Computational protocol: Glycoprotein fucosylation is increased in seminal plasma of subfertile men

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Protocol publication

[…] Digitalized gels and blots were subjected to densitometric analysis with ImageJ Image Processing and analysis software (National Institutes of Health, U.S. Department of Health and Human Services, Washington, DC, USA). Lectin reactivity was expressed in arbitrary units, defined as the optical density (OD) of an AAL-stained band of pooled seminal plasma containing 0.31 μg of protein. Thus, the AAL reactivity of band “x” was calculated as ODx/OD0.31 μg ref.To see whether the possible alterations in the lectin reactivity are related to the increased/decreased protein amount or to the intensity of the glycosylation process itself, the density of fucosylated epitopes was calculated as the ratio of AAL reactivity to protein content in each individual electrophoretic band. The latter was calculated from SyproRuby stained gels. [...] Statistica 10.0 software (StatSoft Inc., Tulsa, OK, USA) was used for the data analysis. The distribution of the results was presented as whisker-box diagrams, showing median values (central point) and 25%–75% quartiles (boxes). Mann–Whitney nonparametric U-test was used to verify statistically significant differences between fertile (F) and subfertile/infertile (IFtot) groups. To evaluate differences among the groups of seminal plasma samples distinguished by means of routine semen analysis (normozoospermic subfertile, oligozoospermic, asthenozoospermic, oligoasthenozoospermic) data, Kruskal–Wallis ANOVA rank test was applied. […]

Pipeline specifications

Software tools ImageJ, Statistica
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Homo sapiens
Diseases Infertility
Chemicals Fucose