Hyaluronan mixed esters of butyric and retinoic acid drive cardiac and endothelial fate in term placenta human mesenchymal stem cells and enhance cardiac repair in infarcted rat hearts.

We have developed a mixed ester of hyaluronan with butyric and retinoic acid (HBR) that acted as a novel cardiogenic/vasculogenic agent in human mesenchymal stem cells isolated from bone marrow, dental pulp, and fetal membranes of term placenta (FMhMSCs). HBR remarkably enhanced vascular endothelial growth factor (VEGF), KDR, and hepatocyte growth factor (HGF) gene expression and the secretion of the angiogenic, mitogenic, and antiapoptotic factors VEGF and HGF, priming stem cell differentiation into endothelial cells. HBR also increased the transcription of the cardiac lineage-promoting genes GATA-4 and Nkx-2.5 and the yield of cardiac markerexpressing cells. These responses were notably more pronounced in FMhMSCs. FMhMSC transplantation into infarcted rat hearts was associated with increased capillary density, normalization of left ventricular function, and significant decrease in scar tissue. Transplantation of HBR-preconditioned FMhM-SCs further enhanced capillary density and the yield of human vWF-expressing cells, additionally decreasing the infarct size. Some engrafted, HBR-pretreated FMhMSCs were also positive for connexin 43 and cardiac troponin I. Thus, the beneficial effects of HBR-exposed FMhMSCs may be mediated by a large supply of angiogenic and antiapoptotic factors, and FMhMSC differentiation into vascular cells. These findings may contribute to further development in cell therapy of heart failure

Multiple large osteolytic lesions in a patient with systemic mastocytosis: a challenging diagnosis

Patients with advanced variants of Systemic Mastocytosis may develop destructive bone lesions when massive mast cell (MC) infiltrates are present. Finding of large osteolyses in indolent systemic mastocytosis, typically characterized by low MC burden, should prompt investigations for an alternative explanation

Three years experience with thermal and excimer lasers in the treatment peripheral artery disease

Laser angioplasty is an effective tool to revascularize peripheral artery disease, but the major limitation is a high restenosis rate. Our experience with the hot tip laser system has shown a high primary success, 59-73% of the arteries were patent at 18 months, although 21% resulted in severe restenosis. The excimer laser seems to have a better long-term patency. Histology of restenosis specimens removed by atherectomy, shows the key role of the smooth muscle cells in this process

NYIT. Memoria del curso académico

Memoria del Curso Académico 2020/2021, impartido por la Unidad Docente Campo Baeza en el New York Institute of Technolog

RECANALIZATION OF OCCLUDED PERIPHERAL ARTERIES BY HOT TIP LASER SYSTEM

Recanalization of long segment arterial occlusion are not feasible with standard angioplasty technique; the use of Hot Tip laser is evaluated in this experience

The TNF-Family Cytokine TL1A/Death Receptor 3 System Reduces Metabolic Activity in Chronic Lymphocytic Leukemia B Cells

Tumor necrosis factor (TNF)-like cytokine 1A (TL1A) is a member of the TNF superfamily, expressed on dendritic cells, macrophages, lymphocytes, and endothelial cells. It is the only described ligand for death receptor 3 (DR3), a death-domain containing receptor of the TNF-receptor superfamily, mainly expressed on lymphocytes, natural killer (NK) cells, and NK-T cells. TL1A–DR3 interaction results in co-stimulatory signaling for activated T cells, leading to amplification of inflammation and immune responses, correlated with greater pathogenicity in diverse autoimmune diseases. In contrast, in activated B cells the TL1A/DR3 system exerts inhibitory effect on cell proliferation, suggesting that TL1A may also have modulatory and homeostatic functions on B-cell expansion. Chronic lymphocytic leukemia (CLL) is the leukemia with the highest incidence among adults in Western countries. It is well documented that several elements within the tumor microenvironment, including antigens, cytokines, adhesion molecules, and surface receptors, play a fundamental role in supporting the growth of CLL. In contrast, little is known on regulatory mechanisms of CLL growth. In this study, we have investigated the possible regulatory role of the TL1A/DR3 system in B cells from CLL patients. CLL patients from the Hematology Unit at the University Hospital of Verona (Italy) were included in this study (n=37). Disease characteristics and demographic variables were collected on all patients. Purified B cells were obtained by negative selection. DR3 expression was measured on peripheral blood B cells by flow cytometry and western blot at baseline and following B cell receptor (BCR) stimulation with anti-human IgM. DR3 expression was confirmed on lymph-node CLL specimens by immunofluorescence. Metabolic activity of CLL B cells was analyzed by MTT assay. Apoptosis was analyzed by Annexin V assay. TL1A serum levels in CLL patients were measured by ELISA. Baseline analysis in vitro showed that DR3 was expressed at low levels in CLL B cells. Cell activation through stimulation of the BCR induced a significant increase of DR3 expression in a fraction of CLL B cells (p<0.001). Higher levels of DR3 expression were associated with early-stage (Rai 0) disease (p=0.019). The relevance of these findings was confirmed by immuofluorescence analysis of B-CLL lymph-node specimens showing that DR3 was expressed at high levels in some CLL B cells in vivo. Treatment of purified CLL B cells with exogenous recombinant TL1A in vitro, in the presence of BCR stimulation, induced a decreased metabolic activity in 3/8 B-CLL cell samples (37.5%). No change in CLL metabolic activity was observed following treatment with TL1A alone, in the absence of BCR stimulation. Treatment of B-CLL samples with TL1A, either in the presence or absence of BCR stimulation, induced no changes in cell viability, thus ruling out that decreased metabolic activity was due to reduced survival. A soluble form of TL1A was detected in the sera of CLL patients and higher serum levels of TL1A were significantly associated with early-stage (Rai 0) disease (p=0.023) and absence of CD38 expressions (p=0.035). In summary, this study shows that B-CLL metabolic activity can be reduced through the activity of the TL1A/DR3 system, in the presence of the BCR stimulation. Furthermore, TL1A and DR3 levels are higher in patients with early-stage disease. Taken together, these findings suggest that the TL1A/DR3 system in vivo, in the presence of antigen stimulation, may modulate leukemic cell metabolism in early-stage CLL, thus influencing the clinical course of disease

EXPRESSION AND FUNCTIONAL ACTIVITY OF THE TNFR-SUPERFAMILY MEMBER DEATH RECEPTOR 3 IN CHRONIC LYMPHOCYTIC LEUKEMIA B CELLS ACTIVATED BY THE BCR

Background: Growing evidences suggest a dynamic balance of B-cell chron- ic lymphocytic leukemia (B-CLL) cells between the blood and lymphoid tissues, which represent permissive niches for cell proliferation and survival. Within lymphoid tissue sites, molecules of the tumor necrosis factor (TNF) superfam- ily have been shown to play a supportive role and contribute to the pathogen- esis of B-CLL. Thereby, investigating expression and functions of novel TNFR- superfamily members in B-CLL would allow us to gain deeper insights into molecular crosstalk within leukemic microenvironments. Death receptor (DR) 3 is a TNFR-superfamily member expressed in lymphocyte-enriched tissues that binds to the TNF-like ligand 1A (TL1A), a TNF superfamily member. The TL1A/DR3 axis is implicated in regulatory mechanisms of adaptive immune response under physiological and pathological settings. Further evidence for the implication of TL1A in regulatory mechanisms arises from our recent data show- ing an inhibitory function of TL1A on B-cell proliferation. In leukemia cells, DR3 expression and functions have not been explored so far. CLL patients and to explore the possible role of TL1A/DR3 axis in the disease. Methods: B cells were purified from PBMC of 37 B-CLL patients by negative selection with magnetic beads. DR3 surface expression was measured by flow cytometry at baseline and following stimulation with F(ab’)2 anti-human IgM con- jugated to latex microspheres. DR3 expression was confirmed by western blot and immunofluorescence analysis on B-CLL lymph nodes. DR3 function was studied by MTT assay and Annexin V assay. TL1A serum levels were measured by ELISA. Results: Under basal conditions CLL B cells in vitro expressed low levels of DR3. Stimulation of the B cell receptor (BCR) with anti-IgM antibodies induced a sta- tistically significant increase of DR3 expression in CLL B cells (p<0.001). Induced DR3 expression showed great variability amongst B-CLL cells (variance (σ2)=6.38). Flow cytometry data were confirmed by Western blot analysis. The relevance of these findings was further confirmed by immuofluorescence analy- sis of B-CLL lymph-node specimens showing that in vivo high levels of DR3 were expressed by a number of B-CLL cells. To assess whether the anti-IgM-induced DR3 molecule was functionally active in CLL B cells, we examined the ability of DR3 to modulate their metabolic activity. Stimulation of DR3 with TL1A in the presence of BCR engagement showed that TL1A induced an equal or greater than 25% decrease of metabolic activity in 37.5% of B-CLL cell samples. In these samples the modulation is not due to reduced survival, as assessed by Annexin V assay. No change in metabolic activity was observed following TL1A treatment, in the absence of anti-IgM. Higher levels of DR3 expression were significantly associated with early-stage (Rai 0) disease (p=0.019) and higher serum levels of TL1A were significantly associated with early-stage (Rai 0) disease (p=0.023) and absence of CD38 expressions (p=0.035). CLL cells activated by the BCR stimulation express DR3 and TL1A reduces meta- bolic activity in some B-CLL cells. Herein, our data show a novel regulatory role for TL1A that, in the presence of antigen stimulation, may modulate leukemic cell metabolism through DR3 in early-stage B-CLL and suggest that homeostatic functions of TL1A may influence the clinical course of B-CLL disease

Epidemiology of mastocytosis in adults based on a multidisciplinary diagnostic approach.

Background: Mastocytosis is a rare disease. Some epidemiological informations are available about its cutaneous form in children but very few studies addressed the issue of its incidence and prevalence in adults. Aims: We evaluated the epidemiology of mastocytosis in the adult (≥18 years) population of the Verona province (Veneto region, Italy), based on the activity of the Multisciplinary Mastocytosis Outpatient Clinic, established in Verona in January 2006. Methods: Patients with suspected mastocytosis are referred to our Clinic from a network of Allergological, Dermatological and Rheumatological Centers in Northern Italy. We focused on all the consecutive adult patients with a confirmed diagnosis of mastocytosis resident in the Verona province. The diagnosis of Cutaneous (CM) or Systemic Mastocytosis (SM) was established according to the World Health Organization Classification. Patients with Urticaria Pigmentosa who did not perform bone marrow evaluation were defined as having Mastocytosis in the Skin (MIS). Written informed consent was obtained from all the enrolled patients. Data about the general population resident in the Verona province from 2008 to 2011 were found at the www.demo.istat.it site. Historical data about the prevalence and incidence of mastocytosis in Veneto were obtained from the Veneto Tumor Registry, Istituto Oncologico Veneto-IRCCS. Results: Seventy-four adults from a total of 293 patients in our Mastocytosis Registry were identified as living in the Verona province. They were equally distributed by gender (M/F=36/38); median age at first observation was 47.5 (range 20-80) years. The reasons for referral were Urticaria Pigmentosa (n=41), anaphylaxis after hymenoptera sting (n=27) or drugs (n=2) with basal serum tryptase >11.4 ng/mL, unexplained osteoporosis (n=2), other mediator-related symptoms (n=1) and non-MC myeloid neoplasia (n=1). The diagnosis was indolent SM (ISM) in 62 (83.8%) patients; the other cases were CM (n=6), MIS (n=4), aggressive SM (n=1) and SM associated with non-MC disease (n=1). Skin involvement was significantly more frequent in females than in males (78.9% vs 47.2%, respectively, p=.007). There was a trend for a higher median basal tryptase in males vs females (19.25 vs 15.95 ng/mL, respectively, p=.054). D816V KIT mutation was found in 56/64 tested cases (87.5%). We did not find any familial case in this adult population. After a median follow-up of 36 (range 2-112) months all patients are alive. The prevalence of mastocytosis in the Verona province adult population was 9.7 per 100,000 inhabitants, without gender differences. The incidence of new cases did not substantially change from 2008 to 2011 and was 0.93-1.34 per 100,000 inhabitants/year. For comparison, the reported prevalence and incidence in Veneto region before the establishment of our activity (2002-2005) were 0.65 per 100,000 inhabitants and 0.12 new cases per 100,000 inhabitants/year, respectively. Summary / Conclusion: Although rare, mastocytosis in adults is at least 10 times more frequent than previously reported. Due to the close collaboration with all the Allergology, Dermatology and Rheumatology units of our province, we can assume that virtually all patients with suspected mastocytosis are referred to us. However, the actual prevalence may be higher because ISM diagnosis is understimated especially when skin lesions and/or mediator symptoms are absent. A multidisciplinary diagnostic approach and a wide network of specialists are needed to improve the awareness about this disease and to define its actual epidemiology