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Biomedical subjects

Navid Esfandiari

Publications and source records attributed to Navid Esfandiari.

18 recordsLinked to original sources

Expression of glycodelin and cyclooxygenase-2 in human endometrial tissue following three-dimensional culture.

PROBLEM: Our previous study showed that in vitro culture of human endometrial tissue in a three-dimensional (3D) fibrin matrix could mimic the early stages of endometriosis with invasion, gland and stroma formation and sprouting of new vessels. The objective of the present study was to evaluate the expression of glycodelin (Gd) and cyclooxygenase-2 (COX-2), two angiogenic factors, to further validate the 3D culture model of endometriosis. METHOD OF STUDY: Human endometrial fragments were obtained from endometrial biopsies and placed in a 3D fibrin matrix culture. Immunohistochemistry with specific antibodies to Gd and COX-2 was used to examine endometrial epithelium and blood vessels, and 4, 6-diamidino-2-phenylindole staining was used for nuclear identification. RESULTS: Three-dimensional culture of human endometrial tissue in the fibrin matrix resulted in the proliferation of endometrial stromal cells, glandular epithelium and angiogenesis. Gd positive glandular epithelium was seen in 85% of wells with developing endometrial glands and COX-2 positive new vessels were seen in 80% of wells with angiogenesis-like structures after 4 weeks of culture. CONCLUSION: Our findings confirm that angiogenesis occurs following the culture of endometrial tissue in the 3D fibrin matrix, and suggests that Gd and COX-2 might play important roles in promoting neovascularization and cell proliferation in the establishment of endometriosis.

Adult↗

Follicular phase dynamics with combined aromatase inhibitor and follicle stimulating hormone treatment.

OBJECTIVE: The objective of this study was to evaluate follicular phase parameters during ovarian stimulation with FSH alone or with the aromatase inhibitor letrozole. METHODS: Two groups of women undergoing intrauterine insemination (IUI): group I (389 patients; mean age 35 +/- 4.3 yr) underwent 630 IUI cycles stimulated with letrozole and FSH; and group II (134 patients; mean age 36.0 +/- 4.6 yr) underwent 166 IUI cycles stimulated with FSH only. Each group was stratified into ovulatory and anovulatory cycles. Patients were monitored by ultrasound for folliculometry and blood sampling for hormonal assay on d 3, 7, 9, or 10 of the cycle, and on the day of human chorionic gonadotropin administration. RESULTS: Group I had a significantly lower follicular count greater than 10 mm on d 7, greater than 12 mm on d 9 or 10, and greater than 15 mm on the day of human chorionic gonadotropin administration compared to group II (P = 0.006, <0.001, and <0.001, respectively). After stratifying patients by diagnosis, this relationship was maintained only for patients with ovulatory infertility (P = 0.003, <0.001, and <0.001, respectively). Serum estradiol (E2) was significantly lower in the group I ovulatory and anovulatory at the last three monitoring visits (P < 0.001). However, the difference in E2 levels decreased in the preovulatory period with similar E2 levels per mature follicle. No premature preovulatory progesterone rise was observed in either group. However, significantly lower progesterone levels were observed in the second half of the follicular phase in group I (P = 0.02 and <0.001). Endometrial thickness was significantly lower in group I at the second and third visits (P < 0.001, 0.01) but was comparable to group II at the last monitoring visit. Although, the pregnancy rates were similar between the two groups, the multiple pregnancy rate was significantly higher in the FSH-only group (P = 0.039). CONCLUSION: The addition of letrozole modifies the follicular, hormonal, and endometrial dynamics of FSH-stimulated cycles with possible positive effects on the overall cycle outcome.

Adult↗

Effect of a statin on an in vitro model of endometriosis.

OBJECTIVE: To determine the inhibitory effect of a statin on angiogenesis in a three-dimensional (3-D) culture of human endometrial fragments in vitro. Angiogenesis has been proposed as an important mechanism in the pathogenesis of endometriosis, and statins (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors) have been shown to have anti-inflammatory and anti-angiogenic activity. DESIGN: Experimental in vitro study of human endometrial biopsies and 3-D culture in fibrin matrix. SETTING: Research laboratory at a university-affiliated infertility center. PATIENT(S): Forty-six normal ovulating women undergoing infertility treatment. INTERVENTION(S): Endometrial samples obtained from the fundus of the uterine cavity were minced, and the fragments were placed in a three-dimensional fibrin matrix culture system. MAIN OUTCOME MEASURE(S): Presence or absence of proliferation of stromal cells and invasion of the fibrin matrix, presence or absence of vessel sprouting, and immunohistochemical characterization of cellular components. RESULT(S): During the 1st week of culture, invasion of stromal cells into the fibrin matrix occurred in the control group and in some wells outgrowths were observed. After 2 weeks, endometrial glands were observed in the outgrowths at a distance from the main tissue and were growing in conjunction with new vessel formation until the end of culture period. A concentration-dependent effect of lovostatin was seen on cell growth and angiogenesis in the experimental groups. In the presence of 5 and 10 microM of statin, angiogenesis was abolished, and cell proliferation was inhibited. In the presence of 1 microM of lovastatin, angiogenesis was reduced, but cell proliferation was not affected. CONCLUSION(S): The statins were shown to be effective in inhibiting the mechanisms of cell proliferation and angiogenesis in an experimental model for the development of endometriosis-like tissue.

Adult↗

Brown oocytes: implications for assisted reproductive technology.

Fertilization, embryo development, and successful pregnancy can be achieved after transfer of embryos derived from brown oocytes at the same rate as embryos from morphologically normal oocytes. Therefore, brown oocytes are probably normal and this morphological criterion does not seem to be indicative of any adverse outcome in IVF.

Adult↗

Live birth of healthy triplets after in vitro fertilization and embryo transfer in an acromegalic woman with elevated growth hormone.

OBJECTIVE: To report a case of successful triplet pregnancy in an acromegaly patient with IVF and embryo transfer. DESIGN: Case report. SETTING: Tertiary center for assisted reproductive technologies. PATIENT(S): A 33-year-old woman with acromegaly and primary infertility. INTERVENTION(S): Controlled ovulation hyperstimulation and IVF/embryo-transfer treatment using gonadotropins. MAIN OUTCOME MEASURE(S): Uncomplicated pregnancy course and successful take-home triplet infant girls. RESULT(S): Successful ovarian stimulation and IVF is possible in acromegalic patients. CONCLUSION(S): There may be an effect of elevated growth hormone resulting in increased ovarian response to gonadotropin, inducing hyperstimulation and high-order multiple pregnancies in these patients.

Acromegaly↗

Effects of peritoneal fluid on preimplantation mouse embryo development and apoptosis in vitro.

To evaluate the effect of peritoneal fluid (PF) on preimplantation mouse embryo development and apoptosis, PF was obtained from women presenting with (n = 7) and without endometriosis (n = 7). Mouse embryos were cultured to the blastocyst stage in human tubal fluid medium alone (control) or 10% PF. Embryo development was assessed and the total cell number per embryo was determined by Hoechst 33258 staining. Allocation of inner cell mass and trophectoderm in blastocysts, and incidence of apoptosis were determined using confocal microscopy. The blastocyst development rate was significantly lower in the presence of 10% PF (P < 0.01). Total cell number and trophectoderm in blastocysts cultured in culture media alone was significantly higher than in the presence of PF (P = 0.034 and P = 0.01 respectively). A higher incidence of apoptosis was seen in blastocysts cultured in culture media alone, compared with those cultured in the presence of PF (P = 0.04). PF decreases the development of early mouse embryos to the blastocyst stage, as well as the incidence of apoptosis in the resulting mouse blastocysts. No difference was seen in the effect of PF from women with and without endometriosis on mouse embryo development and apoptosis.

Animals↗

Successful pregnancy following transfer of embryos from oocytes with abnormal zona pellucida and cytoplasm morphology.

The failed or impaired fertilization in an IVF cycle may be a result of undetected abnormalities in sperm function, poor oocyte quality, or impaired spermatozoon-oocyte interaction. Whether oocyte dysmorphisms have an impact on intracytoplasmic sperm injection (ICSI) outcome, fertilization, and implantation is controversial. A 33-year-old nulligravida female with a 2-year history of primary infertility was referred to the Toronto Centre for Advanced Reproductive Technology for infertility management. The patient underwent several cycles of ovulation induction followed by timed intercourse or intrauterine insemination without a resulting pregnancy. Following the failed insemination cycles, she proceeded to IVF. Six morphologically abnormal oocytes (including three that were cucumber-shaped) were retrieved and all injected with a single spermatozoon. Four oocytes showed normal fertilization and developed to day-3 embryos with abnormal morphology and were transferred. Two weeks after the embryo transfer, the patient had a positive beta-human chorionic gonadotrophin (beta-HCG). The pregnancy was uneventful, and a healthy baby boy was delivered at 41 weeks of age by Caesarean section. Since fertilization, embryo development, and successful pregnancy was achieved in this case, it is recommend that oocytes with extreme morphological abnormalities should not be discarded as ICSI may overcome the barriers to fertilization and cleavage.

Adult↗

Protein supplementation and the incidence of apoptosis and oxidative stress in mouse embryos.

OBJECTIVE: To estimate the effect of protein supplementation of culture media on reactive oxygen species production and incidence of apoptosis in preimplantation mouse embryos. METHODS: A total of 72 two-cell mouse embryos were cultured in human tubal fluid (HTF) alone (HTF-alone, control) and 71 embryos in HTF with protein supplementation (10% serum substitute supplement; HTF-SSS) for 72 hours. Total cell number per embryo was determined by staining with Hoechst 33258. Allocation of inner cell mass and trophectoderm in blastocysts and incidence of apoptosis were determined by confocal microscopy. Levels of reactive oxygen species in culture media were measured by chemiluminescence assay using luminol as probe. RESULTS: Blastocyst development, total cell number, and the inner cell mass/trophectoderm ratio were similar between the 2 groups. The blastocyst hatching rate was significantly higher in the HTF-SSS group than in the HTF-alone group (20% compared with 4%, P = .007). Level of reactive oxygen species was significantly higher in HTF-alone compared with HTF-SSS at 24 hours (median and interquartile range 28 [13, 43] compared with 0 [0, 1], P = .02), 48 hours (24 [21, 26] compared with 2 [1, 2], P = .02), and 72 hours (26 [9, 32] compared with 2 [2, 3], P = .03). The incidence of apoptosis in blastocysts cultured in HTF-SSS was significantly lower than those in HTF-alone group (mean +/- standard deviation 2.38 +/- 0.68 and 5.81 +/- 1.11, respectively, P = .001). CONCLUSION: Protein supplementation of culture media improves the hatching rate and reduces reactive oxygen species levels and the incidence of apoptosis in mouse preimplantation embryos.

Animals↗

Novel association between sperm reactive oxygen species production, sperm morphological defects, and the sperm deformity index.

OBJECTIVE: To examine the relationship between sperm reactive oxygen species (ROS) production and sperm morphology in a group of infertile men and healthy fertile donors. DESIGN: A prospective clinical study. SETTING: Male infertility clinic, Glickman Urological Institute, The Cleveland Clinic Foundation, Cleveland, Ohio, and the Reproductive Medicine Unit, Liverpool Women's Hospital, United Kingdom PATIENT(S): Thirty-nine infertile men and 13 healthy fertile donors (control). INTERVENTION(S): Standard semen analysis, seminal leukocyte concentration, assessment of sperm morphology, and measurement of sperm ROS production. MAIN OUTCOME MEASURE(S): Levels of sperm ROS production, percentages of different sperm morphological abnormalities, and the sperm deformity index (SDI) scores. RESULT(S): A significant negative correlation was observed between sperm ROS production and the proportion of sperm with normal morphology and borderline morphology. Reactive oxygen species production was positively correlated with the proportion of sperm with amorphous heads, damaged acrosomes, midpiece defects, cytoplasmic droplets, tail defects, and SDI scores. Logistic regression analysis identified a two-variable model including SDI and percentage sperm motility, which correctly identified 84% of individuals with high seminal ROS and 85% of individuals with low seminal ROS. The model had an overall accuracy of 85%. CONCLUSION(S): The standard semen analysis to assess sperm motility, sperm morphology, and the SDI scores is a useful tool in identifying infertile men with high seminal ROS in infertility clinics where facilities for measuring levels of seminal ROS are not available.

Female↗

Heterophile antibody blocking agent to confirm false positive serum human chorionic gonadotropin assay.

BACKGROUND: Persistently false positive human chorionic gonadotropin (hCG) test results in the absence of an intrauterine pregnancy may lead to a diagnosis of ectopic pregnancy or gestational trophoblastic neoplasia and unnecessary testing and therapy. CASE: A 38-year-old woman on oral contraceptives and a 53-year-old perimenopausal woman presented with persistently elevated serum beta-hCG levels from 30 to 225 mIU/mL. Both women had a history of working with mice. Urine hCG testing was negative. Serum beta-hCG levels for both patients were negative after pretreatment of their serum with a heterophile antibody blocking agent. CONCLUSION: These cases illustrate that serum heterophile antibodies can interfere with the hCG enzyme immunoassay and result in false positive values. The addition of heterophile blocking agent to the serum can exclude false positives, thereby preventing unnecessary evaluation and treatment.

Adult↗

Total antioxidant capacity and reactive oxygen species in amniotic fluid.

OBJECTIVE: To identify the presence and/or absence of reactive oxygen species and total antioxidant capacity in amniotic fluid and to determine changes in the levels of reactive oxygen species and total antioxidant capacity with advancing gestational age of pregnancy and fetal or neonatal weights. METHODS: Amniotic fluid was collected from a total of 26 nonsmoking patients. Nine specimens were collected in the third trimester of pregnancy, and 17 specimens were collected in the second trimester. Amniotic fluid reactive oxygen species and total antioxidant capacity levels were determined by chemiluminescence and spectrophotometric assays, respectively. Reactive oxygen species and total antioxidant capacity levels were established and then compared for advancing gestational ages and estimated fetal weights or neonatal weights. RESULTS: Reactive oxygen species levels were present in some but not all specimens, and total antioxidant capacity was present in all specimens. Total antioxidant capacity but not reactive oxygen species levels increased from the second to the third trimester (347.0 mmol/L versus 776.0 mmol/L, P <.001). There was a positive Spearman correlation between total antioxidant capacity and gestational age (r = 0.72, 95% confidence interval 0.43, 1.0, P <.001) and between total antioxidant capacity and estimated fetal weights or neonatal birth weights (r = 0.70, 95% confidence interval 0.40, 1.0, P <.001). There was no correlation between reactive oxygen species and advancing gestational age or weight. CONCLUSION: Total antioxidant capacity levels are present in amniotic fluid at least as early as the second trimester and increase with advancing gestational age and fetal or neonatal weights. Reactive oxygen species levels are not necessarily present in amniotic fluid, and they do not appear to be influenced by gestational age or estimated fetal or neonatal weights.

Adult↗

Utility of the nitroblue tetrazolium reduction test for assessment of reactive oxygen species production by seminal leukocytes and spermatozoa.

The purpose of this study was to evaluate the ability of spermatozoa and leukocytes in semen to produce reactive oxygen species (ROS) by using nitroblue tetrazolium (NBT) staining and to examine the association between NBT staining and levels of ROS as measured by chemiluminescence. Twenty-one infertility patients (leukocytospermia; n = 8; nonleukocytospermia, n = 13) and 9 healthy donors were included. Standard semen analysis and density gradient centrifugation were performed to test NBT staining, ROS, and total antioxidant capacity. A ROS-total antioxidant capacity (ROS-TAC) score was calculated by using principal component analysis. In the leukocytospermic group, after separation on a density gradient, the percentage of NBT-positive staining was significantly higher in sperm suspensions contaminated with leukocytes (median [25th, 75th percentiles]; 70% [61%, 79%]) compared to the nonleukocytospermic group (14.5% [9%, 25.5%]; P =.03) and donors (7% [3%, 11%]; P =.02), respectively. A strong positive correlation was seen between levels of ROS in whole ejaculates and NBT-positive staining in leukocytes (r = 0.59; P <.0006) and in leukocyte fractions (r = 0.72; P <.0001) after density gradient separation. Similarly, ROS was positively correlated with excessive cytoplasmic retention in spermatozoa from whole ejaculates and abnormal spermatozoa after separation on density gradients (r = 0.72; P <.0001). The ROS-TAC score was inversely correlated with NBT staining in leukocytes in whole ejaculates (r = -0.960, P <.0007) and in both leukocyte fractions (r = -0.39; P <.04) and spermatozoa with cytoplasmic retention (r = -0.38; P <.04). Our results indicate that the NBT reduction test can be used to assess the contribution of seminal leukocytes and defective spermatozoa towards ROS generation in semen. Levels of ROS assessed by chemiluminescence assay are strongly correlated with the results of NBT staining.

Antioxidants↗

Effects of temperature on sperm motion characteristics and reactive oxygen species.

OBJECTIVE: To evaluate the effect of temperature on sperm motility, sperm motion characteristics, and production of reactive oxygen species (ROS) in semen. METHODS: We collected semen samples from 12 infertile patients and 12 healthy donors and divided each sample into four aliquots just after liquefaction. One aliquot was analyzed immediately after liquefaction and the remaining three were analyzed after one hour of incubation at 4 degrees C, 25 degrees C, and 37 degrees C. Sperm motility and motion characteristics were measured by computer-assisted sperm analysis (CASA). Levels of ROS were determined in washed sperm suspensions using a chemiluminescence assay and results were expressed as x10(6) counted photons per minute (cpm)/20 million sperm/mL. RESULTS: Motility, curvilinear velocity, straight-line velocity, average path velocity, and amplitude of lateral head displacement decreased significantly from baseline values in both the patients and donors after incubation at 4 degrees C. Percentage motility was highest and ROS levels were lowest in the samples that were incubated at 37 degrees C. CONCLUSION: Based on our results, we suggest that semen samples be stored at 37 degrees C after collection and during transportation and processing.

Adult↗

Positive bacterial culture of semen from infertile men with asymptomatic leukocytospermia.

OBJECTIVE: To examine results of semen culture in a population of infertile men with asymptomatic leukocytospermia, and to determine the correlation between culture results and sperm characteristics in these patients. METHODS: Semen samples were collected from a group of infertile men (n = 80) after 2-3 days of sexual abstinence. Standard semen analysis was performed according to World Health Organization (WHO) guidelines. Seminal leukocyte concentrations were determined by a myeloperoxidase staining technique. Culture of semen was performed on enriched and specific culture media. RESULTS: Based on the results of semen culture, the samples were classified into three groups: group 1 (enteric gram-negative bacilli [Enterobacteriaceae], n = 13), group 2 (bacteria other than Enterobacteriaceae, n = 15), and group 3 (negative cultures, n = 52). No correlation was found between positive semen culture and sperm characteristics (concentration, motility, and morphology). However, seminal leukocyte concentrations were negatively correlated with percentages of motile sperm and normal sperm morphology. CONCLUSION: Our data indicate that semen samples from infertile men with asymptomatic leukocytospermia may have bacterial contamination, evidenced by positive bacterial cultures. Lack of correlation between positive semen cultures and sperm characteristics may be indicative of early or mild (subclinical) infection.

Enterobacteriaceae↗

Gestational carrier is an optimal method for pregnancy in patients with vaginal agenesis (Rokitansky syndrome).

OBJECTIVE: To evaluate the results of successful pregnancy for patients with Mayer-Rokitansky-Kuster-Hauser syndrome using a gestational carrier. METHODS: All gestational carrier patients who attended our infertility clinic between 1995 and 2002 were reviewed for this study. Of the patients, 6 women with MRKH syndrome had 12 ovarian stimulation cycles, which resulted in 12 fresh and 5 frozen embryo transfers into six gestational carriers. RESULTS: The mean number (+/-SD) of embryos obtained per cycle was 4.8 (+/-2.9). Either two (n = 15) or three (n = 2) embryos were transferred to the gestational carrier. Three pregnancies were achieved following fresh embryo transfer, which included one clinical pregnancy that ended in spontaneous abortion, a singleton, and a set of twins. The pregnancy rate for fresh embryo transfer was 25% per retrieval and 50% per patient. No pregnancy was achieved following frozen embryo transfer. CONCLUSION: Gestational carrier is shown to be an effective treatment for patients with Mayer-Rokitansky-Kuster-Hauser syndrome.

Adult↗

Complete failed fertilization after intracytoplasmic sperm injection--analysis of 10 years' data.

OBJECTIVE: To investigate incidence and causes of complete failed fertilization after intracytoplasmic sperm injection (ICSI) in a tertiary care facility. METHODS: A total of 1,779 cycles between February 1994 and December 2003 were analyzed. Study parameters were female age, infertility diagnosis, ovarian stimulation protocol, estradiol level on day of hCG administration, number of follicles, number of oocytes retrieved, number of oocytes injected, and semen parameters. RESULTS: Complete failed fertilization occurred in 23 cycles (1.29%) involving a total of 85 oocytes injected. Infertility causes among patients with failed fertilization included unexplained (43.6%), male factor (26%), presence of more than one factor (17.4%), hysterectomy (4.4%), premature ovarian failure (4.3%), and advanced age (4.3%). In 12 cycles (52%), fewer than 5 follicles were present. In three (13%) cycles, no mature (MII) oocyte was available and in 61% (14/23) fewer than 3 MII oocytes were available for ICSI. Immotile sperm was used for ICSI in 5 cycles (21.7%). The source of sperm in 17 (74%) cycles was from ejaculate, in 4 cycles from testicular aspiration (TESA), one from percutaneous epididymal sperm aspiration (PESA) and one from retrograde ejaculation. CONCLUSIONS: Our data indicate that major contributing factors to failed fertilization after intracytoplasmic sperm injection are number of MII oocytes retrieved and availability of viable sperm for injection. Although the incidence of complete failed fertilization is not remarkable, it may increase with increasing patient age and a lower number of follicles.

Adult↗

Seminal hyperviscosity is associated with poor outcome of controlled ovarian stimulation and intrauterine insemination: a prospective study.

OBJECTIVE: To determine the relationship between seminal hyperviscosity and pregnancy outcome in patients undergoing ovarian hyperstimulation and intrauterine insemination (IUI). METHODS: Patients were enrolled in the study between October 2002 and December 2003 at the Toronto Centre for Advanced Reproductive Technology. This was a prospective trial that included 37 infertile couples with abnormal seminal viscosity who underwent 57 insemination cycles as treatment for infertility (group I) and 37 couples undergoing 51 IUI cycles during the same time period, with normal semen viscosity, who served as controls (group II). Cycles were stimulated using either gonadotropin (FSH) only or FSH combined with an aromatase inhibitor, and raw semen processed for intrauterine insemination using swim-up or density gradient. Results-The mean number (+/-SD) of IUI cycles was 1.93 +/- 1.42 per patient (range 1 to 7 cycles) in group I and 2.4 +/- 1.05 (range 1 to 4) in group II. The overall pregnancy rate (PR) was 14% (8/57) and 11.8% (6/51) per cycle, and 21.6% (8/37) and 16.2% (6/37) per patient in group I and in group II, respectively. Among patients where the male had seminal hyperviscosity, five (62.5%) pregnancies miscarried in the first trimester; there was no miscarriage in the control group. CONCLUSION: The high miscarriage rate in couples with semen hyperviscosity may be attributed to biophysical alterations or chemical changes of the ejaculate that could impact sperm quality despite normal sperm parameters on semen analysis.

Adult↗