Protective effect of alpha glucosyl hesperidin (G-hesperidin) on chronic vanadium induced testicular toxicity and sperm nuclear DNA damage in male Sprague Dawley rats.

The study was conducted to evaluate the vanadium-induced testicular toxicity and its effect on sperm parameters, sperm nuclear DNA damage and histological alterations in Sprague Dawley rats and to assess the protective effect of G-hesperidin against this damage. Treatment of rats with vanadium at a dose of 1 mg kg bw(-1) for 90 days resulted in significant reduction in serum testosterone levels, sperm count and motility. Further, a parallel increase in abnormal sperm morphology and adverse histopathological changes in testis was also associated with vanadium administration when compared to normal control. Moreover, sperm chromatin dispersion assay revealed that vanadium induces sperm nuclear DNA fragmentation. A marked increase in testicular malondialdehyde levels and decreased activity of antioxidant enzymes such as superoxide dismutase and catalase indicates vanadium-induced oxidative stress. Co-administration of G-hesperidin at a dose of 25 and 50 mg kg bw(-1) significantly attenuated the sperm parameters and histological changes by restoring the antioxidant levels in rat testis. These results suggested that vanadium exposure caused reduced bioavailability of androgens to the tissue and increased free radical formation, thereby causing structural and functional changes in spermatozoa. G-hesperidin exhibited antioxidant effect by protecting the rat testis against vanadium-induced oxidative damage, further ensures antioxidant potential of bioflavonoids.

Sperm DNA fragmentation assay by sperm chromatin dispersion (SCD): correlation between DNA fragmentation and outcome of intracytoplasmic sperm injection.

PURPOSE: The aim of the present study was to investigate the relationship between sperm DNA fragmentation index (sDFI) and outcome of intracytoplasmic sperm injection (ICSI). METHODS: All the patients were divided into two groups based on sperm DNA fragmentation analysis by the sperm chromatin dispersion (SCD) method. A total of 237 patients were in the DNA fragmentation normal group (sDFI ≤ 30 %), and 140 patients were in the DNA fragmentation abnormal group (sDFI ≥ 30 %). The relationship of sDFI with the outcome of ICSI was analyzed. RESULTS: A significant difference in semen parameters was observed between the DNA fragmentation normal and abnormal groups [count, motility and morphology (p < 0.05)]. However, no significant difference was seen between the number of oocytes retrieved and fertilization rates between the two groups, whereas the number of embryos progressed to day 3 and the blastocyst formation rate in the remaining embryos after transfer were significantly more in the DNA fragmentation normal group (p < 0.05). A significant negative correlation was noted between DFI values of more than 30 % and number of pregnancies and deliveries (p < 0.05). A higher DFI was also associated with increased abortion rates. CONCLUSIONS: In the present study, sperm with DNA fragmentation showed a negative correlation with semen parameters. Further, sperm with damaged DNA have potential adverse effects on embryo progression, clinical pregnancy rate, and ongoing pregnancies.

CASA derived human sperm abnormalities: correlation with chromatin packing and DNA fragmentation.

PURPOSE: The present study was undertaken to evaluate the effects of morphokinetic abnormalities of human spermatozoa on chromatin packing and DNA integrity and possible beneficial effects of sperm selection in ICSI. METHODS: Semen samples from 1002 patients were analysed for morphology and motility using CASA. Protamine status and DNA fragmentation were analysed by chromomycin A3 staining and sperm chromatin dispersion assay respectively. RESULTS: Sperms with elongated, thin, round, pyri, amorphous, micro and macro forms were significantly higher in teratozoospermic and oligoasthenoteratozoospermic groups. Significant difference in chromatin packing and DNA fragmentation index was observed in these abnormal groups compared with normal. Similarly significant correlation was also seen between abnormal motility parameters and DNA fragmentation index in asthenozoospermic group compared with normal. CONCLUSIONS: Specific abnormal morphological forms have higher incidence of chromatin packing abnormalities and DNA fragmentation. Using these sperms in ICSI might have an impact on fertilization, embryo development and abortion rates. These can be selectively avoided during ICSI procedure to improve ART outcome.

Noninsulin-dependent diabetes mellitus: effects on sperm morphological and functional characteristics, nuclear DNA integrity and outcome of assisted reproductive technique.

The aim of the study was to compare the semen characteristics and nuclear DNA fragmentation in spermatozoa of diabetic and nondiabetic men undergoing assisted reproduction and correlate them with pregnancy outcome. Semen characteristics and nuclear DNA fragmentation were analysed using computer-aided semen analysis system and sperm chromatin dispersion assay (SCD), respectively. Spermatozoa from diabetic patients showed significantly lower progressive (Type A) motility (14.64 ± 9.60 versus 17.99 ± 11.51, P < 0.02) and increased nuclear DNA fragmentation (37.05 ± 12.68 versus 21.03 ± 10.13, P < 0.001). Furthermore, a positive correlation was observed in diabetic patients in terms of blastocyst formation rate (38.13% versus 55.46%, P < 0.001), pregnancy rate (28.57% versus 46.34%, P < 0.001) and miscarriage rate (50.0% versus 24.56%, P < 0.001). The higher percentage of sperm DNA damage because of oxidative stress seen in diabetic patients may be responsible for the poor embryonic development and pregnancy outcome in these individuals.

Vitrification of human early cavitating and deflated expanded blastocysts: clinical outcome of 474 cycles.

PURPOSE: The present study was undertaken to evaluate and compare the post thaw survival, implantation and pregnancy rates of vitrified human early cavitating blastocysts with deflated expanded blastocysts. MATERIAL AND METHODS: Supernumerary blastocysts were vitrified in 30% ethylene glycol-dimethyl sulphoxide based solution using cryoloop. Fully expanded blastocysts were deflated by gentle aspiration of the blastocoelic fluid using a micromanipulator until the cavity collapses prior to vitrification. RESULTS: Of the 576 vitrified blastocysts, 545 (94.61%) survived thawing in the early cavitating blastocyst group which was significantly higher than deflated expanded blastocyst group, in which only 370 survived thawing out of 459 (80.62%). However, no significant difference was observed in implantation and pregnancy rates between early cavitating and deflated expanded blastocyst groups. CONCLUSIONS: Early cavitating blastocyst would be the ideal stage for cryopreservation of human blastocysts as it has higher survival rate and avoids additional invasive procedures like deflation of the blastocoele.

Comparison of follitropin-beta administered by a pen device with conventional syringe in an ART programme - a retrospective study.

OBJECTIVES: This study compares the efficacy and patient tolerance of follitropin-beta (recagon) administered using a pen device with conventional syringe in infertile couples undergoing in vitro fertilization/intracytoplasmic sperm injection treatment. METHODS: Data for 481 patients were retrieved retrospectively for the analysis. Conventional syringe group constituted 204 patients with 217 cycles and 265 patients with 294 cycles in the pen-device group. Down-regulation was achieved with GnRH agonist. RESULTS: Comparison of follitropin-beta administered with pen and syringe showed the following data, respectively. A total dose of 1909.38/2100.65 IU (P < 0.001), duration of stimulation, 9.70/10.47 days (P < 0.05), oestradiol levels on the day of human chorionic gonadotropin, 1488.34/1067.63 pg/ml, number of follicles reaching >16-mm size, 9.75/7.34 (P < 0.05), number of oocytes retrieved, 13.84/9.55 (P < 0.001) and number of embryos available for freezing, 4.56/1.30 (P < 0.05), the above data were observed in pen/conventional syringe groups, respectively. The live birth rates per cycle were 28.85% and 30.95% in the conventional syringe/pen-device groups, respectively. Patient tolerance with respect to pain at injection site was better with the pen device (P < 0.025). CONCLUSION: The data show that follitropin-beta administered with pen device is well tolerated and more efficacious with respect to ovarian stimulation outcome compared with the conventional syringe.

Meiotic spindle and zona pellucida characteristics as predictors of embryonic development: a preliminary study using PolScope imaging.

This study assesses meiotic spindle and zona pellucida characteristics using the PolScope, and analyses their relationship to embryonic development potential. A total of 205 matured oocytes retrieved from 25 patients undergoing ovarian stimulation were imaged for meiotic spindle and zona pellucida characteristics using the PolScope. After intracytoplasmic sperm injection, the oocytes were cultured and assessed for progression to blastocysts. Meiotic spindles were visualized in 78.0% of oocytes. Significantly more oocytes with visible spindles fertilized and progressed to blastocysts compared with oocytes without visible spindles. Oocytes with spindle retardance of >3 nm showed a greater progression to blastocysts compared with those with a retardance of 2-3 nm or <2 nm. More blastocysts were obtained from oocytes with spindle lengths of >12 nm than from oocytes with spindle lengths 10-12 nm or <10 nm. A difference in progression to blastocyst was observed in oocytes with a zona inner layer retardance of >3 nm compared with oocytes with retardance of 2-3 nm or <2 nm. Oocytes with an inner layer zona of 10-12 nm thickness showed better progression compared with those with a thickness of 8-10 nm or <8 nm. Quantitative measurement of length and retardance of the meiotic spindle and zona pellucida has a positive predictive value in relation to embryonic development.

Assessment of uterine cavity by hysteroscopy in assisted reproduction programme and its influence on pregnancy outcome.

OBJECTIVE: The aim of the study was to evaluate if the diagnosis and treatment of uterine cavity abnormalities by hysteroscopy in patients undergoing IVF programme is of any value in improving clinical pregnancy outcome. METHODS: 520 patients participated in this prospective randomized study and were classified into two groups. Group I (n = 265) without office hysteroscopy. Group II (n = 255) had office hysteroscopy and was sub classified into Group II a and Group II b. Group II a (n = 160) had normal hysteroscopic findings whereas Group II b (n = 95) had abnormal office hysteroscopy findings, which were corrected at the same time. RESULT: There was no difference in the mean number of oocytes retrieved, fertilization rate, and number of embryos transferred among the patients in different groups. Statistically significant difference was observed in terms of clinical pregnancy rates between Group I and Group II a (26.2 and 44.44%, P < 0.05), and Group I and Group II b (26.2 and 39.55%, P < 0.05), respectively. CONCLUSION: Patients with recurrent IVF embryo transfer failures after normal hysterosalpingography findings should also be reevaluated using hysteroscopy prior to further commencing IVF-embryo transfer cycles in order to enhance the clinical pregnancy rates.

Successful pregnancy with laparoscopic oocyte retrieval and in-vitro fertilisation in mullerian agenesis.

A 28-year-old woman with mullerian agenesis presented with primary infertility and was considered for laparoscopic oocyte retrieval and in-vitro fertilisation. Her 27-year-old younger sister served as a gestational carrier. The patient underwent ovarian stimulation and 11 mature oocytes were retrieved by laparoscopy. After successful in-vitro fertilisation, two embryos were transferred to the gestational carrier. Two weeks after embryo transfer, the pregnancy was confirmed by serum human chorionic gonadotropin levels. Another two weeks later, an ongoing singleton pregnancy with foetal heartbeat was confirmed by transvaginal ultrasonography.

Multicentric, placebo-controlled, randomised double-blind evaluation of a new herbal cream in vagial infections.

Efficacyand safety of a new herbal cream containing aqueous extracts of Azadirachta indica, Curcuma longa, Pongamia glabra, Glycyrrihiza glabra and Santallum album were evaluated in amulticentric, randomized, double-blind, placebo-controlled study. With active drug treatment, there was significant improvement in various signs like redness, oedema and symptoms like itching, burning, discharge and discomfort, compared to placebo treatment. Microscopic examination of smear and culture showed significant reduction of offending organisms after treatment with active drug. In patient's global evaluation, active drug was rated 70% as very good and in investigators evaluation 82% as very effective and effective. The overall efficacy was as high as 76% with active drug as against only 24% with placebo. Both active drug and placebo were well tolerated.