Cardiac progenitor cells and the development of the human heart

Rat Cardiac progenitor cells and their application in cell therapy

cells and the subsequent heart failure. When the pharmacological approach no longer complies with the disease evolution, organ transplantation appears to be the only treatment able to rescue the patient life. Cell therapy promises to be clinically efficient and would allow circumventing many limitations of organ transplantation, such as organ low availability, major surgical procedures, high costs and longterm immunosuppression [1]. We designed porous Poly-Lactic Acid (PLLA) and Fibroin scaffolds to deliver CPCs in the heart, we isolated and characterized CPCs for the expression of c-Kit, MDR-1 and Sca-1 by flow cytometry, we tested their degree of differentiation in vitro studying the expr…

HSP90 and eNOS partially co-localize and change cellular localization in relation to different ECM components in 2D and 3D cultures of adult rat cardiomyocytes.

Background information. Cultivation techniques promoting three-dimensional organization of mammalian cells are of increasing interest, since they confer key functionalities of the native ECM (extracellular matrix) with a power for regenerative medicine applications. Since ECM compliance influences a number of cell functions, Matrigel-based gels have become attractive tools, because of the ease with which their mechanical properties can be controlled. In the present study, we took advantage of the chemical and mechanical tunability of commonly used cell culture substrates, and co-cultures to evaluate, on both two- and three-dimensional cultivated adult rat cardiomyocytes, the impact of ECM c…

EFFECTS OF CIGARETTE SMOKE ON MODULATION OF PROTEOLYTIC ACTIVITY IN LUNG FIBROBLAST:IMPLICATIONS FOR LUNG INFLAMMATORY DISEAS

New emerging potentials for human Wharton's jelly mesenchymal stem cells: immunological features and hepatocyte-like differentiative capacity.

In recent years, human mesenchymal stem cells (MSC) have been extensively studied. Their key characteristics of long-term self-renewal and a capacity to differentiate into diverse mature tissues favour their use in regenerative medicine applications. Stem cells can be found in embryonic and extra-embryonic tissues as well as in adult organs. Several reports indicate that cells of Wharton's jelly (WJ), the main component of umbilical cord extracellular matrix, are multipotent stem cells, expressing markers of bone marrow mesenchymal stem cells (BM-MSC), and giving rise to different cellular types of both connective and nervous tissues. Wharton's jelly mesenchymal stem cells (WJ-MSC) express …

Expression of heat shock proteins in bladder cancers

Oxidative stress upregulates eNOS and myeloperoxidase (MPO) immunoexpression in hydrogen peroxide (H2O2) exposed HUVEC cells.

ROLE OF CHAPERONES IN HEALTHY BOWEL AND IBD.

The chaperoning system is the wole complement of chaperones, co-chaperones and chaperone cofactors of the body that preserves cell and tissue homeostasis. Its structural and/or functional defects can cause pathologic conditions, nemed chaperonopathies. Large bowel homeostasis includes a healthy status of the mucosal tissues and the microbiota. An alteration of one of them may determine, in turn, modifications of the other. Molecular chaperones of bacteria and human origin have been implicated in inflammatory bowel disease (IBD). In IBD chaperone levels usually increase and their cellular and subcellular loclization change. This is considered a physiological stress-response of mucosal cells …

IMPLANTATION OF CARDIAC STEM CELL-LOADED POLY-LACTIC ACID AND FIBRINOIN SCAFFOLDS INTO NUDE MICE TO EVALUATE POTENTIAL FOR CARDIAC MUSCLE TISSUE ENGINEERING

CCL5(RANTES) inhibits TGFbeta1 induced collagen production in human bronchial epithelial cells by action of Smad protein

Airway epithelial cells modulate bronchial remodelling in COPD and asthma. TGF-β1 up-regulation was observed in the bronchial epithelial cells of asthmatics and COPD patients. TGF-β is involved in airways remodelling mainly acting via the Smad pathway. Increased CCL5 was also observed in bronchi of patients with severe COPD. We investigated the effects of TGF-β1 on collagen type I, Smad 3-4 and 7 expression and the effects of CCL5 on TGF-β1-induced collagen production in human bronchial epithelial cells (16HBE).Cells were treated with 10ng/ml of TGF-β1, 10ng/ml of CCL5 and 10ng/ml of both TGF-β1 and CCL5 for 0,3 and 24 hours.TGF-b1 increased Smad3,Smad4 and collagen type I(p=0.0472, p=0.043…

Cigarette smoke exposure inhibits extracellular MMP-2 (gelatinase A) activity in human lung fibroblasts

Abstract Background Exposure to cigarette smoke is considered a major risk factor for the development of lung diseases, since its causative role has been assessed in the induction and maintenance of an inflamed state in the airways. Lung fibroblasts can contribute to these processes, due to their ability to produce proinflammatory chemotactic molecules and extracellular matrix remodelling proteinases. Among proteolytic enzymes, gelatinases A and B have been studied for their role in tissue breakdown and mobilisation of matrix-derived signalling molecules. Multiple reports linked gelatinase deregulation and overexpression to the development of inflammatory chronic lung diseases such as COPD.…

THREE-DIMENSIONAL CULTURES AND CO-CULTURES OF ADULT RAT CARDIOMYOCYTES

Evalutaion of HSP60, Procaspase-3 and P53 expression after oxydative stress in NCI-H292 cells.

Hsp60 expression, new locations, functions and perspectives for cancer diagnosis and therapy.

Hsp60 in eukaryotes is considered typically a mitochondrial chaperone (also called Cpn60) but in the last few years it has become clear that it also occurs in the cytosol, the cell surface, the extracellular space, and in the peripheral blood. Studies with prokaryotic models have shown that Hsp60 plays a role in assisting nascent polypeptides to reach a native conformation, and that it interacts with Hsp10 (which also resides in the mitochondria and is also named Cpn10). In addition to its role in polypeptide folding in association with Hsp10, other functions and interacting molecules have been identified for Hsp60 in the last several years. Some of these newly identified functions are asso…

Extended characterization of human umbilical cord matrix mesenchymal stem cells: expression of novel markers, immunoregulatory molecules, and differentiating potency.

ISOLATION AND PHENOTYPICAL CHARACTERIZATION OF MESENCHYMAL STEM CELLS FROM PRE-TERM HUMAN UMBILICAL CORD MATRIX

The many lives of Hsp10: From Early Pregnancy Factor to Potential Antitumoral Agent. New Proteomic Data and a Review of the Literature Focusing on Its Immunologic Properties

Human recombinant vasostatin-1 may interfere with cell-extracellular matrix interaction

Hsp60 and Hsp10 as antitumour molecular agent.

Hsp60 and Hsp10 as antitumour molecular agents

The molecular chaperones Hsp60 and Hsp10 are, according to recent reports, involved in cancer development and progression. We, for instance, have found that their expression varies with distinctive patterns in different malignancies: they are overexpressed in colorectal, exocervical and prostate carcinogenesis, and colorectal cancer progression, but they are downregulated during bronchial carcinogenesis. There is also evidence showing that Hsp60 and Hsp10 can be used as therapeutic agents, for example in rheumatoid arthritis. In view of these findings we want now to call attention to the potential of Hsp60 and Hsp10 in cancer therapy.

Recombinant human vasostatin-1 in adult rat myocardiocytes: interactions with Heat Shock Protein 90, endothelial nitric oxide synthase expression and extracellular matrix components

Chaperonology: A novel research field for experimental medicine in the XXI century.

Hsp60 levels in the skeletal muscle are fibre-type specific and increase after endurance training

Expression of Heat Shock Protein in bladder cancers.

Expression of Heat Shock Protein in bladder cancers.

Role of different endurance training programs on cancer cachexia:pointing particular attention to the gender and age differences Macaluso

Evidence from recent publications indicates that repeated exercise may enhance the quality of life of cancer patients (Maddocks et al., 2012). Regular physical activity may attenuate the adverse effects of cancer therapy, prevent or reverse cachexia and improve survival, although not all the patients are able or willing to undertake programs currently being offered. The aims of this study were to analyze: i) the effects of a progressive endurance exercise (progressive Training, pTR) on survival and cachexia in sedentary (SED) mice inoculated (I) with a fresh fragment of solid C26 tumor [SED-I-pTR; SED-I-SED]; ii) the effect of different protocols of endurance exercise (Trained for 30 min, T…

Cancer as a “Mitochondriopathy”.

Mitochondria are subcellular organelles, whose well-known function is to produce adenosine triphosphate (ATP) through oxidative phosphorylation (OXPHOS). Alterations in respiratory activity and mtDNA appear to be a general feature of malignant cells. The presence of mtDNA mutations has been reported in various cancer cells, and the abundance of mtDNA damage is consistent with the intrinsic susceptibility to constitutive oxidative stress. Research about the functional aspects of mtDNA mutations in cancer development and therapeutic response is likely to be fruitful and to have significant clinical and prognostic impact. Although many studies to date have been focused on the identification an…

3D cultures as an in vitro model to study human vasostatin-1 effects

HSP10,HSP70 AND HSP90 IMMUNOHISTOCHEMICAL LEVELS CHANGE IN ULCERATIVE COLITIS AFTER THERAPY

Ulcerative colitis (UC) is a form of inflammatory bowel disease (IBD) characterized by damage of large bowel mucosa and frequent extra-intestinal autoimmune comorbidities. The role played in IBD pathogenesis by molecular chaperones known to interact with components of the immune system involved in inflammation is unclear. We previously demonstrated that mucosal Hsp60 decreases in UC patients treated with conventional therapies (mesalazine, probiotics), suggesting that this chaperonin could be a reliable biomarker useful for monitoring response to treatment, and that it might play a role in pathogenesis. In the present work we investigated three other heat shock protein/molecular chaperones:…

Extracellular release of HSP60 from tumor cells occurs via various secretory pathways

HSP60 and HSP10 downregulation predicts bronchial epithelial carcingenesis.

Immunohistochemistry of Human Hsp60 in Health and Disease: From Autoimmunity to Cancer

Hsp60 (also called Cpn60) is a chaperonin with essential functions for cell physiology and survival. Additionally, its involvement in the pathogenesis of a variety of diseases (e.g., some autoimmune disorders and cancer) is becoming evident with new research. For example, the distribution and levels of Hsp60 in cells and tissues have been found altered in many pathologic conditions, and the significance of these alterations is being investigated in a number of laboratories. The aim of this ongoing research is to determine the meaning of these Hsp60 alterations with regard to pathogenetic mechanisms, diagnosis, classification of lesions, and assessing prognosis and response to treatment. Hsp…

Skeletal muscle HSP60 expression is fiber-type specific and increases after endurance training

Heat shock protein (Hsp) 60 is primarily localized inside mitochondrial, plays a key role in the translocation of proteins and cytoprotection, and its levels increase in skeletal muscle upon exercise. The purpose of this study was to examine muscle fiber specificity of HSP60 at rest and after an endurance training program of 6 weeks. Forth-eight young (7-weeks old) healthy mice (BALB/c) were subdivided into sedentary and trained groups. Training was performed over a period of 6 weeks on a rota-rod, at a gradually increasing duration and speed. Eight mice of each group were sacrificed after 15, 30 and 45 days. Two days after the last exercise session all mice were sacrificed by cervical disl…

Hsp60 is actively secreted by human tumor cells

Background Hsp60, a Group I mitochondrial chaperonin, is classically considered an intracellular chaperone with residence in the mitochondria; nonetheless, in the last few years it has been found extracellularly as well as in the cell membrane. Important questions remain pertaining to extracellular Hsp60 such as how generalized is its occurrence outside cells, what are its extracellular functions and the translocation mechanisms that transport the chaperone outside of the cell. These questions are particularly relevant for cancer biology since it is believed that extracellular chaperones, like Hsp70, may play an active role in tumor growth and dissemination. Methodology/Principal Findings S…

Modulation of MMP-2 activity in human lung fibroblasts by sigarette smoke: role in the maintenance of the inflamed state in COPD.

Hsp60 and Hsp10 down-regulation predicts bronchial epithelial carcinogenesis in smokers with chronic obstructive pulmonary disease.

BACKGROUND. The relation between smoking, chronic obstructive pulmonary disease (COPD), and lung cancer (LC) is an open field of investigation. A higher frequency of adenocarcinoma has been reported in patients with COPD. Heat shock proteins (Hsps) are implicated in tumoral cell growth and differentiation. The aim of the present study was to investigate the expression of Hsp60 and Hsp10 in bronchial biopsies from smokers with COPD and in 10 lung cancer patients and to evaluate the association between Hsps expression and carcinogenetic steps of LC. METHODS. An immunohistochemical study was performed for Hsp60 and Hsp10 in bronchial biopsies from 35 COPD (postbronchodilator forced expiratory …

Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit

Background/Aim: Epithelial-mesenchymal communication plays a key role in tissue homeostasis and abnormal signaling contributes to chronic airways disease such as COPD. Most in vitro models are limited in complexity and poorly represent this epithelial-mesenchymal trophic unit. We postulated that cellular outgrowth from bronchial tissue would enable development of a mucosal structure that recapitulates better in vivo tissue architecture. Materials and Methods: Bronchial tissue was embedded in Matrigel and outgrowth cultures monitored using time-lapse microscopy, electrical resistance, light and electron microscopy. Cultures were challenged repetitively with cigarette smoke extract (CSE). Res…

Coronary drainage in theOctopus vulgaris systemic heart

The vascular architecture of the coronary system of the systemic heart of the octopus (Octopus vulgaris) has been studied by means of both the corrosion-cast method and scanning electron microscopy. It is shown that the Octopus systemic heart myocardium is supplied through a very rich capillary network, the complexity of which probably reflects the complex organization of the ventricular wall. Drainage occurs by way of a classic venous system that originates from this capillary network. Morphological evidences are reported indicating that the coronary bed communicates directly with the ventricular lumen. In the isolated perfused systemic heart, the flow through this system is related direct…

Evaluation of hsp60, procaspase-3 and p53 expression after oxydative stress in H292 cells

Extended characterization of hypoimmunogenic mesenchymal stem cells isolated from the subendocardial layer of human hearts from chronic heart failure patients

Purpose: Mesenchymal stem cells (MSC) are pluripotent cells which are known to reside in a number of adult organs. In human heart, various populations of stem/progenitor cells have been isolated by different groups. Several efforts still need to be made to better characterize resident or migrating MSC populations in terms of markers expression and immunogenic potential prior to their use for regenerative medicine applications in heart diseases. Methods: MSC were isolated from the sub-endocardial layer of left ventricle sections of hearts explanted from patients affected by chronic heart failure (CHF). The expression of several markers characteristic of the MSC lineage was assessed at both t…

Medium-Term Culture of Primary Oral Squamous Cell Carcinoma in a Three-Dimensional Model: Effects on Cell Survival Following Topical 5-Fluororacile Delivery by Drug-Loaded Matrix Tablets

Since the activity of several conventional anticancer drugs is restricted by resistance mechanisms and dose-limiting side-effects, the design of formulations for local application on malignant lesions seems to be an efficient and promising drug delivery approach. In this study, the effect of locally applied 5-FU on cell death was evaluated both in a SCC4/HEK001 model and in a newly proposed 3D outgrowth model of oral squamous cell carcinoma (OSCC). Initially, the optimal drug dose was established by delivery of solutions containing different amounts of 5-FU. The solution containing 1% (w/v) of 5-FU resulted effective in inducing cell death with complete eradication of cell colonies. Buccal …

Clinical anatomic, immunomorphologic and molecular anatomic data suggest interplay of thyroidal molecules, autoantibodies and Hsp60 in Hashimoto’s disease

Hsp60 is, typically, a mitochondrial protein, but it also occurs in the cytosol, vesicles, and plasma membrane, and in the intercellular space and biological fluids, e.g., blood. Changes in the levels and distribution of Hsp60 are linked to several pathologies, including cancer and chronic inflammatory and autoimmune disorders. What is the histopathological pattern of Hsp60 in the thyroid of Hashimoto’s patients? Are there indications of a pathogenic role of Hsp60 that may make Hashimoto’s thyroiditis a chaperonopathy? Experiments reported here provide information regarding those questions. We found by various immunomorphological techniques increased levels of Hsp60 in the thyroid from HT p…

Isolation and characterization of pluripotent cells from the subendocardial layer of human hearts from chronic heart failure.

Purpose: Several literature reports indicated the existence of pluripotent cells residing into the heart wall, even if in vivo heart regeneration is still a poorly comprised phenomenon. Post-infarct chronic heart failure is a pathology for which heart transplant remains a key option of clinical intervention. The amount of data collected in recent years forces researchers to better characterize the properties of adult stem cells, especially for those isolated from heart. Methods: We developed a new protocol to obtain, from the subendocardial layer of post-infarct hearts explanted from chronic heart failure patients undergoing heart transplantation, populations of mesenchymal stem cells. The …

Cardiac Stem Cell Research: An Elephant in the Room?

Heart disease is the leading cause of death in the industrialized world, and stem cell therapy seems to be a promising treatment for injured cardiac tissue. To reach this goal, the scientific community needs to find a good source of stem cells that can be used to obtain new myocardium in a very period range of time. Since there are many ethical and technical problems with using embryonic stem cells as a source of cells with cardiogenic potential, many laboratories have attempted to isolate potential cardiac stem cells from several tissues. The best candidates seem to be cardiac "progenitor" and/or "stem" cells, which can be isolated from subendocardial biopsies from the same patient or from…

Role of oxidative and nitrosative stress biomarkers in chronic heart failure

In this review, we present recent insights on chronic heart failure (CHF) and the potential role of tumor necrosis factor (TNF)-alpha, interleukins, myeloperoxidase (MPO), and nitrosative stress in the progression of this disease process. Reactive oxygen species (ROS) are produced as a consequence of aerobic metabolism. Under physiologic conditions, their unfavourable effect in causing oxidative damage is counteracted by antioxidants. An imbalance in favour of oxidants leads to oxidative stress, and contributes to myocyte apoptosis, direct negative inotropic effects, and reduced bioavailability of nitric oxide (NO). Together, these effects lead to impaired vasodilatation of the coronary, pu…

Oxidative stress apoptosis in carcinogenesis: molecular players and interactions.

Giovanni Filippo Ingrassia: A five-hundred year-long lesson.

Giovanni Filippo Ingrassia was born five centuries ago in Regalbuto, a small town in the center of Sicily. After his medical course in Padua, under the guidance of Vesalius and Fallopius, he gained international fame as a physician and was recruited as a Professor of human anatomy in Naples and later in Palermo. He is remembered as "the new Galen" or "the Sicilian Hippocrates." He contributed to the knowledge of human anatomy through the description of single bones rather than the whole skeleton. In particular, he was the first to describe the "stapes," the "lesser wings of the sphenoid" and various other structures in the head (probably the pharyngotympanic tube) as well as in the reproduc…

Isolation, characterization, differentiative properties of human mesenchymal stem cells isolated from the sub-endocardial layer of post-infarct chronic heart failure-affected patients

Mesenchymal stem cells (MSC) are pluripotent cells which reside in several adult organs, including heart, even if heart regeneration in vivo is still a poorly comprised phenomenon. Contrasting literature reports suggest that several efforts should be made to better characterize resident or migrating MSC populations (for both markers expression and immunogenic potential) prior to their effective use for regenerative medicine applications in heart diseases. We developed a new protocol to obtain human sub-endocardial MSC (HSE-MSC) from post-infarct hearts explanted from chronic heart failure (CHF) patients undergoing heart transplantation. We characterized HSE-MSC by immunocytochemistry (ICC) …

HSP60 and HASP10 overexpression in metastatic colon cancer

Structural and Mechanical Characteristics of the Heart of the Icefish Chionodraco hamatus (Lönnberg)

The absence of haemoglobin from the blood of icefish, antarctic teleosts of the family Chaenichthydae, has stimulated important physiological mechanisms which serve to compensate for the absence of the respiratory pigment. In view of the central role played by the circulation in meeting the oxygen demands of the tissues, it is not surprising to find that the cardiocirculatory system has been extensively remodelled in icefish. These adaptations include: (1) increases in blood volume, from two to four times larger than that in many teleosts (Hemmingsen and Douglas 1970; Holeton 1970); (2) increases in blood flow associated with lowered oxygen demands (Hemmingsen and Douglas 1970; Holeton 1970…

Serological, immunomorphological and bioinformatics analyses suggest Hsp60 is involved in Hashimoto’s thyroiditis pathogenesis.

EXTRACELLULAR RELEASE OF HSP60 FROM TUMOR CELLS

A multipronged approach to unveil the emerging role of Hsp60 in chronic obstructive pulmonary disease

Inflammation is a major component of chronic obstructive pulmonary disease (COPD) and its cause and mechanisms are still incompletely understood. For example, the role of heat shock proteins (Hsps), many of which are molecular chaperones, has not been explored in detail in COPD, despite the fact that these molecules are known to participate in inflammation in other diseases. It has been shown that extracellular Hsps can signal certain types of T cells, macrophages, dendritic cells, and neutrophils and, thereby, elicit inflammation and immunity. However, these phenomena have not been investigated in COPD despite: a) the increasing awareness of Hsp participation in inflammation and immunity; …

Vasostatin-1 may bind to an extracellular matrix interacting protein

NUTRITION, OXIDATIVE STRESS AND INTESTINAL DYSBIOSIS: INFLUENCE OF DIET ON GUT MICROBIOTA IN INFLAMMATORY BOWEL DISEASES.

Background: Microbiota refers to the population of microorganisms (bacteria, viruses and fungi) that inhabit the entire gastrointestinal tract, more particularly the colon whose role is to maintain the integrity of the intestinal mucosa and control the proliferation of pathogenic bacteria. Alteration in the composition of the gut microbiota is called dysbiosis. Dysbiosis redisposes to inflammatory bowel diseases such as ulcerative colitis, Crohn disease and indeterminate colitis. Methods: The purpose of this literature review is to elucidate the influence of diet on the composition of the gastrointestinal microbiota in the healthy gut and the role of diet in the development of dysbiosis. Co…

Increased nitrotyrosine plasma levels in relation to systemic markers of inflammation and myeloperoxidase in chronic heart failure

The presence of a reciprocal link between inflammation and oxidative/nitrosative stress has been postulated in chronic heart failure (CHF). We aimed to determine signs of nitrosative stress in serum/plasma of CHF patients. ELISA tests were used for quantification of serum/plasma levels of Nitrotyrosine (NT), H(2)O(2), total NO, nitrite (NO(2)(-)), myeloperoxidase (MPO), Tumor Necrosis Factor-alpha (TNFalpha) and pro-Brain Natriuretic Peptide (proBNP) in 66 CHF patients (9 in NYHA I, 34 NYHA II, 23 NYHA III) and in 14 age-matched healthy subjects. NT levels were higher in NYHA III CHF patients compared to NYHA II (p<0.05), NYHA I (p<0.03) and controls (p<0.02), whereas NO(2)(-) and total NO …

HSP60 expression during carcinogenesis: a molecular “Proteus” of carcinogenesis?

Sir, I read with much interest the comprehensive review by Ciocca and Calderwood (2005) in which they analyze the diagnostic, prognostic, predictive, and treatment values of a number of Hsps in oncology. Concerning the implications of Hsps in cancer diagnosis in particular, they report that since “Hsps are overexpressed in a wide range of malignant cells and tissues … Hsp detection is not useful in diagnostic immunopathology.” Nevertheless, “Hsp expression levels can help indicate the presence of abnormal changes during the process of carcinogenesis.” The authors support this remark by reference to several papers by our group (Cappello et al 2002–2003, 2003a, 2003b, 2003c) in which we demon…

Oxidative Stress Upregulates eNOS and Myeloperoxidase (MPO) immunoexpression in hydrogen peroxide (H202) exposed HUVEC cells

Silk fibroin scaffolds enhance cell commitment of adult rat cardiac progenitor cells.

The use of three-dimensional (3D) cultures may induce cardiac progenitor cells to synthesize their own extracellular matrix (ECM) and sarcomeric proteins to initiate cardiac differentiation. 3D cultures grown on synthetic scaffolds may favour the implantation and survival of stem cells for cell therapy when pharmacological therapies are not efficient in curing cardiovascular diseases and when organ transplantation remains the only treatment able to rescue the patient’s life. Silk fibroin-based scaffolds may be used to increase cell affinity to biomaterials and may be chemically modified to improve cell adhesion. In the present study, porous, partially orientated and electrospun nanometric n…

Stem cell populations in the heart and the role of Isl1 positive cells

Cardiac progenitor cells are multipotent stem cells isolated from both embryonic and adult hearts in several species and are able to differentiate at least into smooth muscle cells, endothelial cells and cardiomyocytes. The embryonic origin of these cells has not yet been demonstrated, but it has been suggested that these cells may derive from the first and secondary heart fields and from the neural crest. In the last decade, two diffe-rent populations of cardiac progenitor or stem cells have been identified and isolated, i.e., the Islet1 positive (Isl1+) and c-Kit positive (c-Kit+)/Stem Cell Antigen-1 positive (Sca-1+) cells. Until 2012, these two populations have been considered two separ…

Clinical Anatomy and information technology.

ANALYSIS OF THE EFFECT OF CSE EXPOSURE ON THE EXPRESSION OF MMPS IN PULMONARY EPITHELIAL CELL LINES

PRENATAL DEVELOPMENT OF TEMPORO-MANDIBULAR JOINTS: STATE OF ART

The aim of this work is to analyze the state of the art of temporo-mandibular joint (MJ) to understand the varoius stage of the development of the same during embryogenesis. Various theories have been analyzed, such as the formation of apoptotic or the important role of growth factors, or the Valencia et studies in which are analyzed to numerous articular diseases in various stage of development. By the aforementioned studies show that many factors, of a different nature, are to be involved in the prenatal developlment of this important joint.

Morphological features of the idiopathic dilated cardiomyopathy.

Evaluation od hsp60, pro-caspase-3 and p53 expression after oxidative stress in NCI-H292 cells

Upon oxidative stress, the antiapoptotic Hsp60/procaspase-3 complex persists in mucoepidermoid carcinoma cells.

Hsp60, a mitochondrial chaperonin highly conserved during evolution, has been found elevated in the cytosol of cancer cells, both in vivo and in vitro, but its role in determining apoptosis during oxidative stress (OS) has not yet been fully elucidated. The aim of the present work was to study the effects of OS on Hsp60 levels and its interactions with procaspase- 3 (p-C3) and p53 in tumor cells. NCI-H292 (mucoepidermoid carcinoma) cells were exposed to various concentrations of hydrogen peroxide (H2O2) for 24 hours. Cell viability was determined by Trypan blue and MTT assays. DNA damage was assessed by the Comet assay, and apoptosis was measured by the AnnexinV cytofluorimetric test. Expos…

A mouse model of alcoholic liver disease reveals protection by Lactobacillus fermentum

The knowledge and treatment of alcoholic liver disease is still plagued with gaps mostly due to the inherent limitations of research with patients. We developed an animal model for studying liver histopathology, Hsp-chaperones involvement, and response to treatment. The system was standardized using mice to which ethanol was orally administered alone or in combination with Lactobacillus fermentum for 4, 8 and 12 weeks and applying a battery of techniques (histology, immunohistochemistry, Western blotting, real-time PCR, immunoprecipitation, 3-nitrotyrosine labeling) to assess liver pathology and Hsp60, iNOS gene expression and protein levels, and Hsp60 post-translational modifications. Stea…

Isolation and phenotypical characterization of mesenchymal stem cells from the Wharton's jelly of pre-term human umbilical cord

Poly-lactic acid and fibrinoin scaffolds as three-dimensional device to differentiate cardiac stem cells: in vitro and in vivo studies

Italian Journal of Anatomy and Embryology, Vol 115, No 1/2 (Supplement) 2010

CD1a down-regulation in primary invasive ductal breast carcinoma may predict regional lymph node invasion and patient outcome.

AIMS: CD1a is a molecule belonging to the highly conserved group of CD1 proteins. Its expression in dendritic cells is related to the presentation of tumour-derived glycolipid antigens to T cells and, consequently, the development of a successful antitumour response. The aim was to investigate the presence of CD1a+ cells in both primary tumours and lymph nodes (LN) of a series of 35 invasive ductal carcinomas by both immunohistochemistry and reverse transcription-polymerase chain reaction. METHODS AND RESULTS: CD1a antigen was more expressed in N0 than N1 breast cancer (P < 0.0001) in both primary lesions and LN metastases and correlated positively and significantly with oestrogen (ER) (P =…

The dissociation of the Hsp60/pro-Caspase-3 complex by bis(pyridyl)oxadiazole copper complex (CubipyOXA) leads to cell death in NCI-H292 cancer cells

Abstract Cell survival and proliferation are central to carcinogenesis, involving various mechanisms among which those that impede apoptosis are important. In this, the role of the molecular chaperone Hsp60 is unclear since it has been reported that it can be both, pro- or anti-apoptotic. A solution to this riddle is crucial to the development of anti-cancer therapies targeting Hsp60. We addressed this question using a tumor cell line, NCI-H292, and [Cu(3,5-bis(2′-pyridyl)-1,2,4-oxadiazole) 2 (H 2 O) 2 ](ClO 4 ) 2 , CubipyOXA , a copper-containing compound with cytotoxic properties. We treated cells with various doses of the compound and measured cell viability; apoptosis indicators; and le…

Exosome Involvment in Hsp60 secretion by tumor cells.

Cell- extracellular matrix interaction: a putative non-conventional target of vasostatin-1

A comparative analysis of the products of GROEL-1 gene fromChlamydia trachomatisserovar D and the HSP60 var1 transcript fromHomo sapienssuggests a possible autoimmune response

Summary Chlamydia trachomatis serovar D produces large quantities of HSP60-1 during infections, which accumulate inside the host cell inducing autoimmunity. We compare the aminoacid sequences of the human HSP60 with the bacterial counterpart to better elucidate how CTHSP60 may simulate HSP60 from human origin during infection and may induce an autoimmune response. As a result of the comparison we suggest several possible epitopes of the CTHSP60, which may induce autoimmunity.

Expression of 60-kD Heat Shock Protein Increases during Carcinogenesis in the Uterine Exocervix

&lt;i&gt;Objectives:&lt;/i&gt; The aim of the present study was to determine the presence and expression of the 60-kD heat shock protein (HSP60) in the dysplasia-carcinoma sequence in the uterine exocervix and to evaluate its diagnostic and prognostic significance. &lt;i&gt;Methods and Results:&lt;/i&gt; We performed Western blot and immunohistochemical analyses on biopsies from 40 cases, consisting of 10 normal exocervical biopsies, 10 low-grade squamous intraepithelial lesions (L-SIL), 10 high-grade squamous intraepithelial lesions (H-SIL) and 10 cancerous exocervices (G2 grade). The immunohistochemical results were quantified by computer-assisted image analysis. Western blot analysis sho…

Exosomal Hsp60 in human colon cancer

Expression of CD1a by Barrett’s metaplasia epithelial cells

Human Wharton's jelly-derived mesenchymal stem cells express several immunomodulatory molecules both in their naïve state and hepatocyte-like differentiated progeny: prospects for their use in liver diseases

Wharton’s jelly (WJ), the main constituent of umbilical cord, is a reliable source of mesenchymal stem cells (MSC). WJ-MSC show unique ability in crossing lineage borders. As other extraembryonic mesenchymal populations (placenta and amnionderived cells), WJ-MSC express several immunomodulatory molecules, essential during the initial phases of human development. Indeed, our recent work pointed out the expression of non-classical HLA molecules as HLA-G in such cells, together with a favorable combination of B7 costimulators. Very few data in literature suggest that some of the immune features of the naïve cells are maintained after performing differentiation. The aim of this work was extendi…

RESEARCH OF UNDIFFERENTIATED CELLS IN ADULT RAT MYOCARDIUM

Multi-chaperone – interactors network (mtcin) in mitochondria: role in neoplasm development and methodology of analysis.

ISOLATION AND CHARACTERIZATION OF CD276+/HLA-E+ HUMAN SUB-ENDOCARDIAL MESENCHYMAL STROMAL CELLS FROM CHRONIC HEART FAILURE PATIENTS: ANALYSIS OF DIFFERENTIATION CAPACITY AND IMMUNOMODULATORY MARKERS EXPRESSION.

Ten kilodalton heat shock protein (HSP10) is overexpressed during carcinogenesis of large bowel and uterine exocervix

In the present study, we evaluated the presence and the level of expression of HSP10 in two carcinogenetic models: the 'adenoma-carcinoma sequence' of large bowel and the 'dysplasia-carcinoma sequence' of uterine exocervix. We found HSP10 was overexpressed during the carcinogenesis of both organs. In particular, HSP10 was overexpressed early in large bowel carcinogenesis, while the expression of this protein in exocervical carcinogenesis gradually increased from normal through dysplastic to neoplastic tissues. The quantitative analysis of immunohistochemistry and the Western blotting confirmed these results. Our previous observations showed overexpression of HSP60 in the same carcinogenetic…

Distinctive patterns of Hsp60 levels and localization in human colon mucosa.

Lymphatic vessels of the dura mater: a new discovery?

Aspelund et al. discover the presence of a lymphatic vessel network in the dura mater of the mouse brain and show that these dural lymphatic vessels are important for the clearance of macromolecules from the brain.

Embryonic and foetal Islet-1 positive cells in human hearts are also positive to c-Kit.

During embryogenesis, the mammalian heart develops from a primitive heart tube originating from two bilateral primary heart fields located in the lateral plate mesoderm. Cells belongings to the pre-cardiac mesoderm will differentiate into early cardiac progenitors, which express early transcription factors which are also common to the Isl-1 positive cardiac progenitor cells isolated from the developing pharyngeal mesoderm and the foetal and post-natal mice hearts. A second population of cardiac progenitor cells positive to c-Kit has been abundantly isolated from adult hearts. Until now, these two populations have been considered two different sets of progenitor cells present in the heart in…

Hsp60 and Hsp10 overexpression in metastatic colon cancer.

Chaperonopathies of senescence and the scrambling of interactions between the chaperoning and the immune systems

Aging entails progressive deterioration of molecules and supramolecular structures, including Hsp chaperones and their complexes, paralleled by functional decline. Recent research has changed our views on Hsp chaperones. They work inside and outside cells in many locations, alone or forming teams, interacting with cells, receptors, and molecules that are not chaperones, in roles that are not typically attributed to chaperones, such as protein folding. Hsp chaperones form a physiological system with a variety of functions and interactions with other systems, for example, the immune system. We propose that chaperone malfunctioning due to structural damage or gene dysregulation during aging ha…

Ultrastructure of the conus arteriosus ofScyliorhinus stellaris

The functional morphology of the fourth chamber of the fish heart, the bulbus arteriosus, differs in teleosts and elasmobranchs. In elasmobranchs, this chamber is referred to as the conus arteriosus because its wall is made up of myocardial tissue in addition to elastic tissue. As a result of this feature, the function of the conus is very specialized. We report the basic ultrastructure of the wall of the conus arteriosus in the elasmobranch Scyllium stellare. The wall consists of three layers: the outer epicardial, the middle myocardial, and the inner endocardial layer. The outer layer is composed of the visceral pericardium consisting of cuboidal mesothelial cells, beneath which numerous …

HSP60 expression during carcinogenesis: where is the pilot?

ANP and CD34 expression in human dilated cardiomiopathies: a new insight

Immunopositivity of heat shock protein 60 as a biomarker of bronchial carcinogenesis

Changes in immunohistochemical levels and subcellular localization after therapy and correlation and colocalization with CD68 suggest a pathogenetic role of Hsp60 in ulcerative colitis.

In an earlier work, the role of heat shock protein (Hsp60) in the pathogenesis of ulcerative colitis (UC) was suggested by its significant increase in the pathological mucosa parallel with an increase in inflammatory cells. More data in this direction are reported in this work. We analyzed by immunohistochemistry biopsies of colon tissue from 2 groups of patients with UC and treated with either 5-aminosalicylic acid (5-ASA) alone or in combination with a probiotic. We looked for inflammatory markers and Hsp60. Both the treatments were effective in reducing symptoms but the group treated with both 5-ASA and probiotics showed better clinical results. Amelioration of symptoms was associated wi…

Chlamydia trachomatis Infection and Anti-Hsp60 Immunity: The Two Sides of the Coin

Chlamydia trachomatis (CT) infection is one of the most common causes of reproductive tract diseases and infertility. CT-Hsp60 is synthesized during infection and is released in the bloodstream. As a consequence, immune cells will produce anti-CT-Hsp60 antibodies. Hsp60, a ubiquitous and evolutionarily conserved chaperonin, is normally sequestered inside the cell, particularly into mitochondria. However, upon cell stress, as well as during carcinogenesis, the chaperonin becomes exposed on the cell surface (sf-Hsp60) and/or is secreted from cells into the extracellular space and circulation. Reports in the literature on circulating Hsp and anti-Hsp antibodies are in many cases short on detai…

ADULT STEM CELLS: THE REAL ROOT INTO THE EMBRYO?

During embryonic development, a pool of cells may become a reserve of undifferentiated cells, the embryo-stolen adult stem cells (ESASC). ESASC may be responsible for adult tissue homeostasis, as well as disease development. Transdifferentiation is a sort of reprogramming of ESASC from one germ layer-derived tissue towards another. Transdifferentiation has been described to take place from mesoderm to ectodermal- or endodermal-derived tissues and viceversa but not from ectodermal- to endodermal-derived tissues. We hypothesise that two different populations of ESASC could exist, the first ecto/mesoblast-committed and the second endo/mesoblast-committed. If confirmed, this hypothesis could le…

Analisi quantitativa del tessuto tendineo di ratto adulto allenato

Hsp10 beyond mitochondria: novel locations predict as yet undescribed roles

A novel copper compound, CuNV110, induces apoptosis in tumor cells by dissociation of the Hsp60- pro-caspase 3 complex

The biological activity of CuNV1110, a novel copper chemical compound, has been recently studied on cancer cells and it has been showed that it reduces the cell viability, in a dose and time dependent manner, and induces cell apoptosis. In this study we evaluated the possible mechanisms by which CuNV1110 induces cell apoptosis. In particular we looked at its effects on Hsp60 levels and caspase 3 activation. We used an in vitro model of a pulmonary mucoepidermoid carcinoma (NCI-H292 cells). We found that CuNV1110 reduces the cell viability and induces cell apoptosis in a dose/time dependent manner. Then, we found that Hsp60 levels decrease with the increasing concentrations of CuNV110; by co…

New frontiers in regenerative medicine in cardiology: the potential of Wharton's jelly mesenchymal stem cells.

Cardiomyopathies are still the first cause of death in the world. The identification of resident stem cells, comprising those derived from sub-endocardial stroma, suggests the possible self regeneration of the heart under autocrine/paracrine modulation in the cardiac microenvironment. Nevertheless, because of the limited in vivo regeneration potential of damaged cardiac tissue, the use of drugs and ultimately cardiac transplantation remain the common treatments of heart diseases and defects. The differentiative potential of embryonic and mesenchymal stem cells (MSCs) derived from different tissues (such as bone marrow and adipose tissue) was extensively explored in cell therapy for regenera…

Research of cardiomyocyte precursors in adult rat heart

Recent reports supported the existence of stem cells in adult hearts. However, phenotype and localization of these cells have not been completely described and it is unknown if cardiac regenerative potential differs from one subject to another. The aims of our work were to identify different populations of cardiac stem cells by the analysis of specific markers and to evaluate the expression variability of these markers in 12 adult rat hearts. The expression of CD9, taube nuss and nanog suggests the presence of stem cells from the earliest stages of embryogenesis in adult myocardium. Their different expression could be associated to the degree of stem cell differentiation. CD34 and c-Kit ant…

CD1a expression in primary breast cancer and lymph nodes: correlation with clinicopathological parameters.

The pathogenetic role of the chaperonin Hsp60 in chronic obstructive pulmonary disease: new data and perspectives

Trafficking and secretion patterns of Hsp60 in tumor cells.

EVALUATION OF HSP60, PROCASPASE-3 AND p53 EXPRESSION AFTER OXYDATIVE STRESS IN NCI-H292 CELLS

The Molecular Anatomy of Human Hsp60 and its Similarity with that of Bacterial Orthologs and Acetylcholine Receptor Reveal a Potential Pathogenetic Role of Anti-Chaperonin Immunity in Myasthenia Gravis.

Heat-shock protein 60 (Hsp60) is ubiquitous and highly conserved being present in eukaryotes and prokaryotes, including pathogens. This chaperonin, although typically a mitochondrial protein, can also be found in other intracellular sites, extracellularly, and in circulation. Thus, it can signal the immune system and participate in the development of inflammation and immune reactions. Both phenomena can be elicited by human and foreign Hsp60 (e.g., bacterial GroEL), when released into the blood by infectious agents. Consequently, all these Hsp60 proteins become part of a complex autoimmune response characterized by multiple cross reactions because of their structural similarities. In this s…

Morphological features of the idiopathic dilated cardiomyopathy

Inhibition of Hsp60 expression by doxorubicin and replicative senescence instauration in mucoepidermoid carcinoma cells

CD1a down-regulation in primary invasive ductal breast carcinoma may predict regional lymph node invasion and patient outcome

Aims: CD1a is a molecule belonging to the highlyconserved group of CD1 proteins. Its expression indendritic cells is related to the presentation of tumour-derived glycolipid antigens to T cells and, consequently,the development of a successful antitumour response.The aim was to investigate the presence of CD1a+ cellsin both primary tumours and lymph nodes (LN) ofa series of 35 invasive ductal carcinomas by bothimmunohistochemistry and reverse transcription-poly-merase chain reaction.Methods and results: CD1a antigen was more expressedin N0 than N1 breast cancer (P < 0.0001) in bothprimary lesions and LN metastases and correlatedpositively and significantly with oestrogen (ER) (P =0.0025) and…

Endurance training increases inflammatory cells in small airways of mice

CD1a and antitumour immune response: expression of CD1a by metaplastic epithelial cells

Cigarette smoke causes caspase-independent apoptosis of bronchial epithelial cells from asthmatic donors

BackgroundEpidemiologic studies have demonstrated important links between air pollution and asthma. Amongst these pollutants, environmental cigarette smoke is a risk factor both for asthma pathogenesis and exacerbation. As the barrier to the inhaled environment, the bronchial epithelium is a key structure that is exposed to cigarette smoke.ObjectivesSince primary bronchial epithelial cells (PBECs) from asthmatic donors are more susceptible to oxidant-induced apoptosis, we hypothesized that they would be susceptible to cigarette smoke-induced cell death.MethodsPBECs from normal and asthmatic donors were exposed to cigarette smoke extract (CSE); cell survival and apoptosis were assessed by fl…

Synthetic scaffolds may be used for the orientation of cardiac stem cells properties, differentiation and extracellular matrix interactions in cardiac tissue engineering: in vitro and in vivo studies.

Chaperonopathies of senescence and the scrambling of interactions between the chaperoning and the immune systems.

Aging entails progressive deterioration of molecules and supramolecular structures, including Hsp chaperones and their complexes, paralleled by functional decline. Recent research has changed our views on Hsp chaperones. They work inside and outside cells in many locations, alone or forming teams, interacting with cells, receptors, and molecules that are not chaperones, in roles that are not typically attributed to chaperones, such as protein folding. Hsp chaperones form a physiological system with a variety of functions and interactions with other systems, for example, the immune system. We propose that chaperone malfunctioning due to structural damage or gene dysregulation during aging ha…

Human Hsp60 with Its Mitochondrial Import Signal Occurs in Solution as Heptamers and Tetradecamers Remarkably Stable over a Wide Range of Concentrations

It has been established that Hsp60 can accumulate in the cytosol in various pathological conditions, including cancer and chronic inflammatory diseases. Part or all of the cytosolic Hsp60 could be naive, namely, bear the mitochondrial import signal (MIS), but neither the structure nor the in solution oligomeric organization of this cytosolic molecule has still been elucidated. Here we present a detailed study of the structure and self-organization of naive cytosolic Hsp60 in solution. Results were obtained by different biophysical methods (light and X ray scattering, single molecule spectroscopy and hydrodynamics) that all together allowed us to assay a wide range of concentrations of Hsp60…

5-Fluororacile-loaded matrix tablets for locoregional delivery: effects on a three-dimensional culture model of primary oral squamous cell carcinoma

MicroRNAs expression in c-Kit+/Sca-1+/MDR-1+ cardiac stem cells in three dimensional cultures

Expression of Heat Shock Proteins Hsp10, Hsp27, Hsp60, Hsp70 and Hsp90 in urothelial carcinoma of urinary bladder

CD1A expression in Barrett’s oesophagus: new role for an old molecule against the metaplastic progression

Oxidative stress induces myeloperoxidase expression in endocardial endothelial cells from patients with chronic heart failure.

Increased oxidative stress has been implicated in the pathogenesis of a number of cardiovascular diseases. Recent findings suggest that myeloperoxidase (MPO) may play a key role in the initiation and maintenance of chronic heart failure (CHF) by contributing to the depletion of the intracellular reservoir of nitric oxide (NO). NO consumption through MPO activity may lead to protein chlorination or nitration, leading to tissue damage. Primary cultures of human endocardial endothelial cells (EEC) obtained at heart transplantation of patients with CHF and human umbilical vein endothelial cells (HUVEC) were subjected to oxidative stress by incubation with hydrogen peroxide at non lethal (60 mic…

Hsp60 response in experimental and human temporal lobe epilepsy due to hyppocampal sclerosis

Hsp60 is widely distributed in the brain, and its alteration has been involved in different neurological disorders. Epilepsy is considered one of the most common neurological disorders and typically involves the hippocampal formation. Compelling evidence describes a role of mitochondria, oxidative stress and both innate and adaptive immunity during epileptogenesis in temporal lobe epilepsy due to hyppocampal sclerosis (TLE-HS). Here, we investigate the Hsp60 involvement in experimental and human epilepsy. Firstly, expression and distribution of Hsp60 in epileptic hippocampi of a rat model of temporal lobe epilepsy (TLE), based on the phenomenon of maximal dentate gyrus activation (MDA), usi…

Wharton's Jelly Mesenchymal Stem Cells and Immune Modulation: Regenerative Medicine Meets Tissue Repair

CD1a immunopositivity could help to address prognosis of intestinal-type Barrett's metaplasia

Recent patents and advances on isolation and cellular therapy applications of mesenchymal stem cells from human umbilical cord Wharton's jelly

In recent years, important advances were made to clarify the biology and potential use of mesenchymal stem cells (MSC) in the therapy of a number of disorders. MSC are present in a number of tissues, ranging from adult bone marrow, to several adult organs, adipose tissue and, in the last years, the fetal-associated (also named as extraembryonic) tissues (e.g. placenta, amniotic membrane, umbilical cord). In particular, research on cells derived from mature umbilical cord, a tissue which is still discarded at birth, showed that mesenchymal stem cells can be successfully isolated from the Wharton’s jelly (WJ), the main constituent of this organ. This review will take in to account the patents…

Hsp60, a Novel Target for Antitumor Therapy: Structure-Function Features and Prospective Drugs Design

Heat shock protein 60 kDa (Hsp60) is a chaperone classically believed to be involved in assisting the correct folding of other mitochondrial proteins. Hsp60 also plays a role in cytoprotection against cell stressors, displaying for example, antiapoptotic potential. Despite the plethora of studies devoted to the mechanism of Hsp60's function, especially in prokaryotes, fundamental issues still remain unexplored, including the definition of its role in cancer. Key questions still unanswered pertain to the differences in structure-function features that might exist between the well-studied prokaryotic GroEL and the largely unexplored eukaryotic Hsp60 proteins. In this article we discuss these …

Extending the usefulness of extraembryonic mesenchymal stem cellsin regenerative medicine: expression of novel markers, immunoregulatory molecules, and novel differentiation capacities.

Mesenchymal stem cells (MSC) are considered promising tools in regenerative medicine applications. Even if they are already applied clinically, their extended characterization is being increasingly viewed as a needed feature, in order to avoid contrasting results when translating “in vitro” experiments to “in vivo” approaches. We recently demonstrated in human MSC isolated from the umbilical cord matrix (HEMSC) the expression of novel markers indicative of their stemness, as well as differentiative and immune properties [1]. HEMSC were cultured and subject to multiple molecular and morphological analyses to determine the expression of markers of interest. Undifferentiated HEMSC expressed im…

Poorly differentiated synovial sarcoma: A case report

Poorly differentiated synovial sarcoma is a rare soft tissue tumor. We studied a case arising in the pleural cavity of a young subject, characterised by the presence of spindle cell, small cell, and large epithelioid cell areas. We performed stains for mucosubstances and analysed the expression of cytokeratins 5/6, 7, 8, 18, 19, CEA, CD34, Ber-Ep4 and calretinin to characterize the phenotype of this neoplasm. We furthermore assessed immunohistochemically the presence of p53, Bcl-2, Bax and caspase 3, four apoptotic markers, to evaluate a relationship between apoptotic activity and the behaviour of this tumor. Our findings showed a strong presence of calretinin, p53 and Bcl-2 in all three ar…

CD1A expression in Barrett's oesophagus: new role for an old molecule against metaplastic progression

Medium-term Culture of Normal Human Oral Mucosa: A Novel Three-dimensional Model to Study the Effectiveness of Drugs Administration

Tissue-engineered oral mucosal equivalents have been developed for in vitro studies for a few years now. However, the usefulness of currently available models is still limited by many factors, mainly the lack of a physiological extracellular matrix (ECM) and the use of cell populations that do not reflect the properly differentiated cytotypes of the mucosa of the oral cavity. For this reason, we have developed a novel three-dimensional culture model reflecting the normal architecture of the human oral mucosa, with the main aim of creating a better in vitro model where to test cellular responses to drugs administration. This novel 3D cell culture model (3D outgrowth) was set up using an arti…

TISSUE ENGINEERING FOR THE DEVELOPMENT OF THREE-DIMENSIONAL INVITRO MODELS OF HUMAN MUCOSAE

Traditional two-dimensional (2D) cell cultures only partially reflect the morpho-molecular pattern of human cells in tissues, and they are also unable to fully mimic the complexity of the in vivo microenvironment. Cell development in in vivo systems differs significantly from classical 2D cell culture models, especially with regard to the morphology, growth kinetics, gene expression and the degree of differentiation. Tissue engineering allows the reproduction of tissues by cell seeding on biocompatible scaffolds, to form a homogenous ...

Nestin as a marker of adult rat cardiac stem cells

Study of axillary lymph node asymmetry in a female population

We analysed a large series of axillary lymph nodes, with and without metastases following radical mastectomy for breast cancer. We found left/right asymmetry in numbers of lymph nodes, and also asymmetry of lymph node dimensions, which could have been the caused by tumoral antigenic stimulation. The distribution of hyperplastic node patterns differed significantly.

Hsp60 and Hsp10 increase in colon mucosa of Crohn's disease and ulcerative colitis.

The purpose of this work was to determine in colon mucosa of Crohn’s disease (CD) and ulcerative colitis (UC) in relapse: a) the levels of the chaperonins Hsp60 and Hsp10; b) the quantity of inflammatory cells; and c) if the levels of chaperonins parallel those of inflammation cells. Twenty cases of CD and UC and twenty normal controls (NC) were studied using immunohistochemistry, Western blotting and immunofluorescence. Immunohistochemically, Hsp60 and Hsp10 were increased in both inflammatory bowel diseases (IBD) compared to NC. These results were confirmed by Western blotting. Hsp60 and Hsp10 occurred in the cytoplasm of epithelial cells in CD and UC but not in NC. Hsp60 and Hsp10 co-loc…

Anatomia dell'uomo

Modulation of MMP-2 activity in human lung fibroblasts by cigarette smoke: role in maintenance of the inflamed state in copd

Expression of CD1a by Barrett’s metaplasia epithelial cells.

Hsp60 from cancer cells can reach near and distant targets: A proposal for a multistage pathway

Cancer cells have means to influence other cells in their vicinity and distant, and in this signal-delivering mechanisms the chaperonin Hsp60 plays a role, which is currently being recognized as potentially crucial for the growth and dissemination of at least certain types of tumors. In order to arrive at its destination, Hsp60, a typical resident of mitochondria in normal and tumor cells, leaves the organelle and reaches the blood. In the latter, Hsp60 can travel and arrive at targets situated far away from its origin. The details of the route followed by Hsp60 and their molecular mechanisms have not yet been fully elucidated. We investigated Hsp60 levels and secretion in normal and tumor …

Two isoelectric variants of HSP10 are down-regulated by cigarette smoke exposure in airway cells: A Proteomic study

Cigarette smoke exposure affects expression and extracellular activity levels of gelatinase A and B in airway epithelial cells

Isolation and characterization of Oct-4+/HLA-G+ mesenchymal stem cells from human umbilical cord matrix: differentiation potential and detection of new markers

The presence of multipotent cells in several adult and embryo-related tissues opened new paths for their use in regenerative medicine. Extraembryonic tissues such as umbilical cord are considered a promising source of stem cells, potentially useful in therapy. The characterization of cells from the umbilical cord matrix (Wharton''s Jelly) and amniotic membrane revealed the presence of a population of mesenchymal-like cells, sharing a set of core-markers expressed by "mesenchymal stem cells". Several reports enlightened the differentiation capabilities of these cells, even if at times the lack of an extensive characterization of surface markers and immune co-stimulators expression revealed h…

Atrial natriuretic peptide and CD34 overexpression in human idiopathic dilated cardiomyopathies.

Idiopathic dilated cardiomyopathy (IDCM) is a primary myocardial disease of unknown cause characterized by ventricular chamber enlargement with impaired contractile function. In familial forms of IDCM, mutations of genes coding for cytoskeletal proteins related to force transmission, such as dystrophin, cardiac actin, desmin, and delta-sarcoglycan, have been identified. Here, we report the data of a retrospective investigation carried out to evaluate the expression of atrial natriuretic peptide (ANP), CD34, troponin T and nestin in the myocardium of patients affected with IDCM. Formalin-fixed and paraffin-embedded consecutive tissue sections from the ventricular wall of 10 human normal hear…

Control of target cell survival in thyroid autoimmunity by T helper cytokines via regulation of apoptotic proteins

After autoimmune inflammation, interactions between CD95 and its ligand (CD95L) mediate thyrocyte destruction in Hashimoto's thyroiditis (HT). Conversely, thyroid autoimmune processes that lead to Graves' disease (GD) result in autoantibody-mediated thyrotropin receptor stimulation without thyrocyte depletion. We found that GD thyrocytes expressed CD95 and CD95L in a similar manner to HT thyrocytes, but did not undergo CD95-induced apoptosis either in vivo or in vitro. This pattern was due to the differential production of TH1 and TH2 cytokines. Interferon gamma promoted caspase up-regulation and CD95-induced apoptosis in HT thyrocytes, whereas interleukin 4 and interleukin 10 protected GD …

60KDa chaperonin (HSP60) is over-expressed during colorectal carcinogenesis

The aim of the present study was to evaluate the expression of the heat shock protein 60 (HSP60), a mitochondrial matrix-associated protein belonging to the chaperonin family, in colorectal adenomas and cancers, comparing them to normal colonic tissues and hyperplastic polyps. We performed both immunohistochemistry and Western blot analysis for HSP60. Immunohistochemistry resulted positive in all tubular adenomas and infiltrating adenocarcinomas. By contrast, normal tissues and hyperplastic polyps were negative. Quantitative analysis showed that tubular adenomas with different levels of dysplasia did not present statistical differences concerning HSP60 positivity. In addition, carcinomas al…

DNA sequence and comparative analysis of homologous HSP60 genes of Chlamydia trachomatis serovar D and Homo Sapiens

Heat shock protein 60 levels in tissue and circulating exosomes in human large bowel cancer before and after ablative surgery

BACKGROUND: Heat shock protein 60 (Hsp60) is a chaperonin involved in tumorigenesis, but its participation in tumor development and progression is not well understood and its value as a tumor biomarker has not been fully elucidated. In the current study, the authors presented evidence supporting the theory that Hsp60 has potential as a biomarker as well as a therapeutic target in patients with large bowel cancer. METHODS: The authors studied a population of 97 subjects, including patients and controls. Immunomorphology, Western blot analysis, and quantitative real-time polymerase chain reaction were performed on tissue specimens. Exosomes were isolated from blood and characterized by electr…

ANP and CD34 Expression in Human Dilated Cardiomiophaties: a new insight.

Role of Adrenomedullin in LSP-mediated lung injury

Distribution of mitochondrial chaperonins in lung cells

Three-Dimensional cultures and co-coltures of adult rat cardiomyocytes

Hsp60 expression in skeletal muscle increase after endurance training

EXPOSURE OF BRONCHIAL EPITHELIAL CELLS TO CIGARETTE SMOKE EXTRATS (CSE) INDUCES CASPASE-3-INDEPENDENT APOPTOSIS

HSP60 and HSP10 overexpression in metastastic colon cancer

Doxorubicin anti-tumor mechanisms include Hsp60 post-translational modifications leading to the Hsp60/p53 complex dissociation and instauration of replicative senescence.

Hsp60 is a pro-carcinogenic chaperonin in certain tumor types by interfering with apoptosis and with tumor cell death. In these tumors, it is not known whether or not doxorubicin anti-tumor effects include a blockage of the pro-carcinogenic action of this protein. We used the human lung mucoepidermoid cell line NCI-H292 and different doses of doxorubicin to measure cell viability, cell cycle progression, cell senescence indicators, Hsp60 levels and its post-translational modifications as well as the release of the chaperonin into the extracellular environment. Cell viability was reduced in relation to doxorubicin dose and this was paralleled by the appearance of cell senescence markers. Con…

A novel isolation technique and extended characte-rization of mesenchymal stem cells from human umbilical cord matrix: disclosing new potentials for regenerative medicine applications.

Oxidative stress induces myeloperoxidase expression by HUVEC and chronic heart failure endocardial endothelial cells. Active contribution of endothelium to an inflammatory heart disease.

Heat-shock protein 60 kDa and atherogenic dyslipidemia in patients with untreated mild periodontitis: a pilot study

Identification of predictors of cardiovascular risk can help in the prevention of pathologic episodes and the management of patients at all stages of illness. Here, we investigated the relationships between serum levels of Hsp60 and dyslipidemia in patients with periodontitis by performing a cross-sectional study of 22 patients with mild periodontitis without any prior treatment for it (i.e., drug naive) and 22 healthy controls, matched for age and body mass index (BMI). All subjects were evaluated for periodontal status, gingival inflammation, and oral hygiene. Levels of circulating Hsp60, C-reactive protein (CRP), and plasma lipids were measured, and small, dense low-density lipoproteins …

Characterization of the immunomodulatory properties of mesenchymal stem cells isolated from Wharton's jelly

In vitro and in vivo detection of end-products of MPO-driven oxidative and nitrosative stress in human endocardial endothelial cells from patients affected by chronic heart failure

Effects of Conjugated Linoleic Acid Associated With Endurance Exercise on Muscle Fibres and Peroxisome Proliferator-Activated Receptor γ Coactivator 1 α Isoforms

Conjugated linoleic acid (CLA) has been reported to improve muscle hypertrophy, steroidogenesis, physical activity, and endurance capacity in mice, although the molecular mechanisms of its actions are not completely understood. The aim of the present study was to identify whether CLA alters the expression of any of the peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) isoforms, and to evaluate the possible existence of fibre-type-specific hypertrophy in the gastrocnemius and plantaris muscles. Mice were randomly assigned to one of four groups: placebo sedentary, CLA sedentary, placebo trained, or CLA trained. The CLA groups were gavaged with 35 μl per day of Tonalin® FFA 8…

Effects of chromogranin a-derived peptides on hsp90 expression in co-cultures of adult rat cardiac myocytes and fibroblasts

Asthmatic Bronchial Epithelium Is More Susceptible to Oxidant-Induced Apoptosis

Abnormal apoptotic mechanisms are associated with disease pathogenesis. Because the asthmatic bronchial epithelium is characteristically damaged with loss of columnar epithelial cells, we postulated that this is due to unscheduled apoptosis. Using an antibody directed toward the caspase cleavage product of poly(ADP-ribose) polymerase, immunohistochemistry applied to endobronchial biopsies showed higher levels of staining in the bronchial epithelium of subjects with asthma as compared with normal control subjects (% epithelial staining [median (range) = 10.5 (1.4-24.5) versus 0.4 (0.0-9.7)]; P < 0.001). Because we were unable to determine whether this difference was due to ongoing inflammati…

Role of immunohistochemical expression of PCNA and p53 in prostate carcinoma

Circulating exosomal Hsp60 as a new marker of colon cancer.

Hsp60 and hsp10 expression in advanced colorectal cancer

Structural analysis of rat patellar tendon in response resistance and endurance training.

Little is known about tendon adaptations induced by mechanical loading. Our goal was to evaluate the effects of two different exercise training protocols on adult rat patellar tendon. Ninety-six male Wistar rats were divided into a sedentary group (control), a resistance-trained group and an endurance-trained group. The examinations were performed after 15, 30 and 45 days of training and after 2 weeks of rest since training was stopped. The content of collagen fibers and the cell nuclei number were quantified on tendon cross sections. In order to assess the training effectiveness, we evaluated the heart/body weight ratio, which was higher in 45 day-trained rats than their controls (P<0.01),…

Human HSP10 variants downregulation after cigarette smoke extract exposure in lung cells

The impact of cigarette-smoke stress (a form of oxidative stress) on human lung fibroblasts and epithelial cells, particularly its effect on Hsp10 expression, has not been characterized despite the fact that a role for mitochondrial chaperonins in the development of lung diseases, ranging from chronic obstructive pulmonary disease to bronchial carcinogenesis, has been suggested (1). We studied the effects of non-lethal doses of cigarette smoke extract (CSE) on the expression of Hsp10 in human lung fibroblasts (HFL-1 line) and epithelial cells (16HBE line). Proteomics was carried out using 2D-IPG, silver stain, western blotting, and mass-spectrometry; mRNA was measured by RT-PCR. Database se…

MEDIUM-TERM CULTURE OF THE NORMAL ORAL MUCOSA: A NOVEL THREE-DIMENSIONAL MODEL TO STUDY THE EFFECTIVENESS OF DRUGS ADMINISTRATION

Isolation and characterization of human extraembryonal mesoderm mesenchymal stem cells from umbilical cord

ALCOHOLIC LIVER DISEASE: A MOUSE MODEL REVEALS PROTECTION BY LACTOBACILLUS FERMENTUM

Objectives Alcoholism is one of the most devastating diseases with high incidence, but knowledge of its pathology and treatment is still plagued with gaps mostly because of the inherent limitations of research with patients. We developed an animal model for studying liver histopathology, Hsp (heat-shock protein)-chaperones involvement, and response to treatment. Methods The system was standardized using mice to which ethanol was orally administered alone or in combination with Lactobacillus fermentum following a precise schedule over time and applying, at predetermined intervals, a battery of techniques (histology, immunohistochemistry, western blotting, real-time PCR, immunoprecipitation, …

SENESCENCE-ASSOCIATED HSP60 EXPRESSION IN NORMAL HUMAN SKIN FIBROBLASTS

HSP90 AND ENOS PARTIALLY CO-LOCALIZE AND CHANGE CELLULAR LOCALIZATION IN RELATION TO DIFFERENT ECM COMPONENTS IN 2D AND 3D CULTURES OF ADULT RAT CARDIOMYOCYTES.

TGFbeta signaling: roles of Smads

Changes in immunohistochemical levels and subcellular localization after therapy and correlation and colocalization with CD68 suggest a pathogenetic role of Hsp60 in ulcerative colitis.

In an earlier work, the role of heat shock protein (Hsp60) in the pathogenesis of ulcerative colitis (UC) was suggested by its significant increase in the pathological mucosa parallel with an increase in inflammatory cells. More data in this direction are reported in this work. We analyzed by immunohistochemistry biopsies of colon tissue from 2 groups of patients with UC and treated with either 5-aminosalicylic acid (5-ASA) alone or in combination with a probiotic. We looked for inflammatory markers and Hsp60. Both the treatments were effective in reducing symptoms but the group treated with both 5-ASA and probiotics showed better clinical results. Amelioration of symptoms was associated wi…

Effect of a pathogenic stressor on human HSP10.

Senescence-associated HSP60 expression in Normal Human skin fibroblasts

Normal mammalian fibroblasts cultured in vitro undergo a limited number of divisions before entering a senescent phase in which they can be maintained for long periods but cannot be induced to divide. Senescent cells become unresponsive to growth-promoting signals and exhibit senescent cell morphology with flattened and enlarged cell shape. Several chaperones have a direct effect on cellular senescence. HSP60 has been largely studied in our laboratories and it has been associated with uncontrolled cell proliferation in tumor cells. Since senescence is firmly regulated during cell cycle progression, we wanted to investigate HSP60 protein level during cellular senescence. Our data show that H…

Cardiac stem cell-loaded poly-lactic acid and fibrinoin scaffolds as devices for cardiac muscle tissue regeneration

The expression of HSP60 and HSP10 in large bowel carcinomas with lymph node metastase

Abstract Background The involvement of Heat Shock Proteins (HSP) in cancer development and progression is a widely debated topic. The objective of the present study was to evaluate the presence and expression of HSP60 and HSP10 in a series of large bowel carcinomas and locoregional lymph nodes with and without metastases. Methods 82 Astler and Coller's stage C2 colorectal cancers, of which 48 well-differentiated and 34 poorly-differentiated, were selected along with 661 lymph nodes, including 372 with metastases and 289 with reactive hyperplasia only, from the same tumours. Primitive tumours and both metastatic and reactive lymph nodes were studied; specifically, three different compartment…

Could tubular interstitium be a source of adult epithelial stem cells?

CD1A AND ANTITUMOUR IMMUNE RESPONSE:EXPRESSION OF CD1A BY METAPLASTIC EPITHELIAL CELLS

HSP60 and HSP10 overespression in metastatic colon cancer

Exposure of bronchial epithelial cells to Cigarette Smoke Extracts (CSE) induces caspase-independent apoptosis.

Oxidative stress-driven expression of myeloperoxidase in endothelial cells results in accumulation of markers of oxidative and nitrosative stress in vitro and in vivo

Endothelial cells (ECs), are able to manage with higher concentrations of ROS as well as reactive nitrogen species, respect to the other cell types. The unbalance between oxidizing species and antioxidant cellular defences has been implicated in the pathogenesis of cardiovascular diseases. While the role of MPO in the oxidative burst is well established, the effects of this enzyme on endothelial biology have only recently come to light. MPO can interact with endothelial –derived NO causing its depletion and favouring its conversion to nitrogen dioxide radical, a compound responsible of protein nitration generating 3-nitrotyrosine. Moreover, MPO is the only enzyme capable to form hypoclorous…

Silibin hepatoprotection occur via NFKB signaling inhibition in murine nonalcholic fatty liver disease

Nestin as a marker of adult rat cardiac cells

Effects of antioxidants on CSE-induced cell death in human asthmatic primary bronchial epithelial cells

The link between cigarette smoke (CS) and lung inflammation is quite strong, however relatively little is still known on the effects of CS on human bronchial epithelial cells survival during asthma. In this study we focused our attention on the apoptotic effects of CS on healthy (HC) and asthmatic (AS) primary bronchial epithelial cells (PBEC) and on the role of antioxidants to protect epithelial cells from CSE-induced apoptosis. Twenty subjects (10 HC and 10 AS) were recruited for this study and PBEC were obtained by bronchoscopy. PBEC were treated with oxidants (H2O), anti-oxidants (GSH and AA) and cigarette smoke extracts (CSE). Early apoptosis (EA) and necrosis were measured by flow cyt…

Gut microbiota imbalance and chaperoning system malfunction are central to ulcerative colitis pathogenesis and can be counteracted with specifically designed probiotics: a working hypothesis

In this work, we propose that for further studies of the physiopathology and treatment for inflammatory bowel diseases, an integral view of the conditions, including the triad of microbiota-heat shock proteins (HSPs)-probiotics, ought to be considered. Microbiota is the complex microbial flora that resides in the gut, affecting not only gut functions but also the health status of the whole body. Alteration in the microbiota's composition has been implicated in a variety of pathological conditions (e.g., ulcerative colitis, UC), involving both gut and extra-intestinal tissues and organs. Some of these pathologies are also associated with an altered expression of HSPs (chaperones) and this is…

ASSOCIATION BETWEEN HELYCOBACTER PYOLRI INFECTION AND PATHOLOGICAL ORAL MANIFESTATIONS

DATA FROM LITERATURE ARE CONTROVERSAL REGARDING THE PRESENCE OF bELICOBACTER PYLORI (HP) IN DENTAL PLAQUE AND ITS ASSOCIATION WITH GASTRIC INFECTION. oNE OF THE POSSIBLE MECHANISMS SUGGETSED FOR RE-INFECTION IS THE RECOLONIZATION WITH HP DENTAL PLAQUE. THE PURPOSE OF THIS REVIEW WAS TO DETERMINE WHETHER DENTAL PLAQUE, POOR ORAL HYGIENE, AND PERIODONTAL DISEASE WERE RISK FACTORS FOR HP INFECTION.

c-Kit positive cells isolated from Adult Rat Myocardium can organze them-selves into a tissue-like cell mass.

Effect of a pathogenetic stressor on human Hsp10.

Tetralogy of Fallot as a Model to Study Cardiac Progenitor Cell Migration and Differentiation During Heart Development

Tetralogy of Fallot (ToF) has long been considered a congenital disorder that occurs due to environmental alterations during gestation. Recently, several mutated genes have been discovered that are thought to be responsible for the malformations observed in ToF. These genetic mutations, which are microdeletions, are sporadic and are frequently also present in trisomy 21 patients. The ToF malformations can be lethal, but for the last 50 years, surgical repairs that place an artificial patch to repair the four features of ToF have improved the survival of patients with ToF. However, 0.5% to 6% of patients who survive after surgical repair of ToF die of sudden cardiac death caused by ventricul…

Mitochondrial chaperones in cancer: From molecular biology to clinical diagnostics

Mitochondria are cell organelles involved in processes of cell life and death, and therefore also in tumoral transformation. Indeed, mitochondria dysfunction is a prominent feature of cancer cells. Mitochondrial proteins and DNA have also been previously studied as markers of tumorigenesis. Heat shock proteins (HSPs) are ubiquitous evolutionary conserved proteins. HSPs enhance their expression in stressed cells and they are involved in gene expression regulation, DNA replication, signal transduction, differentiation, apoptosis, cellular senescence or immortalization. This review reflects recent views on the role of some mitochondrial molecular chaperones as prohibitin, mortalin and HSP60/HS…

Naïve Hsp60, similarly to GroEL, oligomerizes to build heptameric and tetradecameric structures.

CIGARETTE SMOKE EXPOSURE DOWNREGULATES TWO ISOELECTRIC VARIANTS OF HUMAN Hsp10 IN LUNG EPITHELIAL CELLS AND FIBROBLASTS: A PROTEOMIC STUDY.

CD1a: a novel biomarker for Barrett's metaplasia?

educational book. Alexandria: American Society for Clinical Oncology, 2001: 226–44. 49 Hussain M, Smith DC, Al-Sukhun S, et al. Preliminary results of HER-2/neu screening and treatment with trastuzumab (Herceptin) (T), paclitaxel (P), carboplatin (C) and gemcitabine (G) in patients with advanced urothelial cancer. Proc Am Soc Clin Oncol 2002; 21: 800a. 50 Al-Sukhun S, Hussain M. Current understanding of the biology of advanced bladder cancer. Cancer 2003; 97 (8 suppl): 2064–75. 51 Hussain SA, Ganesan R, Hiller L, et al. Pro-apoptotic genes BAX and CD40L are predictors of survival in transitional cell carcinoma of the bladder. Br J Cancer 2003; 88: 586–92. 52 Stein JP, Ginsberg DA, Grossfiel…

Endurance Exercise and Conjugated Linoleic Acid (CLA) Supplementation Up-Regulate CYP17A1 and Stimulate Testosterone Biosynthesis

A new role for fat supplements, in particular conjugated linoleic acid (CLA), has been delineated in steroidogenesis, although the underlying molecular mechanisms have not yet been elucidated. The aims of the present study were to identify the pathway stimulated by CLA supplementation using a cell culture model and to determine whether this same pathway is also stimulated in vivo by CLA supplementation associated with exercise. In vitro, Leydig tumour rat cells (R2C) supplemented with different concentrations of CLA exhibited increasing testosterone biosynthesis accompanied by increasing levels of CYP17A1 mRNA and protein. In vivo, trained mice showed an increase in free plasma testosterone…

Heat shock proteins may have effects on cellular senescence and may lead to neoplastic transformation

Isolation and characterization of pluripotent cells from the subendocardial layer of human hearts from chronic heart failure patients

IMMUNOLOCALIZATION STUDIES OF A NON-CONVENTIONAL RECEPTOR FOR VASOSTATIN-1 ON THREE-DIMENSIONAL PRIMARY CULTURES OF ADULT RAT MYOCARDIOCYTES

Epithelial-Mesenchymal communication following Rhinovirus infection of asthmatic bronchial epithelial cells.

Hydrogen peroxide in endothelium: multifaceted roles in cellular stress and signalling.

MicroRNA and Cardiac Stem Cell Therapy

Cardiac Progenitor Cells (CPCs) are multipotent cells of the myocardium. They are located inside niches of the heart muscle, can be isolated, characterized and used for cardiac regeneration in stem cell therapy. Actually, CPCs may be isolated by tissue digestion with or without cell sorting, but it is difficult to achieve the maximum level of differentiation when these cells are implanted into a damaged myocardium. The knowledge recently acquired on small molecules of non-coding RNAs, microRNA (miRNA), may improve the use of these cells in stem cell therapy. In fact, these small molecules may be attached to devices or adminstered as they are or in combination with nanoparticles in order to …

Cigarette smoke exposure downregulates two isoelectric variants of human HSP10 in airway epithelial cells: A Proteomic study

Non-classical type I HLAs and B7 costimulators revisited: analysis of expression and immunomodulatory role in undifferentiated and differentiated MSC isolated from human umbilical cord Wharton's jelly

Introduction. Wharton’s jelly (WJ), the main constituent of umbilical cord, emerged as a reliable and uncontroversial source of mesenchymal stem cells (MSC). WJ-MSC show unique ability in crossing lineage borders, therefore being capable to trans-ifferentiate towards mature cytotypes derived from the three germ layers. As other fetal-associated cells, WJ-MSC express several immunomodulatory molecules, essential during the initial phases of human development and for the processes linked to the tolerance of the mother to the semiallogeneic embryo. Very few data are present in literature on the maintenance of the immune privilege of the naïve cells after performing differentiation. Our previou…

CD1a expression by Barrett's metaplasia of gastric type may help to predict its evolution towards cancer

As emerging in the recent literature, CD1a has been regarded as a molecule whose expression may reflect tumour evolution. The aim of the present work was to investigate the expression of CD1a in a series of Barrett's metaplasia (BM), gastric type (GTBM), with and without follow-up, in order to analyse whether its expression may help to diagnose this disease and to address the outcome. Indeed, GTBM may be confused sometimes with islets of ectopic gastric mucosa and its evolution towards dysplasia (Dy) or carcinoma (Ca) could not be foreseen. We showed a significant higher expression of CD1a in GTBM than in both Dy and Ca; nevertheless, the number of positive GTBM was significantly lower in t…

Cell-extracellular matrix interaction: a putative non-conventional target of Vasostatin-1. A confocal analysis

CD1a and antitumour immune response

Primary immune response is based on the capacity of local professional antigen-presenting cells (whose prototype is represented by dendritic cells, DCs) to take up and present antigens to selected clones of T cells, but also to non-specific effector cells such as macrophages or natural killer cells. The four CD1 proteins, all of which share a limited homology to class I MHC proteins, are differently expressed in various cell types, of both mesenchymal and, as recently described, epithelial lineage. Regarding the role of CD1 molecules in the anti-tumour response, it has been reported that CD1+ dendritic cells are involved in the first steps of the primary immune response in a number of malig…

Wharton’s jelly mesenchymal stem cells differentiation into hepatocyte-like cells: functional characterization and expression of immunomodulatory molecules

Mesenchymal stem cells derived from Wharton’s jelly (WJ-MSCs) recently emerged as promising tools for cellular therapy due to their ability to differentiate into diverse cell types and their immunomodulatory features. Little is known on the expression of immunomodulatory molecules in mature cells differentiated from WJ-MSCs, therefore we aimed to characterize the extent of maintenance of the naive traits of these cells also in a highly specialized differentiated counterpart. WJ-MSCs were differentiated into hepatocyte-like cells (HLCs) with a four weeks protocol. RT-PCR, flow cytometry, IHC and ICC were performed to assess expression of key markers in both undifferentiated and differentiate…

INDUCIBLE NITRIC OXIDE SINTHASE (iNOS) PROTECTS THE HUMAN BRONCHIAL EPHITELIUM EXPOSED TO OXIDANTS BY APOPTOSIS

Foreign body response to subcutaneously implanted scaffolds for cardiac tissue engineering.

Silk fibroin scaffolds enhance cell commitment of adult rat cardiac progenitor cells

The use of three-dimensional (3D) cultures may induce cardiac progenitor cells to synthesize their own extracellular matrix (ECM) and sarcomeric proteins to initiate cardiac differentiation. 3D cultures grown on synthetic scaffolds may favour the implantation and survival of stem cells for cell therapy when pharmacological therapies are not efficient in curing cardiovascular diseases and when organ transplantation remains the only treatment able to rescue the patient's life. Silk fibroin-based scaffolds may be used to increase cell affinity to biomaterials and may be chemically modified to improve cell adhesion. In the present study, porous, partially orientated and electrospun nanometric n…

Hsp60 response in experimental and human temporal lobe epilepsy

The mitochondrial chaperonin Hsp60 is a ubiquitous molecule with multiple roles, constitutively expressed and inducible by oxidative stress. In the brain, Hsp60 is widely distributed and has been implicated in neurological disorders, including epilepsy. A role for mitochondria and oxidative stress has been proposed in epileptogenesis of temporal lobe epilepsy (TLE). Here, we investigated the involvement of Hsp60 in TLE using animal and human samples. Hsp60 immunoreactivity in the hippocampus, measured by Western blotting and immunohistochemistry, was increased in a rat model of TLE. Hsp60 was also increased in the hippocampal dentate gyrus neurons somata and neuropil and hippocampus proper …

CD1A IMMUNOPOSITIVITY COULD HELP TO DISTINGUISH BARRETT's METAPLASIA FROM HETEROTOPIC GASTRIC MUCOSA

OPLA scaffold, collagen I, and horse serum induce a higher degree of myogenic differentiation of adult rat cardiac stem cells

In the last few years, a major goal of cardiac research has been to drive stem cell differentiation to replace damaged myocardium. Several research groups have attempted to differentiate potential cardiac stem cells (CSCs) using bi- or three-dimensional systems supplemented with growth factors or molecules acting as differentiating substances. We hypothesize that these systems failed to induce a complete differentiation because they lacked an architectural space. In the present study, we isolated a pool of small proliferating and fibroblast-like cells from adult rat myocardium. The phenotype of these cells was assessed and the characterized cells were cultured in a collagen I/OPLA scaffold …

Medium-term culture of embryonic lungs: a novel three-dimensional model

Modulation of MMP-2 activity in human lung fibroblasts by cigarette smoke: role in the manteinance of the inflamed state in COPD

Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer

Large bowel carcinogenesis involves accumulation of genetic alterations leading to transformation of normal mucosa into dysplasia and, lastly, adenocarcinoma. It is pertinent to elucidate the molecular changes occurring in the pre-neoplastic lesions to facilitate early diagnosis and treatment. Heat shock proteins (Hsps), many of which are molecular chaperones, are implicated in carcinogenesis, and their variations with tumor progression encourage their study as biomarkers. There are many reports on Hsps and cancer but none to our knowledge on their systematic quantification in pre-neoplastic lesions of the large bowel. We performed immunohistochemical determinations of Hsp10, Hsp60, Hsp70, …

Heat Shock Protein-60 and Risk for Cardiovascular Disease

Cardiovascular disease (CVD) is a leading cause of morbidity and mortality worldwide. There is growing evidence that molecularchaperones, many of which are heat shock proteins HSPs, are involved in CVD pathogenesis. In this review we focus on HSP60,the human mitochondrial chaperone that also displays extramitochondrial and extracellular functions. HSP60 is typically cytoprotectivebut a number of stress conditions determine its conversion to a potentially toxic molecule for cells and tissues. We present illustrative examplesof specific subtypes of CVD where HSP60 is implicated in the initiation and/or progression of disease. The data not only indicatea pathogenic role for HSP60 but also its …

Stem Cell Populations and Regenerative Potential in Chronic Inflammatory Lung Diseases

Several acute and chronic inflammatory pathologies of the lung are accompanied by structural modifications of airway mucosa that vary depending on the severity, duration and type of the disease. These morphological changes, that determine organ dysfunction, are not always reversible. Indeed, the cycle of injury and repair, influencing airway wall re- generation, may sometimes break off and an exacerbation of the pathology may occur. The mechanisms at the base of airway remodelling during inflammation have been widely studied and numerous evidences indicate that the molecular dialogue among the cells of the mucosa has an essential role in orchestrating cell differentiation and tissue repair.…

Characterization of a novel three-dimensional outgrowth model of human bronchial mucosa

Recently it has been hypothesized that a reactivation of the epithelial-mesenchymal trophic unit (EMTU) of the respiratory mucosa, caused by extensive epithelial damage, may be responsible for the pathogenesis of asthma. However, most research on this subject so far has been limited by the unavailability of in vitro models correctly mimicking the three-dimensional architecture of the properly differentiated human bronchial mucosa. For this reason, we have developed a novel 3D outgrowth model, obtained from bronchial biopsies and composed of both human bronchial epithelial cells (goblet and columnar cells) and fibroblasts with their basement membrane. Here we present the preliminary data obt…

Is chlamydial heat shock protein 60 a risk factor for oncogenesis?

Heat shock protein 60 (HSP60) plays an important role in the protein folding of prokaryotic and eukaryotic cells. Most of the papers published on chlamydial HSP60 concern its role in immune response during infection. In the last decade, exposure to Chlamydia trachomatis has been consistently associated with the development of cervical and ovarian cancer. Moreover, it has been suggested that chlamydial HSP60 may have an anti- apoptotic effect during persistent infection. We hypothesize that the accumulation of exogenous chlamydial HSP60 in the cytoplasm of actively replicating eukaryotic cells may interfere with the regulation of the apoptotic pathway. The concomitant expression of viral onc…

Silybin enhances mitochondrial function and inhibits NFkB activation in murine nonalcoholic fatty liver disease

Italian Journal of Anatomy and Embryology, Vol 115, No 1/2 (Supplement) 2010

Inducible nitric oxide synthase (iNOS) protects the human bronchial epithelium exposed to oxidants by apoptosis

Human Hsp10 and Early Pregnancy Factor (EPF) and their relationship and involvement in cancer and immunity: current knowledge and perspectives.

This article is about Hsp10 and its intracellular and extracellular forms focusing on the relationship of the latter with Early Pregnancy Factor and on their roles in cancer and immunity. Cellular physiology and survival are finely regulated and depend on the correct functioning of the entire set of proteins. Misfolded or unfolded proteins can cause deleterious effects and even cell death. The chaperonins Hsp10 and Hsp60 act together inside the mitochondria to assist protein folding. Recent studies demonstrated that these proteins have other roles inside and outside the cell, either together or independently of each other. For example, Hsp10 was found increased in the cytosol of different t…

Embryonic and foetal Islet-1 positive cells in human hearts are also positive to c-Kit.

During embryogenesis, the mammalian heart develops from a primitive heart tube originating from two bilateral primary heart fields located in the lateral plate mesoderm. Cells belongings to the pre-cardiac mesoderm will differentiate into early cardiac progenitors, which express early transcription factors which are also common to the Isl-1 positive cardiac progenitor cells isolated from the developing pharyngeal mesoderm and the foetal and post-natal mice hearts. A second population of cardiac progenitor cells positive to c-Kit has been abundantly isolated from adult hearts. Until now, these two populations have been considered two different sets of progenitor cells present in the heart in…

Hsp60 and human aging: Les liaisons dangereuses 

Stressors can cause abnormal intracellular accumulation of Hsp60 and its localization in extramitochondrial sites, secretion, and circulation, with immune system activation. Dysfunction of chaperones associated with their quantitative and qualitative decline with aging (chaperonopathies of aging) characterizes senescence and is a potential causal factor in the physiological deterioration that occurs with it. The role of Hsp60 in aging is not easy to elucidate, because aging is accompanied by pathologies (e.g., cardiovascular and neurodegenerative disorders, osteoporosis, diabetes, cancer, etc.) in which Hsp60 has been implicated but, although those disorders are more frequent in the elderly…

Effects of cigarette smoke on modulation of proteolytic activity in lung fibroblasts: implications for lung inflammatory diseases

Effects of Conjugated Linoleic Acid Associated With Endurance Exercise on Muscle Fibres and Peroxisome Proliferator-Activated Receptor γ Coactivator 1 α Isoforms

Conjugated linoleic acid (CLA) has been reported to improve muscle hypertrophy, steroidogenesis, physical activity, and endurance capacity in mice, although the molecular mechanisms of its actions are not completely understood. The aim of the present study was to identify whether CLA alters the expression of any of the peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) isoforms, and to evaluate the possible existence of fibre-type-specific hypertrophy in the gastrocnemius and plantaris muscles. Mice were randomly assigned to one of four groups: placebo sedentary, CLA sedentary, placebo trained, or CLA trained. The CLA groups were gavaged with 35 μl per day of Tonalin® FFA 8…

Heat shock protein 10 and signal transduction: a “capsula eburnea” of carcinogenesis?

To date, little is known either about the physical interactions of heat shock protein 10 (Hsp10) with other proteins within the cell or its involvement in signal transduction pathways. Hsp10 has been considered mainly as a partner of Hsp60 in the Hsp60/10 protein folding machine. Only recently, Hsp10 was reported to interact with proteins involved in deoxyribonucleic acid checkpoint inactivation, termination of M-phase, messenger ribonucleic acid export, import of nuclear proteins, nucleocytoplasmic transport, and pheromone signaling pathways. At the same time, Hsp10 expression can be up-regulated in cancer cells, because it accumulates as the cell transformation progresses. Recent data sug…

IFN-gamma-induced protein 10 is a novel biomarker of rhinovirus-induced asthma exacerbations

BACKGROUND: Rhinovirus-induced acute asthma is the most frequent trigger for asthma exacerbations. OBJECTIVE: We assessed which inflammatory mediators were released from bronchial epithelial cells (BECs) after infection with rhinovirus and then determined whether they were also present in subjects with acute virus-induced asthma, with the aim to identify a biomarker or biomarkers for acute virus-induced asthma. METHODS: BECs were obtained from bronchial brushings of steroid-naive asthmatic subjects and healthy nonatopic control subjects. Cells were infected with rhinovirus 16. Inflammatory mediators were measured by means of flow cytometry with a cytometric bead array. Subjects with acute a…

Fibroin and poly-lactic acid scaffolds may be used in cardiac tissue engineering to drive the differentiation of cardiac progenitor cells: in vitro and in vivo studies.

Skeletal muscle Heat shock protein 60 increases after endurance training and induces peroxisome proliferator-activated receptor gamma coactivator 1 α1 expression

AbstractHeat shock protein 60 (Hsp60) is a chaperone localizing in skeletal muscle mitochondria, whose role is poorly understood. In the present study, the levels of Hsp60 in fibres of the entire posterior group of hindlimb muscles (gastrocnemius, soleus and plantaris) were evaluated in mice after completing a 6-week endurance training program. The correlation between Hsp60 levels and the expression of four isoforms of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC1α) were investigated only in soleus. Short-term overexpression of hsp60, achieved by in vitro plasmid transfection, was then performed to determine whether this chaperone could have a role in the activa…

Role of endothelial cell stress in the pathogenesis of chronic heart failure.

Endothelial cells are key modulators of diverse physiological processes, and their impaired function is a cause of numerous cardiovascular diseases. Under physiologic condition, the reactive oxygen and nitrogen mediators in endothelia lead to the signal propagation of the initial stimulus, by forming molecules with a longer half-life like hydrogen peroxide. Hydrogen peroxide is the focus of growing attention in endothelial biology, and consequently the enzymes involved in its generation and clearance are viewed as novel mediators of great importance. In particular, among peroxidases, myeloperoxidase is recognized as a key enzyme, capable of impairing intracellular NO reservoirs as well as p…

Medium-Term Culture of Primary Oral Squamous Cell Carcinoma in a Three- Dimensional Model: Effects on Cell Survival Following Topical 5-Fluororacile Delivery by Drug-Loaded Matrix Tablets

Since the activity of several conventional anticancer drugs is restricted by resistance mechanisms and dose-limiting side-effects, the design of formulations for local application on malignant lesions seems to be an efficient and promising drug delivery approach. In this study, the effect of locally applied 5-FU on cell death was evaluated both in a SCC4/HEK001 model and in a newly proposed 3D outgrowth model of oral squamous cell carcinoma (OSCC). Initially, the optimal drug dose was established by delivery of solutions containing different amounts of 5-FU. The solution containing 1% (w/v) of 5-FU resulted effective in inducing cell death with complete eradication of cell colonies. Buccal …

Hsp60/procaspase 3 complex formation in human cells: impact on apoptosis and carcinogenesis.

Isolation and Characterization of CD276+/HLA-E+ Human Subendocardial Mesenchymal Stem Cells from Chronic Heart Failure Patients: Analysis of Differentiative Potential and Immunomodulatory Markers Expression

Mesenchymal stem cells (MSCs) are virtually present in all postnatal organs as well as in perinatal tissues. MSCs can be differentiated toward several mature cytotypes and interestingly hold potentially relevant immunomodulatory features. Myocardial infarction results in severe tissue damage, cardiomyocyte loss, and eventually heart failure. Cellular cardiomyoplasty represents a promising approach for myocardial repair. Clinical trials using MSCs are underway for a number of heart diseases, even if their outcomes are hampered by low long-term improvements and the possible presence of complications related to cellular therapy administration. Therefore, elucidating the presence and role of MS…

Data mining-based statistical analysis of biological data uncovers hidden significance: clustering Hashimoto’s thyroiditis patients based on the response of their PBMC with IL-2 and IFN-γ secretion to stimulation with Hsp60

The pathogenesis of Hashimoto's thyroiditis includes autoimmunity involving thyroid antigens, autoantibodies, and possibly cytokines. It is unclear what role plays Hsp60, but our recent data indicate that it may contribute to pathogenesis as an autoantigen. Its role in the induction of cytokine production, pro- or anti-inflammatory, was not elucidated, except that we found that peripheral blood mononucleated cells (PBMC) from patients or from healthy controls did not respond with cytokine production upon stimulation by Hsp60 in vitro with patterns that would differentiate patients from controls with statistical significance. This "negative” outcome appeared when the data were pooled and ana…

Convergent sets of data from in vivo and in vitro methods point to an active role of Hsp60 in chronic obstructive pulmonary disease pathogenesis.

BackgroundIt is increasingly clear that some heat shock proteins (Hsps) play a role in inflammation. Here, we report results showing participation of Hsp60 in the pathogenesis of chronic obstructive pulmonary diseases (COPD), as indicated by data from both in vivo and in vitro analyses.Methods and resultsBronchial biopsies from patients with stable COPD, smoker controls with normal lung function, and non-smoker controls were studied. We quantified by immunohistochemistry levels of Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90, and HSF-1, along with levels of inflammatory markers. Hsp10, Hsp40, and Hsp60 were increased during progression of disease. We found also a positive correlation between th…

Hsp60 molecular anatomy and role in colorectal cancer diagnosis and treatment

Quantitative changes in Hsp60 during the development of some tumors suggest that this chaperonin plays a role in carcinogenesis. A description of the specific role(s) of Hsp60 in tumor-cell growth and proliferation is still incomplete, but it is already evident that monitoring its levels and distribution in tissues and fluids has potential for diagnosis and staging, and for assessing prognosis and response to treatment. Although Hsp60 is considered an intramitochondrial protein, it has been demonstrated in the cytosol, cell membrane, vesicles, cell surface, extracellular space, and blood. The knowledge that Hsp60 occurs at all these locations opens new avenues for basic and applied research…

Exposure of bronchial epithelial cells to Cigarette Smoke Extract (CSE) induces caspase-indipendent apoptosis.

Effects of oxidative stress on primary human endothelial cells. Potential active contribution of endothelium to inflammatory heart diseases

New perspectives on the roles of proteinases and lung structural cells in the pathogenesis of chronic obstructive pulmonary disease.

Establishment of a novel three-dimensional in vitro model to study the alterations of the epithelial-mesenchymal trophic unit (emtu) in asthma

Endurance training increases infiammatori cells in small airways of mice.

Characterization of c-kit positive cardiac stem cells subpopulations obtained from adult rat myocardium

Chaperonology: a novel research field for experimental medicine in XXI century.

Senescence-associated HSP60 expression in normal human skin fibroblasts

Normal mammalian fibroblasts cultured in vitro undergo a limited number of divisions before entering a senescent phase in which they can be maintained for long periods but cannot be induced to divide. Senescent cells become unresponsive to growth-promoting signals and exhibit senescent cell morphology with flattened and enlarged cell shape. Several chaperones have a direct effect on cellular senescence. HSP60 has been largely studied in our laboratories and it has been associated with uncontrolled cell proliferation in tumor cells. Since senescence is firmly regulated during cell cycle progression, we wanted to investigate HSP60 protein level during cellular senescence. Our data show that H…

Involvement of caspase-3 and GD3 ganglioside in ceramide-induced apoptosis in Farber disease.

Farber's disease (FD) is a rare genetic disorder caused by ceramidase deficiency, which results in ceramide accumulation in lung, liver, colon, skeletal muscle, cartilage, and bone. Although this disease has been symptomatically characterized, little is known about its molecular pathogenetic process. Because recent studies reported that ceramide accumulation induces GD3 ganglioside formation and apoptosis, we investigated, in tissue obtained via colonoscopy from seriously involved patients, the possible involvement of ceramide in FD colonocyte destruction. Histochemical and TUNEL analyses of paraffin-embedded sections revealed that 45 ± 4.3% of FD colonocytes showed morphological signs of …

Elevated blood Hsp60, its structural similarities and cross-reactivity with thyroid molecules, and its presence on the plasma membrane of oncocytes point to the chaperonin as an immunopathogenic factor in Hashimoto's thyroiditis.

The role Hsp60 might play in various inflammatory and autoimmune diseases is under investigation, but little information exists pertaining to Hashimoto’s thyroiditis (HT). With the aim to fill this gap, in the present work, we directed our attention to Hsp60 participation in HT pathogenesis. We found Hsp60 levels increased in the blood of HT patients compared to controls. The chaperonin was immunolocalized in thyroid tissue specimens from patients with HT, both in thyrocytes and oncocytes (Hurthle cells) with higher levels compared to controls (goiter). In oncocytes, we found Hsp60 not only in the cytoplasm but also on the plasma membrane, as shown by double immunofluorescence performed on …

Role of CD1A and HSP60 in the antitumoral response of oesophageal cancer

Oesophageal cancer (OC) is one of the most common and severe forms of tumor. A wider knowledge of molecular mechanisms which lead to a normal epithelium becoming a neoplasm may reveal new strategies to improve treatment and outcome of this disease. In this review, we report recent findings concerning molecular events which take place during carcinogenesis of the oesophagus. In particular, we focus on the role of two molecules, CD1a and Hsp60, which are overexpressed in oesophageal and many other types of tumor. Both molecules may present tumor antigens and promote in situ the stimulation of an antitumoral immune activity. We suggest there is a synergistic action between these molecules. Fur…

Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit

Background/Aim: Epithelial-mesenchymal communication plays a key role in tissue homeostasis and abnormal signaling contributes to chronic airways disease such as COPD. Most in vitro models are limited in complexity and poorly represent this epithelial-mesenchymal trophic unit. We postulated that cellular outgrowth from bronchial tissue would enable development of a mucosal structure that recapitulates better in vivo tissue architecture. Materials and Methods: Bronchial tissue was embedded in Matrigel and outgrowth cultures monitored using time-lapse microscopy, electrical resistance, light and electron microscopy. Cultures were challenged repetitively with cigarette smoke extract (CSE). Res…

IDENTIFICATION OF C-KIT/CD105 AND ISL-1 CELLS IN HUMAN FETAL AND INFANT HEARTS

During embryogenesis, the mammalian heart develops from a primitive heart tube, which derives from two bilateral primary heart fields located in the lateral plate mesoderm. Later on in the development process, the atrioventricular (A-V) canal and the sinu-atrial segment, at the venous pole, and the conotruncus, at the arterial pole, are added to the heart tube just prior to tube looping. In 2001 Waldo and colleagues and Kelly and colleagues demonstrated the presence of a secondary or anterior heart field in the ventral pharyngeal mesoderm. This region contains a pool of NKX2.5 and GATA-4 positive precardiac cells which migrate to the arterial pole of the primary heart tube. Isl-1 is a marke…

HSP90 AND ENOS PARTIALLY CO-LOCALIZE AND CHANGE CELLULAR LOCALIZATION IN RELATION TO DIFFERENT ECM COMPONENTS IN 2D AND 3D CULTURES OF ADULT RAT CARDIOMYCYTES

BACKGROUND INFORMATION: Cultivation techniques promoting three-dimensional organization of mammalian cells are of increasing interest, since they confer key functionalities of the native ECM (extracellular matrix) with a power for regenerative medicine applications. Since ECM compliance influences a number of cell functions, Matrigel-based gels have become attractive tools, because of the ease with which their mechanical properties can be controlled. In the present study, we took advantage of the chemical and mechanical tunability of commonly used cell culture substrates, and co-cultures to evaluate, on both two- and three-dimensional cultivated adult rat cardiomyocytes, the impact of ECM c…

The role of biomaterials in the direction of cardiac stem cells properties, differentiation and extracellular matrix interactions in cardiac tissue engineering

Introduction: One of the main problems in the rapid translation of preclinical cell‐based therapy to restore damaged myocardium is to find the best delivery route and the best time of cell injection into the myocardium. Intramyocardial injection of stem cells is by far the mostused delivery technique in preclinical studies. Three‐dimensional scaffolds may be used to deliver a limited number of stem cells in their undifferentiated state, but many biomaterials may cause a foreign body reaction on their own. We have recently demonstrated that c‐Kit positive cardiac progenitor cells are able to organize themselves into a tissue‐like cell mass in collagen I three‐dimensional cultures within 72h …

Endurance-type exercise induces mouse heart angiogenesis: a morphological study.