Male pronuclear formation and embryo development following intracytoplasmic injection of ovine pretreated sperm

Background: Failure of Male Pronucleus [MPN] formation is a major concern in the success of Intracytoplasmic Sperm Injection [ICSI] in some species. Despite the conducted unsuccessful efforts to improve ICSI efficiency in ovine, the present study was aimed to improve MPN formation and embryo development in ovine ICSI procedure through accompaniment of sperm pretreatment with co-injection of some compounds

Methods: In experiment 1, sperm were treated with either 2-mercaptoethanol [2ME], glutathione [GSH], or DTT [dithiothreitol] in combination with Heparin [Hep]. Following DNA integrity and fragmentation assessments, the best sperm pretreatment approach in induction of sperm head decondensation was applied for the second and third experiments. In experiment 2, in vitro matured oocytes were subjected to ICSI using pretreated sperm with or without GSH and Sperm Extract [SE] co-injection. In experiment 3, the procedure was followed as experiment 2 with acrosome reacted sperm

Results: The highest percentages of oocyte activation were observed in Hep+GSH and Hep+2ME groups. The greatest MPN formations were also observed in the same groups when ICSI procedure was accompanied with GSH co-injection. Despite the higher percentage of MPN formation and oocyte activation in Hep+GSH and Hep+2ME groups, none of the employed strategies could increase the cleavage and blastocyst rates compared to the control

Conclusion: In our study condition, despite the lack of significant increase in embryo development in treated groups, the significant increase in MPN formation in Hep+GSH+co.GSH and Hep+2ME+co.GSH groups indicates the lower chance of parthenote formation that means a higher chance of normal fertilization compared with control

psychometric characteristics of the exercise benefits/barriers scale among Iranian elderly

The objective of this study was to investigate the validity and reliability of the Farsi version of the perceived benefits/barriers scale of physical activity in Iranian elderly. Overall, 388 elderly subjects [60 yr and over] completed the demographic characteristics questionnaire, the Exercise Benefits/Barriers Scale [EBBS] and the Yale physical activity scale. Data were analyzed through of exploratory factor analysis, using Varimax rotation, Cronbach's alpha and Pearson correlation. The matrix table of rotated elements of Farsi version of EBBS showed ten components, and 41 items predicted 61.83% of variance. 28 items in 5 components for benefits of, and 13 items in 5 components for barriers to physical activity were identified. Cronbach's alpha for internal consistency in the whole scale and its subscales was 0.83, 0.94 and 0.68. In addition, positive and significant correlation was found between overall benefits and their sub-scales as well as between overall barriers and their subscales. Moreover, there was significant and positive correlation between physical activity and the benefits [r=0.209, P=0.005] and significant negative correlation between physical activity and the barriers [r=-0.231, P=0.001]. The results showed acceptable reliability and validity of the Farsi version of Exercise Benefits/Barriers Scale among Iranian elderly

effect of macromolecule source and type of media during in vitro maturation of sheep oocytes on subsequent embryo development

Oocyte maturation and subsequent in vitro production [IVP] of embryos are affected by diverse groups of chemicals in maturation medium which are needed for successful mammalian oocyte maturation during which the dramatic cytoplasmic and nuclear reprogramming events take place. This study was designed to evaluate the effects of protein source [fetal bovine serum, FBS, and bovine serum albumin, BSA] as well as two different maturation media during in vitro maturation of ovine oocytes on subsequent embryo development. Cumulus oocyte complexes were recovered from ovaries obtained from slaughter house and cultured for 24 hr in either TCM-199 or SOFaa maturation medium supplemented with 10% [v/v] FBS or 0.8% [w/v] BSA. Data were analyzed by one-way ANOVA using Sigma Stat [Ver. 2]. A p-value smaller than 0.05 was considered statistically significant. The proportions of cleavage and total blastocyst [evaluated on days 3 and 6, respectively] were significantly higher in FBS than BSA supplemented groups, though no differences were observed between the two used different maturation media. The cryotolerance of blastocysts was negatively influenced by the presence of FBS rather than BSA during IVM. The quality of produced embryos, however, was affected neither by the source of macromolecules nor the maturation medium in terms of hatching rate, total blastocyst cells and inner cell mass/total cell ratio. The rate of oocyte development was improved by the presence of FBS, though the cryosurvival of resulting blastocysts was negatively influenced by the presence of the serum during in vitro production of sheep oocytes

Congenital anomalies in infants conceived by assisted reproductive techniques

Many studies show that congenital defects in infants conceived by assisted reproductive techniques [ART] are more than infants of normal conception [NC]. The aim of this study is to determine the frequency of congenital anomalies in ART infants from Royan Institute and to compare congenital anomalies between two ART techniques. In a cross-sectional descriptive study, 400 ART infants from Royan Institute who resided in Tehran were selected by non-random, consecutive sampling. Infants were examined twice [until 9 months of age] by a pediatrician. Infants' congenital anomalies were described by each body system or organ and type of ART. Data were analyzed by SPSS version 16 and Fisher's exact test. The frequency of different organ involvement in the two examinations were: 40 [10%] skin, 25 [6.2%] urogenital system, 21 [5.2%] gastrointestinal tract, 13 [3.2%] visual, and 8 [2%] cardiovascular system. Major congenital defects in infants conceived by in vitro fertilization [IVF] and intracytoplasmic sperm injection [ICSI] were hypospadiasis, inguinal hernia, patent ductus arteriosus plus ventricular septal defect [PDA + VSD], developmental dysplasia of the hip, lacrimal duct stenosis during the first year of life, hydronephrosis and urinary reflux over grade III, undescending testis, ureteropelvic junction stenosis, and torticoli. Two-thirds of ART infants had no defects. A total of 7% of IVF and ICSI infants had one of the major abovementioned congenital anomalies. This rate was higher than NC infants [2%-3%]. There was no difference between the ICSI and IVF group

effect of biopsy during precompacted morula stage on post vitrification development blastocyst derived bovine embryos

Improvements on embryo micromanipulation techniques led to the use of embryo biopsy in commercial embryo transfer programs for genetic analysis of preimplantation bovine embryos. The aim of this study was to evaluate the quality of bovine blastocyst derived from embryos biopsied at different precompacted morulae stages by assessment of cryosurvivability of the resulting blastocysts. The in vitro produced bovine embryos were subjected to biopsy at days 2,3, and 4 post-isemination with different cell numbers [4 to 16-cells]. Embryo cell biopsy was carried out in a 100 micro l drop of H-SOF following pronase drilling by aspiration of one blastomrere. The biopsied embryos were then cutured in SOFFaaBSA co-cultured with oviduct cells-monolayer until blastocyst formation. The blastocysts were cryopreserved at room temperature after exposure of equilibration [glycerol 1.4 M for 5 min and then glycerol 1.4 M and ethylene glycol 3.6 M for 5 min] and vitrification solutions [3.4 M glycerol and 4.6 M ethylene glycol]. The blastocysts were loaded into the center of 0.25 ml straws separated by air bubbles from 2 columns of sucrose 0.5 M and plunged immediately into liquid nitrogen. There was no significant difference in cryosurvivability of vitrified-warmed blastocysts derived from biopsied embryos at different pre-compacted morula stages. The quality of biopsy derived blastocysts was identical to that of non-biopsy derived ones in terms of post vitrification survival and hatching rates. In conclusion there was no preference between different times of embryo biopsy at precompacted morula stages in term of cryosurvivability of biopsy derived bovine blastocysts

Effects of timing on cell biopsy from pre-compacted morula stage bovine embryos on subsequent embryonic development

Embryo biopsy has potential applications in molecular research processes in domestic animals, besides its application in sex determination in embryo transfer programs. The objective of the present study was to assess the in vitro development of bovine embryos biopsied on different days of precompacted morula stage. Slaughterhouse-derived oocytes were matured in vitro, fertilized [Day-0] by frozen-thawed, Percol-separated spermatozoa and cultured on oviductal cell monolayer. The embryos were subjected to cell biopsy on Days 2, 3, and 4 postinsemination at 4-16-cell stages. The data were analyzed using ANOVA and Chi-squared tests [SigmaStat, version 2]. A p-value < 0.05 was considered significant. Biopsies carried out at 16-cell stage [Day-4] resulted in 94% of embryos developing to the blastocyst stage, which was significantly higher [p < 0.05] than the ones biopsied at 8-cell stage on Day-4 [64%], and those undergoing the procedure on Day-3 [49% and 46% at 4-cell and 8-cell stages, respectively] and Day-2 [39% and 33% at 4-cell and 8-cell stages, respectively]. No significant differences were observed between biopsied and non-biopsied embryos on a given day. The total cell number in biopsy-derived blastocysts ranged between 103 and 135. The difference in the number of total cells, dead cells and cell allocation to trophectoderm and inner cell mass between non-biopsied and biopsy-derived blastocysts was insignificant. Biopsy of bovine embryos at 4-16-cell stages had no adverse effects on in vitro developmental potentials and the 16-cell stage embryos, biopsied on Day-4 was the best stage for blastomere removal

effect of the duration of in vitro maturation [IVM] on parthenogenetic development of ovine Oocytes

The aim of this study was to compare the effect of time of parthenogenetic activation [22 hr versus 27 hr after In Vitro Maturation-IVM] on in vitro development of ovine oocytes using either single [lonomycin 5 microM for 5 minor Ethanol 7% for 7 m/n] or combined [ionomycin and ethanol with 6-DMAP 2 mM for 3 hr] activation treatments. The abattoir-derived in vitro matured activated oocytes were cultured in modified synthetic oviductal fluid and assessed for the cleavage, blastocyst, and hatching rates. The single-activated oocytes had a reduction in cleavage, blastocyst and hatching rates compared to the combined-activated oocytes [except for the cleavage at 27 hr]. In single-treated groups the rates of cleavage and blastocyst were increased as the maturation time was extended from 22 hrto 27 hr. The numbers of total cells and Inner Cell Mass [ICM], though insignificant, were greater in combined-treated groups compared to the single treatment. The number of ICM in Eth+6-DMAP group activated at 27 hr was lower than 22 hr. Nonetheless, irrespective of the activation protocol, development to the blastocyst stage, the numbers of total cell, ICM, and cell allocation [ICM/total cells] were significantly lower in parthenogenetic than fertilized embryos. In conclusion, though the cleavage and blastocyst rates in single-treated groups were positively influenced by the extension of duration of IVM [27 hr], there was a trend of decreased numbers of total cells and ICM in slightly aged oocytes. Moreover, developmental potential of ovine parthenotes, especially in young oocytes, was improved by the addition of 6-DMAP to the activation regimen