This volume aims to provide a range of methods and protocols for studying tumor angiogenesis in vitro and in vivo to reflect advances in the field. The chapters in this book cover topics such as: morphological aspects of tumor angiogenesis, aortic ring assay and its use for the study of tumor angiogenesis, ex vivo tissue culture model for anti-angiogenic drug testing, transgenic zebrafish, orthotopic models of ovarian cancer, and uncovering metabolic effects of anti-angiogenic therapy in tumors by induced metabolic bioluminescence imaging. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting edge and thorough, Tumor Angiogenesis Assays: Methods and Protocols is a valuable resource for anyone interested in tumor angiogenesis assay research.

This book traces significant aspects of the history of immunology, exploring the immune system and immunodeficiency. The author recounts human hematopoietic development, and how a distinction of the immune system into thymus-dependent and thymus-independent components has been demonstrated in different animal species, including amphibians, birds, and mammals. Other themes explored in this book include discoveries about the role of the thymus of the Bursa of Fabricius in the development of immunologic competence, and observations on the changes in the lymphoid organs after bursectomy and thymectomy in chickens. Readers will discover how the bursa provides a unique microenvironment for the proliferation and differentiation of B cells, while thymectomized and irradiated animals were deficient in lymphocytes that mediated inflammatory responses, as assessed by skin graft rejection, delayed-type hypersensitivity, and graft versus host reaction. A clear perspective for understanding several diseases and also the entire lymphoid system emerges through the experiments and extensive histopathological studies of patients with primary immunodeficiency diseases that are described in these chapters. Researchers in the life sciences, in biomedicine and the history of medicine will all find something of value in this highly engaging work. It will also appeal to those with an interest in public health and neurobiology.

"Vascular Morphogenesis: Methods and Protocols" provides a range of techniques for studying vascular morphogenesis in vivo and in vitro, reflecting advances in the field. Endothelial cell signaling is currently believed to promote fundamental cues for cell fate specification, embryo patterning, organ differentiation and postnatal tissue remodeling. Understanding the concept of vascular bed specificity represents a major challenge for future investigations. Indeed, one of the most interesting theoretical perspectives and practical applications of endothelial cell signaling is the possibility for these cells to maintain their inductive potential during adult life. Written in the highly successful "Methods in Molecular Biology" series format, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.

Practical and authoritative, "Vascular Morphogenesis: Methods and Protocols" will serve as a instructive and useful reference for cell biologists, anatomists, pathologists and physiologists, as well as all those interested in this area of biological study.

Mast cells are versatile, tissue-homing secretory cells, which were first described by Paul Ehrlich in 1878. Mast cells have long been implicated in the pathogenesis of allergic reactions and certain protective responses to parasites. Their functional role, however, has been discovered to be increasingly complex and multifarious. Mast cells have been implicated in various cell-mediated immune reactions, being found in tissues from multiple disease sites, and as a component of the host reaction to bacteria, parasite, and even virus infections. They have also been shown to participate to angiogenic and tissue repair processes after injury. The importance of a possible functional link between chronic inflammation and cancer has long been recognized. As most tumours contain inflammatory cell infiltrates, which often include plentiful mast cells, the question as to the possible contribution of mast cells to tumour development has progressively been emerged. In this book, the general biology of these cells, their development, anatomical distribution and phenotype as well as their secretory products will first be discussed. The biology of tumour cells, their structural and molecular characteristics, the specificity of the tumour microenvironment and the development of a vascular network in the tumour context will be analyzed. The involvement of mast cells in tumour biology and tumour fate will then be considered, with particular emphasis on the capacity of these cells to stimulate tumour growth by promoting angiogenesis and lymphangiogenesis. The last chapter suggest that mast cells may serve as a novel therapeutic target for cancer treatment.

In spite of their relatively low prevalence, systemic vasculitides have been the object of intensive basic and clinical investigations over the last few years. As a consequence, important advancements have been achieved: from updated diagnostic and classification criteria and a more rational nomenclature to the recognition of an expanding spectrum of clinical manifestations and potentially devastating complications; from the recognition of the viral etiology of conditions such as HCV-related cryoglobulinemic vasculitis and HBV-associated polyarteritis nodosa to newly named variable vessel vasculitis exemplified by Behcet's disease; from single-organ vasculitis such as central nervous system vasculitis to the more recently emerging features of the IgG4-related, immune-mediated diseases that are showing remarkable clinical heterogeneity. In addition, intriguing pathogenetic hypotheses are being reported for certain chronic, relapsing vasculitides that are improving our understanding of their biology and basic pathophysiology. New avenues are being explored that will hopefully allow a deeper comprehension of the relationships between certain virus-driven vasculitides and lymphoproliferation, and possibly lead to the identification of novel biomarkers that may be used to single out patients at an increased risk of relapse. This explosion of knowledge is obviously resulting in state-of-the-art, personalized treatments of systemic vasculitides. This book is a collection of reviews on the major vasculitides, written by scientists and clinicians with a multi-year experience in this field. We hope it will provide the reader with a stimulating container of new advances in scientific knowledge and more rational therapeutic approaches to this fascinating chapter of pathology.

In 1971, J. Folkman published in the New England Journal of Medicine a hypothesis that tumor growth is angiogenesis-dependent. Folkman introduced the concept that tumors probably secrete diffusible molecules that could stimulate the growth of new blood vessels toward the tumor and that the resulting tumor neovascularization could conceivably be prevented or interrupted by angiogenesis inhibitors. Solid and haematological tumors consist of an avascular and a subsequent vascular phase. Assuming that this depends on the release of angiogenic factors, acquisition of angiogenic capability can be seen as an expression of progression from neoplastic transformation to tumor growth and metastasis.

Beginning in the 1980 s, the biopharmaceutical industry began exploiting the field of antiangiogenesis for creating new therapeutic compounds for modulating new blood vessels in tumor growth. In 2004, Avastin (Bevacizumab), a humanized anti-VEGF monoclonal antibody, was the first angiogenesis inhibitor approved by the Food and Drug Administration for the treatment of colorectal cancer. At present, it has been estimated that over 20,000 cancer patients worldwide have received experimental form of antiangiogenic therapy.

This book offers a historical account of the relevant literature. It also emphasizes the crucial and paradigmatic role of angiogenesis as a biological process and the significance of antiangiogenic approach for the treatment of tumors."

It has been generally accepted that angiogenesis is involved in the pathogenesis of hematological malignancies, like acute and chronic leukemia, lymphoma, myelodysplastic syndromes, myeloproliferative neoplasms and multiple myeloma. The extent of angiogenesis in the bone marrow has been correlated with disease burden, prognosis and treatment outcome. Reciprocal positive and negative interactions between tumor cells and bone marrow stromal cells, namely hematopoietic stem cells, fibroblasts, osteoblasts/osteoclasts, endothelial cells, endothelial progenitor cells, T cells, macrophages and mast cells, mediated by an array of cytokines, receptors and adhesion molecules, modulate the angiogenic response in hematological tumors. More recently, it has been emphasized the pro-angiogenic role of the so called "vascular niche," indicating a site rich in blood vessels where endothelial cells and mural cells such as pericytes and smooth muscle cells create a microenvironment that affects the behavior of several stem and progenitor cells, in hematological malignancies.

* Several in vivo assays are currently used in the study of angiogenesis and antiangiogenesis * The chick embryo chorioallantoic membrane is one of the most common and versatile assay to study angiogenesis and antiangiogenesis in vivo * Angiogenesis plays a critical role in tumor progression and metastasis * Antiangiogenesis is one of most promising approach to the treatment of cancer and metastasis The chick embryo chorioallantoic membrane (CAM) is an extraembryonic membrane which serves as a gas exchange surface and its function is supported by a dense capillary network. Because of its extensive vascularization and easy accessibility, the CAM has been broadly used to study the morpho-functional aspects of the angiogenesis process in vivo and to investigate the efficacy and mechanisms of action of pro-angiogenic and anti-angiogenic natural and synthetic molecules. The CAM is a suitable site for transplanting tissues, which can survive and develop in the CAM by peripheral anastomoses between graft and original CAM vasculature or by new angiogenic vessels grown from the CAM that invade the graft. While the formation of peripheral anastomoses between host and pre-existing donor vessels is the main, and the most common, mechanism involved in the revascularization of embryonic grafts, the growth of CAM-derived vessels into the graft is only stimulated in tumor grafts. The CAM has long been a favored system for the study of tumor angiogenesis and metastasis, because at this stage the chick immunocompetence system is not fully developed and the conditions for rejection have not been established. Tumors remain avascular for 72 h, after which they are penetrated by new blood vessels and begin a phase of rapid growth. Also, delivery of tumor cells onto the CAM allows the fine study of the effects of tumor derived angiogenic growth factors on blood vessel structure and functionality. The CAM may also used to verify the ability to inhibit th

The study of medical history is interesting in itself and may help to modify the view sometimes expressed that medical students and doctors are lacking in culture of any sort. Moreover, some historical perspective is often advantageous when one is considering the multitude of advances that are now taking place in the theory and practice of medicine.

This book, containing a series of collected papers concerning immunology and pathology and vascular biology and angiogenesis, drives us through scientific milestones in the history of medicine in the course of the past two centuries and highlights the contribution of pioneering scientists whose discoveries have paved the way to many researchers working in the fields of cell biology, developmental biology, immunology, pathology, and oncology.

This book will serve as a resource for scientists, historians of medicine and philosophers of science and medicine.

This work describes the importance of tumor microenvironment in favouring tumor progression and angiogenesis. Under physiological conditions, angiogenesis is dependent on the balance of positive and negative angiogenic modulators within the vascular microenvironment and requires the functional activities of a number of molecules, including angiogenic factors, extracellular matrix proteins, adhesion molecules and proteolytic enzymes. In normal tissues, vascular quiescence is maintained by the dominant influence of endogenous angiogenesis inhibitors over angiogenic stimuli. Tumor angiogenesis is linked to a switch in the balance between positive and negative regulators, and mainly depends on the release by inflammatory or neoplastic cells of specific growth factors for endothelial cells, that stimulate the growth of the blood vessels of the host or the down-regulation of natural angiogenesis inhibitors. In particular, the inflammatory infiltrate may contribute to tumor angiogenesis, and there are many reports of associations between tumor inflammatory infiltrate, vascularity and prognosis. New therapeutic approaches have been developed with the aim to control tumor angiogenesis through targeting of different components of tumor microenvironment.

This detailed new edition aims to provide a range of methods and protocols for studying vascular morphogenesis in vivo and in vitro to reflect up-to-date advances in the field. Of note, the book explores endothelial cell signaling is currently understood to promote fundamental cues for cell fate specification, embryo patterning, organ differentiation, and postnatal tissue remodeling. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and timely, Vascular Morphogenesis: Methods and Protocols, Second Edition will serve as an ideal guide to this area of study for a broad audience amongst cell biologists, anatomists, pathologists, and physiologists.

This second edition provides new and updated methods and protocols for studying tumor angiogenesis in vitro and in vivo. Chapters detail morphological aspects of tumor angiogenesis, aortic ring, ex vivo tissue culture model for anti-angiogenic drug testing, transgenic zebrafish, orthotopic models of ovarian cancer, and uncovering metabolic effects of anti-angiogenic therapy in tumors by induced metabolic bioluminescence imaging.Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, Tumor Angiogenesis Assays: Methods and Protocols, Second Edition is a valuable resource for all researchers interested in learning more about this important and developing field.

Blood vessels of tumors display many structural and functional abnormalities. Their unusual leakiness, potential for rapid growth and remodeling, and expression of distinctive surface molecules mediate the dissemination of tumor cells in the bloodstream and maintain the tumor microenvironment. Like normal blood vessels, they consist of endothelial cells, mural cells and their enveloping basement membrane. Common features, irrespective of their origin, size and growth pattern, are absent hierarchy, formation of large-caliber sinusoidal vessels, markedly heterogeneous density, increased permeability, decreased and abnormal pericyte-endothelial cell adhesion, irregular basement membrane structure, and the incorporation of bone-marrow-derived endothelial progenitor cells in the microvasculature. A number of specific tumor endothelial markers have been identified, as well as chromosomal abnormalities. These markers may be used to deliver drugs specifically and selectively to the tumor microvasculature.

Advances in Genetics serial, Volume 110 highlights new advances in the field, with this new volume presenting interesting chapters on a variety of timely topics, all written by an international board of authors.

This volume aims to provide a range of methods and protocols for studying tumor angiogenesis in vitro and in vivo to reflect advances in the field. The chapters in this book cover topics such as: morphological aspects of tumor angiogenesis, aortic ring assay and its use for the study of tumor angiogenesis, ex vivo tissue culture model for anti-angiogenic drug testing, transgenic zebrafish, orthotopic models of ovarian cancer, and uncovering metabolic effects of anti-angiogenic therapy in tumors by induced metabolic bioluminescence imaging. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting edge and thorough, Tumor Angiogenesis Assays: Methods and Protocols is a valuable resource for anyone interested in tumor angiogenesis assay research.

In spite of their relatively low prevalence, systemic vasculitides have been the object of intensive basic and clinical investigations over the last few years. As a consequence, important advancements have been achieved: from updated diagnostic and classification criteria and a more rational nomenclature to the recognition of an expanding spectrum of clinical manifestations and potentially devastating complications; from the recognition of the viral etiology of conditions such as HCV-related cryoglobulinemic vasculitis and HBV-associated polyarteritis nodosa to newly named variable vessel vasculitis exemplified by Behcet's disease; from single-organ vasculitis such as central nervous system vasculitis to the more recently emerging features of the IgG4-related, immune-mediated diseases that are showing remarkable clinical heterogeneity. In addition, intriguing pathogenetic hypotheses are being reported for certain chronic, relapsing vasculitides that are improving our understanding of their biology and basic pathophysiology. New avenues are being explored that will hopefully allow a deeper comprehension of the relationships between certain virus-driven vasculitides and lymphoproliferation, and possibly lead to the identification of novel biomarkers that may be used to single out patients at an increased risk of relapse. This explosion of knowledge is obviously resulting in state-of-the-art, personalized treatments of systemic vasculitides. This book is a collection of reviews on the major vasculitides, written by scientists and clinicians with a multi-year experience in this field. We hope it will provide the reader with a stimulating container of new advances in scientific knowledge and more rational therapeutic approaches to this fascinating chapter of pathology.

This work describes the importance of tumor microenvironment in favouring tumor progression and angiogenesis. Under physiological conditions, angiogenesis is dependent on the balance of positive and negative angiogenic modulators within the vascular microenvironment and requires the functional activities of a number of molecules, including angiogenic factors, extracellular matrix proteins, adhesion molecules and proteolytic enzymes. In normal tissues, vascular quiescence is maintained by the dominant influence of endogenous angiogenesis inhibitors over angiogenic stimuli. Tumor angiogenesis is linked to a switch in the balance between positive and negative regulators, and mainly depends on the release by inflammatory or neoplastic cells of specific growth factors for endothelial cells, that stimulate the growth of the blood vessels of the host or the down-regulation of natural angiogenesis inhibitors. In particular, the inflammatory infiltrate may contribute to tumor angiogenesis, and there are many reports of associations between tumor inflammatory infiltrate, vascularity and prognosis. New therapeutic approaches have been developed with the aim to control tumor angiogenesis through targeting of different components of tumor microenvironment.

It has been generally accepted that angiogenesis is involved in the pathogenesis of hematological malignancies, like acute and chronic leukemia, lymphoma, myelodysplastic syndromes, myeloproliferative neoplasms and multiple myeloma. The extent of angiogenesis in the bone marrow has been correlated with disease burden, prognosis and treatment outcome. Reciprocal positive and negative interactions between tumor cells and bone marrow stromal cells, namely hematopoietic stem cells, fibroblasts, osteoblasts/osteoclasts, endothelial cells, endothelial progenitor cells, T cells, macrophages and mast cells, mediated by an array of cytokines, receptors and adhesion molecules, modulate the angiogenic response in hematological tumors. More recently, it has been emphasized the pro-angiogenic role of the so called “vascular niche”, indicating a site rich in blood vessels where endothelial cells and mural cells such as pericytes and smooth muscle cells create a microenvironment that affects the behavior of several stem and progenitor cells, in hematological malignancies.

Vascular Morphogenesis: Methods and Protocols provides a range of techniques for studying vascular morphogenesis in vivo and in vitro, reflecting advances in the field. Endothelial cell signaling is currently believed to promote fundamental cues for cell fate specification, embryo patterning, organ differentiation and postnatal tissue remodeling. Understanding the concept of vascular bed specificity represents a major challenge for future investigations. Indeed, one of the most interesting theoretical perspectives and practical applications of endothelial cell signaling is the possibility for these cells to maintain their inductive potential during adult life. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, Vascular Morphogenesis: Methods and Protocols will serve as a instructive and useful reference for cell biologists, anatomists, pathologists and physiologists, as well as all those interested in this area of biological study.

The study of medical history is interesting in itself and may help to modify the view sometimes expressed that medical students and doctors are lacking in culture of any sort. Moreover, some historical perspective is often advantageous when one is considering the multitude of advances that are now taking place in the theory and practice of medicine. This book, containing a series of collected papers concerning immunology and pathology and vascular biology and angiogenesis, drives us through scientific milestones in the history of medicine in the course of the past two centuries and highlights the contribution of pioneering scientists whose discoveries have paved the way to many researchers working in the fields of cell biology, developmental biology, immunology, pathology, and oncology. This book will serve as a resource for scientists, historians of medicine and philosophers of science and medicine.

As pointed out by Auerbach in 1991 "Perhaps the most consistent limitation to progress in angiogenesis research has been the availability of simple, reliable, reproducible, quantitative assays of the angiogenesis response." In vitro angiogenesis assays, based on endothelial cell cultures or tissue explant, focus on isolated endothelial cell functions (e.g., endothelial cell proliferation, migration, or invasion) and do not examine the coordination of cell functions required for a successful angiogenic response (Jain et al., 1997; Auerbach et al., 2000). Although in vitro angiogenesis assays have been useful for identi cation of potential molecular targets to block endothelial cell responses and preliminary screening of novel pharmacological agents, they frequently cannot be correlated with in vivo angiogenesis measurements. This is most likely the result of the c- plex and multiple cellular mechanism evoked during new blood vessel formation in vivo. In vitro assays cannot be considered conclusive and the activity of a compound must be con rmed in other assays of increasing complexity, including in vivo assays of angiogenesis, angiogenic-dependent tumor growth, and metastasis.

Blood vessels of tumors display many structural and functional abnormalities. Their unusual leakiness, potential for rapid growth and remodeling, and expression of distinctive surface molecules mediate the dissemination of tumor cells in the bloodstream and maintain the tumor microenvironment. Like normal blood vessels, they consist of endothelial cells, mural cells and their enveloping basement membrane. Common features, irrespective of their origin, size and growth pattern, are absent hierarchy, formation of large-caliber sinusoidal vessels, markedly heterogeneous density, increased permeability, decreased and abnormal pericyte-endothelial cell adhesion, irregular basement membrane structure, and the incorporation of bone-marrow-derived endothelial progenitor cells in the microvasculature. A number of specific tumor endothelial markers have been identified, as well as chromosomal abnormalities. These markers may be used to deliver drugs specifically and selectively to the tumor microvasculature.

Mast cells are versatile, tissue-homing secretory cells, which were first described by Paul Ehrlich in 1878. Mast cells have long been implicated in the pathogenesis of allergic reactions and certain protective responses to parasites. Their functional role, however, has been discovered to be increasingly complex and multifarious. Mast cells have been implicated in various cell-mediated immune reactions, being found in tissues from multiple disease sites, and as a component of the host reaction to bacteria, parasite, and even virus infections. They have also been shown to participate to angiogenic and tissue repair processes after injury. The importance of a possible functional link between chronic inflammation and cancer has long been recognized. As most tumours contain inflammatory cell infiltrates, which often include plentiful mast cells, the question as to the possible contribution of mast cells to tumour development has progressively been emerged. In this book, the general biology of these cells, their development, anatomical distribution and phenotype as well as their secretory products will first be discussed. The biology of tumour cells, their structural and molecular characteristics, the specificity of the tumour microenvironment and the development of a vascular network in the tumour context will be analyzed. The involvement of mast cells in tumour biology and tumour fate will then be considered, with particular emphasis on the capacity of these cells to stimulate tumour growth by promoting angiogenesis and lymphangiogenesis. The last chapter suggest that mast cells may serve as a novel therapeutic target for cancer treatment.

In 1971, J. Folkman published in the New England Journal of Medicine a hypothesis that tumor growth is angiogenesis-dependent. Folkman introduced the concept that tumors probably secrete diffusible molecules that could stimulate the growth of new blood vessels toward the tumor and that the resulting tumor neovascularization could conceivably be prevented or interrupted by angiogenesis inhibitors. Solid and haematological tumors consist of an avascular and a subsequent vascular phase. Assuming that this depends on the release of angiogenic factors, acquisition of angiogenic capability can be seen as an expression of progression from neoplastic transformation to tumor growth and metastasis.

Beginning in the 1980 s, the biopharmaceutical industry began exploiting the field of antiangiogenesis for creating new therapeutic compounds for modulating new blood vessels in tumor growth. In 2004, Avastin (Bevacizumab), a humanized anti-VEGF monoclonal antibody, was the first angiogenesis inhibitor approved by the Food and Drug Administration for the treatment of colorectal cancer. At present, it has been estimated that over 20,000 cancer patients worldwide have received experimental form of antiangiogenic therapy.

This book offers a historical account of the relevant literature. It also emphasizes the crucial and paradigmatic role of angiogenesis as a biological process and the significance of antiangiogenic approach for the treatment of tumors."