(-)-Epigallocatechin-3-gallate Modulates the Differential Expression of Survivin Splice Variants and Protects Spermatogenesis During Testicular Torsion

The anti-apoptotic effect of (-)-epigallocatechin-3-gallate (EGCG) during unilateral testicular torsion and detorsion (TT/D) was established in our previous study. In mice, the smallest inhibitor of apoptosis, survivin, is alternatively spliced into three variants, each suggested to have a unique function. Here, we assessed how EGCG exerts its protective effect through the expression of the different survivin splice variants and determined its effect on the morphology of the seminiferous tubules during TT/D. Three mouse groups were used: sham, TT/D+vehicle and TT/D treated with EGCG. The expression of the survivin variants (140 and 40) and other apoptosis genes (p53, Bax and Bcl-2) was measured with semi-quantitative RT-PCR. Histological analysis was performed to assess DNA fragmentation, damage to spermatogenesis and morphometric changes in the seminiferous tubules. In the TT/D+vehicle group, survivin 140 expression was markedly decreased, whereas survivin 40 expression was not significantly different. In parallel, there was an increase in the mRNA level of p53 and the Bax to Bcl-2 ratio in support of apoptosis induction. Histological analyses revealed increased DNA fragmentation and increased damage to spermatogenesis associated with decreased seminiferous tubular diameter and decreased germinal epithelial cell thickness in the TT/D+vehicle group. These changes were reversed to almost sham levels upon EGCG treatment. Our data indicate that EGCG protects the testis from TT/D-induced damage by protecting the morphology of the seminiferous tubules and modulating survivin 140 expression.

effect of prostate tissue inflammation in benign prostatic hyperplasia on enhancer of zeste homolog 2 ribonucleic acid expression

Enhancer of zeste homolog 2 [EZH2] has been recently found to regulate several genes involved in immunoresponse and autocrine inflammation network. The aim of the study was to quantitate EZH2 messenger ribonucleic acid [mRNA] expression, evaluate its relation to conditions of prostatitis associated with benign prostatic hyperplasia [BPH], and correlate it with the levels of the inflammatory marker interlukin 6 [IL-6]. Cross-sectional study in Middle Eastern men with BPH and prostatitis or BPH only. Transrectal ultrasound-guided prostate biopsies were collected from 106 patients suspected of having prostate cancer; however, the histology revealed BPH. Upon further pathological examination, 56 of these cases were identified as BPH with prostatitis and classified as: acute prostatitis [n=13]; active chronic prostatitis [n=32]; and, chronic inactive prostatitis [n=12]. Serum IL-6 levels and EZH2 mRNA expression were measured and compared between patient groups. EZH2 mRNA was overexpressed in BPH with prostatitis patients compared to BPH only patients [P<.0001]. BPH with active chronic prostatitis had higher EZH2 expression than BPH with acute or chronic inactive prostatitis compared to BPH only [P=.05 and .73, respectively]. EZH2 mRNA expression showed a negative correlation with IL-6 concentrations in BPH with prostatitis patients [rs=-0.31, P=.02]. EZH2 overexpression was associated with an increased risk of having BPH with prostatitis [crude odds ratio 0.20, 95% CI 0.06-0.65, P=.0076]. EZH2 mRNA expression correlates positively with prostatitis conditions associated with BPH and negatively with serum IL-6 levels. This supports the possible involvement of EZH2 mRNA overexpression in the development of prostate inflammation, and its new regulatory role in suppressing the expression of some inflammatory network genes

Urinary survivin mRNA expression and urinary nuclear matrix protein 22 BladderChek[R] and urine cytology in the detection of transitional cell carcinoma of the bladder

To compare the diagnostic performance of urine cytology [UC], survivin mRNA expression, and the NMP22 BladderChek[R] [NMP22BC] test for the detection, grading and staging of transitional cell carcinoma [TCC] of the bladder. Voided urine samples collected from 25 healthy controls and 80 patients diagnosed with TCC of the bladder were subjected to UC, the NMP22BC test and reverse-transcription real-time PCR for survivin mRNA expression. Survivin mRNA expression showed the highest sensitivity [87.5%] followed by the NMP22BC test [61.3%] while UC exhibited the lowest sensitivity [40%]. All three urine markers had a similar specificity of 96% [95% CI 80.5-99.3%]. Survivin mRNA expression was the only urine marker that showed a significant difference in relation to tumour histological grade [X[2] 8.5, p = 0.015]. None of the three urine markers was significantly related to tumour pathological stages. The diagnostic sensitivity of urinary urviving mRNA expression was superior to that of UC and the NMP22BC test and correlates with tumour pathological grade but not stage

Survivin downregulation is associated with vasectomy-induced spermatogenic damage and apoptosis

To evaluate the expression of the apoptotic genes survivin, Bax and Bcl-2 in vasectomized rabbits and to determine their relation with vasectomy-induced spermatogenic impairment and germ cell apoptosis. Twelve adult rabbits [6-12 months old] were divided into three groups: sham control, unilateral vasectomy or bilateral vasectomy. Six months after vasectomy, testicular tissue was analyzed for germ cell apoptosis and DNA fragmentation by the TUNEL assay and gel electrophoresis, respectively. Spermatogenesis was assessed using the Johnsen score. The relative gene expression of survivin, Bax and Bcl-2 was measured using reverse transcription followed by reAl time PCR. Compared to sham animals, a significant decrease in testicular survivin mRNA levels was measured in the two vasectomy animal groups [p<0.05]. This was accompanied by a significant increase in the Bax:Bcl-2 ratio in the vasectomized animals [p<0.05]. In addition, these data showed positive correlation with enhanced apoptotic index, damage to spermatogenesis and DNA fragmentation after vasectomy. These findings demonstrate that vasectomy-induced damage to spermatogenesis due to testicular apoptosis may be associated with survivin downregulation and Bax overexpression