Introduction: Disorders in the expression of any GENE effective in spermatogenic pathway is known as a probable cause of non-obstructive azoospermia and male infertility. The way responsible GENEs for sperm motility are expressed can considerably affect male fertility. Recent studies show that TSGA10 GENE is effective in the natural process of spermatoGENEsis as protein produced by this GENE in mouse results in the production of the main structure of sperm tail. Up to now, no comprehensive studies have been done on the way this GENE is expressed in the infertile's testical tissue.Materials & Methods: In this study, TSGA10 mRNA expression in testicular samples of 84 patients with non-obstructive azoospermia was investigated by semi-quantitative nested RT-PCR in Avesina Infertility Clinic during 2005-6. Moreover, expression levels of TSGA10 during spermatoGENEsis were evaluated using Johnsen's method for histopathologic scoring of the samples. For statistical analysis, SPSS software (Version 11.2) was used. The difference between GENE expressions was done based on quantitative variables by the use of t-test and covariance analysis and α<0.05 was regarded as a statistically significant value.Results: Testicular TSGA10 mRNA expression was observed in 31 patients, (36.9%), with non-obstructive azoospermia which it had a statistically significant correlation with spermatoGENEsis progress (p<000.0). Histopathologically, the GENE had been expressed in patients with higher Johnsen's score of spermatoGENEsis while a lack of expression was seen in all of those with Johnsen's score less than 4.5.Conclusion: The findings indicate that TSGA10 is expressed in human TESTIS and it is restricted to germ cells. It seems that lack of TSGA10 expression may have negative effects on spermatoGENEsis and on male fertility. On the other hand, determination of the timing of GENE expression in a certain level of spermatoGENEsis may also be used to determine levels of spermatoGENEsis in azoospermic patients alongside histopathological findings.

Introduction: TESTIS-SPECIFIC GENE antigen 10 (TSGA10) is a less-known GENE, which is involved in the vague biological paths of different cancers. Here, we investigated the TSGA10 expression using different concentrations of glucose under hypoxia and also its interaction with the hypoxia-inducible factor 1 (HIF-1). Methods: The breast cancer MDA-MB-231 and MCF-7 cells were cultured with different concentrations of glucose (5. 5, 11. 0 and 25. 0 mM) under normoxia/hypoxia for 24, 48, and 72 hours and examined for the HIF-1α expression and cell migration by Western blotting and scratch assays. The qPCR was employed to analyze the expression of TSGA10. Three-dimensional (3D) structure and the energy minimization of the interacting domain of TSGA10 were performed by MODELLER v9. 17 and Swiss-PDB viewer v4. 1. 0/UCSF Chimera v1. 11. The UCSF Chimera v1. 13. 1 and Hex 6. 0 were used for the molecular docking simulation. The Cytoscape v3. 7. 1 and STRING v11. 0 were used for protein-protein interaction (PPI) network analysis. The HIF-1a related hypoxia pathways were obtained from BioModels database and reconstructed in CellDesigner v4. 4. 2. Results: The increased expression of TSGA10 was found to be significantly associated with the reduced metastasis in the MDA-MB-231 cells, while an inverse relationship was seen between the TSGA10 mRNA level and cellular migration but not in the MCF-7 cells. The C-terminal domain of TSGA10 interacted with HIF-1α with high affinity, resulting in PPI network with 10 key nodes (HIF-1α , VEGFA, HSP90AA1, AKT1, ARNT, TP53, TSGA10, VHL, JUN, and EGFR). Conclusions: Collectively, TSGA10 functional expression alters under the hyper-/hypo-glycemia and hypoxia, which indicates its importance as a candidate bio-target for the cancer therapy.

BACKGROUND: In vitro GENEration of germ cells introduces a novel approach to male infertility and provides an effective system in GENE tracking studies, however many aspects of this process have remained unclear. We aimed to promote mouse embryonic stem cells (mESC) differentiation into germ cells and evaluate its effectiveness with tracking the expression of the Tsga10 during this process. METHODS: mESCs were differentiated into germ cells in the presence of Retinoic Acid. Based on developmental schedule of the postnatal TESTIS, samples were taken on the 7th, 12th, and 25th days of the culture and were subjected to expression analysis of a panel of germ cell SPECIFIC GENEs. Expression of Tsga10 in RNA and protein levels was then analyzed.RESULTS: Transition from mitosis to meiosis occurred between 7th and 12th days of mESC culture and post-meiotic GENE expression did not occur until the 25th day of the culture. Results showed low level of Tsga10expression in undifferentiated stem cells. During transition from meiotic to post-meiotic phase, Tsga10 expression increased in 6.6 folds. This finding is in concordance with in vivo changes during transition from pre-pubertal to pubertal stage. Localization of processed and unprocessed forms of the related protein was similar to those in vivo as well.CONCLUSIONS: Expression pattern of Tsga10, as a GENE with critical function in spermatoGENEsis, is similar during in vitro and in vivo germ cell GENEration. The results suggest that in vitro derived germ cells could be a trusted model to study GENEs behavior during spermatoGENEsis.

TESTIS SPECIFIC GENE antigen 10 (TSGA10) is a protein which has roles in spermatoGENEsis and cancers so that deletion or mutation in the TSGA10 GENE resulted in non-obstructive infertility and aberrant expression of this protein, was detected in solid tumors and leukemia. Despite the crucial roles of TSGA10 in tumoriGENEsis and infertility, yet it is not obvious how various nsSNPs of its GENE impress the structure and function of the TSGA10. Therefore, it is worthwhile to investigate the potential highly deleterious nsSNPs by several in-silico tools before launching costly experimental approaches. In the current study, we employed several different machine learning algorithms in a two-step screening procedure to analyze single nucleotide substitutions of TSGA10 GENE. Prediction tools were included SIFT, PROVEAN, PolyPhen-2, SNAP2, SNPs & GO, PhD-SNP for the first step and the second step included predictive tools such as I-mutant 3.0, MUpro, SNPeffect 4.0 (LIMBO, WALTZ, TANGO, FoldX), MutationTaster and CADD. Also, the 3D models of significantly damaging variants were built by Phyre2. The results elucidated 15 amino acid alterations as the most deleterious ones. Among these S563P, E578K, Q580P, R638L, R638C, R638G, R638S, L648R, R649C, R649H were located in a domain which is approved to has interaction with the HIF1-A protein and D62Y, R105G, D106V and D111Y were located on phosphodiesterase domain. In sum, these predicted mutations significantly influence the function of TSGA10 and they could be used for precise study of this protein in infertility and cancer experimental investigations.

TESTIS cancer is a relatively rare cancer in men. Because TESTIS cancer has not studied since long time ago in Mashad University of Medical Sciences, this study was performed. In our study TESTIS cancer made 1.8% of all cancers in men. In this study 96 patient with TESTIS cancer followed up. The peak incidence age of for seminoma was 20-40 years and for non-seminoma 20-30 years. In 66.5% of patients tumor was in right side and 33.5% in left. 14% of patients was cryptorchide. In 9% tumor developed in cryptorchide TESTIS and in 5% in contralateral TESTIS, the average delay time from beginning of symptoms was 6 months. In our patients 91.5% of seminomas were classic and 9.5% anaplastic. Tow year survival in patient with TESTIS tumor was 77% (89% in seminoma and 62% in non-seminoma). Five year survival in our patients was 69% (77% in seminoma and 60% in non-seminoma). The best result of therapy found in patients with non-seminoma who got RPLND (with 2 years survival of 100% and 5 year survival of 85%). This result may'be too low due to high stage of disease at the time of diagnosis.

Introduction: Production of antibodies against SPECIFIC proteins of TESTIS germ cells is of great significance for the investigation of processes involved in spermatoGENEsis, study of infertility problems and determination of the probable role of these proteins as cancer-TESTIS antigens. Murine TESTIS SPECIFIC Recombinant Protein 101 (mTEX101) is a 38kDa, GPI-anchored protein which is expressed in TESTIS germ cells of adult mice but it seems to be absent in other tissues. The structure and function of mTEX101 is not completely understood yet, but it is speculated that it may transducer biochemical signals into the cytoplasm since mTEX101 does not have an intracellular domain but the precise mechanisms are still ambiguous.Materials and Methods: RNA was extracted from three adult mice TESTIS. The RNA was used in RT-PCR, employing a pair of SPECIFIC primers for mTEX101 ORF region. TA-cloning technique was performed by the insertion of mTEX101 into a pGEM-T Easy Vector, followed by its sub cloning into a His-tagged expression vector, pET-28a (+). The recombinant mTEX101 was then produced by transfection of the expression vector into BL 21 (DE3) E. coli strain. Results: A recombinant protein, weighing 27kDa, was produced upon IPT Ginduction of the bacterial host. The presence of mTEX101 protein was detected through Western blot analysis by anti-mTEX101 peptide antibodies. Conclusion: We produced mTEX101 recombinant protein that could be used for the production of mono and polyclonal antibodies.

Introduction: Phosphorylation reactions highlight the essential role of protein kinases in sperm motility. TSSK6 protein is thought to play a crucial role in this process. Indeed, sperm obtained from mice with the Tssk6-/- genotype are unable to carry out fertilization and exhibit decreased motility. This leads us to investigate the causes of asthenozoospermia by searching for polymorphisms in the TSSK6 GENE. Therefore, the objective of this study is to identify polymorphisms in the TSSK6 GENE in men affected by asthenozoospermia. Material & Methods: The methodology involved direct sequencing of spermatozoa DNA. Thirty ejaculates were analyzed, including 20 from asthenozoospermic men and 10 from normozoospermic men. Spermograms were performed according to WHO procedures. DNA extraction was carried out using the phenol/chloroform method followed by conventional PCR. The amplicons were sequenced using the Sanger method, and the sequences were analyzed with BioEdit software. The data were analyzed using Fisher and Mann-Whitney tests. Results: The results revealed mutations in the TSSK6 GENE in both normospermic and asthenozoospermic ejaculates. The synonymous mutations c.690T>C (p.Tyr230Tyr) and c.372C>A (p.Arg124Arg) were the most frequent, occurring at rates of 50% and 33.33%, respectively. Analysis of the mutations using PolyPhen-2 indicated that all mutations observed in normozoospermic samples are benign and would not affect sperm quality. However, only the mutations described in asthenozoospermic samples are predicted to be damaging to the protein. Conclusion: In conclusion, mutations in the TSSK6 GENE were observed in infertile men. Deleterious mutations in the TSSK6 protein are associated with asthenozoospermia

Cancer/TESTIS antigens (CTAs) as a group of tumor antigens are the novel subjects for developing cancer vaccine and immunotherapy approaches. They aberrantly express in tumors with highest normal expression in TESTIS, and limited or no expression in normal tissues.There are important similarities between the processes of germ-cell and cancer cell development SpermatoGENEsis begins at puberty when expression of novel cell-surface antigens occurs when the immune system has been refined the ability to distinguish self from non-self. Whereas macrophage and lymphocytes are commonly found within interstitial spaces of the TESTIS, these antigen-presenting cells are rarely seen within the seminiferous tubules. These observations have led to the concept of the immune privileged site for TESTIS. Localized normal expression of the CT GENEs in TESTIS that makes them immunogenic for immune system, in one side, and their abnormal expression in different kinds of cancer cells, in the other side, has make them as promising target for developing cancer vaccines and new cancer therapeutics approaches. In malignancies, GENE regulation is disrupted which results aberrant expression of CT antigen in a proportion of tumors of various types. For some CTAs, data support their fundamental role in tumoriGENEsis. Several authors believe it is not clear whether they have an essential role in tumoriGENEsis or they are by-products of chromatin variations in cancer. There is a growing list of CTAs within them advanced clinical trials are running by using some of them in cancers like lung cancer, malignant melanoma and neuroblastoma. In this review we discuss the GENE TSGA10 as an example of CT GENEs. TSGA10 expresses in its highest levels in elongating spermatids and localized in the fibrous sheath of mature sperm. This GENE is proposed as a serological biomarker in cutaneous lymphoma. Its abnormal expression has been reported in different cancers such as acute lymphoblastic leukemia, breast, brain, gastrointestinal and a range of other cancers either in mRNA or protein levels. It has an important role in angioGENEsis in cancer tumors because of its effects in the GENE hypoxia-inducible factor (HIF1). Absence or lack of TSGA10 expression has been reported in ascosporic infertile men.