Research Article Artificial Neural Network to Predict Varicocele Impact on Male Fertility through Testicular Endocannabinoid Gene Expression Profiles Davide Perruzza , 1 Nicola Bernabò , 1 Cinzia Rapino , 2 Luca Valbonetti, 1 Ilaria Falanga, 1 Valentina Russo, 1 Annunziata Mauro , 1 Paolo Berardinelli, 1 Liborio Stuppia, 3 Mauro Maccarrone , 4,5 and Barbara Barboni 1 1 Faculty of Biosciences and Technology for Food, Agriculture and Environment, University of Teramo, 64100 Teramo, Italy 2 Faculty of Veterinary Medicine, Agriculture and Environment, University of Teramo, 64100 Teramo, Italy 3 Department of Psychological, Health and Territorial Sciences, School of Medicine and Health Sciences, University “G. d’Annunzio” of Chieti and Pescara, 66100 Chieti, Italy 4 Department of Medicine, Campus Bio-Medico University of Rome, 00128 Rome, Italy 5 European Center for Brain Research, IRCCS Santa Lucia Foundation, 00164 Rome, Italy Correspondence should be addressed to Davide Perruzza; davideperruzza86@gmail.com Received 22 June 2018; Accepted 1 November 2018; Published 13 November 2018 Academic Editor: Pradeep Tyagi Copyright © 2018 Davide Perruzza et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Te relationship between varicocele and fertility has always been a matter of debate because of the absence of predictive clinical indicators or molecular markers able to defne the severity of this disease. Even though accumulatedevidence demonstrated that the endocannabinoid system (ECS) plays a central role in male reproductive biology, particularly in the testicular compartment, to date no data point to a role for ECS in the etiopathogenesis of varicocele. Terefore, the present research has been designed to investigate the relationship between testicular ECS gene expression and fertility, using a validated animal model of experimental varicocele (VAR), taking advantage of traditional statistical approaches and artifcial neural network (ANN). Experimental induction of VAR led to a clear reduction of spermatogenesis in lef testes ranging from a mild (Johnsen score 7: 21%) to a severe (Johnsen score 4: 58%) damage of the germinal epithelium. However, the mean number of new-borns recorded afer two sequential matings was quite variable and independent of the Johnsen score. While the gene expression of biosynthetic and degrading enzymes of AEA (NAPE- PLD and FAAH, respectively) and of 2-AG (DAGL and MAGL, respectively), as well as their binding cannabinoid receptors (CB 1 and CB 2 ), did not change between testes and among groups, a signifcant downregulation of vanilloid (TRPV1) expression was recorded in lef testes of VAR rats and positively correlated with animal fertility. Interestingly, an ANN trained by inserting the lef and right testicular ECS gene expression profles (inputs) was able to predict varicocele impact on male fertility in terms of mean number of new-borns delivered (outputs), with a very high accuracy (average prediction error of 1%). Te present study provides unprecedented information on testicular ECS gene expression patterns during varicocele, by developing a freely available predictive ANN model that may open new perspectives in the diagnosis of varicocele-associated infertility. 1. Introduction Varicocele is considered among the most common causes of male infertility and afects approximately 15-20% of the general male population. It is reported in 19-41% of men with primary infertility, as well as in up to 80% of men with secondary infertility [1]. Since the frst evidence supporting a link between varicocele and infertility was pub- lished [2, 3], an extensive research efort has been addressed to elucidate underling mechanisms and their relationship with varicocele-associated testicular dysfunction and male fertility outcome. Varicocele is functionally related to an abnormal dilation and tortuosity of the pampiniform plexus veins, which are Hindawi BioMed Research International Volume 2018, Article ID 3591086, 15 pages https://doi.org/10.1155/2018/3591086