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Rationale. Human amniotic fluid-derived (hAF) stem cells are considered a novel class of multipotent stem cells, sharing characteristics of both embryonic and adult stem cells. In fact, they proliferate rapidly, are able to differentiate into cells of all the embryonic germ layers, but do not form teratoma. It has been already reported that the embryoid bodies (EBs) obtained from hAFs have a cardiac potential, but it has not been described a functional terminal differentiation in cardiomyocytes (CMs) yet. Objective. Aim of this study was to foster the cardiomyogenic potential of hAFs in order to obtain a cellular population with morphological and functional features of CMs. Methods and Results. AFCs were exposed sequentially to inducing factors (Ascorbic Acid, 5-Azacytidine, BMP4, ActivinA, VEGF) up to 15 days and differentiation was monitored. Only the hAF samples expressing the multipotency markers SSEA4, OCT4 and CD90 (CardiopoieticAF) responded to the differentiation process loosing their stemness and increasing the cardiac nuclear factors NKX2.5 and GATA4. After the differentiation cells expressed high levels of the sarcomeric proteins (cTnT, αMHC and αSA), the gap junction marker Connexin43 and both atrial and ventricular markers; moreover, up to 90% of the cells was positive for CACNA1C and SERCA2a, cardiac calcium pumps involved in the excitation/contraction coupling, and about 30% of the CardiopoieticAF-derived cells presented spontaneous intracellular Ca2+ waves and Ca2+ fluctuation in response to caffeine or adrenergic stimulation. Some spontaneous beating foci were also observed. Conclusion. Our results demonstrate that CardiopoietichAFs can fully differentiate into a homogenous population of CM-like cells, characterized by cardiac-specific molecular, structural, and functional properties. Thus, CardiopoietichAFs can hold great promise for the development of in vitro models of cardiac genetic disorders, for drug discovery and testing, and for the emerging field of cardiovascular regenerative medicine., Italian Journal of Anatomy and Embryology, Vol. 122, No. 1 (Supplement) 2017

Purpose: To evaluate the effect of walking-training on serum angiogenic potential in postmenopausal women. Materials and Methods: Thirty-four postmenopausal women (56.18 ± 4.24 yr) participated in a 13 weeks program of walking-training. Anthropometric measures, VEGF, IL-1α, IL-1ß, IL-2, IL-8, IL-10, IL-12p70, TNF-α, C-reactive protein, insulin, IGF-1, cortisol, DHEA-S, leptin, visfatin, resistin and adiponectin were evaluated before and after training. Moreover, serum samples were tested for their ability to chemo – attract endothelial cells and to support the in vitro formation of capillary – like structures. Results: After training, the levels of IL-8, TNF-α, leptin and resistin were significantly lower, levels of DHEA-S and adiponectin increased, serum angiogenic properties improved, whereas no changes in anthropometric parameters or VEGF were detected. Conclusions: Walking training reduces inflammatory status and leads to a significant improvement in serum angiogenic properties in the absence of modifications in body composition and VEGF level., Italian Journal of Anatomy and Embryology, Vol. 122, No. 1 (Supplement) 2017

The concept of “the Core” describes the complex of anatomical components of the trunk, pelvis and shoulder girdle that are responsible for maintaining the stability of the spine and pelvis and are critical for the transfer of energy from larger torso to the smaller extremities, during many sport and daily-living activities (1). This concept rooted in sport science and rehabilitation and recently Core Training (CT) became very popular as a method to prevent injuries and improve sport performance and physical fitness. It consists in the progressive training of the musculature of the Core with special emphasis in posture and lumbar spine stability. This aspect requires a fine coordination and body awareness that often are poor developed or regressed after an injury. Then, it is important to include exercises of Sensorial Refinement (SR) that may stimulate the refinement of perceptually neglected areas (2). The aim of the study was to evaluate the effect of combined CT and SR on physical performance and to compare these effects with traditional core training. Furthermore, the effect on retention after 4 weeks of detraining was evaluated. Two groups of participants were recruited (age >30 < 50) and assigned to experimental (CT and SR: EXP, n = 9) or control (CT: CON, n = 9) group. Both groups trained ten weeks, with a frequency of two sessions per week. Training consisted in 10’ of warm up, 40’ of workout and 10’ of cool down. Workout of EXP group consisted in 20’ of SR and 20’ of CT whereas CON group performs 40’ of CT. Participants where tested by: Star Excursion Balance Test (SEBT) for the dynamic balance of lower body, Upper Quarter Y Balance Test (YBT-UQ) to assess upper extremities function in a closed-chain position and McGill test to evaluate muscular endurance of the core. Both groups improved core endurance after training and worsen it at follow up; moreover, both groups ameliorate upper an lower body control (SEBT and YBT-UQ) after training but only EXP group improved or maintained it at follow up. Since the information about the movements of the body are elaborated in the somatomotor cortex for fine coordination, the combination of CT and SR should better promote the consolidation of motor memory and long-term body control., Italian Journal of Anatomy and Embryology, Vol. 121, No. 1 (Supplement) 2016

Introduction: With menopause women face many changes that may lead to the loss of health related fitness, especially if sedentary. In particular, the estrogens deficiency affects the endothelial function thus increasing the incidence of the cardiovascular diseases. The cardio-protective effects of physical exercise is at least partially due to its ability to improve the health of arterial walls by influencing the endothelial function. Nonetheless, a direct angiogenetic of the physical exercise cannot be ruled out. VEGF is an important modulator of vascular growth but there are contrasting results about its response to physical exercise. Aim of our study was to compare the effects of two aerobic training on the VEGF levels and on the angiogenesis in postmenopausal women. Material and Methods. 34 Postmenopausal women underwent a 13 weeks training. In order to analyse the angiogentic effects, plasmatic VEGF levels were analysed before (T0) and after the training (T1). Moreover, the ability of the T0 and T1 serum to chemoattract endothelial cells and to induce them to form tubes was analyzed. Results: All post-menopausal women increase VEGF (P=0,014) after training. In vitro tests evidenced that when cultured in presence of T1 serum, HUVEC cells improved their ability to form tube (P, Italian Journal of Anatomy and Embryology, Vol 119, No 1 (Supplement) 2014

Purpose. Human amniotic fluid-derived stem cells (hAFSC) are multipotent stem cells sharing characteristics of both embryonic and adult stem cells. It has been already reported that hAFSC can differentiate toward cardiac lineage though the embryonic body (EB) formation, but the 3D structure and cellular heterogeneity of EB represent an important limitation. Aim of this study was to fully differentiate the hAFSC overcoming the EB limitations. Methods. hAFSC were obtained from normal amniocentesis. Cells cultured in monolayer were exposed sequentially to Ascorbic Acid, 5-Azacytidine, BMP4, ActivinA, VEGF up to 20 days. Differentiation was evaluated monitoring by Western Blot, immunofluorescent and cytometric analyses the expression of CD90, as mesenchymal stem cell marker, and of Nkx2.5, Gata4, sarcomeric α-actinin (αSA), α cardiac myosin heavy chain (αMHC), cardiac T-troponin (TnT)and Connexin43 as cardiac markers. Results. During the differentiation cultures cells underwent a progressive decrease of CD90 accompanied by the induction cardiac markers. After 15 days we evidenced that almost the entire cell population was positive for αMHC, αSA cTnT and Connexin 43 expressions (Table I); moreover, even if the % of Gata4+ and Nkx2.5+ cells did not varied during the culture, a significant increase of Nkx2.5 nuclear translocation (9.1±0.9% vs 18.0±1.8% Nkx2.5 nuclear positive cells in hAFSC and differentiated cells respectively, p, Italian Journal of Anatomy and Embryology, Vol 119, No 1 (Supplement) 2014

It is well known that menopause is associated with an increase of cardiovascular (CV) risk factors, including changes on metabolic profile, body composition, haemdynamic loads and cardiovascular remodelling. The latter starts with an increase of arterial elastance and early concentric left ventricular remodeling, manifested by an increment of the relative wall thickness. We investigated morphological and functional effects of short-term aerobic exercise at moderate intensity without diet restriction on cardiovascular apparatus in post-menopause women. Seventy-six post-menopausal healthy sedentary postwomen (56± 4 yrs) underwent clinical history, physician and anthropometric exam, 12-lead electrocardiography. Echocardiography was used to assess LV geometry and systolic and diastolic functions. Relative diastolic wall thickness (RWT), midwall fractional shortening (MFS) and arterial elastance (AE) were calculated to evaluate the LV concentric remodeling, the intrinsic systolic function and the vascular load, respectively. Data were collected before and after 14 weeks of moderate aerobic exercise training (four time per week). After the intervention program, systolic and diastolic arterial pressure, heart rate and hip circumference were significantly decreased (P = 0,01; P = 0,00; P = 0,00; P = 0,01, respectively). Left ventricular mass/height 2.7, ejection fraction and MFS were higher (p< 0,00; P= 0,00; P =0,00), while relative wall thickness (RWT) and AE were lower (P, Italian Journal of Anatomy and Embryology, Vol 118, No 2 (Supplement) 2013

Purpose. Cardiomyocytes express IL-6 and its signal transducer, 130-kDa glycoprotein (gp130) that influence cell growth, apoptosis, differentiation and survival by an autocrine pathway. High levels of circulating IL-6 have been reported in patients with congestive heart failure (HF) and after myocardial infarction and IL-6-gp130-signaling participates in HF prevention and compensatory hypertrophy, influencing remodeling processes and inducing Protein kinase C (PKC)-dependent apoptosis. Recent studies demonstrated the presence of precursor cells able to differentiate into cardiomyocytes in the adult heart through signaling pathways similar to those in the embryonic heart thus participating to the physiologic repair of the injured heart. In this line, the study of the embryonic signaling may represent an important cue to understand the molecular bases of the regenerative capacity myocardial progenitors. Aim of this study was to investigate the signal transduction pathways evoked by IL-6 treatment on cardiomyoblasts and its possible differentiating effects. Methods. H9c2 cells cultured in medium supplemented with 1% FBS (differentiation-promoting medium) in presence of IL-6 (10 ng/ml) up to three days. Results. Western Blot and Immunofluorence analysis demonstrated that Il-6 after 3 days of treatment induced a marked increase of the expression of α-myosin heavy chain (α-MHC; terminal cardiac differentiation marker) together with a microfilament reorganization and morphological modifications, that included mono-nucleated cells elongation and fusion into multinucleated tubes. This process was accompanied by an evident nuclear translocation of Nkx2.5 (early myocardial development transcription factor), and by a sub-cellular redistribution of gp130. Results suggest that the biological effects evoked by IL6 were at least partially mediated by PI3K through Akt and PKCzeta pathways modulation, as confirmed by the varied phosphorylated PI3K level observed after 30 min of IL-6 treatment and PI3K induction of downstream PKCzeta phosphorilation, but not PKCalpha and PKCdelta, whereas Akt de-phosphorylation was detected. Conclusions. Our observations evidenced that PI3K is a key regulator of IL-6- induction of embryonic cardiomyocytes differentiation by PKCzeta and Akt-signalling pathways. These data suggest a possible role of IL-6 dependent pathways in regenerative capacity of myocardial cells following injuries., Italian Journal of Anatomy and Embryology, Vol 116, No 1 (Supplement) 2011

Italian Journal of Anatomy and Embryology, Vol 115, No 1/2 (Supplement) 2010

Italian Journal of Anatomy and Embryology, Vol 115, No 1/2 (Supplement) 2010