Differential response to hepatic differentiation stimuli of amniotic epithelial cells isolated from four regions of the amniotic membrane

Human Amniotic Epithelial Cells (hAEC) isolated from term placenta are a promising source for regenerative medicine. However, it has long been debated whether the hAEC population consists of heterogeneous or homogeneous cells. In a previous study, we investigated the characteristics of hAEC isolated from four different regions of the amniotic membrane finding significant heterogeneity. The aim of this study was to evaluate the hepatic differentiation capability of hAEC isolated from these four regions. Human term placentae were collected after caesarean section and hAEC were isolated from four regions of the amniotic membrane (R1-R4, according to their relative distance from the umbilical cord) and treated in hepatic differentiation conditions for 14 days. hAEC-derived hepatocyte-like cells showed marked differences in the expression of hepatic markers: R4 showed higher levels of Albumin and Hepatocyte Nuclear Factor (HNF) 4α whereas R1 expressed higher Cytochrome P450 enzymes, both at the gene and protein level. These preliminary results suggest that hAEC isolated from R1 and R4 of the amniotic membrane are more prone to hepatic differentiation. Therefore, the use of hAEC from a specific region of the amniotic membrane should be taken into consideration as it could have an impact on the outcome of therapeutic applications

Ultrastructural Patterns of Cell Damage and Death Following Gamma Radiation Exposure of Murine Erythroleukemia Cells

Radiation causes damage to cell surface membranes, cytoplasmic organelles, and the nuclear process of DNA synthesis and repair, and this eventually results in different modes of cell death. In this study we examined murine erythroleukemia (MEL) cells, exposed to 15 and 60 Gy of 10 MeV photonic energy, and left in culture for up to 96 hours. Electron microscopical analysis was performed on conventionally embedded samples and freeze-fracture replicas, in order to detect ultrastructural patterns of cell damage and death. Of interest was the observation of chromatin condensates, nuclear membrane associations and nuclear pore redistribution during early apoptosis. Pronounced rearrangements of transmembrane particles during late stages of cellular necrosis were also found. The morphological damage induced by both doses of radiation as a function of time after exposure was only quantitatively but not qualitatively different

Ionizing Radiation-Induced Apoptosis and DNA Repair in Murine Erythroleukemia Cells

A morphological study of DNA repair and apoptotic patterns in relationship with cell cycle events was performed on murine erythroleukemia cells. The presence and distribution of DNA replicon sites were evaluated through the BrdU-anti BrdU immunofluorescence and immunogold techniques in light and electron microscopy. Different patterns of labelling and percentages of BrdU positive cells were observed depending on irradiation dose (up to 60 Gy) and time in post-irradiation culture (up to 24 hours). An enlargement of the S phase of the cell cycle was evidenced 18 hours post-irradiation as determined by flow cytometry analysis. The high resolution approach showed that, in spite of several morphological alterations, BrdU labelling was present even in cells displaying early and late apoptotic features

Replacement of hematopoietic system by allogeneic stem cell transplantation in myelofibrosis patients induces rapid regression of bone marrow fibrosis

Bone marrow fibrosis is a hallmark of primary and post ET/PV myelofibrosis. To investigated the impact of replacement of the hematopoietic system in myelofibrosis patients by allogeneic stem cell transplantation on bone marrow fibrosis, we studied bone marrow fibrosis on bone marrow samples from 24 patients with myelofibrosis before and after dose-reduced conditioning followed by allogeneic stem cell transplantation from related or unrelated donor. Using the European Consensus on Grading Bone Marrow Fibrosis, before allografting all patients had advanced fibrosis MF-2 (n = 13) or MF-3 (n = 11). After transplantation, a complete (MF-0) or nearly complete (MF-1) regression of bone marrow fibrosis was seen in 59 % at day +100, in 90 % at day +180, and in 100 % at day +360. No correlation between occurrence of acute graft-versus-host disease, and fibrosis regression on day +180 was seen. We conclude that dose-reduced conditioning, followed by allogeneic stem cell transplantation, resulted in a rapid resolution of bone-marrow fibrosis suggesting the bone marrow fibrogenesis is a highly dynamic rather than static process in patients with myelofibrosis

TNF-alpha-induced apoptosis in Daudi cells: multiparametric analysis

Tumour necrosis factor alpha (TNF-alpha) is a cytokine that induces physiological and pathophysiological effects in the immune system. In this study we analyzed its action on a human lymphoma cell line (Daudi cells) after 1 h, 6 h and 24 h of incubation. Using vital DNA stains, DNA gel electrophoresis, in situ nick translation, transmission electron microscopy and flow cytometry we showed that as early as after 6 h of treatment, target cells were able to undergo death by apoptosis. This was associated with cell shrinkage, chromatin condensation, apoptotic bodies in the cytoplasm without the typical DNA fragmentation into low molecular weight nucleosomes. Of interest was the observation of a significant number (60%) of cells positive to the nick translation in specimens treated for 6 h, decreasing to 40% in samples treated for 24 h, when most of the cells were in late apoptosis. In addition, no subdiploid peak was evident in flow cytometry regardless of the time of incubation with TNF. Our study on Daudi cells clearly supports the existence of alternative forms of apoptosis in which DNA degradation does not result only in oligonucleosomal fragmentation

Comunicazione medica scritta: costruiamo informazioni di qualit\ue0 per il paziente? : Una valutazione della qualit\ue0 del materiale informativo a disposizione dei pazienti asmatici e diabetici negli ambulatori milanesi

OBJECTIVES: 1) To identify the extent and the nature of information pamphlets available for asthmatic and diabetic patients in secondary care hospitals in Milan; 2) To investigate what patients think about the quality of pamphlets using a questionnaire developed ad hoc. METHODS: Pamphlets have been identified by contacting 8 specialised ambulatory hospitals. The questionnaire was developed to achieve a quality rate in 3 dimensions of the materials (Style and readability, The condition and Treatment options). Patients were recluted in the same ambulatory hospital. RESULTS: 4 ambulatory hospitals and 40 patients agreed to participate in our survey. We identified 8 pamphlets (5 diabetes, 3 asthma). 6 of these have been promoted byfarmaceutical companies. The mean length was 31 pages and the issue most covered was treatment options (33% of pages dedicated). 30 pages (12% of the total) presented with publicity advertisements (6% direct and 6% indirect). The dimension with the best satisfaction rate was Condition, followed by Style and readability. CONCLUSIONS: In Milan, in secondary care asthmatic and diabetic patients, are often invited to consult pamphlets. They positively judged this material. During the study ambulatory physicians were poorly attentive about problems concerning patient access to information

Lymphocyte binding to K562 cells: effect of target cell irradiation andcorrelation with ICAM-1 and LFA-3 expression

Tumor irradiation induces modifications in the interaction of surviving target cells with immune cells. This interaction is mediated by adhesion molecules, whose expression can be strongly altered by radiation treatment. Here the probably of K562 tumor cells for lymphocyte binding was studied after exposure of target cells to different doses of gamma-radiation. Results were correlated to the expression of ICAM-1 and LFA-3 adhesion molecules on target cells. Radiation treatment enhanced the expression of both ICAM-1 and LFA-3 on the surface of target cells in a dose and time of culture-dependent fashion, reaching a maximum 24 hrs postirradiation, when also lymphocyte binding was increased. 10-30 Gy irradiation of K562 cells in vitro induces after 24 hrs, an up-regulation of ICAM-1 and LFA-3 expression that, in turn, increase lymphocyte binding, making tumor cells more exposed to cytotoxic attack. The progressive morphological damage induced by radiation, documented by the scattering singlas in flow cytometry and by electron microscopy analysis of irradiated K562 cells, induced, particularly at delayed times of culture in high doses irradiated cells, alterations of the target cell surface that might prevent the correct interaction with immune cells

Curcuma longa is able to induce apoptotic cell death of pterygium-derived human keratinocytes

Copyright © 2017 Silvia Sancilio et al. Pterygium is a relatively common eye disease that can display an aggressive clinical behaviour. To evaluate the in vitro effects of Curcuma longa on human pterygium-derived keratinocytes, specimens of pterygium from 20 patients undergoing pterygium surgical excision were collected. Pterygium explants were put into culture and derived keratinocytes were treated with an alcoholic extract of 1.3% Curcuma longa in 0.001% Benzalkonium Chloride for 3, 6, and 24 h. Cultured cells were examined for CAM5.2 (anti-cytokeratin antibody) and CD140 (anti-fibroblast transmembrane glycoprotein antibody) expression between 3th and 16th passage to assess cell homogeneity. TUNEL technique and Annexin-V/PI staining in flow cytometry were used to detect keratinocyte apoptosis. We showed that Curcuma longa exerts a proapoptotic effect on pterygium-derived keratinocytes already after 3 h treatment. Moreover, after 24 h treatment, Curcuma longa induces a significant increase in TUNEL as well as Annexin-V/PI positive cells in comparison to untreated samples. Our study confirms previous observations highlighting the expression, in pterygium keratinocytes, of nuclear VEGF and gives evidence for the first time to the expression of nuclear and cytoplasmic VEGF-R1. All in all, these findings suggest that Curcuma longa could have some therapeutic potential in the treatment and prevention of human pterygium