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Danieli P.,Coronary Care Unit and Laboratory of Clinical and Experimental Cardiology | Danieli P.,Laboratory of Experimental Cardiology for Cell and Molecular Therapy | Malpasso G.,Coronary Care Unit and Laboratory of Clinical and Experimental Cardiology | Malpasso G.,Laboratory of Experimental Cardiology for Cell and Molecular Therapy | And 22 more authors.
Stem Cells Translational Medicine | Year: 2015

The paracrine properties of human amniotic membrane-derived mesenchymal stromal cells (hAMCs) have not been fully elucidated. The goal of the present study was to elucidate whether hAMCs can exert beneficial paracrine effects on infarcted rat hearts, in particular through cardio-protection and angiogenesis. Moreover, we aimed to identify the putative active paracrine medi-ators.hAMCswereisolated,expanded,andcharacterized.Invitro,conditionedmediumfromhAMC (hAMC-CM) exhibited cytoprotective and proangiogenic properties. In vivo, injection of hAMC-CM into infarcted rat hearts limited the infarct size, reduced cardiomyocyte apoptosis and ventricular remodeling, andstrongly promoted capillary formation at theinfarct border zone. Genearray analysis led to the identification of 32 genes encoding for the secreted factors overexpressed by hAMCs. Among these, midkine and secreted protein acidic and rich in cysteine were also upregulated at the protein level. Furthermore, high amounts of several proangiogenic factorswere detected in hAMC-CMbycytokine array.Ourresultsstronglysupport theconcept thattheadministrationof hAMC-CM favors the repair process after acute myocardial infarction. © Alpha Med Press 2015.


Fusco C.,Immunohematology and Transfusion Service | Guerini F.R.,Foundation Medicine | Nocera G.,Italian National Cancer Institute | Ventrella G.,University of Naples Federico II | And 9 more authors.
Journal of Neuroimmunology | Year: 2010

Killer Immunoglobulin-like Receptor (KIR) genes may affect both resistance and susceptibility to autoimmune disorders, but their role in the pathogenesis of Multiple Sclerosis (MS) is still unclear. To evaluate the involvement of KIRs and their HLA ligands in the development of MS we performed genotyping of HLA -A, -B, -Cw, -DRB1 and KIRs loci in 121 RRMS patients and 103 healthy controls (HC). Results evidenced a possible protective role of the activating KIR2DS1 gene (py=0.001; OR:0.38), enhanced in the presence of its ligand group HLA-C2 (py=0.0001; OR:0.23). Our data suggest that the presence of functional compounds of activating KIR receptors together with their HLA ligands, allowing the immunomodulatory function of NK cells, may have a protective role against the disease. © 2010 Elsevier B.V.


Soria A.,University of Milan Bicocca | Guerini F.R.,Foundation Medicine | Bandera A.,University of Milan Bicocca | Bolognesi E.,Foundation Medicine | And 8 more authors.
PLoS ONE | Year: 2011

Background: In HIV-infected individuals, mechanisms underlying unsatisfactory immune recovery during effective combination antiretroviral therapy (cART) have yet to be fully understood. We investigated whether polymorphism of genes encoding immune-regulating molecules, such as killer immunoglobulin-like receptors (KIR) and their ligands class I human leukocyte antigen (HLA), could influence immunological response to cART. Methods: KIR and HLA frequencies were analyzed in 154 HIV-infected and cART-treated patients with undetectable viral load divided into two groups: 'immunological non responders' (INR, N = 50, CD4 + T-cell count <200/mm 3) and full responders (FR, N = 104, CD4 + T-cell count >350/mm 3). Molecular KIR were typed using polymerase chain reaction-based genotyping. Comparisons were adjusted for baseline patient characteristics. Results: The frequency of KIR2DL3 allele was significantly higher in FR than in INR (83.7% vs. 62%, P = 0.005). The functional compound genotype HLA-C1 +/KIR2DL3 +, even at multivariable analysis, when adjusted for nadir CD4 + T-cell count, was associated with reduced risk of INR status: odds ratio (95% Confidence Intervals) 0.34 (0.13-0.88), P = 0.03. Conclusions: Reduced presence of the inhibitory KIR2DL3 genotype detected in INR might provoke an imbalance in NK function, possibly leading to increased immune activation, impaired killing of latently infected cells, and higher proviral burden. These factors would hinder full immune recovery during therapy. © 2011 Soria et al.


Pisano F.,Coronary Care Unit | Pisano F.,Laboratory of Experimental Cardiology for Cell and Molecular Therapy | Altomare C.,Laboratory of Experimental Cardiology for Cell and Molecular Therapy | Cervio E.,Coronary Care Unit | And 20 more authors.
Stem Cells | Year: 2015

Several studies have demonstrated that miRNA are involved in cardiac development, stem cell maintenance, and differentiation. In particular, it has been shown that miRNA133, miRNA1, and miRNA499 are involved in progenitor cell differentiation into cardiomyocytes. However, it is unknown whether different miRNA may act synergistically to improve cardiac differentiation. We used mouse P19 cells as a cardiogenic differentiation model. miRNA499, miRNA1, or miRNA133 were transiently over-expressed in P19 cells individually or in different combinations. The over-expression of miRNA499 alone increased the number of beating cells and the association of miRNA499 with miRNA133 exerted a synergistic effect, further increasing the number of beating cells. Real-time polymerase chain reaction showed that the combination of miRNA499-‰+-‰133 enhanced the expression of cardiac genes compared with controls. Western blot and immunocytochemistry for connexin43 and cardiac troponin T confirmed these findings. Importantly, caffeine responsiveness, a clear functional parameter of cardiac differentiation, was increased by miRNA499 in association with miRNA133 and was directly correlated with the activation of the cardiac troponin I isoform promoter. Cyclic contractions were reversibly abolished by extracellular calcium depletion, nifedipine, ryanodine, and IP3R blockade. Finally, we demonstrated that the use of miRNA499-‰+-‰133 induced cardiac differentiation even in the absence of dimethyl sulfoxide. Our results show that the areas spontaneously contracting possess electrophysiological and pharmacological characteristics compatible with true cardiac excitation-contraction coupling. The translational relevance of our findings was reinforced by the demonstration that the over-expression of miRNA499 and miRNA133 was also able to induce the differentiation of human mesenchymal stromal cells toward the cardiac lineage. © 2015 Alpha Med Press.


Rattazzi M.,University of Padua | Rattazzi M.,Ca Foncello Hospital | Villalta S.,Ca Foncello Hospital | Galliazzo S.,Ca Foncello Hospital | And 15 more authors.
Clinical Science | Year: 2013

The relationship between MetS (metabolic syndrome), levels of circulating progenitor/immune cells and the risk of VTE (venous thromboembolism) has not yet been investigated. We studied 240 patients with previous VTE and 240 controls. The presence of MetS was identified according to NCEP ATP III guidelines and flow cytometry was used to quantify circulating CD34+ cells. VTE patients showed higher BMI (body mass index), waist circumference, triacylglycerol (triglyceride) levels, blood glucose, hs-CRP (high-sensitivity C-reactive protein) and lower HDL-C (high-density lipoprotein cholesterol) levels. The prevalence of MetS was significantly higher in VTE (38.3%) than in control individuals (21.3%) with an adjusted OR (odds ratio) for VTE of 1.96 (P=0.002). VTE patients had higher circulating neutrophils (P<0.0001), while the CD34+ cell count was significantly lower among patients with unprovoked VTE compared with both provoked VTE (P=0.004) and controls (P=0.003). Subjects were also grouped according to the presence/absence of MetS (MetS+ or MetS- ) and the level (high/low) of both CD34+ cells and neutrophils. Very high adjusted ORs for VTE were observed among neutrophils_high/MetS+ (OR, 3.58; P<0.0001) and CD34+ _low/MetS+ (OR, 3.98; P<0.0001) subjects as compared with the neutrophils_low/MetS- and CD34+ _high/MetS- groups respectively. In conclusion, low CD34+ blood cell count and high circulating neutrophils interplay with MetS in raising the risk for venous thromboembolic events. © The Authors Journal compilation 2013 Biochemical Society.


Zanconato G.,University of Verona | Cavaliere E.,University of Verona | Iacovella C.,University of Verona | Vassanelli A.,Immunohematology and Transfusion Service | And 3 more authors.
Journal of Maternal-Fetal and Neonatal Medicine | Year: 2012

Objective: To assess prevalence and causes of severe acute maternal morbidity cases and evaluate their impact on feto-maternal wellbeing and on facility resources. Study Design: Observational retrospective study adopting management-based criteria in a tertiary care public hospital during a 5-year period. Criteria adopted were: intensive care unit admission, blood transfusion≥ 4 units, emergency peripartum hysterectomy and arterial embolization at any time during pregnancy. Results: A total of 80 cases were identified, most of them (97.5%) through a combination of two criteria, ICU admission and blood transfusion. Commonest severe obstetric morbidities were major obstetric haemorrhage (48.8%) and hypertensive disorders (27.5%). Immigrant status (OR 1.68, 95% CI 1.032.7), pre-term birth (OR 4.15, 95% CI 2.56.8), Caesarean section (OR 7.74,95% CI 4.214.3) were factors significantly associated with SAMM cases. Major abdominal surgery was necessary in 26 women (32.5%), with emergency peripartum hysterectomy in 11 (13.5%). These events led to an average blood consumption per woman of 6.5±12.8 units and a mean hospital stay of 8.9±5.0 days, significantly longer (p<0.001) than the average duration of post-delivery care. Maternal mortality to morbility ratio was 1:80. Conclusions: An integrated intervention-based approach proved to be effective in finding severe acute maternal morbidity cases. Information on underlying causes and associated risk factors may improve prevention and treatment of obstetric morbidities, thus reducing feto-maternal adverse effects and hospital expenditures. © 2012 Informa UK, Ltd.


Ghidini C.,Laboratory of Biotechnologies | Sottini A.,Laboratory of Biotechnologies | Zanotti C.,Laboratory of Biotechnologies | Serana F.,University of Brescia | And 4 more authors.
Minerva Gastroenterologica e Dietologica | Year: 2010

A genotyping assay was setup to assess the prevalence, in the population of a Northern Italian city, of the C/T-13910 single nucleotide polymorphism, closely associated to lactose malabsorption in many world areas including Sardinia. The results were compared to published Italian data, in order to evaluate the worth of a future validation of the assay for use in routine practice. DNA was extracted from blood samples of 123 randomly chosen healthy blood donors coming from the same city area, and was analyzed by a real-time polymerase chain reaction (PCR) genotyping assay; the frequency of the hypolactasia-associated CC-geno-type was compared to the weighted average of results extracted from studies reporting the frequency of hypolactasic phenotype or geno-type in nearby or distant Italian regions. Sixty-five percent of donors carried the CC-geno-type, a percentage similar to other northern Italian cities, but significantly higher than what previously determined in surrounding Italian regions at the phenotype level, i.e. by breath test This discrepancy parallels recent reports of non concordance between results of geno-typing and hypolactasic phenotype in some world areas, including a neighbouring Northern Italian city. A north-south gradient of CC-prevalence was also observed. These results reinforce the notion of wide inter-regional variations in the frequency of C/T-13910 polymorphism and of incostant concordance with hypolactasic phenotype, even in subjects from the same country. Given the unsatisfactory results recently obtained from validation of a related assay in a neighbouring city, the authors decided not to proceed further and keep the assay only as a diagnostic aid in special situations.


Manfroi S.,Immunohematology and Transfusion Service | Scarcello A.,Immunohematology and Transfusion Service | Pagliaro P.,Immunohematology and Transfusion Service
Transfusion and Apheresis Science | Year: 2015

Molecular genetic studies on Duffy blood group antigens have identified mutations underlying rare FY*Null and FY*X alleles. FY*Null has a high frequency in Blacks, especially from sub-Saharan Africa, while its frequency is not defined in Caucasians. FY*X allele, associated with Fy(a-b + w) phenotype, has a frequency of 2-3.5% in Caucasian people while it is absent in Blacks. During the project of extensive blood group genotyping in patients affected by hemoglobinopathies, we identified FY*X/FY*Null and FY*A/FY*Null genotypes in a Caucasian thalassemic family from Sardinia. We speculate on the frequency of FY*X and FY*Null alleles in Caucasian and Black people; further, we focused on the association of FY*X allele with weak Fyb antigen expression on red blood cells and its identification performing high sensitivity serological typing methods or genotyping. © 2015 Elsevier Ltd.

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