Barbara Franceschini

Quantitative evaluation and modeling of two-dimensional neovascular network complexity: the surface fractal dimension

BackgroundModeling the complex development and growth of tumor angiogenesis using mathematics and biological data is a burgeoning area of cancer research. Architectural complexity is the main feature of every anatomical system, including organs, tissues, cells and sub-cellular entities. The vascular system is a complex network whose geometrical characteristics cannot be properly defined using the principles of Euclidean geometry, which is only capable of interpreting regular and smooth objects that are almost impossible to find in Nature. However, fractal geometry is a more powerful means of quantifying the spatial complexity of real objects.MethodsThis paper introduces the surface fractal dimension (Ds) as a numerical index of the two-dimensional (2-D) geometrical complexity of tumor vascular networks, and their behavior during computer-simulated changes in vessel density and distribution.ResultsWe show that Dssignificantly depends on the number of vessels and their pattern of distribution. This demonstrates that the quantitative evaluation of the 2-D geometrical complexity of tumor vascular systems can be useful not only to measure its complex architecture, but also to model its development and growth.ConclusionsStudying the fractal properties of neovascularity induces reflections upon the real significance of the complex form of branched anatomical structures, in an attempt to define more appropriate methods of describing them quantitatively. This knowledge can be used to predict the aggressiveness of malignant tumors and design compounds that can halt the process of angiogenesis and influence tumor growth.

Usefulness of cancer-testis antigens as biomarkers for the diagnosis and treatment of hepatocellular carcinoma

Despite advances in our cellular and molecular knowledge, hepatocellular carcinoma (HCC) remains one of the major public health problems throughout the world. It is now known to be highly heterogeneous: it encompasses various pathological entities and a wide range of clinical behaviors, and is underpinned by a complex array of gene alterations that affect supra-molecular processes.Four families of HCC tumour markers have been recently proposed: a) onco-fetal and glycoprotein antigens; b) enzymes and iso-enzymes; c) cytokines and d) genes. A category of tumour-associated antigens called cancer-testis (CT) antigens has been identified and their encoding genes have been extensively investigated. CT antigens are expressed in a limited number of normal tissues as well as in malignant tumors of unrelated histological origin, including the liver. Given that cancers are being recognized as increasingly complex, we here review the role of CT antigens as liver tumour biomarkers and their validation process, and discuss why they may improve the effectiveness of screening HCC patients and help in determining the risk of developing HCC.

Immunolocalization of Sperm Protein 17 in Human Testis and Ejaculated Spermatozoa

Sperm protein 17 (Sp17) is a highly conserved mammalian protein whose primary function is still poorly understood. Immunohistochemistry (IHC) in the human testis reveals the presence of Sp17 in some spermatocytes and abundantly in spermatids. All spermatogonia, Sertoli cells, and Leydig cells appear to be immunonegative for Sp17, whereas some interstitial cells are immunopositive. IHC recognized two distinct populations (immunopositive or not for Sp17) in the ejaculated spermatozoa. Although it will be necessary to clarify why some ejaculated spermatozoa do not contain Sp17, its distribution suggests that this protein may be associated with some phases of germinal cell differentiation.

The Complex Functions of Mast Cells in Chronic Human Liver Diseases

Mast cells (MCs) are multifunctional effector cells of the immune system. MCs were originally thought to be involved in IgE-associated immediate hypersensitivity and allergic disorders, but it is now known that they contain or elaborate an array of mediators with a multitude of effects on many other cells. A number of studies have found that MCs are involved in various liver diseases. Although still controversial, they seem to be involved in the liver’s fibrotic response to chronic inflammation and parasitic infection. Hepatic fibrosis is the most frequent liver response to toxic, infectious, or metabolic agents. During the establishment of this pathological condition, there is an increase in the components of the basement membrane that leads to continuous basement membrane-like structures being raised within Disse’s space and a decrease in the number of sinusoid endothelial fenestrae. This leads to a complex process called “sinusoidal capillarization.” At the cellular level, liver fibrogenesis is initiated by hepatocyte necrosis, which induces the recruitment of a large number of inflammatory cells, including MCs, which can be considered the primary effectors of the process changing sinusoidal endothelial cells into capillary-type endothelial cells. We review the roles played by MCs in hepatic chronic diseases and describe a biopsy section of hepatic tissue taken from a patient with chronic C virus-related hepatitis showing diffuse sinusoidal capillarization and a high density of MCs. This observation has led us to hypothesize a relationship between these highly specialized cells and sinusoidal capillarization.

Sperm protein 17 is expressed in human nervous system tumours

BackgroundHuman sperm protein 17 (Sp17) is a highly conserved protein that was originally isolated from a rabbit epididymal sperm membrane and testis membrane pellet. It has recently been included in the cancer/testis (CT) antigen family, and shown to be expressed in multiple myeloma and ovarian cancer. We investigated its immunolocalisation in specimens of nervous system (NS) malignancies, in order to establish its usefulness as a target for tumour-vaccine strategies.MethodsThe expression of Sp17 was assessed by means of a standardised immunohistochemical procedure [(mAb/antigen) MF1/Sp17] in formalin-fixed and paraffin embedded surgical specimens of NS malignancies, including 28 neuroectodermal primary tumours (6 astrocytomas, 16 glioblastoma multiforme, 5 oligodendrogliomas, and 1 ependymoma), 25 meningeal tumours, and five peripheral nerve sheath tumours (4 schwannomas, and 1 neurofibroma),.ResultsA number of neuroectodermal (21%) and meningeal tumours (4%) were found heterogeneously immunopositive for Sp17. None of the peripheral nerve sheath tumours was immunopositive for Sp17. The expression pattern was heterogeneous in all of the positive samples, and did not correlate with the degree of malignancy.ConclusionThe frequency of expression and non-uniform cell distribution of Sp17 suggest that it cannot be used as a unique immunotherapeutic target in NS cancer. However, our results do show the immunolocalisation of Sp17 in a proportion of NS tumour cells, but not in their non-pathological counterparts. The emerging complex function of Sp17 makes further studies necessary to clarify the link between it and immunopositive cells.

Fractal analysis of two-dimensional vascularity in primary prostate cancer and surrounding non-tumoral parenchyma

Prostate cancer is the fifth most frequent cancer in the world. However, none of the actual prognostic factors provide a valid index for predicting patient outcome. Here, we evaluate the two-dimensional vascularity in primary prostate tumors and surrounding non-tumoral parenchyma by means of fractal geometry, and assess any correlations between the results and some clinical and pathological parameters of prostate carcinoma. Prostate sections from 27 carcinoma patients were treated with CD34 antibodies. Two >10mm(2) areas of tumoral and surrounding non-tumoral parenchyma were digitized using an image analysis system that automatically quantified the fractal dimension of the vascular surface. Data were correlated with patients age, PSA level, clinical and pathological stage, Gleason score, tumor volume, vascular invasion, surgical margins, and biochemical relapse. Two groups of patients were distinguished on the basis of whether the fractal dimension of their tumoral vascular surface was higher (group 1) or lower (group 2) than that of the surrounding non-tumoral parenchyma. Statistically significant between-group differences were found in terms of serum PSA levels (p=0.0061), tumor volume (p=0.0017), and biochemical relapse (p=0.031). The patients in group 2 had a poorer outcome. Our findings suggest a group of prostate cancer patients with a poor outcome, and the vascular surface fractal dimension as a helpful geometrical index in clinical practice.

Some Remarks on the Somatic Expression of Sperm Protein 17

Fabio GRIZZI, Barbara FRANCESCHINI, Paul L. HERMONAT, Yong LIU and Maurizio CHIRIVA-INTERNATI* Scientific Direction, Istituto Clinico Humanitas, Rozzano, Milan, Italy Foundation “Michele Rodriguez”, Institute for Quantitative Methods in Medicine, Milan, Italy Division of Obstetrics and Gynecology, University of Arkansas Medical Sciences, Little Rock, AK, USA Texas Tech University Health Science Center, Department of Microbiology and Immunology, Lubbock, TX, USA

Vascular architecture: is it a helpful histopathological biomarker for hepatocellular carcinoma?

Hepatocellular carcinoma (HCC) remains one of the major public health problems throughout the world. Although originally associated with tumorigenic processes, liver angiogenesis has also been observed in the context of different liver inflammatory, fibrotic, and ischemic conditions. Here we investigate the fractal dimension as a quantitator of non-Euclidean two-dimensional vascular geometry in a series of paired specimens of primary HCC and surrounding non-tumoral tissue, and discuss why this parameter might provide additional information regarding cancer behavior. The application of fractal geometry to the measurement of liver vascularity and the availability of a computer-aided quantitative method can eliminate errors in visual interpretation, and make it possible to obtain closer-to-reality numerals that are compulsory for any measurement process.

Sperm protein 17 and AKAP-associated sperm protein cancer/testis antigens are expressed in ciliated hepatic foregut cysts

Ciliated hepatic foregut cysts (CHFCs) are retained benign lesions of the liver. However, a case of squamous cell metaplasia and five cases of squamous cell carcinoma arising from a CHFC have been described. The potential of malignant transformation makes the identification of new biomarkers necessary. As the cancer/testis antigen sperm protein 17 (Sp17) has been detected in oral and oesophageal squamous cell carcinomas, the aim of this study was to investigate the expression of Sp17 and AKAP‐associated sperm protein (ASP), which has a shared N‐terminal sequence with Sp17, in four surgically resected CHFCs.

The Metrizer: an innovative device for achieving virtual hepatic biopsies

The last few years have brought rapid growth in the number of Virtual Microscopes (VM) and the promotion of digital histology. In this digital era, it is natural to assist in the transportation of histology imaging into computer technology support. We have also assisted in a quick and precise race towards improved technology capable of acquiring detailed digital images which realistically report the original slide for easy consultation [1,2]. The advanced technology herein proposed aims at being a sophisticated imaging archive. Digitally scanning slides however, does not give additional information to the vision of tissue structures, even with high resolution or improved colour and image precision. We can now say that high enlargement microscopic observation can only give same details to structures visible to the eye directly through the microscope. This means that when VMs are used in a correct manner they are capable of giving significant progress to highlighting, not quantifying, a critical field in applying technology specific to computerised measuring. The potential to improve precision and objectivity of measurements can be achieved with additional technical equipment. The aim of this study is to present a machine invented with a calculation potential to facilitate the work of the observer in a medical practice, not only in terms of easy retrieval of images but also as an instrument for the automatic analysis of digital histology. The “Metrizer” aims at supplying precise and objective descriptions and measurements of the specimen under observation. These descriptions do not substitute the pathologist but they should assist him/her to assert with objectivity and safety the entity of the observed pathology [3-8]. Material and methods The “Metrizer” (Figure 1) is an automatic, compact machine composed of a lens for microscopic observation, digital cameras, a movement device and a computer complete with software to control machine movement and image analysis. With these components the machine facilitates consultation of histological preparations (virtual microscop e-s lide scanner function) and supplies objective numeric data regarding the state of the tissue harbouring diverse diseases without causing fatigue or human error. Using the Slide Scanner function it enables to automatically digitalise the entire histological slide in high definition and makes it easily accessible in the digital archive.