Erythropoietin Effect on Testicular Germinal Epithelium Cells in Undescended Testis Mice Model.

BACKGROUND: Undescended testis is an absence of testis in the scrotum, the incidence was 15 cases per 1000 from 1974 to 1996 in Europe. At Saiful Anwar Regional Hospital East Java, from January 2015 to July 2019 there were 60 boys diagnosed with undescended testis. A temperature rise of testis located in the abdominal triggers production of reactive oxygen species, causing impairment of the testicular epithelial germ cells and spermatogenesis, leading to many complications. Erythropoietin is a glycoprotein hormone that circulates in the body and has a positive effect on ischemic injury/gonadal reperfusion. OBJECTIVE: To find out ROS involvement in undescended testis and efficacy of EPO as an additional therapy for undescended testis. METHODS: This study is an experimental study with a post-test only control group design, using 18 male Wistar mice conditioned to be undescended testis for 7 days and underwent orchidopexy and some are given additional erythropoietin 1000iu/Kg 3 times a week. RESULTS: Before and after the intervention, the mean body weight of mice did not experience a significant difference, meanwhile testicular volume showed a significant difference between the orchidopexy and EPO groups (p = 0.005 and 0.001). Johnsen's score were found significant in the EPO group. Malone dialdehyde level in EPO and orchidopexy group showed significant difference p = 0.01 and 0.009 when compared to undescended testis group. CONCLUSION: There was the involvement of ROS in undescended testis and additional EPO improve impairment of germinal epithelial cells and spermatogenesis process due to undescended testis.

Germline Mutations and Polymorphisms of Androgen Receptor in Prostate Cancer Patients: Frequency and Results of in Silico Analysis

Background: Germline and somatic polymorphisms and mutations of the Androgen Receptor (AR) gene are known to be associated with the incidence of prostate cancer (PCa) in different populations. In this study we assessed germline AR polymorphisms and mutations in PCa patients with prediction of pathogenicity of the identified mutations by in silico analysis. Methods: Diagnosis of PCa was based on histopathology of prostate tissue (Gleason Score criteria) and serum prostate-specific antigen (PSA) levels. Genomic DNA was extracted from peripheral blood of 38 patients. All exons and exon-intron boundaries of AR were amplified using polymerase chain reactions (PCR) followed by Sanger sequencing. In silico analysis was performed using Polyphen-2 and Mutation Taster®. Results: Two polymorphisms, CAG repeat sequence (13-34 repeats in length) and p.Pro214Glu (MAF: 0.0789) located in exon 1 were identified. A missense mutation (c.47C>A/p.Pro146Glu) and in-frame deletion of a CAG sequence leading to loss of Arginine at codon 85 (c.252_254delCAG/p.Arg85-) were identified in a 70 year old patient with a Gleason Score and PSA level of 2 and 2.4ng/dL, respectively. His PSA level decreased to < 0.5 ng/dL after 9 months of androgen deprivation therapy. Identified mutations were predicted to be non-disease causing by Polyphen-2 and Mutation Taster®. Conclusion: Our data demonstrated that the frequency of germline mutations of AR was low in PCa patients in Indonesia (5.26%: 2/38 alleles), so that they are not likely to be major etiological factors. The in silico analysis of identified AR mutations in this study corroborated the clinopathology features of the patient.

Changes to signal peptide and the level of transforming growth factor- ß1 due to T869C polymorphism of TGF ß1 associated with lupus renal fibrosis.

Lupus Nephritis (LN) is a serious manifestation of lupus that can lead to End Stage Renal Disease (ESRD). Fibrosis is the main feature of ESRD, and it is likely influenced by Transforming Growth Factor Beta1 (TGFß1). The T869C gene polymorphism of TGFß1 is assumed to change the signal peptide, that has potential to interfere the urine production and renal protein expression of TGFß1. The influence of T869C gene polymorphism on TGFß1 production and renal fibrosis was evaluated in this study. Subjects were 45 patients LN with renal fibrosis and 45 participants without renal fibrosis as control, that were recruited from 2011 to 2013.Their urinary TGFß1 levels and TGFß1 gene polymorphisms were examined. All lupus patients underwent renal biopsy to assess their protein expression of TGFß1 in the renal tissue by immunohistochemistry and their renal fibrosis by morphometry and chronicity index. Changes in the signal peptide interaction with Signal Recognition Particle (SRP) and translocon of endoplasmic reticulum were analyzed by Bioinformatics. Levels of urinary and protein expression of TGFß1 increased in the LN with renal fibrosis group. There were significant differences in levels of urinary TGFß1 in T, C allele and TT, TC, CC genotypes between case and control groups. Furthermore, patients with C allele are 3.86 times more at risk of renal fibrosis than T allele. The C allele encodes proline, which stabilizes the interaction of the TGFß1 signal peptide with SRP and translocon, resulting in elevation of TGFß1 secretion. Our results indicated that T869C gene polymorphism of TGFß1 changes the signal peptide, that contributes to the production of urinary TGFß1 and affects renal fibrosis in lupus nephritis.

The Potential effect of G915C polymorphism in regulating TGF-ß1 transport into Endoplasmic Reticulum for cytokine production.

The TGF-ß1 cytokine concentration is known to be higher in nephritis with implied Lupus Nephritis severity. The production of TGF-ß1 cytokine is associated with G915C polymorphism. Therefore, it is of interest to study G915C polymorphism. The G915C polymorphism changes codon 25 which encodes arginine into proline in the signal peptide of TGF-ß1. The amino acid substitution affects signal peptide properties that may inhibit the transport of TGF-ß1 into the endoplasmic reticulum and eventually decline the cytokine production. Hence, the effect of G915C polymorphism on the properties of the signal peptide, the ability of TGF-ß1 transport into the endoplasmic reticulum and the concentrations of urinary TGF-ß1 in Lupus Nephritis patients was studied. The arginine substitution into proline decreased the polarity of the signal peptide for TGF-ß1. The increased hydrophobicity with increased binding energy of the signal peptide for TGF-ß1 to Signal Recognition Particle (SRP) and translocon is shown. This implies decreased protein complex stability in potentially blocking the transport of TGF-ß1 into the endoplasmic reticulum. This transport retention possibly hampers the synthesis and maturation of TGF-ß1 leading to decreased cytokine production.