International Journal of Reproductive BioMedicine
Volume:2 Issue: 1, May 2004

Since its first clinical application in early 90s, preimplantation genetic diagnosis (PGD) hasbecame a powerful diagnostic procedure in clinical practice for avoiding the birth of an affectedchild as well as increasing the assisted reproductive technologies (ART) outcome. The techniqueinvolves the screening of preimplantation embryos for chromosomal abnormalities in certainindications such as advanced maternal age, repeated abortions and translocations, or for single genedefects, the majority of which are cystic fibrosis and thalassaemias. In this context, it becomes analternative option for traditional prenatal diagnosis. So far, more than 1000 unaffected babies havebeen born after PGD, indicating that the procedure is safe and effective in prevention of geneticdefects as well as increasing the ART outcome. Besides its diagnostic value and expandingindications such as cancer predisposition, dynamic mutations and late onset disorders, a newfeature, namely preimplantation human leuckocyte antigen (HLA) typing also demonstrates itsnovel therapeutic role in contemporary medicine. This article summarizes the recent status of PGDand discusses the current limitations and future perspectives associated with PGD techniques.

Proper collections of human ejaculates are necessary for semen analysis and infertilitytreatment purposes. The objective of this study was to assess the seminal characteristics ofejaculates collected by patients via masturbation and coitus interruptus. Thirty individualsproduced one sample via masturbation and one via incomplete coitus during a 3-days interval.The semen parameters were compared and analyzed with student t-test and Nemar test. Theresults showed that mean values for progressive motility of spermatozoa were increased from46.81+15.7% to 58.76+13.5% in coitus interruptus and masturbation, respectively (P<0.01).Also, the mean values for normal sperm morphology was 54.03+25.1% in coitus interruptusand 63.36+13.4% in samples collected via masturbation (P<0.01). In addition, spermconcentration was significantly improved in ejaculates collected with masturbation (P<0.05).Although, insignificant, the concentration of round cells were lower in specimens collectedvia masturbation than coitus interruptus. Therefore, via masturbation method, better semencharacteristics were yielded which subsequently may improve the infertility treatmentoutcome.

The cytokine of granulocyte macrophage colony stimulating factor (GM-CSF) is aglycoprotein, which is synthesized in the female reproductive tract and has embryonic trophiceffect in mammals. The objective of this study was to examine the optimal dosage of GMCSFto improve the mouse embryo development in vitro. To collect two and eight cellsembryos, the pregnant NMRI mice were sacrificed by cervical dislocation at 48 h and 72 hpost hCG injections, respectively. The embryos were cultured randomlly in T6 mediumsupplemented with 5 mg /ml bovine serum albumin (BSA) and 0, 2, and 10 ng / ml humanrGM-CSF. The data of blastocyst formation and hatching in different groups of embryoculture were compared by chi-square analysis. The results showed that the developmentalrates of 2 and 8 cells embryos to hatching blastocyst in the presence of 2 ng/ml of GM-CSFtheir control groups (51.5% and 49.7%, respectively) were more than those in the othergroups, but insignificant. It seems more researches are necessary to confirm this suggestionthat the GM-CSF with 2 ng/ml concentration may have a better potential, not only to enhancethe developmental rates of 2 and 8 cells embryos but also for decreasing the degeneration ofthose embryos.

The luteal phase defect is a common event following the ovarian stimulation.The aim of the present study was to evaluate the use of human chorionic gonadotropine (hCG)and progesterone hormones to improve the luteal phase defect.

60 mice were superovulated routinely with human menopausalgonadotropin (hMG) (7.5U) and hCG (10U). The mice were mated and divided into 3groups: 1- control (n=20) 2- hCG treatment (n= 20), and 3-Progesterone treatment (n=20).Each group was divided again into two subgroups. The mice (10 from each group) had noinjection in group one and were injected intraperiteneal (IP) by hCG (5U/day) andprogesterone (1mg/day) subcutaneously (sc) in groups 2 and 3, respectively for four days. Onthe day 5, the animals were killed by cervical dislocation and the uterus were flushed to countthe number of blastocyst and their quality. The above treatment were carried out for 12 daysin the other 10 mice in each group. Similarly group one had no injection and groups 2 and 3were injected by hCG and progesterone for 12 days respectively by the same manner asmention above. The animals were killed on day 13 and the implanted embryos were counted.The uterus and ovary were processed on days 5 and 13 of pregnancy for histological studies.

The mean number of blastocysts per mouse were: 12.2%, 2.6% and 3% in group 1 to3, respectively. The nomber of implanted embryos were 29 as: 13 living fetus in one mouseand 16 resorption fetus in the other. The morphology of uterus on day 5 was as follow: nodevelopment in the stroma and endometrial gland in control group, the stroma andendometrial gland so developed to form the saw teeth appearance which indicated onreceptivity of uterus in hCG treated group similar to progesterone treated group, but withoutthe saw teeth appearance. The continuation of hCG injection maintained the receptivity ofuterus; while, the continuation in progesterone caused metaplesia of epithelium. Themorphology of ovaries in all three groups showed no changes in corpus luteum size on day 5,and showed the following changes on day 13: increasing the number of primary andsecondary follicles in control group; while, reducing the size of corpus luteum in hCG group.

Progesterone did not improve the uterus and implantation rate. The prolongedusage of progesterone can change the morphology of uterus to more abnormal state inconterast to the prolonged usage of hCG.

Enzyme Linked ImmunoSorbent Assay (ELISA) has been described as analternative to radioimmunoassay for the mammalian and nonmammalian steroids detection. Inthis study, a simple and rapid ELISA is described and validated for 4-pregnen-3,20, dione(progesterone).

A general procedure for preparation of the acetylcholinesteraselabelled steroid is described which is applicable to any steroid. Use of acetylcholinesterasetracer increased the sensitivity of assay so that reliable measurements of each steroid could beachieved with only 10 μl of plasma.

Typical standard curves for progesterone steroids showed a workable range(detection limit) from 0.8 to 400 pg/well and the sensitivity of the assay taken as theconcentration of steroid that induced 90% of B/B0, was 1.5 pg. Inter-assay variations that gaveapproximately 50% displacement was 9.2% for 10 replicates and intra-assay co-efficient ofvariation was less than 10% over the central part of the standard curve between 3 and 200pg/well. There was a strong positive correlation (r>0.999) between the amount of steroidadded to plasma and the amount measured.

Method described here was applied to measure progesterone in plasma and thismethodology could be of great interest to researchers measuring steroid hormones.

The retrieval of good quality oocytes that is accomplished with selection of thebest induction ovulation protocol on the basis of patients condition, age and cause ofinfertility, is one of the most important aspects of ART cycles. The objective was to evaluatethe efficacy of low dose, long acting GnRH-a (Decapeptyle) for pituitary desensitization andoutcome of ART compared to long protocol of short acting GnRH-a (Busereline).

In this randomized clinical trial that was performed at Yazd IVFCenter, 60 patients with 61 cycles of ART were included. Patients with endometriosis or age> 40 were excluded in this study. Using COH-ET, patients were randomly divided into twogroups. In group one, 30 patients received a single half dose of Decapeptyle (1.87mg) in midlutealphase. In the other group, 31 patients received Buserelin daily (0.5mg), starting fromprevious mid-luteal phase. This was reduced to 0.25mg from gonadotropin administration dayand was continued until the day of hCG injection. In these groups, the number of oocytes, thefertilization, cleavage, pregnancy and cancellation rates were compared.

In two groups, there was no case of cancellation due to premature LH surge. Ingroup I, the mean number of gonadotropins was 27.5+4.2 ampoules while in the secondgroup, it was 28.4±2.8 ampoules (P>0.05). 312 oocytes from group I and 294 oocytes fromgroup II were retrieved. Oocyte quality in group II was better than group I (84.3% vs 77.2%,P<0.05). In long-acting GnRH-a group fertilization rate was 81.9% versus 71.1%in group II(P<0.01). However, embryo development in Group I (85.6% vs 94.1%, P<0.05) was lowerthan group II. Although, pregnancy rate was 20% in Group I which was higher than group II(12.6%) but, there was no significant difference in cancellation, pregnancy rate andgonadotropins dose in two groups.

The low dose long acting GnRH-a is a useful method for pituitary suppression.Low dose GnRH-a combined with gonadotropins permitted the retrieval of good qualityoocytes and had no effect on oocytes. The fertilization and pregnancy rates with this methodare acceptable and its cost and tolerance is valuable for patients.

It has been proposed that oxidative stress plays an important role in maleinfertility. The aims of this study were to compare seminal plasma levels of 15-F2t-isoprostane(8-iso-PGF2α), malondialdehyde (MDA), and total (sum of free and bound) homocysteine(tHcy) in normozoospermic vs. asthenozoospermic men, and to examine the relationshipsbetween tHcy and lipid peroxidation products.

The study was a case-control study with a simple random sampling.The case group consisted of 15 asthenozoospermic males. This group was compared with 15normozoospermic men. Seminal plasma levels of 15-F2t-isoprostane and tHcy were measuredusing commercially available enzyme immunoassay (EIA) kits. MDA levels were determinedby the thiobarbituric acid (TBA) assay. The Mann-Whitney U test was used to compare twogroups. Coefficients of correlation were calculated using Spearman’s correlation analysis. Allhypothesis tests were two-tailed with statistical significance assessed at the p value <0.05level.

MDA levels were lower in asthenozoospermic subjects than in control subjects(0.72±0.06 μM vs. 0.40±0.06 μM; p<0.05). No differences were seen in 15-F2t-isoprostanelevels in asthenozoospermic subjects and controls (65.00±3.20 pg/ml vs. 58.17±4.12 pg/ml;p>0.05). Interestingly, tHcy levels were slightly higher in asthenozoospermic subjects than incontrols (6.18±1.17 μM vs. 4.8±0.52μM). Sperm motility was inversely correlated withseminal plasma 15-F2t-isoprostane and MDA levels, respectively (p<0.05).

Seminal plasma levels of 15-F2t-isoprostane and tHcy showed no significantdifferences between normozoospermic and asthenozoospermic men. Sperm motilitycorrelated inversely with seminal plasma levels of 15-F2t-isoprostane and MDA. Norelationship was found between tHcy and lipid peroxidation. However, higher sample size isrequired to confirm these findings.

Pelvic inflammatory disease (PID) is a rare complication of transvaginal oocyte retrieval. Itmay result in failure of assisted reproductive techniques (ART). During a 7 years period, 5958transvaginal ultrasound-guided oocyte retrievals resulted in 10 cases of acute PID. Eight out of 10patients were diagnosed infertile because of endometriosis. Two patients had mild ovarian, 3 hadstage III, and 2 had stage IV endometriosis. One patient had a 3-4 cm ovarian endometrioma.After treatment, no mortality was encountered among the 10 patients, although none of themconceived. This observation supports the previous reports that endometriosis can raise the risk ofPID after oocyte retrieval. More vigorous antibiotic prophylaxis and better vaginal preparationare recommended when oocyte pickup is performed in patients with endometriosis.