Antioxidant enzyme profile and lipid peroxidation products in semen samples of testicular germ cell tumor patients submitted to orchiectomy

<div><p>ABSTRACT Purpose To determine enzymatic antioxidant and lipid peroxidation levels in seminal plasma of patients orchiectomized for testicular tumors. Materials and Methods The study included 52 patients: 26 control men and 26 orchiectomized patients for testicular tumor, of which 12 men had seminoma tumor and 14 men non-seminoma tumor. After semen analysis performed according to the WHO guidelines, an aliquot of semen was centrifuged and the seminal plasma was collected. Lipid peroxidation was performed by thiobarbituric acid reactive substances (TBARS) assay and antioxidant profile was assessed by analyzing catalase, glutathione peroxidase (GPx) and superoxide anion (SOD) activities using colorimetric assays with a standard spectrophotometer. Data were tested for normality and compared using one-way ANOVA (p<0.05). Results Seminoma and non-seminoma groups presented lower sperm concentration and morphology when compared to control group (p=0.0001). Both study groups (seminoma and non-seminoma) presented higher TBARS levels when compared to control group (p=0.0000013). No differences were observed for SOD (p=0.646) andGPx (p=0.328). It was not possible to access the enzymatic activity of catalase in any group. Conclusion Patients with testicular tumor present increased semen oxidative stress, but no differences were observed in antioxidant levels, even after orchiectomy. This indicates that most likely an increased generation of oxidative products takes place in these patients.</p></div

Sperm traits on <i>in vitro</i> production (IVP) of bovine embryos: Too much of anything is good for nothing

<div><p>Sperm samples used on fertilization strongly influence the <i>in vitro</i> production (IVP) rates. However, sperm traits behind this effect are not stated consistently until now. This study aimed to evaluate the isolated and combined effect of some sperm traits (MB: total motility before Percoll^® gradient, MA: total motility after Percoll^® gradient, AI: acrosome integrity, MI: membrane integrity, MP: mitochondrial membrane potential, and CR: chromatin resistance) on IVP rates. This is the first study focusing on the isolated effect of distinct traits. For this purpose, the experiment was divided in three steps. In first step, to study behavior of traits sperm samples (n = 63 batches) were analyzed and ranked based on each trait. In second step, samples ranked were selected from target ranks regions and allocated in groups of four to five batches, creating Higher and Lower groups, according to two different approaches. One aimed to form groups that differed to all sperm traits simultaneously (effect of combined traits). The other aimed to form groups that differed only to a single sperm trait while no differences were observed for the remaining traits (effect of each isolated trait). In third step, for each group successfully formed in step 2, sperm samples were individually and prospectively used for IVP. Cleavage, embryo development and blastocyst rates were recorded and compared between Higher and Lower of respective trait groups. Surprisingly, evaluation of isolated effects revealed that lower levels of MB, AI and MP resulted in higher embryo development and blastocyst rates (p<0.05), which was not observed on cleavage rate. We conclude that sperm traits strongly influence embryo development after <i>in vitro</i> fertilization (IVF), affecting the zygote competence to achieve blastocyst stage. Individually, levels of MB, AI or MP could be some of the key traits that may define IVP efficiency on current systems of embryo production.</p></div

Comparison of sperm profile between groups.

<p>Comparison of motility before Percoll^® (MB), motility after Percoll^® (MA), acrosome integrity (AI), membrane integrity (MI), mitochondrial membrane potential (MP) and chromatin resistance (CR) between higher groups and lower groups of isolated and combined effects.</p

Acronyms of higher and lower groups composed to each effect studied.

<p>Acronyms of higher and lower groups composed to each effect studied.</p

Comparison of IVP yields between groups.

<p>Comparison of cleavage, blastocyst and embryo development between higher groups and lower groups of isolated and combined effects.</p

Comparison of cleavage, blastocyst, and embryo development rates between higher (HMP; n = 17) and lower (LMP; n = 16) mitochondrial membrane potential.

<p>HMP, higher mitochondrial membrane potential; LMP, lower mitochondrial membrane potential.</p

Comparison of cleavage, blastocyst, and embryo development rates between higher (HAI; n = 21) and lower (LAI; n = 20) acrosome integrity.

<p>HAI, higher acrosome integrity; LAI, lower acrosome integrity.</p

Correlations coefficient (Rho) among percentages of sperm motility before Percoll^®, motility after Percoll^®, acrosome integrity, membrane integrity, mitochondrial membrane potential and chromatin resistance (Step 1; n = 252).

<p>Correlations coefficient (Rho) among percentages of sperm motility before Percoll^®, motility after Percoll^®, acrosome integrity, membrane integrity, mitochondrial membrane potential and chromatin resistance (Step 1; n = 252).</p

Mean of coefficient of variation among four replicates of 63 batches.

<p>Mean of coefficient of variation among four replicates of 63 batches.</p

Comparison of cleavage, blastocyst, and embryo development rates between higher (HMB; n = 20) and lower (LMB; n = 20) motility before Percoll^®.

<p>HMB, higher motility before Percoll^®; LMB, lower motility before Percoll^®.</p