Expressed proteins and activated pathways in conditioned embryo culture media from IVF patients are diverse according to infertility factors.

OBJECTIVE: Given that the embryo culture medium secretome reflects the embryo development, we hypothesize that protein profiles are affected according to infertility factors, which can be responsible for detrimental embryonic developmental competence. The aim of this study was to screen the protein profile of conditioned embryo culture media in patients presenting deep infiltrating endometriosis (ENDO) and polycystic ovarian syndrome (PCOS) undergoing IVF, by proteomics approaches. The control group was constituted by tubal factor patients. METHODS: Patients underwent in vitro fertilization (IVF) treatment as routine and oocytes were fertilized by ICSI. The embryos were group cultured until day 3 of development, and after transfer the culture media were collected. For the proteomics analysis, two pools of samples were prepared for groups CONTROL and PCOS, and 4 pools of samples for group DIE. Samples were prepared to deplete high abundant proteins and followed evaluated by high throughput proteomics approach. RESULTS: The embryonic organ and tissue development were physiological functions activated, based on proteins identified in the 3 study groups of samples. The samples coming from DIE patients presented a high calcium activity and on the other hand, embryos coming from PCOS patients showed a decreased calcium action. Other pathways as grow factors through the EGF signaling pathway overexpressed in ENDO culture medium and protein kinase A in PCOS were also observed. CONCLUSIONS: Proteomic embryonic secretome will advance our knowledge of early embryogenesis and additionally could lead to improved selection of embryos for transfer warrants further investigation.

Prognostic value of DNA and mRNA e6/e7 of human papillomavirus in the evolution of cervical intraepithelial neoplasia grade 2.

OBJECTIVE: This study aimed at evaluating whether human papillomavirus (HPV) groups and E6/E7 mRNA of HPV 16, 18, 31, 33, and 45 are prognostic of cervical intraepithelial neoplasia (CIN) 2 outcome in women with a cervical smear showing a low-grade squamous intraepithelial lesion (LSIL). METHODS: This cohort study included women with biopsy-confirmed CIN 2 who were followed up for 12 months, with cervical smear and colposcopy performed every three months. RESULTS: Women with a negative or low-risk HPV status showed 100% CIN 2 regression. The CIN 2 regression rates at the 12-month follow-up were 69.4% for women with alpha-9 HPV versus 91.7% for other HPV species or HPV-negative status (P < 0.05). For women with HPV 16, the CIN 2 regression rate at the 12-month follow-up was 61.4% versus 89.5% for other HPV types or HPV-negative status (P < 0.05). The CIN 2 regression rate was 68.3% for women who tested positive for HPV E6/E7 mRNA versus 82.0% for the negative results, but this difference was not statistically significant. CONCLUSIONS: The expectant management for women with biopsy-confirmed CIN 2 and previous cytological tests showing LSIL exhibited a very high rate of spontaneous regression. HPV 16 is associated with a higher CIN 2 progression rate than other HPV infections. HPV E6/E7 mRNA is not a prognostic marker of the CIN 2 clinical outcome, although this analysis cannot be considered conclusive. Given the small sample size, this study could be considered a pilot for future larger studies on the role of predictive markers of CIN 2 evolution.