New Technique for Aortic Valve Functional Annulus Reshaping Using a Handmade Prosthetic Ring

Background. Despite a wide development in aortic leaflets repair techniques, aortic valve annuloplastic procedures are still poorly investigated. We present our aortic valve annuloplastic system consisting of a handmade prosthetic ring with 2 components for reshaping the aortic annulus and sinotubular junction (STJ) and illustrates our surgical approach and clinical results. Methods. Since February 2003, 45 patients with aortic valve regurgitation underwent aortic annuloplasty using the new ring. Mean patient age was 58 16 years (range, 46 to 76 years). The ring has 2 components: a circular ring used to undersize the circumference of the aortoventricular junction and 3-crown-like shape ring used for STJ remodeling. The circular ring was sutured into the left ventricular outflow tract in the aortic subvalvular position and the STJ ring was sutured from the outside of the aortic root just at the level of the STJ. The 3 vertical bands of the STJ ring were fixed to the underlying circular ring to stabilize the continuity between the STJ and nadir of the aortic valve. Results. No in-hospital death occurred. Two patients had residual trivial aortic valve regurgitation postoperatively, and 3 patients required treatment for residual aortic regurgitation (more than mild). Mean length of the coaptation surface was 10 2 mm. The mean clinical follow-up (100% complete) was 22 16 months. All patients were free from cardiac and valve-related events, and no complications due to ring implantation occurred. Conclusions. This ring reshapes the functional aortic annulus and stabilizes all components of the native aortic valve stent to improve long-term results of valve repair

Toll-like receptor-4 signaling pathway in aorta aging and diseases: "its double nature"

Recent advances in the field of innate immunity have revealed a complex role of innate immune signaling pathways in both tissue homeostasis and disease. Among them, the Toll-like receptor 4 (TLR-4) pathways has been linked to various pathophysiological conditions, such as cardiovascular diseases (CVDs). This has been interrogated by developing multiple laboratory tools that have shown in animal models and clinical conditions, the involvement of the TLR-4 signaling pathway in the pathophysiology of different CVDs, such as atherosclerosis, ischemic heart disease, heart failure, ischemia-reperfusion injury and aorta aneurysm. Among these, aorta aneurysm, a very complex pathological condition with uncertain etiology and fatal complications (i.e. dissection and rupture), has been associated with the occurrence of high risk cardiovascular conditions, including thrombosis and embolism. In this review, we discuss the possible role of TLR-4signaling pathway in the development of aorta aneurysm, considering the emerging evidence from ongoing investigations. Our message is that emphasizing the role of TLR-4signaling pathway in aorta aneurysm may serve as a starting point for future studies, leading to a better understanding of the pathophysiological basis and perhaps the effective treatment of this difficult human disease

Lavoro, organizzazione e cura

Il volume raccoglie contributi sul significato del lavoro nei contesti organizzativi e nei contesti di cura

New molecular markers for the evaluation of gamete quality

Purpose: Only 30 % of IVF cycles result in a pregnancy, so that multiple embryos need to be replaced, per treatment cycle, to increase pregnancy rates, resulting in a multiple gestation rate of 25 %. The use of new markers in the gamete selection, could reduce the number of the oocytes to be fertilized and embryos to be produced, but the tools to evidence the gamete competence remain unavailable and more studies are needed to identify bio-markers to select the best oocyte and sperm to produce embryos with higher implantation potentiality. Methods: To define oocyte competence, the apoptosis of the surrounding cumulus cells and the oxygen consumption rates for individual oocytes before fertilization seems to provide a non-invasive marker of oocyte competence and hence a quantitative assessment of the reproductive potential for the oocyte. The chromatin integrity seems to be used also as biological marker of sperm competence, together with the morphological evaluation of large vacuoles in the head. Results: The apoptosis rate of cumulus cells lower than 25 % and an higher oxygen consumption could be an evidence of an overall metabolic activity, related to a better fertilization ability and embryo cleavage quality. The apoptosis rate of the sperm chromatin, evaluated by direct Tunel in situ analysis, seems to be, also for the male gamete, a marker of competence and implantation potentiality, in particular when it is lower than 20 %. The evaluation of the presence of large vacuoles in the sperm head prior to perform ICSI seems to increase the implantation rate, but it is not associated to chromatin integrity. Conclusions: The biological concept of competence appears unrelated to any morphological parameters, so that it is necessary to investigate new molecular markers in the gamete selection. Apoptosis of cumulus cells in the oocytes and spermatozoa, revealing the presence of large vacuoles, could help to determine the competence of the gamete to be fertilize. © 2013 Springer Science+Business Media New York

AKT, p-AKT, ERK1/2 and p-ERK1/2 in Mural Granulosa Cells Are Not Correlated to Different Ovarian Stimulation Protocols in Patients Undergoing Assisted Reproductive Treatment

(1) Background: In this paper we aim to study the relationship between the expression levels of molecules involved in apoptotic/survival pathways, considered as molecular markers of oocyte competence (i.e., AKT, p-AKT, ERK1/2, and p-ERK1/2) in mural granulosa cells (MGCs) and the administration of r-FSH alone or combined with exogenous r-LH, in ovarian stimulation protocol. Moreover, we aim to evaluate oocyte competence by comparing normally cleaved embryos that were transferred in the uterus, with embryos that were arrested during in vitro culture. (2) Methods: The study included 34 normo-responder women undergoing ICSI procedures. All subjects were divided into two groups. Group A consisted of 18 women stimulated with r-FSH and used as a control group; Group B consisted of 14 women stimulated with r-FSH combined with r-LH. The MGCs were obtained from individual follicles. Immunoblot analyses were carried out to analyze the AKT, p-AKT, ERK1/2, and p-ERK1/2 levels in MGCs and to correlate them with the ovarian stimulation protocol. Furthermore, the oocyte competence was evaluated, for each follicle, according to the development of the embryo during in vitro culture and the pregnancy outcome. (3) Results: We found no significant difference in the levels of molecules in isolated MGCs between groups A and B. These results, in light of our previous research, suggest for the first time, to our knowledge, that cumulus cells and mural granulosa cells in the same follicle show different expression levels of molecules involved in the apoptotic mechanism. (4) Conclusions: Our results could clarify some controversial data in the literature where cumulative cell pools of cumulus and granulosa were analyzed, described as ovarian follicle cells, and used as markers of oocyte competence. In this paper, we found evidence that cumulus and granulosa cells need to be analyzed separately

Oxidative Stress in the Pathogenesis of Aorta Diseases as a Source of Potential Biomarkers and Therapeutic Targets, with a Particular Focus on Ascending Aorta Aneurysms

: Aorta diseases, such as ascending aorta aneurysm (AsAA), are complex pathologies, currently defined as inflammatory diseases with a strong genetic susceptibility. They are difficult to manage, being insidious and silent pathologies whose diagnosis is based only on imaging data. No diagnostic and prognostic biomarkers or markers of outcome have been known until now. Thus, their identification is imperative. Certainly, a deep understanding of the mechanisms and pathways involved in their pathogenesis might help in such research. Recently, the key role of oxidative stress (OS) on the pathophysiology of aorta disease has emerged. Here, we describe and discuss these aspects by revealing some OS pathways as potential biomarkers, their underlying limitations, and potential solutions and approaches, as well as some potential treatments

Are Endothelial Progenitor Cells the Real Solution for Cardiovascular Diseases? Focus on Controversies and Perspectives

Advanced knowledge in the field of stem cell biology and their ability to provide a cue for counteracting several diseases are leading numerous researchers to focus their attention on \u201cregenerative medicine\u201d as possible solutions for cardiovascular diseases (CVDs). However, the lack of consistent evidence in this arena has hampered the clinical application. The same condition affects the research on endothelial progenitor cells (EPCs), creating more confusion than comprehension. In this review, this aspect is discussed with particular emphasis. In particular, we describe biology and physiology of EPCs, outline their clinical relevance as both new predictive, diagnostic, and prognostic CVD biomarkers and therapeutic agents, discuss advantages, disadvantages, and conflicting data about their use as possible solutions for vascular impairment and clinical applications, and finally underline a very crucial aspect of EPCs \u201ccharacterization and definition,\u201d which seems to be the real cause of large heterogeneity existing in literature data on this topic

APOPTOTIC ANALYSIS OF CUMULUS CELLS FOR THE SELECTION OF COMPETENT OOCYTES TO BE FERTILIZED BY INTRACYTOPLASMIC SPERM INJECTION (ICSI)

Oocyte quality is one of the main factors for the success of in vitro fertilization protocols. Apoptosis is known to affect oocyte quality and may impair subsequent embryonic development and implantation. The aim of this study was to investigate the apoptosis rate of single and pooled cumulus cells of cumulus cell\u2013oocyte complexes (COCs), as markers of oocyte quality, prior to intracytoplasmatic sperm injection (ICSI).We investigated the apoptosis rate by TUNEL assay (DNA fragmentation) and caspase-3 immunoassay of single and pooled cumulus cells of COCs. The results showed that DNA fragmentation in cumulus cells was remarkably lower in patients who achieved a pregnancy than in those who did not. Cumulus cell apoptosis rate could be a marker for the selection of the best oocytes to be fertilized by intracytoplasmatic sperm injection

SELECTION OF THE BEST OOCYTES FOR INTRACYTOPLASMICSPERM INJECTION (ICSI) USING APOPTOTIC ANALYSIS OF CUMULUS CELLS

Introduction: We studied the apoptosis rate of the cumulus cells of individual cumulus-oocyte complex (COC), to verify a relationship with clinical outcomes, in terms of pregnancy and implantation rates. Usually oocytes are selected using morphological criteria. We tried to verify if cumulus cell apoptotic rate could be used as molecular criteria in selecting oocytes with higher implantation potentiality (1;2). Materials and Methods: The study design consisted in two different trials: in the first, we investigated apoptosis rate in cumulus cells of the three selected oocytes, to be fertilized by intracytoplasmic sperm injection (ICSI); in a second trial, average apoptosis rate of the cumulus cells coming from the three selected oocytes to be fertilized by ICSI and the pooled remaining oocytes were compared, when more than 5 COCs were aspirated. In a first trial we included 22 consecutive couples undergoing ICSI cycles, 20 in a second one, for a total of 42 patients. We selected the three oocytes for (ICSI) on the basis of the morphological appearance of the cumulus, according to Veek’s criteria. The cumulus cells of each COC were submitted to apoptotic assays (3). The patients were classified, on the basis of pregnancy success, in A Group (pregnant patients) and B Group (patients with negative βhCG). Results: Both trials showed that apoptosis in the cumulus cells was remarkably lower in the A Group if compared with B Group. The apoptosis rate in the selected COCs was similar to pooled COCs for each patient, confirming that apoptosis rate in cumulus cells is characteristic for patient. Out of 22 patients involved in the first trial, 8 were pregnant (36.3% A Group) and 14 were not pregnant (B Group). In the second trial 4 of a total of 20 patients were pregnant (20%). In the first trial a total of 58 metaphase II oocytes and 56 in the second trial were studied. In the second trial 38 oocytes where pooled to compare apoptosis rate with the three selected oocytes pools. In the first trial the incidence of DNA fragmentation, evaluated by TUNEL assay (fig. 1), of the cumulus cells from individual treated oocytes, was lower in A Group than in B Group (6.7% ranging between 2.2–13.3 vs 13.19% ranging between 6.2–34.9 respectively, p<0.05). To confirm if DNA fragmentation was related to apoptosis process, we performed caspase-3 immunoassay in the same cells (fig. 2). Data showed a lower capase-3 activity in cumulus cells of pregnant than in those of non-pregnant patients (5.2% ranging between 1.2–8.6 vs 11.8% ranging between 5.6–14.8, p<0.05). It is noteworthy to underline that pregnant patients usually exhibited, at least, one COC with a DNA fragmentation rate (TUNEL) less than 10% and caspase-3 activity rate less than 7%. Four (A Group) of 20 patients involved in the second trial were pregnant but two aborted at 8–9 weeks. The low number of pregnant patients did not allow us to have a powerful statistical analysis of apoptotic rate in cumulus cells, but it seems evident that a higher apoptotic rate in cumulus cells is associated to the pregnancy failure (B Group) and in aborted patients of A Group, ranging from 10 to 60.3%. Conclusion: The data seem to demonstrate that apoptosis may be a marker for the selection of the best oocytes to be submitted to ICSI treatment. All pregnant patients showed a lower apoptosis rate in cumulus cells if compared with patients with pregnancy failure. 86° CONGRESSO NAZIONALE SIBS - PALERMO 24-25 OTTOBRE 2013 72 References 1. Ruvolo G, Bosco L, Pane A, Morici G, Cittadini E, Roccheri MC. Lower apoptosis rate in human cumulus cells after administration of recombinant luteinizing hormone to women undergoing ovarian stimulation for in vitro fertilization procedures. Fertil Steril. 2007 Mar; 87(3):542-6. Epub 2006 Nov 27. 2. Host E, Gabrielsen A, Lindenberg S, Smidt-Jensen S 2002 Apoptosis in human cumulus cells in relation to zona pellucida thickness variation, maturation stage, and cleavage of the corresponding oocyte after intracytoplasmic sperm injection. Fertility and Sterility 77, 511-515. 3. Bosco L, Ruvolo G, Morici G, Manno M, Cittadini E, Roccheri MC. Apoptosis in human unfertilized oocytes after intracytoplasmic sperm injection. Fertil Steril. 2005 Nov; 84(5):1417- 23. FIGURE 1. Apoptosis evaluation using TUNEL assay in human cumulus cells. (A1, A2, A3) A group; (B1, B2, B3) B group; (C1, C2, C3) positive control for TUNEL assay. (A1, B1, C1) fragmented DNA; (A2, B2, C3) propidium iodide staining; (A3, B3, C3) merge. Scale bar = 15 μm. FIGURE 2. Apoptosis evaluation using Cleaved caspase 3 immunofluorescence in situ assay in human cumulus cells. (A1, A2, A3) A group; (B1, B2, B3) B group; (A1, B1) Cleaved caspase 3; (A2, B2) propidium iodide staining; (A3, B3) merge. Scale bar = 15 μm