1. Relationship between seminal plasma levels of anandamide congeners palmitoylethanolamide and oleoylethanolamide and semen quality 
Akwasi Atakora Amoako, B.Sc., M.B., Ch.B., M.R.C.O.G., Ph.D., Timothy Hywel Marczylo, Ph.D., Janine Elson, M.D., F.R.C.O.G., Anthony Henry Taylor, Ph.D., Jonathon M. Willets, Ph.D., Justin Chi Konje, M.D., F.R.C.O.G. 
Fertility and Sterility 
Volume 102, Issue 5, Pages (November 2014)
DOI: /j.fertnstert
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Relationship between seminal plasma levels of palmitoylethanolamide (PEA) and oleoylethanolamide (OEA) and semen quality. Seminal plasma levels of (A) PEA and (B) OEA were quantified by ultra-performance-liquid chromatography-tandem electrospray-ionization-mass spectrometry in men with normozoospermia, asthenozoospermia, teratozoospermia, azoospermia, and oligoasthenoteratozoospermia. Data are reported as median and interquartile range and all samples were processed as duplicates. Comparison between groups were made using Kruskal-Wallis one-way analysis of variance (ANOVA) followed by Dunn's ad hoc post test. ∗P, ∗∗P, ∗∗∗P represents data significantly (P<.05, P<.01, and P<.001 respectively) and ns represents data not significantly lower than normozoospermia. NOR = normozoospermia (n = 45); AST = asthenozoospermia (n = 11); TER = teratozoospermia (n = 8); AZO = azoospermia (n = 4); OAT = oligoasthenoteratozoospermia (n = 22).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Effect of palmitoylethanolamide (PEA) and oleoylethanolamide (OEA) on sperm motility. Human spermatozoa were incubated in Biggers, Whitten Whittingham (BWW) medium at 37°C in a 5% CO2 incubator and stimulated with (blue) or without (red) (A) PEA or (B) OEA (1 μM) for up to 90 minutes. Aliquots of 10 μL were taken at 15, 30, 45, 60, and 90 minutes and sperm motility was evaluated by means of phase contrast microscopy. Data are expressed as mean ± SEM (n = 4 separate experiments). Significant differences were determined by two-way analysis of variance (ANOVA). Pairwise comparison between treated and time-matched controls were determined by paired Student's t test; ∗P<.05, ∗∗P<.01, ∗∗∗P<.001. Human spermatozoa were incubated in BWW medium at 37°C and stimulated with or without increasing concentrations of PEA (C) or OEA (D) for up to 30 minutes. Aliquots of 10 μL were taken and sperm motility was evaluated by means of phase contrast microscopy and expressed as a percentage of motile sperm. Columns are mean ± SEM of six independent experiments performed in duplicate. Significant differences were determined by one-way ANOVA and Dunnett's post hoc test; ∗∗P<.01, ∗∗∗P<.001 versus control.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Effect of palmitoylethanolamide (PEA) and oleoylethanolamide (OEA) on sperm viability and mitochondria activity. Human spermatozoa were incubated in Biggers, Whitten Whittingham (BWW) medium at 37°C in a 5% CO2 incubator and stimulated with or without increasing concentrations of (A) PEA or (B) OEA (10 nM–10 μM) for up to 6 hours. Sperm viability was measured by the Live/Dead sperm viability assay and fluorescence microscopy and expressed as percentage of viable sperm. Columns are mean ± SEM of six independent experiments performed in duplicate. Significant differences were determined by one-way analysis of variance (ANOVA) and Dunnett's post hoc test. ∗∗P<.01, ∗∗∗P<.001 versus 0 nM PEA or OEA. Human spermatozoa were incubated in BWW medium at 37°C in a 5% CO2 incubator and stimulated with or without increasing concentrations of (C) PEA or (D) OEA (10 nM–1 μM) for up to 30 minutes. Aliquots of sperm suspension were stained with fluorescence dye JC-1 and mitochondria activity monitored by flow cytometry. The graphs indicate the percentage of sperm cells exhibiting orange fluorescence, indicating the higher mitochondrial activity, in the presence of the indicated doses of PEA or OEA. Data are expressed as mean ± SEM of four separate experiments, each run in duplicates.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions