1,721,412 research outputs found
SELECTION OF THE BEST OOCYTES FOR INTRACYTOPLASMICSPERM INJECTION (ICSI) USING APOPTOTIC ANALYSIS OF CUMULUS CELLS
Full text linkIntroduction: 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
A relationship between human sperm morphology and DNA fragmentation in infertile patients
No full textApoptosi in cellule del cumulo come possibile bio-marcatore molecolare di competenza ovocitaria: risultati di uno studio pilota condotto su pazienti sottoposte a diversi protocolli di stimolazione ovarica.
No full textL'apoptosi come criterio di valutazione comparativa tra attività LH e hCG
Il narcisismo e la Polis. La lezione di Alcibiade e lo scacco di Socrate
No full textL'articolo riprende l'analisi di un testo classico dei dialoghi platonici, L'Alcibiade, per riflettere sul narcisismo e il suo rapporto con la cultura del liberismo contemporane
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
Full text link: 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
- …
