Cumulus cell gene expression in luteal-phase-derived oocytes after double stimulation in one menstrual cycle.

RESEARCH QUESTION: Does double stimulation (DuoStim) affect cumulus cell gene expression in luteal-phase-derived oocytes? DESIGN: This prospective observational study included 39 patients with reduced ovarian reserve. Fifteen patients (group 1) underwent IVF with a gonadotrophin-releasing hormone antagonist in the follicular phase and 24 patients (group 2) underwent DuoStim. A total of 149 cumulus cell samples were divided into two groups according to the phase of the cycle: group 1 included 55 follicular-phase-derived oocytes and group 2 included 94 luteal-phase-derived oocytes. The expression levels of the following genes were assessed using quantitative polymerase chain reaction: HAS2, VCAN, ALCAM, PTGS2, GREM1, ITPKA, TRPM7, SDC4, CALM2, SPSB2, TP53I3, PGR and PFKP. RESULTS: The expression of 10 out of 13 genes in cumulus cells was similar between DuoStim luteal-phase-derived oocytes and follicular-phase-derived oocytes. A significant increase in the mRNA levels of VCAN (15.542 ± 6.8 versus 20.353 ± 10.58; P = 0.001), SDC4 (1.016 ± 0.65 versus 1.318 ± 0.97; P = 0.013), and TP53I3 (0.185 ± 0.09 versus 0.270 ± 0.11; P = 1.19E-05) was observed in group 2. The number of oocytes collected (5.57 ± 2.3 versus 5.7 ± 2.7; P > 0.05) and the number of blastocysts were comparable between the groups (2.1 ± 2.1 versus 2.7 ± 2.2; P > 0.05). CONCLUSIONS: The DuoStim approach leads to changes in the follicular environment. It affects the expression levels of VCAN, SDC4, and TP53I3 in the cumulus cells of luteal-phase-derived oocytes. These results, however, did not correlate with oocyte maturation, embryo quality and pregnancy rate.

Cell and genetic predictors of human blastocyst hatching success in assisted reproduction.

The aim was to identify cell and genetic predictors of human blastocyst hatching success in assisted reproduction programmes via a prospective case-control study. Blastocysts, donated by couples in assisted reproduction programmes were used. Hatching success assessment was performed after 144-146 h post-fertilization. The mRNA expression levels of cathepsin V (CTSV), GATA-binding protein 3 (GATA3) and human chorionic gonadotropin beta subunit 3, 5, 7 and 8 (CGB) genes were detected by quantitative real-time polymerase chain reaction. The odds ratio (OR) of hatching due to zona pellucida (ZP) thickness, oocyte and sperm quality, embryo quality and mRNA expression of CTSV, GATA3 and CGB genes in blastocysts was determined. From 62 blastocysts included in the study, 47 (75.8%) were unable to hatch spontaneously. The ZP thickening, and oocyte and sperm quality did not affect human blastocyst ability to hatch, except the combination of cytoplasmic and extracytoplasmic oocyte dysmorphisms (OR = 1.25; 95% confidence interval = 1.08, 1.45). Hatching-capable blastocysts had higher Gardner scale grade and mRNA expression of CTSV, GATA3 and CGB genes than hatching-incapable blastocysts. The human blastocyst hatching success depends on the blastocyst Gardner grade, but not on ZP and gamete quality. Blastocyst development was regulated by CTSV, GATA3 and CGB gene expression.

Cytokine gene expression in the skin and peripheral blood of atopic dermatitis patients and healthy individuals.

OBJECTIVES: Atopic dermatitis (AD) is an increasingly common, chronic, relapsing, inflammatory skin disease characterized by impaired epidermal barrier function and cutaneous inflammation. The prevalence of AD has steadily increased during the past few decades. The aim of this study was to comparatively investigate cytokine gene expression in the skin and peripheral blood of atopic dermatitis patients and healthy individuals. RESULTS: In the skin of patients with AD, a significant increase of the level of gene expression was observed for interleukin (IL)-2r (p < 0.0023), IL-5 (p = 0.002), IL-6 (p < 0.0023), IL-8 (p = 0.01), IL-12B (p < 0.0023), IL-10 (p < 0.0023), IL-23 (p = 0.002), IL-29 (p < 0.0023), and transforming growth factor beta (tGFbeta) (p < 0.0023) as compared to healthy individuals. In contrast, no difference between AD patients and healthy donors was detected with respect to cytokine gene expression in the peripheral blood. METHODS: Samples of skin and peripheral blood from 48 severe AD patients (SCORAD = 78.5 [57;89], IGA = 4.2 [3,9;4,7]) at the age of 17 to 45 years and 20 healthy donors aged from 19 to 32 years were analyzed for gene expression of cytokines using real-time reverse transcription polymerase chain reaction (RT-PCR). CONCLUSIONS: Activity of markers of chronic inflammation and Th1 immune response in severe AD, namely IL-2r, IL-8, IL-12B, IL-23, IL-29 and TGFbeta, as well as activity of anti-inflammatory IL-5 were predominant in the skin but not in the blood of AD patients.