Molecular investigation of mutations in androgen receptor and 5-alpha-reductase-2 genes in 46,XY Disorders of Sex Development with normal testicular development.

In this study, we aimed to determine androgen receptor (AR) and SRD5A2 gene mutations in 45 patients characterised by 46,XY Disorders of Sex Differentiation (DSD) signs with normal testicular development referred to the Children's Medical Center from February 2015 to September 2017. Karyotype and sex hormone analyses were performed. Cytogenetic investigation showed that seven patients were 46,XX DSD, six patients were chromosomal DSD and 32 patients were 46,XY DSD. Eight exons of the AR gene and five exons of the SRD5A2 gene were amplified. Two cases were affected with androgen insensitivity syndrome (AIS) (missense mutation on exon 7, position c.3637 G>A: p.R841H and position c.3610 G>A: p.R832Q), one case was affected with 5-alpha-reductase deficiency type 2 (missense mutation at c.578A>G: p.N193S on exon 4), and 22 cases (88%) did not demonstrate AIS or 5α-RD2 gene abnormality. Due to the great impact of these disorders on human lifestyle, evaluation of genes involved can improve genetic counselling and therapeutic management. We focused on the AR and SRD5A2 genes in patients with 46,XY DSDs with normal testicular development referred to the Children's Medical Center from all over the country to eventually culminate in a reliable prenatal diagnosis protocol at this major referral centre giving service to a great number of families with consanguineous marriages.

Flow cytometric evaluation of sperm superoxide anion production in rats with experimental varicocele.

OBJECTIVES: To determine whether intracellular superoxide anion production changes in the sperm of rats with experimental varicocele and to study mitochondrial membrane potential (MMP), sperm viability, antioxidant activity, Bax and Bcl-2 changes, and sperm count and motility in these rats. METHODS: The study groups consisted of 32 male rats divided into the following 4 groups: control, sham, varicocele-1, and varicocele-2. Experimental varicocele was established by partial ligation of the left renal vein in the latter 2 groups. Animals were killed 2 and 6 months after surgery, and dilation of the internal spermatic veins was observed. Then, superoxide anion production, MMP, antioxidant activity, and sperm characteristics were evaluated. Flow cytometry was used to study sperm superoxide anion production, assessed by dihydroethidium and MMP with rhodamine 123. RESULTS: Our results showed intracellular superoxide anion production significantly increased, and MMP, sperm viability, sperm count, and motility decreased in rats with experimental left varicocele. More Bax-positive cells were seen in the varicocele groups. However, there was no significant difference for seminal plasma antioxidant activity among all groups. CONCLUSIONS: These findings suggest that a main source of reactive oxygen species production in varicocele condition is intracellular.

Beneficial effects of MnTBAP, a broad-spectrum reactive species scavenger, in rat renal ischemia/reperfusion injury.

BACKGROUND: In recent years, several lines of evidence have implicated reactive species as contributors to renal ischemia/reperfusion injury (I/R). This study was designed to investigate the effect of Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP), a broad-spectrum reactive species scavenger, in the prevention of renal I/R injury. METHODS: Experiments were performed on rats anesthetized with pentobarbital. After tracheotomy, the right femoral artery was cannulated and the mean arterial pressure and heart rate were recorded. A midline laparatomy was performed, and the renal arteries were carefully separated from surrounding tissues. After surgery and a stabilization period (60 min), the animals were randomly assigned to four groups: sham-operated; sham+MnTBAP; I/R; I/R+MnTBAP. In I/R groups, the rats were subjected to bilateral renal artery occlusion for 40 min followed by 6 h reperfusion. Other groups underwent the surgery protocol but did not undergo renal artery occlusion, and were maintained under anesthesia for the duration of the experiment. Rats were administered either MnTBAP (10 mg kg(-1), i.v. bolus, 15 min prior to I/R) or saline. Renal function was assessed by plasma creatinine (Cr), blood urea nitrogen (BUN), and aspartate aminotransferase (AST) measurements. The fractional excretion of Na(+) (FE(Na+)) and urinary N-acetyl-beta-D-glucosaminidase (NAG) activities were also measured. Renal section damage was evaluated by light microscopy, and oxidative stress status was evaluated by measurements of plasma and renal vitamin E levels. RESULTS: We found that MnTBAP significantly reduced the I/R-mediated increases in plasma Cr, BUN, AST, FE(Na+), and NAG and improved the renal tissue histology. CONCLUSION: Our results showed that MnTBAP was effective in preventing the development of I/R-induced renal injury.