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Summary The aim of this investigation was to determine the presence of abnormal sperm chromatin packaging in spermatozoa with large nuclear vacuoles (LNV) selected via high magnification by analysing the pattern of chromomycin A3 (CMA3) staining. A prospective observational study was designed to analyse semen samples obtained from 66 men undergoing infertility diagnosis and treatment. The numbers of cells with normal (dull yellow staining of the sperm head/CMA3‐negative) and abnormal (bright yellow fluorescence of the sperm head/CMA3‐positive) chromatin packaging were determined on slides with normal and LNV spermatozoa. The presence of bright yellow fluorescence (CMA3‐positive) was significantly higher (p < 0.0001) in spermatozoa with LNV than in normal spermatozoa (719/1351; 53.2% vs. 337/835; 40.3%, respectively), reflecting a higher percentage of abnormal chromatin packaging in spermatozoa with large LNV. Our data support the hypothesis that the presence of LNV reflects the presence of abnormal chromatin packaging, which may facilitate sperm DNA damage. As sperm nuclear vacuoles are evaluated more precisely at high magnifications using motile sperm organelle morphology examination (MSOME), the present results support the use of high‐magnification sperm selection for intracytoplasmic sperm injection (ICSI).

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A total of 150 patients were submitted to in-vitro fertilization (IVF) and endometrial pattern and thickness were assessed on the day of administration of human chorionic gonadotrophin (HCG). Ovarian stimulation was performed with gonadotrophins [human menopausal (HMG) or follicle stimulating hormone (FSH)] after down-regulation with leuprolide acetate. The endometrium was evaluated by vaginal ultrasound and classified into two groups: pattern I (a 'triple line' multilayer) and pattern II (fully homogeneous and hyperechogenic in relation to myometrial tissue). Pattern I was detected in 129 cycles (86%) and pattern II in 21 cycles (14%). The clinical pregnancy rates per cycle were similar (P = 0.79) for pattern I (29.4%) and pattern II (33.3%). There was no significant difference (P = 0.40) in the number of miscarriages between patients with pattern I and those with pattern II. Endometrial thicknesses were similar (10.3 +/- 2.0 mm and 11.2 +/- 3.1 mm) (P = 0.25). The thicknesses were similar (P = 0.14) for pregnant (10.8 +/- 2.1 mm) and non-pregnant (10.2 +/- 2.2 mm) women, but no pregnancies occurred when thickness was <7.0 mm. However, there was a significant difference (P = 0.04) between pregnant (10.8 +/- 1.9 mm) and non-pregnant (10.0 +/- 1.9 mm) women who showed pattern I. The conclusions from these data are that endometrial ultrasound in terms of pattern and thickness is of no prognostic value in IVF cycles on the day of administration of HCG. However, a minimum thickness has to be achieved for pregnancy to occur (7.0 mm). The presence of pattern I appears more likely to favour pregnancy.