International Journal of Reproductive BioMedicine
Volume:5 Issue: 5, Apr 2007

Fertilization involves direct interaction of the sperm and oocyte, fusion of the cellmembranes and union of male and female gamete genomes. The completion of thisprocess and subsequent embryo development depend in part on the inherent integrity ofthe sperm DNA. Sperm genome quality has been emphasized for several years asplaying a major role in early embryogenesis. There is clinical evidence showing thathuman sperm DNA damage may adversely affect reproductive outcomes and thatspermatozoa of infertile men possess substantially more DNA damage than dospermatozoa of fertile men. Testing DNA integrity may help selecting spermatozoa withintact DNA or with the least amount of DNA damage for use in assisted reproductivetechniques (ARTs). This review will focus on how sperm DNA is organized, whatcauses sperm DNA damage and what impact this damage may have on reproductiveoutcome.

Determination of transgenic embryos from non transgenic embryos sibling is animportant step in producing homozygous transgenic mice. These steps need by PCR or southernblotting followed extraction of DNA, but both techniques require skill and consume time.

The aim of this study was simulation of high accuracy method using novelenhanced green fluorescent protein (EGFP) gene cassette to eliminate some consumetime in livestock industry to assay high quality embryos and morphological plasticity.

We modified pQE-Tri systemic vector with EGFP and IRESsequence to trace out coming planning of molecular farming transgene using coinjectionmethod.

The combination of these sequences successfully showed the faint and normalexpression of transgene in mouse pre-implantation stage embryos. The low rate ofsurviving green positive embryos as compare as only medium and physical treatmentscould be partly from the physical damage caused by microinjection and gene integration.Furthermore, application of the enhanced GFP marker facilitated subtle detection ofasymmetrical division appeared in some of transgenic embryos.

The results of current mouse simulation model imply that an efficientproduction and propagation of transgenic livestock can be done by co-injection of everyeconomical gene with this novel transgene and also it can be suitable gene cassette fornumerous experiments and study of protein behaviors in living cells.

Preparation of oocytes is one of the critical factors that determine thedevelopmental competence of embryos produced by in vitro fertilization (IVF).

In this study, the effect of cysteamine, type of media and glutathione (GSH)level on blastocysts development after in vitro maturation of mouse oocytes wereinvestigated.

Premature female mice were primed with pregnant marestimulating gonadotrophin (PMSG), and germinal vesicle (GV) stage oocytes wereobtained 45 hr later. GV oocytes were cultured in presence of 0, 50, 100, 200 and 500μm cysteamine in TCM199 and MEME media. After IVM, MII oocytes were in vitrofertilized (IVF) and in vitro cultured (IVC) in order to observe embryo development. Agroup of In Vivo Ovulated (IVO) oocytes after priming with PMSG and HCG also wereincluded in this study. 5,5-Dithio-bis (2nitrobenzoic acid) DTNB-recycling protocolwas used for GSH assay.

Rate of IVM and IVF were improved in all oocytes treated with cysteamine inthe two medium except 500 μm (81% MII rate in TCM and 64% MII in MEME). Rateof blastocyst in 100 μm cysteamine in TCM1199 and 200 μm in MEME was highercompared to control groups (In TCM 45% and in MEME 35%). In vivo MII and GVoocytes represented the highest and lowest GSH level respectively.

Our results revealed that the media and concentration of cysteamine canaffects on IVM, IVF and rate of blastocysts development on dose dependant manner.

Vitrification is assumed to be a promising method to cryopreserve humanoocytes but still needs optimizing.

The aim of this study was to improve the single step and step-wisevitrification effects on maturing mouse GVBD oocytes by ethylene glycol (EG) inconventional straws.

Oocytes with compact cumulus cells were cultured for 3hr inTCM199 supplemented with 10% fetal bovine serum (FBS) in 5% CO2 in air. GVBDoocytes were randomly allocated into three groups. (1) Control (non-vitrified group), (2)exposed to single-step vitrification (contained of EG 20%+0.5M sucrose), (3) exposedto step-wise vitrification (2%, 5%, 10%, 20%EG +0.5M sucrose). In vitrificationgroups,oocytes were thawed and underwent additional 21 hr maturation. Viability ofoocytes and maturation to MII stage were analyzed using inverted microscope andadditionally by staining of propidium iodide and Hoechst 33342.

All non-vitrified oocytes were viable after 24 hr; however, viability of vitrifiedsamples in single-step group was significantly lower than that of the step-wise andcontrol Groups. Also, the maturation rate in the step-wise group was significantly higher(p < 0.05) compared to single-step.

These results suggest that step-wise vitrification of GVBD oocytes ascompared to single step vitrification was better in the rate of survival and in vitromaturation of oocytes.

Spinal cord injury (SCI) occurs most often to young men at the peak oftheir reproductive health. Only 10% of SCI men can father children without medicalassistance due to potential impairments in ejaculation and sperm quality.

The main objective of this experimental study was to evaluate theepididymal necrospermia- sperm death, after chronic SCI in rat.

Forty-five adult Wistar rats were divided into 3 groups ofSCI, sham, and control. Following laminectomy, SCI was induced onto exposed duramatter (T10) of anesthetized rats. Sham group underwent laminectomy of T10 only;while, control rats were not exposed to any type of injury or medication. Thespermatozoa from cauda epididymis were aspirated after 50 days for analysis ofnecrospermia with three assays of Eosin-Y staining, Hypo-osmotic swelling (HOS), andHoechst 33258 fluorescent dye.

The rate of necrospermia in SCI rats was significantly increased whencompared with other groups (p<0.05). Also, the rates of necrspermia in SCI sampleswere similar with application of 3 assays (Eosin-Y: 46.11±9.41; HOS: 45.88±8.89;Hoechst: 46.76±9.31). Total necrospermia was not observed in any of the epididymalsamples.

The results showed that chronic SCI is associated with high rate ofepididymal necrospermia in mammals such as rats. It is, therefore, recommended that aneffective laboratory technique, such as Hoechst 33258 should be used for separation oflive and motile sperms from necrospermic ones for assisted reproduction program.

During the detorsion of a torsioned ovary, oxidant agents are released andmelatonin as an antioxidant can reduce ischemia. We studied the histopathologicalchanges after using melatonin on experimental torsioned ovary in cat.

The aim of this experimental study was to investigate the effects ofMelatonin on histopathological changes in torsion – detorsion injury in cat ovary.

An adnexal torsion – detorsion model was created by using 20adult cats randomly divided equally in to 2 groups of Saline and Melatonin. Ischemia wasinduced by iathrogenic 360° clockwise torsion of the cat adnex for 3 hr. Reperfusionwas achieved for 3 hr. Melatonin or saline were injected intra peritoneally (10mg/kg) 30min before ovarian detorsion in both groups. After 3 hr of ovarian detorsion, ovariantissue was removed and fixed in 10% formalin solution, embedded in paraffin andevaluated for ischemic indices.

Histological examination showed a significant improvement in ovarianmorphology in the melatonin treated cats. Edema and vasoconstriction in saline groupwere more severe than Melatonin group (p-value = 0.009). Hemorrhage and leukocyteinfiltration were also more obvious in saline group (p-value 0.0018)

Our results demonstrated that Melatonin administration reduced ovarianhistopathological damage due to oxidative injury associated with torsion.

Severe preeclampsia is a quite well-known entity with high incidence ofboth maternal and fetal morbidity and mortality. Although little is known about itsetiology, inherited disorders of hemostasis and antiphospholipid syndrome have beenpostulated as common causes. The present study was conducted to evaluate theassociation of these two entities with preeclampsia in a group of Iranian patients.

A case-control study was performed on 26 parturients withsevere preeclampsia and 26 healthy pregnant women who were matched according tothe age, parity, gestational age and previous history of abortion. A 10cc blood samplewas obtained and the following factors were measured: factor V Leiden, protein S,protein C, antithrombin III, anticardiolipin antibodies (IgM and IgG) and the presenceof the lupus anticoagulant antibody.

We have not found any significant difference in the values of factor V Leiden,antithrombin III, protein C, protein S, and anticardiolipin-IgG between preeclamptic(case) and non-preeclamptic (control) parturients. Meanwhile, lupus anticoagulantantibody was detected in one case and one control. However, anticardiolipin IgM wasshown to be significantly higher in the preeclamptic patients. Severe preeclampticparturients were 4.4 times more likely to develop elevated levels of IgM (OR=4.4, 95%CI=1.9-10, p<0.05).

Our results failed to reveal any significant association betweenpreeclampsia and indices of inherited disorders of hemostasis, except for anticardiolipinIgM. Thus, routine screening of these indices are not recommended due to highexpenses and shortness of reliability.

The value of serial measurement of serum ß subunit of human chorionicgonadotropin (ßHCG) and ultrasonography in the early diagnosis of ectopic pregnancyhas well established.

The objective of this study was to explore the diagnostic value of raisinglevel of serum ßHCG, single measurement of progesterone (P) and estradiol (E2) inearly diagnosis of ectopic pregnancy.

Serum levels of ßHCG and estradiol were measured by RadioImmuno Sorbent Assay (RIA) and progesterone level was measured by Enzyme LinkedImmuno Sorbent Assay (ELISA) techniques in 43 symptomatic women with ectopicpregnancy and 42 women with normal intrauterine pregnancy in Alzahra Hospital,Tabriz, Iran. These values were compared by T-test. By determining cut-off levels ofthese parameters the efficiency and sensitivity of them in prediction of ectopicpregnancy was estimated.

The mean serum levels of ßHCG, estradiol and progesterone in patients withectopic pregnancies (940 ± 552 mlu/ml, 593 ± 237 pg/ml, 5.83 ± 3.41 ng/ml,respectively) were significantly lower than these levels in normal intrauterinepregnancies (4620 ± 2030 mlu/ml, 1627 ± 435 pg/ml, 24.8 ± 6.08 ng/ml, respectively).The average rate of ßHCG rising was 8.2% for 24 hours in patients with ectopicpregnancy (EP) and 32.8% in normal intrauterine pregnancies (NIUP).Conclusions In this study single measurement of serum progesterone level has thegreatest sensitivity (100%) and specificity (98%) in the diagnosis of ectopic pregnancy.

Group B streptococcus is regarded as a potential factor for adverseoutcomes of pregnancy such as preterm birth.

To study the association of maternal vaginal colonization with group Bstreptococcus (GBS) and preterm labor.

From April 2005 to May 2006, vaginal culture for GBS wereconducted in 101 laboring women with a gestational age of 24-37 weeks and 105women admitted for term delivery at maternity center of Afzalipour Hospital in Kerman,Iran. Student`s t test and Chi square test were used to compare continuous andcategorical data between the groups. Using multivariate logistic regression theassociation between GBS colonization and preterm labor was analyzed. P-values<0.05were considered as significant.

Colonization was detected in 9.2% of all mothers. Although GBS colonizationwas found more frequently in preterm than term patients (12 v/s 7 cases), the differencewas not statistically significant. However, GBS positivity was roughly associated withpreterm labor. Age was also a risk factor for GBS colonization. No case of perinatalsepsis occurred during the study period.

Maternal colonization for GBS is relatively low in our center. Increasingage enhances the risk of colonization. Vaginal colonization of GBS is relativelyassociated with preterm labor.

Smoking has negative effects on reproductive process. Exposing tocigarette smoking (passive smoking) may exert some effects as the direct smoking.

The aim of this study was to evaluate the correlation between ovarianresponse and passive smoking in women who underwent ART cycles.

One hundred-sixty patients who underwent ICSI between2000 and 2001 were studied in a prospective cohort study. The case group includedwomen whose husbands smoked at least 5 cigarettes daily for 1 year or more. Thecontrol group included women with nonsmoking husbands. Women with high FSHlevel (>12 IU/ml) were excluded. Long standard protocol with GnRH agonist and HMGwere used in all patients. In vitro fertilization and embryo transfer was carried out in astandard fashion.

Eighty one women were in case group and 82 in control group. Ovarianresponse variables were not significantly different between two groups but there was asignificant relation between passive smoking and fertilization (RR= 1.18, 95% CI: 1.07-1.31). However pregnancy rate was not significantly different between two groups.Moreover there were no significant differences between heavy and light smokers inovarian response outcomes.

This study showed no correlation between ovarian response parametersand passive smoking in women underwent ART cycles, whereas fertilization rate issignificantly lower in this group compared to control group. It may be related to spermquality than oocytes. Assessment of nicotin in follicular fluid and cytogenetic evaluationof embryo before transfer are recommended for more information and confirmation.