We determine pore occupancy during a displacement sequence that involves waterflooding, gas injection and water re-injection. In the water-wet sample considered, brine occupied the smallest pores, gas the biggest, while oil occupied pores of intermediate size and is displaced by both water and gas. Double displacement events have been observed, where gas displaces oil that displaces water or vice versa. The thickness of water and oil layers have been quantified, as have the contact angles between gas and oil, and oil and water. These results are used to explain the nature of trapping in three-phase flow, specifically how oil preferentially traps gas in the presence of water

Determination of contact angles for three-phase flow in porous media using an energy balance

HYPOTHESIS: We define contact angles, θ, during displacement of three fluid phases in a porous medium using energy balance, extending previous work on two-phase flow. We test if this theory can be applied to quantify the three contact angles and wettability order in pore-scale images of three-phase displacement. THEORY: For three phases labelled 1, 2 and 3, and solid, s, using conservation of energy ignoring viscous dissipation (Δa1scosθ12-Δa12-ϕκ12ΔS1)σ12=(Δa3scosθ23+Δa23-ϕκ23ΔS3)σ23+Δa13σ13, where ϕ is the porosity, σ is the interfacial tension, a is the specific interfacial area, S is the saturation, and κ is the fluid-fluid interfacial curvature. Δ represents the change during a displacement. The third contact angle, θ13 can be found using the Bartell-Osterhof relationship. The energy balance is also extended to an arbitrary number of phases. FINDINGS: X-ray imaging of porous media and the fluids within them, at pore-scale resolution, allows the difference terms in the energy balance equation to be measured. This enables wettability, the contact angles, to be determined for complex displacements, to characterize the behaviour, and for input into pore-scale models. Two synchrotron imaging datasets are used to illustrate the approach, comparing the flow of oil, water and gas in a water-wet and an altered-wettability limestone rock sample. We show that in the water-wet case, as expected, water (phase 1) is the most wetting phase, oil (phase 2) is intermediate wet, while gas (phase 3) is most non-wetting with effective contact angles of θ12≈48° and θ13≈44°, while θ23=0 since oil is always present in spreading layers. In contrast, for the altered-wettability case, oil is most wetting, gas is intermediate-wet, while water is most non-wetting with contact angles of θ12=134°±~10°,θ13=119°±~10°, and θ23=66°±~10°