Association of angiotensin-converting enzyme (ACE) I/D variation with biochemical parameters and oxidative stress markers in systemic lupus erythematosus patients in west of Iran.

PURPOSE: We aimed to study insertion/deletion (I/D) variation (rs4646994) of ACE gene in a group of SLE patients in west of Iran and its possible relationship with oxidative stress. METHOD AND RESULTS: Genotypes and allele frequencies related to ACE (I/D) variation were determined in 108 SLE patients and 110 gender and age-matched healthy controls using PCR. Neopterin, malondialdehyde (MDA), and serum lipid concentrations were determined by HPLC and enzyme assay respectively. The overall distribution of ACE I/D genotypes in SLE patients was different from that of the control group (P = 0.005). DD genotype compared to ID genotype increased the risk of SLE (OR = 2.57, 95% CI 1.4-4.8, P = 0.003). ID genotype compared to the II genotype decreased the risk of disease (OR = 0.45, 95% CI 0.2-0.99, p = 0.042). SLE patients with DD, ID, and II genotypes had lower paraoxonase (PON) activity and higher serum levels of MDA and neopterin versus control patients. We also detected a significant protective effect against SLE in presence of ACE I alleles and lack of angiotensin II receptor, type 1 (AGTR1) A1166C (NCBI reference SNP id: rs5186), C alleles in this study (OR = 0.31, 95% CI 0.14-0.68, P = 0.002). CONCLUSIONS: Carriers of the DD genotype of ACE gene with higher serum concentrations of neopterin and MDA, and lower PON activity had a high risk to develop SLE, while ID genotype decreased the risk of disease development by 2.22 times compared to II genotype.

Ceratonia siliqua (Carob) extract improved in vitro development of vitrified-warmed mouse germinal vesicle oocytes: assessment of possible mechanism.

Oocyte banking is a vital step for safekeeping and spreading genetic resources of animals. It is also used for fertility preservation of human. Oocyte vitrification is closely related to the lower developmental competence which includes the cryo-injury arisen during vitrification. The aim of the present study was to evaluate the maturation, embryonic development and production of reactive oxygen species (ROS) of mice oocytes following the supplementation vitrification media with different concentrations of Ceratonia siliqua (carob) extracts. In this experimental study, germinal vesicle oocytes collected from 8 to 10 week-old female NMRI mice (30-40 gr) were randomly divided into six groups of vitrification media supplemented with 0 (control), 5, 10, 20, 30 and 50 µg/ml C. siliqua. After thawing, oocytes were put in an in vitro maturation medium (IVM) (α-MEM: Alpha Minimum Essential Medium). 3-4 and 24 h (hr) later, the oocyte nuclear maturity was checked. Standard in vitro fertilization was performed on the matured oocytes (MII), and embryonic development was followed. Extra- and intra-cellular ROS was measured in IVM medium after 24 h of oocyte incubation. The addition of 20 and 30 µg/ml C. siliqua extract to vitrification media improved normal morphology of warmed germinal vesicle (GV) oocytes, rate of germinal vesicle break down (GVBD), and metaphase 2 (MII) oocyte formation significantly (p < 0.05). Fertilization rate, (embryonic development to 2 cells stage, 4-8 cells stage, and > 8 cells stage increased in the 30 µg/ml C. siliqua group significantly (p < 0.05). Furthermore, supplementation of 30 µg/ml C. siliqua in vitrification media significantly decreased extra- and intra-cellular of ROS as well as embryonic fragmentation (p < 0.05). In conclusion, supplementation of GV oocyte vitrification media with carob extract improved maturation, fertilization, and embryonic development rate and decreased extra- and intra-cellular ROS levels.

Does supplementation of sperm freezing/thawing media with Ceratonia siliqua improve detrimental effect of cryopreservation on sperm parameters and chromatin quality in normozoospermic specimens?

Human sperm banking is an important procedure in the assisted reproductive technique centers. It entails sperm damage. The aim of this study was to investigate beneficial effect of Ceratonia siliqua (C. siliqua) supplement in freezing/thawing media on post thaw sperm parameters and sperm chromatin quality in normozoospermic samples. Forty normozoospermic specimens were included in this prospective study. Each sample was divided into ten groups. In groups one to five, 0 (as control group) 5, 10, 20 and 30 µg/ml C. siliqua were added to freezing medium and in groups six to ten, similar concentration of C. siliqua were added to thawing medium for 30 min incubation. Sperm concentration, progressive motility, normal morphology, viability, aniline blue (AB), toluidine blue (TB) and sperm chromatin dispersion (SCD) staining tests were evaluated before vitrification and after thawing. The results showed that 10 and 20 µg/ml supplementation of C. siliqua in freezing/thawing media significantly increased progressive motility, normal morphology and viability of sperm (p < 0.05) as well as decreased AB, TB and SCD (p < 0.05). Also, 20 µg/ml had significantly higher improvement compared to 10 µg/ml C. siliqua (p < 0.05). The present study showed that C. siliqua supplemented freezing/thawing media can improve sperm quality of normozoospermic samples after freezing/thawing.