Previously, we reported apolipoprotein A-I (apoA-I), the major protein component of high-density lipoprotein (HDL), has potent anti-melanoma activity. We used DNA microarray and bioinformatics to interrogate gene expression profiles of tumors from apoA-I expressing (A-I Tg+/-) versus apoA-I-null (A-I KO) animals to gain insights into mechanisms of apoA-I tumor protection. Differential expression analyses of 11 distinct tumors per group with > 1.2-fold cut-off and a false discovery rate adjusted p < 0.05, identified 176 significant transcripts (71 upregulated and 105 downregulated in A-I Tg+/- versus A-I KO group). Bioinformatic analyses identified the mevalonate and de novo serine/glycine synthesis pathways as potential targets for apoA-I anti-tumor activity. Relative to A-I KO, day 7 B16F10L melanoma tumor homografts from A-I Tg+/- exhibited reduced expression of mevalonate-5-pyrophosphate decarboxylase (Mvd), a key enzyme targeted in cancer therapy, along with a number of key genes in the sterol synthesis arm of the mevalonate pathway. Phosphoglycerate dehydrogenase (Phgdh), the first enzyme branching off glycolysis into the de novo serine synthesis pathway, was the most repressed transcript in tumors from A-I Tg+/-. We validated our mouse tumor studies by comparing the significant transcripts with adverse tumor markers previously identified in human melanoma and found 45% concordance. Our findings suggest apoA-I targets the mevalonate and serine synthesis pathways in melanoma cells in vivo, thus providing anti-tumor metabolic effects by inhibiting the flux of biomolecular building blocks for macromolecule synthesis that drive rapid tumor growth.

Aims: To understand the molecular pathways involved in oxidative stress (OS)-mediated sperm dysfunction against a hypoxic and hyperthermic microenvironment backdrop of varicocele through a proteomic approach. Results: Protein selection (261) based on their role in redox homeostasis and/or oxidative/hyperthermic/hypoxic stress response from the sperm proteome data set of unilateral varicocele (UV) in comparison with fertile control displayed 85 to be differentially expressed. Upregulation of cellular oxidant detoxification and glutathione and reduced nicotinamide adenine dinucleotide (NADH) metabolism accompanied with downregulation of protein folding, energy metabolism, and heat stress responses were observed in the UV group. Ingenuity pathway analysis (IPA) predicted suppression of oxidative phosphorylation (OXPHOS) (validated by Western blotting [WB]) along with augmentation in OS and mitochondrial dysfunction in UV. The top affected networks indicated by IPA involved heat shock proteins (HSPs: HSPA2 and HSP90B1). Their expression profile was corroborated by immunocytochemistry and WB. Hypoxia-inducible factor 1A as an upstream regulator of HSPs was predicted by MetaCore. Occurrence of reductive stress in UV spermatozoa was corroborated by thiol redox status. Innovation: This is the first evidence of a novel pathway showing aberrant redox homeostasis against chronic hypoxic insult in varicocele leading to sperm dysfunction. Conclusions: Upregulation of antioxidant system and dysfunctional OXPHOS would have shifted the redox balance of biological redox couples (GSH/GSSG, NAD+/NADH, and NADP+/NADPH) to a more reducing state leading to reductive stress. Chronic reductive stress-induced OS may be involved in sperm dysfunction in infertile men with UV, where the role of HSPs cannot be ignored. Intervention with antioxidant therapy warrants proper prior investigation.

Background and challenges to implementation In India, more than 275 million adults use tobacco, which kills 1.2 million of these every year. More than one-quarter of India´s youth begin tobacco use before they are 16 years old and 85% of all users begin before they are 18, the legal age of tobacco use. A diversity of tobacco products spurs India´s hidden epidemic. In May 2003, the Indian Parliament passed a landmark tobacco control legislation - the Cigarettes and other Tobacco Products Act (COTPA). Following this the Government of India launched the National Tobacco Control Programme (NTCP) in 2007, with the aim to protect children and youth and to enforce the tobacco control legislation. Intervention or response We assess the progress of tobacco control efforts in the states by analysing the compliance towards provisions of COTPA and achieving goals of The NTCP. We use third party compliance studies, analyses done by civil societies, orders and action taken reports of state governments and district administration, and media report from states. We prepare a semi-structured report card based on compliance towards criteria specified under NTCP and COTPA. Results and lessons learnt The achievement in tobacco control have been mixed. We will present a state-wise, updated assessment of performance of tobacco control efforts in India, and identify drivers for their success and reasons for their limited progress. We find that political and administrative commitment and leadership are critical to get tobacco control started at sub-national level. Conclusions and key recommendations Sustainable enforcement and adoption of evidence-based strategies, investing in developing skilled human resources, and financial commitments (especially co-investments from sub-national sources) are key factors in achieving goals under National Tobacco Control Programme and ensuring long-term gains that advance tobacco control.

PURPOSE:Varicocele may disrupt testicular microcirculation and induce hypoxia-ischemia related degenerative changes in testicular cells and spermatozoa. Superoxide production at low oxygen concentration exacerbates oxidative stress in men with varicocele. Therefore, the current study was designed to study the role of mitochondrial redox regulation and its possible involvement in sperm dysfunction in varicocele associated infertility.MATERIALS AND METHODS:We identified differentially expressed mitochondrial proteins in 50 infertile men with varicocele and in 10 fertile controls by secondary liquid chromatography-tandem mass spectroscopy data driven in silico analysis. Identified proteins were validated by Western blot and immunofluorescence. Seminal oxidation-reduction potential was measured.RESULTS:We identified 22 differentially expressed proteins related to mitochondrial structure (LETM1, EFHC, MIC60, PGAM5, ISOC2 and import TOM22) and function (NDFSU1, UQCRC2 and COX5B, and the core enzymes of carbohydrate and lipid metabolism). Cluster analysis and 3-dimensional principal component analysis revealed a significant difference between the groups. All proteins studied were under expressed in infertile men with varicocele. Liquid chromatography-tandem mass spectroscopy data were corroborated by Western blot and immunofluorescence. Impaired mitochondrial function was associated with decreased expression of the proteins (ATPase1A4, HSPA2, SPA17 and APOA1) responsible for proper sperm function, concomitant with elevated seminal oxidation-reduction potential in the semen of infertile patients with varicocele.CONCLUSIONS:Impaired mitochondrial structure and function in varicocele may lead to oxidative stress, reduced ATP synthesis and sperm dysfunction. Mitochondrial differentially expressed proteins should be explored for the development of biomarkers as a predictor of infertility in patients with varicocele. Antioxidant therapy targeting sperm mitochondria may help improve the fertility status of these patients.

The seminal plasma protein profile is a reflection of the spermatozoa homeostasis. This study aimed to validate, using Western Blot (WB), potential biomarkers identified in the seminal plasma of men with primary or secondary infertility in comparison with fertile donors. This study includes 18 sperm samples from proven fertile men, men with primary infertility and men with secondary infertility (n=6/group). Samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the differently expressed proteins (DEPs) in the infertile group and key DEPs were validated using WB. Proteomic profiles of seminal plasma from fertile men, men with primary or secondary infertility were obtained by mass-spectrometry. Based on the functional bioinformatics analysis using IPA, Metacore and public annotation databases key DEPs associated with infertility related processes were selected for validation by WB. For primary infertility, the chosen proteins were annexin A2, transferrin, cell division control protein 42, CD63, peroxiredoxin 2, semenogelin 1 and 2. For secondary infertility, we selected annexin A2, complement C4, complement C3, heat shock protein 70, semenogelin 1 and amyloid precursor protein. The statistical analysis of WB results was performed in the MedCalc using a Mann-Whitney test (p<0.05). Our validation using WB showed an overexpression of annexin A2 (p=0.03) in men with primary infertility when compared to fertile donors, which was concordant with the mass spectrometry results. In men with secondary infertility, we observed an underexpression of complement C3 (p=0.03) relative to fertile men. However, this result did not match with the proteomic analysis that indicated an overexpression of complement C3. Annexin A2, which is involved in sperm binding, may be a potential biomarker for the diagnosis of men with primary infertility. Further validation of additional proteins is necessary to identify other candidate proteins that may also be implicated in male infertility.

POTENTIAL BIOMARKERS FOR MEN WITH PRIMARY INFERTILITY Ana Dias Martins1,2, Ashok Agarwal1, Luna Samanta3, Rakesh Sharma1, Banu Gopalan4, Belinda Willard5, Zhihong Cui6, Damayanthi Durairajanayagam7, Edmund Sabanegh8 1American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, U.S.A. 2Dep. Microscopia Lab. de Biologia Celular and Unit for Multidisciplinary Research in Biomedicine, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, Portugal. 3Department of Zoology, Ravenshaw University, Odisha, India. 4Bioinformatics, Yorg Corporation, Plano, U.S.A. 5Research Core Services Proteomics, Cleveland Clinic, Cleveland, U.S.A. 6College of Pharmaceutical Sciences, Southwest University, Chongqing, China. 7Discipline of Physiology-Faculty of Medicine, Universiti Teknologi MARA, Selangor, Malaysia. 8Urology, Cleveland Clinic, Cleveland, U.S.A.

PROTEINS IN SEMINAL PLASMA OF INFERTILE MEN WITH UNILATERAL VARICOCELE Manesh Kumar Panner Selvam1, Ashok Agarwal1, Luna Samanta2, Rakesh Sharma1, Sajal Gupta1, Damayanthi Durairajanayagam3, Zhihong Cui4, Ahmet Ayaz5, Banu Gopalan6, Belinda Willard7, Edmund Sabanegh8 1American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, U.S.A. 2Department of Zoology, Ravenshaw University, Odisha, India. 3Discipline of Physiology-Faculty of Medicine, Universiti Teknologi MARA, Selangor, Malaysia. 4College of Pharmaceutical Sciences, Southwest University, Chongqing, China. 5Department of Urology, Tulane Medical Center, New Orleans, U.S.A. 6Bioinformatics, Yorg Corporation, Plano, U.S.A. 7Research Core Services Proteomics, Cleveland Clinic, Cleveland, U.S.A. 8Urology, Cleveland Clinic, Cleveland, U.S.A.

Non-small cell lung cancer (NSCLC) is the major form of lung cancer, with adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC) being its major subtypes. Smoking alone cannot completely explain the lung cancer etiology. We hypothesize that altered lung microbiome and chronic inflammatory insults in lung tissues contribute to carcinogenesis. Here we explore the microbiome composition of LUAD samples, compared to LUSC and normal samples. Extraction of microbiome DNA in formalin-fixed, paraffin-embedded (FFPE) lung tumor and normal adjacent tissues was meticulously performed. The 16S rRNA product from extracted microbiota was subjected to microbiome amplicon sequencing. To assess the contribution of the host genome, CD36 expression levels were analyzed then integrated with altered NSCLC subtype-specific microbe sequence data. Surprisingly phylum Cyanobacteria was consistently observed in LUAD samples. Across the NSCLC subtypes, differential abundance across four phyla (Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes) was identified based on the univariate analysis (p-value < 6.4e-4 to 3.2e-2). In silico metagenomic and pathway analyses show that presence of microcystin correlates with reduced CD36 and increased PARP1 levels. This was confirmed in microcystin challenged NSCLC (A427) cell lines and Cyanobacteria positive LUAD tissues. Controlling the influx of Cyanobacteria-like particles or microcystin and the inhibition of PARP1 can provide a potential targeted therapy and prevention of inflammation-associated lung carcinogenesis.

To identify proteomic markers in semen samples of fertile men with high reactive oxygen species (ROS) levels. Normozoospermic men (WHO 2010) who fathered a child in the last two years were divided into two groups; control (N=10; ROS < 93 RLU/s/106 sperm) and ROS+ (N=10; ROS > 93 RLU/s/106 sperm). Spermatozoa and seminal plasma were analyzed separately for proteomics and Western blot (WB) analysis. Semen samples were collected after 2-5 days of abstinence from 20 proven fertile men after Institutional Review Board approval. ROS levels were evaluated by a luminol-based chemiluminescence assay. Liquid chromatography-tandem mass spectrometry (LC/MS-MS) was used for quantitative proteomic analysis to identify the differentially expressed proteins (DEPs). Using Ingenuity Pathways Analysis, DEPs involved in molecular pathways related to redox balance were selected and validated by WB (N=5/group). WB results were analyzed by Mann-Whitney test and considered significant when P < 0.05. The DEPs identified in spermatozoa (371) and seminal plasma (44) are primarily involved in oxireductase, endopeptidase inhibitor, and antioxidant activities. Among the selected DEPs, the mitochondrial NADH:Ubiquinone Oxidoreductase Core Subunit S1 (NDUFS1) was underexpressed (P=0.01), while several antioxidant proteins including superoxide dismutase 1 (SOD1; P=0.03) and peroxiredoxin 4 (PRDX4; P=0.04) were overexpressed in spermatozoa of ROS+ group as revealed by both proteomics and WB. Similarly, seminal plasma proteomic data showed that haptoglobin, S100 calcium-binding protein A9, PRDX4, and Serpin B6 were overexpressed in ROS+ group. However, no significant difference was observed in their expression level when analyzed by WB. Preservation of fertility in ROS+ men may be facilitated by the overexpression of antioxidant proteins. Proteins SOD1 and PRDX4 may serve as potential biomarkers for the management of infertile men with high ROS level from different etiologies.

Increasing evidence suggests that hyperphosphorylation and aggregation of microtubule-associated protein tau (MAPT or tau) correlates with the development of cognitive impairment in Alzheimer's disease (AD) and related tauopathies. While numerous attempts have been made to model AD-relevant tau pathology in various animal models, there has been very limited success for these models to fully recapitulate the progression of disease as seen in human tauopathies. Here, we performed whole genome gene expression in a genomic mouse model of tauopathy that expressed human MAPT gene under the control of endogenous human MAPT promoter and also were complete knockout for endogenous mouse tau [referred to as 'hTau MaptKO(Duke)' mice]. First, whole genome expression analysis revealed 64 genes, which were differentially expressed (32 up-regulated and 32 down-regulated) in the hippocampus of 6-month-old hTau MaptKO(Duke) mice compared to age-matched non-transgenic controls. Genes relevant to neuronal function or neurological disease include up-regulated genes: PKC-alpha (Prkca), MECP2 (Mecp2), STRN4 (Strn4), SLC40a1 (Slc40a1), POLD2 (Pold2), PCSK2 (Pcsk2), and down-regulated genes: KRT12 (Krt12), LASS1 (Cers1), PLAT (Plat), and NRXN1 (Nrxn1). Second, network analysis suggested anatomical structure development, cellular metabolic process, cell death, signal transduction, and stress response were significantly altered biological processes in the hTau MaptKO(Duke) mice as compared to age-matched non-transgenic controls. Further characterization of a sub-group of significantly altered genes revealed elevated phosphorylation of MECP2 (methyl-CpG-binding protein-2), which binds to methylated CpGs and associates with chromatin, in hTau MaptKO(Duke) mice compared to age-matched controls. Third, phoshpho-MECP2 was elevated in autopsy brain samples from human AD compared to healthy controls. Finally, siRNA-mediated knockdown of MECP2 in human tau expressing N2a cells resulted in a significant decrease in total and phosphorylated tau. Together, these results suggest that MECP2 is a potential novel regulator of tau pathology relevant to AD and tauopathies.

Among infertile men, a diagnosis of unilateral varicocele is made in 90% of varicocele cases and bilateral in the remaining varicocele cases. However, there are reports of under-diagnosis of bilateral varicocele among infertile men and that its prevalence is greater than 10%. In this prospective study, we aimed to examine the differentially expressed proteins (DEP) extracted from spermatozoa cells of patients with bilateral varicocele and fertile donors. Subjects consisted of 17 men diagnosed with bilateral varicocele and 10 proven fertile men as healthy controls. Using the LTQ-orbitrap elite hybrid mass spectrometry system, proteomic analysis was done on pooled samples from 3 patients with bilateral varicocele and 5 fertile men. From these samples, 73 DEP were identified of which 58 proteins were differentially expressed, with 7 proteins unique to the bilateral varicocele group and 8 proteins to the fertile control group. Majority of the DEPs were observed to be associated with metabolic processes, stress responses, oxidoreductase activity, enzyme regulation, and immune system processes. Seven DEP were involved in sperm function such as capacitation, motility, and sperm-zona binding. Proteins TEKT3 and TCP11 were validated by Western blot analysis and may serve as potential biomarkers for bilateral varicocele. In this study, we have demonstrated for the first time the presence of DEP and identified proteins with distinct reproductive functions which are altered in infertile men with bilateral varicocele. Functional proteomic profiling provides insight into the mechanistic implications of bilateral varicocele-associated male infertility.

Retinopathy of prematurity (ROP) causes 100,000 new cases of childhood blindness each year. ROP is initiated by oxygen supplementation necessary to prevent neonatal death. We used organ systems pharmacology to define the transcriptomes of mice that were cured of oxygen-induced retinopathy (OIR, ROP model) by hypoxia-inducible factor (HIF) stabilization via HIF prolyl hydroxylase inhibition using the isoquinolone Roxadustat or the 2-oxoglutarate analog dimethyloxalylglycine (DMOG). Although both molecules conferred a protective phenotype, gene expression analysis by RNA sequencing found that Roxadustat can prevent OIR by two pathways: direct retinal HIF stabilization and induction of aerobic glycolysis or indirect hepatic HIF-1 stabilization and increased serum angiokines. As predicted by pathway analysis, Roxadustat rescued the hepatic HIF-1 knockout mouse from retinal oxygen toxicity, whereas DMOG could not. The simplicity of systemic treatment that targets both the liver and the eye provides a rationale for protecting the severely premature infant from oxygen toxicity.

OBJECTIVE To compare the sperm protein profile between infertile men with unilateral varicocele and infertile men with bilateral varicocele.METHODS This prospective study investigated 50 infertile patients with clinical varicocele (33 unilateral and 17 bilateral) seeking fertility workup between March 2012 and April 2014. Routine sperm parameters, reactive oxygen species, total antioxidant capacity, and sperm deoxyribonucleic acid fragmentation were assessed in their semen. Sperm protein profile was characterized only in pooled samples of 5 unilateral and 3 bilateral varicocele samples, respectively, using sodiumdodecyl sulfate-polyacrylamide gel electrophoresis and an Linear Trap Quadrupole-Orbitrap Elite hybrid mass spectrophotometer system. Differences in protein expressionwere analyzed using gel analysis software, followed by protein identification using mass spectroscopy analysis. Differentially expressed proteins and their abundance were quantified by comparing spectral counts, followed by bioinformatics analysis.RESULTS Unique expression of 64 proteins in the bilateral group and 31 proteins in the unilateral group was obtained. Core functions of the top protein interaction networks were post-translational modification (w122 proteins associated with acetylation), protein folding, free-radical scavenging, cell death, and survival. The top molecular and cellular functions were protein degradation, free radical scavenging, and post-translational modifications, whereas the top pathways were protein ubiquitination and mitochondrial dysfunction. Major biological pathways for the 253 differentially expressed proteins were metabolism, apoptosis, and signal transduction.CONCLUSION Functional proteomic profiling helps identify the differential processes or pathways that are affected based on the nature of varicocele (bilateral or unilateral) and provide insights into the mechanistic implications of varicocele-associated male infertility. (C) 2015 Elsevier Inc.

Background: The etiology of varicocele, a common cause of male factor infertility, remains unclear. Proteomic changes responsible for the underlying pathology of unilateral varicocele have not been evaluated. The objective of this prospective study was to employ proteomic techniques and bioinformatic tools to identify and analyze proteins of interest in infertile men with unilateral varicocele.Methods: Spermatozoa from infertile men with unilateral varicocele (n = 5) and from fertile men (control; n = 5) were pooled in two groups respectively. Proteins were extracted and separated by 1-D SDS-PAGE. Bands were digested and identified on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Bioinformatic analysis identified the pathways and functions of the differentially expressed proteins (DEP).Results: Sperm concentration, motility and morphology were lower, and reactive oxygen species levels were higher in unilateral varicocele patients compared to healthy controls. The total number of proteins identified were 1055, 1010 and 1042 in the fertile group, and 795, 713 and 763 proteins in the unilateral varicocele group. Of the 369 DEP between both groups, 120 proteins were unique to the fertile group and 38 proteins were unique to the unilateral varicocele group. Compared to the control group, 114 proteins were overexpressed while 97 proteins were underexpressed in the unilateral varicocele group. We have identified 29 proteins of interest that are involved in spermatogenesis and other fundamental reproductive events such as sperm maturation, acquisition of sperm motility, hyperactivation, capacitation, acrosome reaction and fertilization. The major functional pathways of the 359 DEP related to the unilateral varicocele group involve metabolism, disease, immune system, gene expression, signal transduction and apoptosis. Functional annotations showed that unilateral varicocele mostly affected small molecule biochemistry and post-translational modification proteins. Proteins expressed uniquely in the unilateral varicocele group were cysteine-rich secretory protein 2 precursor (CRISP2) and arginase-2 (ARG2).Conclusions: The expression of these proteins of interest are altered and possibly functionally compromised in infertile men with unilateral varicocele. If validated, these proteins may lead to potential biomarker(s) and help better understand the mechanism involved in the pathophysiology of unilateral varicocele in infertile men.

Meeting abstracts Myeloid-derived suppressor cells (MDSC), including undifferentiated, monocytic, and granulocytic subsets, are known to play a role in tumor progression via promotion of tumor invasion and angiogenesis, and suppression of Type 1 T cell responses. Elevated peripheral blood

Varicocele the most common correctable cause of male factor infertility is observed in 35% to 50% of adult men with primary infertility. Oxidative stress plays an important role in the pathogenesis of sperm DNA damage in patients with varicocele. The objective was to examine if proteomic analysis and bioinformatic tools can help in identifying proteins of interest in infertile men with unilateral varicocele. This prospective study analyzed spermatozoa proteins from infertile men with unilateral varicocele (n = 5) and healthy fertile men (n = 5) who recently established a pregnancy to identify the proteins of interest involved in the pathophysiology of varicocele and male infertility. Spermatozoa were obtained from infertile men with unilateral varicocele and healthy men of proven fertility. Proteins were extracted and separated by 1-D gel. Bands were digested and on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Functional annotations of proteins were obtained using bioinformatics tools and pathway databases. A total of 1035 proteins were identified in the spermatozoa of fertile group and 757 in the unilateral varicocele group. 369 proteins were differentially expressed. Of these, 120 were unique to the fertile group and 38 to the unilateral varicocele group. 114 were overexpressed and 97 were underexpressed in the unilateral varicocele group. Apoptosis, post-translational modification, protein ubiquitination, mitochondrial dysfunction, oxidative stress response were some of the major functional categories observed for the differentially expressed proteins. Additionally, we have identified about 30 proteins of interest that play a role in sperm motility, capacitation, acrosome reaction and fertilization and may be compromised or altered in infertile men with unilateral varicocele. We identified significant differences in the distribution of differentially expressed proteins (DEP) in fertile men and infertile men with unilateral varicocele. Distinct proteins are involved in various biological processes, molecular function and cellular location. These proteins may serve as potential biomarkers that are involved in the pathophysiology of varicocele and male infertility.

Varicoceles appear to affect later stages of spermatogenesis and cause scrotal hyperthermia, hypoxia, hormonal imbalances, and re-flow of metabolites from renal and/or adrenal glands leading to oxidative stress. The objective was to study the major differences in the distribution of spermatozoa proteins in infertile men with varicocele compared to fertile men. his prospective proteomic study analyzed proteins in spermatozoa from infertile men with unilateral (n=5) or bilateral (n=3) varicocele and men of proven fertility (n=5) to study the proteins involved in the pathophysiology of varicocele-associated male infertility. Spermatozoa proteins were extracted from infertile men with unilateral and bilateral varicoceles and men with proven fertility. Proteins were separated by 1-D gel electrophoresis and bands were digested and identified on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Bioinformatics tools were used to identify pathways and functions of proteins of interest. Of the 99 proteins that were differentially expressed in the varicocele group, 9 proteins were uniquely expressed in the fertile group compared to 2 proteins that were unique to the varicocele group. 12 proteins were overexpressed and 76 were underexpressed in the varicocele group. In the varicocele group, the top networks were energy production, lipid metabolism, post-translational modification and protein folding. While proteins such as outer dense fiber protein 2 isoform 3 and tektin-3, that are known to play a role in sperm motility were overexpressed in the varicocele group, a majority of proteins, such as, acrosin binding protein precursor, calmegin precursor that are involved in spermatid differentiation, spermatid development, spermatogenesis, reproductive cellular processes, and fertility were observed to be underexpressed. Proteomics and bioinformatic analysis are powerful tools in understanding the pathology of varicocele associated male infertility. We have identified key proteins that are altered or modified in the presence of varicocele and may result in male infertility. Infertile men with unilateral or bilateral varicocele have a large number of spermatozoal proteins that are differentially expressed compared to those of fertile men. These protein alterations may be a contributing factor in male infertility.