Papers by Paolo Romagnoli
Virology journal, Sep 20, 2018
Torquetenovirus (TTV) belongs to Anelloviridae family, infects nearly all people indefinitely wit... more Torquetenovirus (TTV) belongs to Anelloviridae family, infects nearly all people indefinitely without causing overt disease establishing a fine and successful interaction with the host. Increasing evidence have shown some human viruses exploit extracellular vesicles thereby helping viral persistence in the host. Here, the presence of TTV in extracellular vesicles circulating in human plasma was investigated. TTV DNA was quantified in plasma-derived exosomes from 122 samples collected from 97 diseased patients and 25 healthy donors. Exosomes enriched vesicles (EEVs) were extracted from plasma and characterized by Nanoparticle tracking analysis, by western blot for presence of tetraspanin CD63, CD81 and annexin II protein and, finally, by electron microscopy (EM). Presence and quantitation of TTV DNA were assessed with an universal single step real-time TaqMan PCR assay. Preliminary investigation showed that the human plasma extracted extracellular vesicles exhibited a main size of 70...
Nanomedicine (London, England), 2017
The spontaneous adsorption of proteins on nanoparticles (NPs) in biological media is exploited to... more The spontaneous adsorption of proteins on nanoparticles (NPs) in biological media is exploited to prepare complexes of NPs and proteins from cancer cells' lysates for application in cancer immunotherapy. Gold (Au) and silica NPs were synthesized, incubated with cancer cells' lysates and characterized. Dendritic cells (DCs) were challenged with protein-coated NPs, their maturation, viability and morphology were evaluated and lymphocytes T proliferation was determined. Silica and Au NPs bound different pools of biomolecules from lysates, and are therefore promising selective carriers for antigens. When incubated with immature DCs, NPs were efficiently endocytosed without cytotoxicity. Finally, protein-coated AuNPs promoted DC maturation and DC-mediated lymphocyte proliferation, at variance with lysate alone and protein-coated silica NPs, that did not promote DCs maturation. These results demonstrate that the spontaneous formation of protein coronas on NPs represents a possible...
Amer J Dermatopathol, 1992
Histopathology, 1986
Dendritic cells in the dermal infiltrate of Sezary syndrome The dermal infiltrates of four patien... more Dendritic cells in the dermal infiltrate of Sezary syndrome The dermal infiltrates of four patients with the SCzary syndrome were studied by electron microscopy and the data were evaluated quantitatively. The nuclear contour index of lymphocytes was calculated, and many turnour cells had an index greater than 6.5. Dendritic cells were found in all cases. The dendritic cells contained smooth and rough endoplasmic reticulum, moderately well-developed Golgi apparatus, scanty lysosomes and many thin and intermediate filaments; their surface was scalloped with numerous vesicles. Birbeck granules were not found in the cytoplasm of dendritic cells. Dendritic cells comprised 24% of infiltrating cells and were interspersed with lymphocytes; 75% of the lymphocytes were in contact with dendritic cells; 35% of the lymphocytes in contact with dendritic cells had a nuclear contour index higher than 6.5 and 76% had a nuclear contour index higher than 5. The data strongly suggest a functional relationship between lymphocytes and dendritic cells in the dermal infiltrate of SCzary syndrome. They are discussed in relation to the hypothesis that the disease is a consequence of chronic immune stimulation.
Italian Journal of Anatomy and Embryology, 2014
Oral surgery, oral medicine, and oral pathology, 1988
During a prospective investigation of oral lesions of 120 consecutive patients positive for human... more During a prospective investigation of oral lesions of 120 consecutive patients positive for human immunodeficiency virus, belonging to the intravenous drug abuser risk group and other risk categories, we observed hairy leukoplakia (HL) in 23 cases (19%). The median age of the patients was 27 years (range, 20 to 50 years). Twenty patients were men and three were women. All but two of the twenty three patients used intravenous drugs for a median period of 6 years (range, 5 to 18 years) and were involved in several episodes of needle sharing. Eight men were also bisexual, one man was homosexual, and one man was hemophiliac and bisexual. Eleven patients had asymptomatic infection, five had lymphadenopathy syndrome, six had AIDS-related complex, and one had acquired immunodeficiency syndrome. In all patients, HL lesions were localized on the lateral borders of the tongue. In twelve patients, the lesion was unilateral, and in eleven patients, it was bilateral. Microscopically, hyperparake...
Basic Mechanisms of Physiologic and Aberrant Lymphoproliferation in the Skin, 1994
Transplantation, 1990
Cyclosporine (CsA), administered to rats at daily doses of 10 and 50 mg/kg body weight, for 21 da... more Cyclosporine (CsA), administered to rats at daily doses of 10 and 50 mg/kg body weight, for 21 days, influenced negatively the structures involved in the synthesis, storage and secretion of digestive enzymes in pancreatic acinar cells. A dose-related, significant reduction in basophilic cell regions, secretion granule content, and overall size of acinar cells was appreciable by light microscopy and morphometry. By electron microscopy, the acinar cells of the rats given 10 mg/kg/day CsA were similar to the controls, whereas with the higher dose most cells showed reduction in the size of nucleoli, increase in the number of lysosomes, and evidence of autophagy. In only a few cells was autophagy particularly severe and involved almost the entire cytoplasm. Nine weeks after withdrawal from CsA treatment, the structural recovery of acinal cells was complete, and features indicating enhanced protein synthesis and mitochondrial multiplication were observed by electron microscopy. In conclusion, prolonged administration of CsA to rats induces changes in the acinar cells indicating a depression of their activity, without substantial impairment in the viability of the most of them, even at high doses. This accounts for complete restoration of the acinar tissue upon withdrawal.
Pancreas, 1988
A morphometrical investigation was carried out on rat pancreatic lobules, which were incubated fo... more A morphometrical investigation was carried out on rat pancreatic lobules, which were incubated for 10 min in MEM. Some lobules were treated with carbamylcholine chloride (CCh) at three different concentrations; those not treated were defined as controls. Upon stimulation, the apical plasma membrane surface area and the number of zymogen granules less than 20 nm away from the apical plasma membrane significantly increased and were directly correlated with increases in secretion. The diameter of zymogen granules decreased at 10(-6) M CCh concentration, but increased at 10(-7) and 10(-5) M CCh concentrations as compared with controls. These results seem to indicate that exocytosis, rather than transmembrane flow of individual molecules, is enhanced by and correlated with protein secretion from the exocrine pancreas.
Journal of the European Academy of Dermatology and Venereology, 2001
Histopathology, 1986
Dendritic cells in the dermal infiltrate of Sezary syndrome The dermal infiltrates of four patien... more Dendritic cells in the dermal infiltrate of Sezary syndrome The dermal infiltrates of four patients with the SCzary syndrome were studied by electron microscopy and the data were evaluated quantitatively. The nuclear contour index of lymphocytes was calculated, and many turnour cells had an index greater than 6.5. Dendritic cells were found in all cases. The dendritic cells contained smooth and rough endoplasmic reticulum, moderately well-developed Golgi apparatus, scanty lysosomes and many thin and intermediate filaments; their surface was scalloped with numerous vesicles. Birbeck granules were not found in the cytoplasm of dendritic cells. Dendritic cells comprised 24% of infiltrating cells and were interspersed with lymphocytes; 75% of the lymphocytes were in contact with dendritic cells; 35% of the lymphocytes in contact with dendritic cells had a nuclear contour index higher than 6.5 and 76% had a nuclear contour index higher than 5. The data strongly suggest a functional relationship between lymphocytes and dendritic cells in the dermal infiltrate of SCzary syndrome. They are discussed in relation to the hypothesis that the disease is a consequence of chronic immune stimulation.
Frontiers in Neuroscience, 2013
The muscle coat of the human lower oesophageal sphincter and stomach was studied 5 cm above and 4... more The muscle coat of the human lower oesophageal sphincter and stomach was studied 5 cm above and 4 cm below the gastro-oesophageal junction. Four subjects were operated on for motility disorders of the esophagus, two for a hypertensive lower oesophageal sphincter and two for an epiphrenic diverticulum; six subjects were operated on for oesophageal or gastric carcinomas. Specimens were fixed in phosphate-buffered OsO 4 , embedded in Epon, contrasted with uranyl acetate and lead citrate and observed under a Siemens Elmiskop Ia electron microscope. Both the oesophageal and gastric muscle cells, which showed features typical of this cell type, were innervated by multiple varicosities that were rich in synaptic vesicles; these varicosities were generally rarely encountered at distances less than 1000 Å from muscle cells. Only a very few, close neuromuscular junctions were detected. Special cells, which correspond to the "interstitial cells of Cajal" as reported by other authors, were discerned at the periphery of muscle cell bundles. These cells were characterized by an elongated cell body with many thin branches and an oval, sometimes indented nucleus. Some pinocytotic vesicles were located at the cell periphery. These cells were surrounded by a discontinuous basal lamina and were seen in close contact with each other and with muscle cells; the close contact areas were often very wide. The cytoplasm contained variable amounts of mitochondria, a well-developed smooth endoplasmic reticulum and a Golgi complex. As a characteristic feature, bundles of thin filaments were located at the cell periphery and were attached to electron-dense areas of the cell membrane. Morphologically, these filaments resembled myofilaments; they were present in variable amounts and were sometimes very numerous. The observation that the cytoplasmic organelles and filaments varied in number, is probably related to the different functional properties of these cells. Interstitial cells were richly innervated by varicose nerve fibers that were densely packed with synaptic vesicles; many close junctions to nerve endings were also detected. These morphological data lead us to assume that the interstitial cells demonstrated by the electron microscope do not correspond to the cells initially identified by Cajal and cannot even be considered connective tissue cells. We propose that they are specialized smooth muscle cells that are involved in generating spontaneous, myogenic electrical activity in the gastrointestinal tract.
Blood, 2011
CD133 is a hallmark of primitive myeloid progenitors. We have addressed whether human cord blood ... more CD133 is a hallmark of primitive myeloid progenitors. We have addressed whether human cord blood cells selected for CD133 can generate dendritic cells, and Langerhans cells in particular, in conditions that promote that generation from CD34+ progenitors. Transforming growth factor-β1 (TGF-β1) and anti–TGF-β1 antibody, respectively, were added in some experiments. With TGF-β, monocytoid cells were recognized after 7 days. Immunophenotypically immature dendritic cells were present at day 14. After 4 more days, the cells expressed CD54, CD80, CD83, and CD86 and were potent stimulators in mixed lymphocyte reaction; part of the cells expressed CD1a and langerin, but not Birbeck granules. Without TGF-β, only a small fraction of cells acquired a dendritic shape and expressed the maturation-related antigens, and lymphocytes were poorly stimulated. With anti–TGF-β, the cell growth was greatly hampered, CD54 and langerin were never expressed, and lymphocytes were stimulated weakly. In conclus...
Endocrine-Related Cancer
Pheochromocytomas (Pheos) and paragangliomas (PGLs) are neuroendocrine tumors. Approximately 30–4... more Pheochromocytomas (Pheos) and paragangliomas (PGLs) are neuroendocrine tumors. Approximately 30–40% of Pheos/PGLs are due to germline mutations in one of the susceptibility genes, including those encoding the succinate dehydrogenase subunits A-D (SDHA-D). Up to 2/3 of patients affected by SDHB mutated Pheo/PGL develop metastatic disease with no successful cure at present. Here, for the first time, we evaluated the effects of SDHB silencing in a three dimension (3D) culture using spheroids of a mouse Pheo cell line silenced or not (wild type/wt/control) for the SDHB subunit. We investigated the role of the microenvironment on spheroid growth and migration/invasion by co-culturing SDHB-silenced or wt spheroids with primary cancer-activated fibroblasts (CAFs). When spheroids were co-cultured with fibroblasts, SDHB-silenced cells showed a significant increase in matrigel invasion as demonstrated by the computation of the migratory areas (P < 0.001). Moreover, cells detaching from the...
Cancers
Metabolic interplay between the tumor microenvironment and cancer cells is a potential target for... more Metabolic interplay between the tumor microenvironment and cancer cells is a potential target for novel anti-cancer approaches. Among stromal components, adipocytes and adipose precursors have been shown to actively participate in tumor progression in several solid malignancies. In adrenocortical carcinoma (ACC), a rare endocrine neoplasia with a poor prognosis, cancer cells often infiltrate the fat mass surrounding the adrenal organ, enabling possible crosstalk with the adipose cells. Here, by using an in vitro co-culture system, we show that the interaction between adipose-derived stem cells (ASCs) and the adrenocortical cancer cell line H295R leads to metabolic and functional reprogramming of both cell types: cancer cells limit differentiation and increase proliferation of ASCs, which in turn support tumor growth and invasion. This effect associates with a shift from the paracrine cancer-promoting IGF2 axis towards an ASC-associated leptin axis, along with a shift in the SDF-1 ax...
Mast cell histochemistry has been proposed in addition to classic histological methods to estimat... more Mast cell histochemistry has been proposed in addition to classic histological methods to estimate the course of traumatic events before and after death. We have addressed the utility of this approach on nine victims of different types of trauma. Sections of wounded skin were stained with hematoxylin and eosin and with fluorescent avidin to tag mast cells. Mast cell numbers were evaluated by both direct and digitalized counts. Intact skin was used as control. The results on mast cells implemented the findings upon hematoxylin and eosin stain and helped to put the wounds and death in chronological sequence. Digitalized morphometry allowed to reduce intra-and inter-observer variation. We conclude that combined histological and histochemical analyses can be of practical use in forensic pathology, that a preliminary setting of the reference values is needed for each laboratory, and that image analysis can be of help for the quantification of the results. The problem of the ''vitality'' of lesions, that is, whether trauma had occurred on a living victim or a deceased body, is a major one in forensic medicine and has been addressed for a long time by routine histological methods (1). Attempts to apply either enzyme histochemistry for disparate enzyme activities (2) or immunohisto-chemistry for disparate extracellular matrix antigens (3) have not been followed by widespread application to forensic medicine. We have previously shown that the number of mast cells (MCs) on the border of skin lesions increases significantly between 5 min and 3 h after trauma and decreases afterward, reaching basal values between 3 and 6 h and further decreasing until 24 h (4); also, these cells increase the expression of TNF-alpha very early during wound healing (5). Oemichen et al. (6) have recently confirmed that MC degranulate at the border of skin wounds within 60 min, as estimated by enzyme histochemistry for a granule-bound ester-ase (7). We have now used the 95% confidence limits, derived from our previously published data on MC counts in early stages after wounding (4,5,8), to assist in the analysis of cases of forensic pathology and see whether the consequent information could be of help to solve those cases or was irrelevant. While the previous studies dealt with skin wounds with known survival time and the variable to analyze was the number of MCs and the degree of inflammatory cell infiltration, in the present study we started from MC and inflammatory infiltration grading to address the survival time. MCs were demonstrated by fluorescent avidin staining of their granules. Avidin tags MCs specifically and with high sensitivity; the binding of the molecule is attributable to electrostatic attraction; hence, it has the same meaning as basophilia but with much higher sensitivity than conventional microscopic dyes as toluidine blue or Giemsa (9,10). This staining has the advantage of being intense hence sensitive, specific, and more simple to achieve than another sensitive method to show degranulation, that is, the enzyme histo-chemical demonstration of naphthol AS-D chloroacetate esterase, which furthermore labels macrophages besides MCs (7). Methods Nine victims were selected as representative of our experience. They were eight men and one woman aged 25–51 years, mean 35.5 years. Autopsies were performed at the Department Section of Forensic Medicine of the University of Florence (Italy). The research was approved by the Department ethical committee and was performed in the respect of the Italian law. Corpses were routinely kept at +4°C from the moment they arrived at the morgue until autopsy. The time between death and autopsy was 24–48 h. A short description of the cases is reported below. Four victims had been found dead with severe multiple weapon wounds: three subjects had knife wounds and one had gun wounds; the analyses were directed at assessing the time sequence of the wounds and their respective role in death. Two victims had been killed in car accidents for which there was circumstantial evidence of multiple hits: in one case, a second car had hit the victim of a first accident, who laid on the ground; in the other case, a second car had hit the victim's car, with the subject inside, which had previously collided with a fixed obstacle.
Journal of Dermatological Science, 1998
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Inflammation & Allergy - Drug Targets, 2010
Mast cells are bone marrow derived cells capable of secreting many active molecules: mediators st... more Mast cells are bone marrow derived cells capable of secreting many active molecules: mediators stored in specific granules, such as histamine and heparin; small molecules produced immediately upon stimulation, such as lipid derivatives and nitric oxide; and many constitutively secreted, pleiotropic cytokines. Thanks to these secretion products and perhaps direct cell-cell interactions, mast cells play roles in inflammation and tissue repair, angiogenesis and fibrosis. Mast cells themselves respond to many mediators of their own, giving rise to autocrine loops. Successful anti-allergic therapies have typically targeted the receptors for mast cell secretory products, particularly those for histamine. Among agents directly affecting mast cells, disodium chromoglycate and glucocorticoids are known since some time, while new pharmacological approaches may stem from the recognition of an interference with mast cell growth and differentiation by cyclosporine A, monoclonal antibodies, interferons, and JAK3 inhibitors. The action of agents that affect mast cell differentiation and function is considered here from a cell and tissue biological perspective as a premise to the application of these agents to the clinics, therefore special attention has been paid to references pertaining to humans.
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Papers by Paolo Romagnoli