Antonino di Stefano - Academia.edu (original) (raw)

Papers by Antonino di Stefano

Research paper thumbnail of Novel in vitro and in vivo data on the cellular localization of Hsp10 in smokers affected by COPD and in lung-derived cell lines exposed to cigarette smoke extract as stressor

Italian journal of anatomy and embryology, 2013

Cigarette smoke is a potent stressor for the respiratory system, contributing to pathogenesis, fo... more Cigarette smoke is a potent stressor for the respiratory system, contributing to pathogenesis, for instance in chronic obstructive pulmonary disease (COPD), but its effects on the expression, function, and cellular localization of mitochondrial chaperonins are still largely unknown. We studied in vivo (airways biopsies) the localization of Hsp10 and Hsp60 in patients (smokers and non-smokers) affected by mild-moderate COPD, and characterized the effects of non-lethal doses of cigarette smoke extract (CSE) on the expression of these molecules in two human cell lines: lung fibroblasts (HFL-1) and bronchial epithelial (16HBE). We applied various in vitro methods: immunohistochemistry (IHC), subcellular fractionation analyses (SFA), Western blotting (WB), immunocytochemistry (ICC), and transmission electron microscopy (TEM) immunogold, and used bioinformatics and databases searches to gather structural in silico data for interpreting and complementing the in vitro results. IHC showed th...

Research paper thumbnail of High serum levels of tumour necrosis factor-? and interleukin-8 in severe asthma: markers of systemic inflammation?

Clin Experiment Allergy, 2006

Research paper thumbnail of Bronchial Inflammation in Chronic Obstructive Pulmonary Disease: Relationship With Emphysema and and Small Airways Disease

Chronic obstructive pulmonary disease (COPD) is a chronic-progressive disease with high prevalenc... more Chronic obstructive pulmonary disease (COPD) is a chronic-progressive disease with high prevalence characterized by progressive chronic airflow limitation not fully reversible, its classifications is based on spirometric values and symptoms. Cigarette smoking represents the most important risk factor for the development of COPD. Pathological studies show inflammation in lungs of patients with COPD. The inflammation in the lungs can lead to different pathological alterations and this to a different subtypes of disease: A) Chronic bronchitis, B) Emphysema, C) small airways disease. Current pathogenetic hypothesis for COPD explains chronic airflow limitation with an abnormal inflammatory response to particles and inhaled gases. Radiological tests like High resolution Computerized Tomography can give macroscopic information about lung parenchyma and airways morphology and moreover can estimate the subtype of COPD.

Research paper thumbnail of Increased MCP-1 and MIP-1beta in bronchoalveolar lavage fluid of chronic bronchitics

The European respiratory journal, 1999

CC-chemokines are chemotactic factors expressed in a wide range of cell types and tissues. The ai... more CC-chemokines are chemotactic factors expressed in a wide range of cell types and tissues. The aim of this study was to evaluate the involvement of CC-chemokines in the airways inflammation of patients affected by chronic bronchitis. The study evaluated, with an immunoassay, the concentrations of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1alpha (MIP-1alpha) and macrophage inflammatory protein-1beta (MIP-1beta), in the bronchoalveolar lavage fluid (BALF) of 12 smokers affected by chronic bronchitis and 14 smoking, 15 nonsmoking and six exsmoking healthy subjects. MCP-1 was significantly increased in patients with chronic bronchitis ((mean+/-SD) 10.75+/-4.04 pg x mL(-1)) and in the smoker control group (12.39+/-5.87 pg x mL(-1)) compared with healthy exsmokers: (7.12+/-1.60 pg x mL(-1), p=0.035 and p=0.045, respectively) and nonsmokers (6.41+/-3.87 pg x mL(-1), p=0.003 and p=0.006, respectively). MIP-1alpha concentrations were undetectable. A significant ...

Research paper thumbnail of Prognostic value of angiopoietin-2 in patients with chronic heart failure

International Journal of Cardiology, 2016

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

International Journal of Cardiology, Jul 10, 2009

The presence of a reciprocal link between inflammation and oxidative/nitrosative stress has been ... more 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.

Research paper thumbnail of Recent Patents on Oxidative Stress-Related Biomarkers in Chronic Heart Failure: The Central Role of Endothelium and Myeloperoxidase

Recent Patents on Biomarkers, 2013

ABSTRACT Endothelial cells are key modulators of multiple physiological processes, and their impa... more ABSTRACT Endothelial cells are key modulators of multiple physiological processes, and their impairment may result in the generation of endothelial dysfunction and ultimately cardiovascular diseases. Under physiologic conditions, reactive oxygen and nitrogen mediators of endothelial cells act to propagate signals driven by different stimuli, by forming molecules with a longer half-life like hydrogen peroxide. Reactive oxygen species (ROS) are constantly produced as a consequence of aerobic metabolism. Under physiologic conditions, their tendency to cause oxidative damage is counterbalanced by the action of antioxidants or oxidant-scavenging enzymes. An imbalance in favour of oxidants leads to oxidative stress, which can result in cardiomyocyte apoptosis and reduced bioavailability of vascular nitric oxide (NO). Together, these processes may lead to altered vasodilatation of the coronary, pulmonary and peripheral vascular beds. Myeloperoxidase is a key enzyme, capable of impairing intracellular NO reservoirs as well as producing oxidized amino acids such as 3-chlorotyrosine or 3-nitrotyrosine. Recent literature data have shown that apart neutrophils, this enzyme may be expressed by other cell types, and remarkably by endothelial cells both in vitro and in vivo, as a consequence of the exposure to oxidative stress. In this review we analyze recent literature and patents related to the detection, quantification and role of myeloperoxidase as a biomarker in patients affected by chronic heart failure or other cardiovascular diseases. A particular focus is given to analytical approaches aimed to detect early signals of variations in ROS or related enzymes or their by-products, via serological or other low-invasivity assays. The developments in this field may constitute a key improvement of the ability to manage and treat patients suffering from CHF as well as allowing to elucidate the molecular mechanisms behind the clinical phenomena.

Research paper thumbnail of Downregulation of IL-27 bronchial epithelial expression by heat-shock protein-60

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

Histochemistry Cell Biol, 2009

The presence of multipotent cells in several adult and embryo-related tissues opened new paths fo... more 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 hidden pitfalls when in vivo transplantation was performed. The present work describes a novel isolation protocol for obtaining mesenchymal stem cells from the umbilical cord matrix. These cells are clonogenic, retain long telomeres, can undergo several population doublings in vitro, and can be differentiated in mature mesenchymal tissues as bone and adipose. We describe for the first time that these cells, besides expressing all of the core-markers for mesenchymal stem cells, feature also the expression, at both protein and mRNA level, of tolerogenic molecules and markers of all the three main lineages, potentially important for both their differentiative potential as well as immunological features.

Research paper thumbnail of Expression of vascular remodeling markers in relation to bradykinin receptors in asthma and COPD

European Respiratory Journal, Sep 1, 2013

Number: 1552 Publication Number: 4863 Abstract Group: 5.1. Airway Pharmacology and Treatment

Research paper thumbnail of Phospho-p38 MAPK expression in COPD bronchi and in oxidative and inflammatory challenged bronchial epithelium

Italian Journal of Anatomy and Embryology, 2014

The role of MAPK kinases in inducing the inflammatory response in the airways of chronic obstruct... more The role of MAPK kinases in inducing the inflammatory response in the airways of chronic obstructive pulmonary diseases (COPD) patients is incompletely studied. Objectives: To investigate the expression of activated MAPK kinases in bronchial biopsies of COPD patients and the MAPK kinase bronchial epithelial response to oxidative and inflammatory stimuli related to COPD. Expression of phospho(p)-p38, p-JNK1 and p-ERK1/2 was measured in the bronchial mucosa using immunohistochemistry in patients with mild/moderate (n=17), severe/very severe (n=16) stable COPD, control smokers (n=16), control non smokers (n=9) and in a group with mild asthma (n=9). 16HBE cells, challenged with oxidative and inflammatory stimuli, were also studied for IL-8 and MAPK kinases mRNA production. P-p38 was the most expressed MAPK kinase in the bronchial mucosa of all subjects. No significant differences were observed for immune-expression of p-p38, p-JNK and p-ERK1/2 between COPD and control subjects. 16HBE cells treated with H 2 O 2 , cytomix (TNFα+IL-1β+IFNγ) and Lipopolysaccharide (LPS) up-regulated IL-8 mRNA production at 1h or 2h after treatments. P38α mRNA was significantly increased after H 2 O 2 and LPS. JNK1 and ERK1 mRNA were not significantly increased after H 2 O 2 , cytomix or LPS treatments. Blocking p38α activity IL-8 mRNA production was not changed at 1h, 2h and 4h after H 2 O 2 or LPS challenge. P-p38 immune-positivity is prevalent in the bronchial mucosa of COPD and asthmatic patients and p38 mRNA is increased after bronchial epithelial challenges suggesting a relevant role for this MAPK kinase in the induction of bronchial inflammation in COPD and asthma.

Research paper thumbnail of Oxidative stress induces myeloperoxidase expression in endocardial endothelial cells from patients with chronic heart failure

Basic Research in Cardiology, 2009

j Abstract Increased oxidative stress has been implicated in the pathogenesis of a number of card... more j Abstract 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 lM) dose for different exposure times (3 and 6 h). Treated and control cells were tested by immunohistochemistry and RT-PCR for MPO and 3-chlorotyrosine expression. Both endothelial cell types expressed myeloperoxidase following oxidative stress, with higher levels in EEC. Moreover, 3-chlorotyrosine accumulation in treated cells alone indicated the presence of MPO-derived hypochlorous acid. Immunohistochemistry on sections from post-infarcted heart confirmed in vivo the endothelial positivity to MPO, 3-chlorotyrosine and, to a minor extent, nitrotyrosine. Immunohistochemical observations were confirmed by detection of MPO mRNA in both stimulated EEC and HUVEC cells. This study demonstrates for the first time that EEC can express MPO after oxidative stress, both in vitro and in vivo, followed by accumulation of 3-chlorotyrosine, an end product of oxidative stress. Deregulation of endothelial functions may contribute to the development of a number of cardiovascular diseases, including CHF. The results also highlight the notion that endothelium is not only a target but also a key player in oxidative-driven cardiovascular stress. j

Research paper thumbnail of HSP60 activity on 16HBE cells after oxidative and pro-inflammatory stimuli

European Respiratory Journal, Sep 1, 2014

Research paper thumbnail of Immunoregulation and FoxP3 expression in the bronchial mucosa of stable COPD patients

European Respiratory Journal, Sep 1, 2012

Number: 1905 Publication Number: P4589 Abstract Group: 3.3. Mechanisms of Lung Injury and Repair

Research paper thumbnail of Hsp60 and Hspl0 as antitumor molecular agents

Cancer Biology Therapy, May 1, 2007

The molecular chaperones Hsp60 and Hsp10 are, according to recent reports, involved in cancer dev... more 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.

Research paper thumbnail of Bradykinin b2 receptor expression in the bronchial mucosa of allergic asthmatics: the role of NF-kB

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, Jan 20, 2015

Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-... more Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-kB) is potentially involved in BK B2 receptor (B2R) regulation. In this observational cross-sectional study B2R and NF-kB expression was evaluated in bronchial biopsies from mild asthmatics (after diluent/allergen challenge) and healthy controls examining the role of NF-kB in B2R expression in primary human fibroblasts from normal and asthmatic subjects (HNBFb and HABFb). B2R and NF-kB (total and nuclear) expression was analyzed by immunohistochemistry in biopsies from 10 mild intermittent asthmatics (48 hours after diluent/allergen challenge) and 10 controls undergoing bronchoscopy. B2R co-localization in 5B5(+) and αSMA(+) mesenchymal cells was studied by immunofluorescence/confocal microscopy, and B2R expression in HABFb/HNBFb incubated with interleukin (IL)-4/IL-13 with/without BK, and after NF-kB inhibitor, by Western blotting. Bronchial mucosa B2R and nuclear NF-kB expression was h...

Research paper thumbnail of Chaperone patterns in vernal keratoconjunctivitis are distinctive of cell and Hsp type, and are modified by inflammatory stimuli

Allergy, Jan 27, 2015

Vernal keratoconjunctivitis (VKC) is a severe ocular allergy with pathogenic mechanism poorly und... more Vernal keratoconjunctivitis (VKC) is a severe ocular allergy with pathogenic mechanism poorly understood and no efficacious treatment. The aims of the study were to determine quantities and distribution of Hsp-chaperones in the conjunctiva of VKC patients and assess their levels in conjunctival epithelial and fibroblast cultures exposed to inflammatory stimuli. Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90, Hsp105, and Hsp110 were determined in conjunctiva biopsies from 9 patients and 9 healthy age-matched normal subjects, using immunomorphology and qPCR. Conjunctival epithelial cells and fibroblasts were cultured and stimulated with IL-1β, histamine, IL-4, TNF-α, or UV-B irradiation, and changes in Hsp levels were determined by Western blotting. Hsp27, Hsp40, Hsp70, and Hsp90 levels increased in the patients' conjunctiva, whereas Hsp10, Hsp60, Hsp100, and Hsp105 did not. Double immunofluorescence demonstrated co-localization of Hsp27, Hsp40, Hsp70, and Hsp90 with CD68 and tryptase. ...

Research paper thumbnail of Hsp27 and Hsp70 in chronic obstructive pulmonary disease: certainties vs doubts

Cell stress & chaperones, 2015

Research paper thumbnail of Role of interleukin-8 in the pathogenesis and treatment of COPD

Research paper thumbnail of Phospho-p38 MAPK Expression in COPD Patients and Asthmatics and in Challenged Bronchial Epithelium

Respiration, 2015

patients with mild/moderate (n = 17), severe/very severe (n = 16) stable COPD, control smokers (n... more patients with mild/moderate (n = 17), severe/very severe (n = 16) stable COPD, control smokers (n = 16), control nonsmokers (n = 9), in mild asthma (n = 9) and in peripheral airways from COPD patients (n = 15) and control smokers (n = 15). Interleukin (IL)-8 and MAPK mRNA was measured in stimulated 16HBE cells. Results: No significant differences in p-p38 MAPK, p-JNK or p-ERK1/2 expression were seen in bronchial biopsies and peripheral airways between COPD and control subjects. Asthmatics showed increased submucosal p-p38 MAPK expression compared to COPD patients (p < 0.003) and control non-smokers (p < 0.05). Hydrogen peroxide (H 2 O 2 ), cytomix (tumour necrosis factor-α + IL-1β + interferon-γ) and lipopolysaccharide (LPS) upregulated IL-8 mRNA at 1 or 2 h. p38 MAPKα mRNA was significantly increased after H 2 O 2 and LPS treatment. JNK1 and ERK1 mRNA were unchanged after H 2 O 2 , cytomix or LPS treatments. Conclusion: p-p38 MAPK expression is similar in stable COPD and control subjects but increased in the bronchi of mild

Research paper thumbnail of Novel in vitro and in vivo data on the cellular localization of Hsp10 in smokers affected by COPD and in lung-derived cell lines exposed to cigarette smoke extract as stressor

Italian journal of anatomy and embryology, 2013

Cigarette smoke is a potent stressor for the respiratory system, contributing to pathogenesis, fo... more Cigarette smoke is a potent stressor for the respiratory system, contributing to pathogenesis, for instance in chronic obstructive pulmonary disease (COPD), but its effects on the expression, function, and cellular localization of mitochondrial chaperonins are still largely unknown. We studied in vivo (airways biopsies) the localization of Hsp10 and Hsp60 in patients (smokers and non-smokers) affected by mild-moderate COPD, and characterized the effects of non-lethal doses of cigarette smoke extract (CSE) on the expression of these molecules in two human cell lines: lung fibroblasts (HFL-1) and bronchial epithelial (16HBE). We applied various in vitro methods: immunohistochemistry (IHC), subcellular fractionation analyses (SFA), Western blotting (WB), immunocytochemistry (ICC), and transmission electron microscopy (TEM) immunogold, and used bioinformatics and databases searches to gather structural in silico data for interpreting and complementing the in vitro results. IHC showed th...

Research paper thumbnail of High serum levels of tumour necrosis factor-? and interleukin-8 in severe asthma: markers of systemic inflammation?

Clin Experiment Allergy, 2006

Research paper thumbnail of Bronchial Inflammation in Chronic Obstructive Pulmonary Disease: Relationship With Emphysema and and Small Airways Disease

Chronic obstructive pulmonary disease (COPD) is a chronic-progressive disease with high prevalenc... more Chronic obstructive pulmonary disease (COPD) is a chronic-progressive disease with high prevalence characterized by progressive chronic airflow limitation not fully reversible, its classifications is based on spirometric values and symptoms. Cigarette smoking represents the most important risk factor for the development of COPD. Pathological studies show inflammation in lungs of patients with COPD. The inflammation in the lungs can lead to different pathological alterations and this to a different subtypes of disease: A) Chronic bronchitis, B) Emphysema, C) small airways disease. Current pathogenetic hypothesis for COPD explains chronic airflow limitation with an abnormal inflammatory response to particles and inhaled gases. Radiological tests like High resolution Computerized Tomography can give macroscopic information about lung parenchyma and airways morphology and moreover can estimate the subtype of COPD.

Research paper thumbnail of Increased MCP-1 and MIP-1beta in bronchoalveolar lavage fluid of chronic bronchitics

The European respiratory journal, 1999

CC-chemokines are chemotactic factors expressed in a wide range of cell types and tissues. The ai... more CC-chemokines are chemotactic factors expressed in a wide range of cell types and tissues. The aim of this study was to evaluate the involvement of CC-chemokines in the airways inflammation of patients affected by chronic bronchitis. The study evaluated, with an immunoassay, the concentrations of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1alpha (MIP-1alpha) and macrophage inflammatory protein-1beta (MIP-1beta), in the bronchoalveolar lavage fluid (BALF) of 12 smokers affected by chronic bronchitis and 14 smoking, 15 nonsmoking and six exsmoking healthy subjects. MCP-1 was significantly increased in patients with chronic bronchitis ((mean+/-SD) 10.75+/-4.04 pg x mL(-1)) and in the smoker control group (12.39+/-5.87 pg x mL(-1)) compared with healthy exsmokers: (7.12+/-1.60 pg x mL(-1), p=0.035 and p=0.045, respectively) and nonsmokers (6.41+/-3.87 pg x mL(-1), p=0.003 and p=0.006, respectively). MIP-1alpha concentrations were undetectable. A significant ...

Research paper thumbnail of Prognostic value of angiopoietin-2 in patients with chronic heart failure

International Journal of Cardiology, 2016

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

International Journal of Cardiology, Jul 10, 2009

The presence of a reciprocal link between inflammation and oxidative/nitrosative stress has been ... more 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.

Research paper thumbnail of Recent Patents on Oxidative Stress-Related Biomarkers in Chronic Heart Failure: The Central Role of Endothelium and Myeloperoxidase

Recent Patents on Biomarkers, 2013

ABSTRACT Endothelial cells are key modulators of multiple physiological processes, and their impa... more ABSTRACT Endothelial cells are key modulators of multiple physiological processes, and their impairment may result in the generation of endothelial dysfunction and ultimately cardiovascular diseases. Under physiologic conditions, reactive oxygen and nitrogen mediators of endothelial cells act to propagate signals driven by different stimuli, by forming molecules with a longer half-life like hydrogen peroxide. Reactive oxygen species (ROS) are constantly produced as a consequence of aerobic metabolism. Under physiologic conditions, their tendency to cause oxidative damage is counterbalanced by the action of antioxidants or oxidant-scavenging enzymes. An imbalance in favour of oxidants leads to oxidative stress, which can result in cardiomyocyte apoptosis and reduced bioavailability of vascular nitric oxide (NO). Together, these processes may lead to altered vasodilatation of the coronary, pulmonary and peripheral vascular beds. Myeloperoxidase is a key enzyme, capable of impairing intracellular NO reservoirs as well as producing oxidized amino acids such as 3-chlorotyrosine or 3-nitrotyrosine. Recent literature data have shown that apart neutrophils, this enzyme may be expressed by other cell types, and remarkably by endothelial cells both in vitro and in vivo, as a consequence of the exposure to oxidative stress. In this review we analyze recent literature and patents related to the detection, quantification and role of myeloperoxidase as a biomarker in patients affected by chronic heart failure or other cardiovascular diseases. A particular focus is given to analytical approaches aimed to detect early signals of variations in ROS or related enzymes or their by-products, via serological or other low-invasivity assays. The developments in this field may constitute a key improvement of the ability to manage and treat patients suffering from CHF as well as allowing to elucidate the molecular mechanisms behind the clinical phenomena.

Research paper thumbnail of Downregulation of IL-27 bronchial epithelial expression by heat-shock protein-60

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

Histochemistry Cell Biol, 2009

The presence of multipotent cells in several adult and embryo-related tissues opened new paths fo... more 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&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s Jelly) and amniotic membrane revealed the presence of a population of mesenchymal-like cells, sharing a set of core-markers expressed by &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;mesenchymal stem cells&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;. 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 hidden pitfalls when in vivo transplantation was performed. The present work describes a novel isolation protocol for obtaining mesenchymal stem cells from the umbilical cord matrix. These cells are clonogenic, retain long telomeres, can undergo several population doublings in vitro, and can be differentiated in mature mesenchymal tissues as bone and adipose. We describe for the first time that these cells, besides expressing all of the core-markers for mesenchymal stem cells, feature also the expression, at both protein and mRNA level, of tolerogenic molecules and markers of all the three main lineages, potentially important for both their differentiative potential as well as immunological features.

Research paper thumbnail of Expression of vascular remodeling markers in relation to bradykinin receptors in asthma and COPD

European Respiratory Journal, Sep 1, 2013

Number: 1552 Publication Number: 4863 Abstract Group: 5.1. Airway Pharmacology and Treatment

Research paper thumbnail of Phospho-p38 MAPK expression in COPD bronchi and in oxidative and inflammatory challenged bronchial epithelium

Italian Journal of Anatomy and Embryology, 2014

The role of MAPK kinases in inducing the inflammatory response in the airways of chronic obstruct... more The role of MAPK kinases in inducing the inflammatory response in the airways of chronic obstructive pulmonary diseases (COPD) patients is incompletely studied. Objectives: To investigate the expression of activated MAPK kinases in bronchial biopsies of COPD patients and the MAPK kinase bronchial epithelial response to oxidative and inflammatory stimuli related to COPD. Expression of phospho(p)-p38, p-JNK1 and p-ERK1/2 was measured in the bronchial mucosa using immunohistochemistry in patients with mild/moderate (n=17), severe/very severe (n=16) stable COPD, control smokers (n=16), control non smokers (n=9) and in a group with mild asthma (n=9). 16HBE cells, challenged with oxidative and inflammatory stimuli, were also studied for IL-8 and MAPK kinases mRNA production. P-p38 was the most expressed MAPK kinase in the bronchial mucosa of all subjects. No significant differences were observed for immune-expression of p-p38, p-JNK and p-ERK1/2 between COPD and control subjects. 16HBE cells treated with H 2 O 2 , cytomix (TNFα+IL-1β+IFNγ) and Lipopolysaccharide (LPS) up-regulated IL-8 mRNA production at 1h or 2h after treatments. P38α mRNA was significantly increased after H 2 O 2 and LPS. JNK1 and ERK1 mRNA were not significantly increased after H 2 O 2 , cytomix or LPS treatments. Blocking p38α activity IL-8 mRNA production was not changed at 1h, 2h and 4h after H 2 O 2 or LPS challenge. P-p38 immune-positivity is prevalent in the bronchial mucosa of COPD and asthmatic patients and p38 mRNA is increased after bronchial epithelial challenges suggesting a relevant role for this MAPK kinase in the induction of bronchial inflammation in COPD and asthma.

Research paper thumbnail of Oxidative stress induces myeloperoxidase expression in endocardial endothelial cells from patients with chronic heart failure

Basic Research in Cardiology, 2009

j Abstract Increased oxidative stress has been implicated in the pathogenesis of a number of card... more j Abstract 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 lM) dose for different exposure times (3 and 6 h). Treated and control cells were tested by immunohistochemistry and RT-PCR for MPO and 3-chlorotyrosine expression. Both endothelial cell types expressed myeloperoxidase following oxidative stress, with higher levels in EEC. Moreover, 3-chlorotyrosine accumulation in treated cells alone indicated the presence of MPO-derived hypochlorous acid. Immunohistochemistry on sections from post-infarcted heart confirmed in vivo the endothelial positivity to MPO, 3-chlorotyrosine and, to a minor extent, nitrotyrosine. Immunohistochemical observations were confirmed by detection of MPO mRNA in both stimulated EEC and HUVEC cells. This study demonstrates for the first time that EEC can express MPO after oxidative stress, both in vitro and in vivo, followed by accumulation of 3-chlorotyrosine, an end product of oxidative stress. Deregulation of endothelial functions may contribute to the development of a number of cardiovascular diseases, including CHF. The results also highlight the notion that endothelium is not only a target but also a key player in oxidative-driven cardiovascular stress. j

Research paper thumbnail of HSP60 activity on 16HBE cells after oxidative and pro-inflammatory stimuli

European Respiratory Journal, Sep 1, 2014

Research paper thumbnail of Immunoregulation and FoxP3 expression in the bronchial mucosa of stable COPD patients

European Respiratory Journal, Sep 1, 2012

Number: 1905 Publication Number: P4589 Abstract Group: 3.3. Mechanisms of Lung Injury and Repair

Research paper thumbnail of Hsp60 and Hspl0 as antitumor molecular agents

Cancer Biology Therapy, May 1, 2007

The molecular chaperones Hsp60 and Hsp10 are, according to recent reports, involved in cancer dev... more 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.

Research paper thumbnail of Bradykinin b2 receptor expression in the bronchial mucosa of allergic asthmatics: the role of NF-kB

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, Jan 20, 2015

Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-... more Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-kB) is potentially involved in BK B2 receptor (B2R) regulation. In this observational cross-sectional study B2R and NF-kB expression was evaluated in bronchial biopsies from mild asthmatics (after diluent/allergen challenge) and healthy controls examining the role of NF-kB in B2R expression in primary human fibroblasts from normal and asthmatic subjects (HNBFb and HABFb). B2R and NF-kB (total and nuclear) expression was analyzed by immunohistochemistry in biopsies from 10 mild intermittent asthmatics (48 hours after diluent/allergen challenge) and 10 controls undergoing bronchoscopy. B2R co-localization in 5B5(+) and αSMA(+) mesenchymal cells was studied by immunofluorescence/confocal microscopy, and B2R expression in HABFb/HNBFb incubated with interleukin (IL)-4/IL-13 with/without BK, and after NF-kB inhibitor, by Western blotting. Bronchial mucosa B2R and nuclear NF-kB expression was h...

Research paper thumbnail of Chaperone patterns in vernal keratoconjunctivitis are distinctive of cell and Hsp type, and are modified by inflammatory stimuli

Allergy, Jan 27, 2015

Vernal keratoconjunctivitis (VKC) is a severe ocular allergy with pathogenic mechanism poorly und... more Vernal keratoconjunctivitis (VKC) is a severe ocular allergy with pathogenic mechanism poorly understood and no efficacious treatment. The aims of the study were to determine quantities and distribution of Hsp-chaperones in the conjunctiva of VKC patients and assess their levels in conjunctival epithelial and fibroblast cultures exposed to inflammatory stimuli. Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90, Hsp105, and Hsp110 were determined in conjunctiva biopsies from 9 patients and 9 healthy age-matched normal subjects, using immunomorphology and qPCR. Conjunctival epithelial cells and fibroblasts were cultured and stimulated with IL-1β, histamine, IL-4, TNF-α, or UV-B irradiation, and changes in Hsp levels were determined by Western blotting. Hsp27, Hsp40, Hsp70, and Hsp90 levels increased in the patients' conjunctiva, whereas Hsp10, Hsp60, Hsp100, and Hsp105 did not. Double immunofluorescence demonstrated co-localization of Hsp27, Hsp40, Hsp70, and Hsp90 with CD68 and tryptase. ...

Research paper thumbnail of Hsp27 and Hsp70 in chronic obstructive pulmonary disease: certainties vs doubts

Cell stress & chaperones, 2015

Research paper thumbnail of Role of interleukin-8 in the pathogenesis and treatment of COPD

Research paper thumbnail of Phospho-p38 MAPK Expression in COPD Patients and Asthmatics and in Challenged Bronchial Epithelium

Respiration, 2015

patients with mild/moderate (n = 17), severe/very severe (n = 16) stable COPD, control smokers (n... more patients with mild/moderate (n = 17), severe/very severe (n = 16) stable COPD, control smokers (n = 16), control nonsmokers (n = 9), in mild asthma (n = 9) and in peripheral airways from COPD patients (n = 15) and control smokers (n = 15). Interleukin (IL)-8 and MAPK mRNA was measured in stimulated 16HBE cells. Results: No significant differences in p-p38 MAPK, p-JNK or p-ERK1/2 expression were seen in bronchial biopsies and peripheral airways between COPD and control subjects. Asthmatics showed increased submucosal p-p38 MAPK expression compared to COPD patients (p < 0.003) and control non-smokers (p < 0.05). Hydrogen peroxide (H 2 O 2 ), cytomix (tumour necrosis factor-α + IL-1β + interferon-γ) and lipopolysaccharide (LPS) upregulated IL-8 mRNA at 1 or 2 h. p38 MAPKα mRNA was significantly increased after H 2 O 2 and LPS treatment. JNK1 and ERK1 mRNA were unchanged after H 2 O 2 , cytomix or LPS treatments. Conclusion: p-p38 MAPK expression is similar in stable COPD and control subjects but increased in the bronchi of mild