Biomedical subjects

R S Jeyendran

Publications and source records attributed to R S Jeyendran.

At least 19 recordsLinked to original sources

PubMed2007

Discordance among blastomeres renders preimplantation genetic diagnosis for aneuploidy ineffective.

PURPOSE: To investigate the contribution of discordance among blastomeres from the same embryo in the interpretation of blastomeres biopsied from day 3 embryos. METHODS: 228 IVF embryos had two blastomeres removed and fluorescent in situ hybridization (FISH) was used to detect aneuploidy of chromosomes 13, 15, 16, 18, 21, 22, X and Y. Of the 228 embryos, 102 had complete FISH results for both blastomeres. RESULTS: When the 2 blastomeres of 102 embryos with successful FISH results were compared, 26 (25.5%) were concordant for all 8 chromosomes and 76 (74.5%) were discordant for one or more chromosomes. Among the 102 embryos, 12 (12%) were disomy in both blastomeres and 37 (36%) were disomic in all 8 chromosomes in one of the two blastomeres. CONCLUSION: Discordance among blastomeres from the same embryo appears to present a significant problem in interpreting results of embryos biopsied on day 3 and analyzed by FISH especially when most PGD's are done on single blastomeres.

Adult↗

PubMed2006

p53 tumour suppressor gene polymorphism is associated with recurrent implantation failure.

Implantation failure is the most frequent cause of lack of pregnancy after IVF and embryo transfer. Successful implantation requires trophoblastic growth into the endometrium to stimulate its own blood supply. The p53 tumour suppressor gene has been shown to regulate cell growth and induce angiogenesis. Therefore, the prevalence of p53 codon 72 polymorphism was investigated among women with a history of recurrent implantation failure to determine whether this polymorphism may serve as a risk factor for implantation failure. DNA was extracted from the buccal swabs obtained from 70 women experiencing implantation failure and polymerase chain reaction amplification of p53 arginine (Arg) 72 and proline (Pro) 72 variants were performed. The frequency of homozygous Arg and Pro or heterozygous Arg/Pro genotypes was compared with frequencies among 205 women experiencing recurrent pregnancy loss and 20 control women. The frequency of Pro72 was significantly higher (P=0.003) among women experiencing recurrent implantation failure compared with women experiencing recurrent miscarriage and with control women. It is concluded that p53 codon 72 polymorphisms may serve as a susceptibility factor affecting the chances of recurrent implantation after IVF or embryo transfer.

Abortion, Habitual↗

PubMed2006

Multiple thrombophilic gene mutations are risk factors for implantation failure.

While the role of inherited thrombophilia has been accepted as a cause of recurrent late pregnancy complications, the contribution of mutated thrombophilic genes to implantation failure has not been studied. Proteins involved in fibrinolysis are necessary for trophoblast invasion into the endometrium. This study compared the prevalence of 10 thrombophilic gene mutations among 42 women with a history of recurrent implantation failure after IVF-embryo transfer with 20 fertile control women. Buccal swabs were taken from all of the women for DNA analyses. Women with a history of implantation failure after IVF-embryo transfer displayed a higher prevalence of PAI-1 4G/5G mutations than controls (P = 0.007). No differences in the frequency of the other specific gene mutations were detected. However, the prevalence of total gene mutations among patients with implantation failure was significantly higher than among controls. More than three gene mutations among the 10 genes studied were observed in 74% of women with implantation failure and 20% of controls (P = 0.0004). It is concluded that inherited thrombophilias are associated with implantation failure. This association is manifest by total number of mutations as well as with PAI-1 mutations.

Adult↗

PubMed2006

Role of p53 codon 72 polymorphism in recurrent pregnancy loss.

Recurrent pregnancy loss (RPL) has been associated with low expression of apoptosis-and angiogenesis-related genes. The p53 tumour suppressor gene has been shown to induce apoptosis and angiogenesis. Recently, a low increased frequency of a p53 codon 72 polymorphism has been reported among women experiencing RPL. This study investigated the prevalence of p53 codon 72 polymorphism among women with a history of RPL, to determine whether this polymorphism may serve as a risk factor for miscarriage. Buccal swabs were obtained from 205 women with a history of two or more consecutive spontaneous abortions and 25 women with at least two live births and not more than one elective abortion. DNA was extracted from the buccal swabs and PCR amplification of p53 arginine(Arg)72 and proline(Pro)72 variants was performed. The frequency of homozygous Arg and Pro or heterozygous Arg/Pro genotypes among RPL patients and controls were not significantly different. No significant differences in allelic frequencies of p53 were observed. In addition, the allelic frequencies between controls and those previously reported were the same. It is concluded that p53 codon 72 polymorphisms do not serve as a susceptibility factor affecting the chances of miscarriage in an unselected population.

Abortion, Habitual↗

PubMed2001

A new CentriSwim procedure to increase the recovery of motile spermatozoa.

A prospectively controlled in vitro study was performed to compare sperm concentration, sperm motility and progressive sperm motility recovered following the standard swim-up procedure and a new CentriSwim procedure. The CentriSwim procedure involves creating a centrifugal force to counteract the force of gravity during sperm swim-up procedure. Two aliquots of semen from 12 normozoospermic ejaculates and 12 laboratory-induced oligoasthenozoospermic specimens were diluted, centrifuged, and 1.0 ml of media layered over the sperm pellet. One aliquant was processed by standard swim-up technique. The other aliquant was processed by CentriSwim procedure involving centrifugation at 200 rpm on a 2-cm radius upward-directing arm, at an angle of 60 degrees for 10 min, creating roughly 0.8 g centrifugal force at room temperature (22-24 degrees C) to counteract the force of gravity. The numbers of spermatozoa recovered from the upper 0.5 ml of the medium following CentriSwim from the normozoospermic ejaculates and laboratory-induced oligoasthenozoospermic specimens were significantly higher than following standard swim-up procedure. No statistical differences in the recovery of percentage sperm motility and progressive sperm motility between the two techniques were observed. In conclusion, the CentriSwim procedure yields higher numbers of motile spermatozoa than the standard swim-up technique.

Cell Separation↗

PubMed1999

Increased recovery of swim-up spermatozoa by application of "antigravitational" centrifugation.

OBJECTIVE: To compare the quantity and quality of sperm recovered following the 10-minute application of an antigravitational force during the swim-up procedure with the standard 60-minute swim-up (SSU) procedure. DESIGN: Prospectively controlled in vitro study. SETTING: Private andrology laboratory and hospital-based infertility practice. INTERVENTION(S): Equal aliquots of semen were evaluated following various intervals of antigravitational centrifugation swim-up (ACSU). ACSU and SSU were then compared. PATIENT(S): Thirty-eight men undergoing therapeutic testing. MAIN OUTCOME MEASURES: Sperm concentration, sperm motility, and progressive sperm motility. RESULTS: The number of sperm recovered from the ACSU procedure was significantly higher than from the SSU procedure. No significant differences in percent motile sperm and progressive motile sperm recovery between the two procedures were observed. CONCLUSION: The ACSU procedure yields a higher number of motile spermatozoa in a much shorter time.

Cell Separation↗

PubMed1999

Epidermal growth factor and leukemia inhibitory factor levels in follicular fluid. Association with in vitro fertilization outcome.

OBJECTIVE: To determine the levels of epidermal growth factor (EGF) and leukemia inhibitory factor (LIF) in follicular fluid, if any, and to assess the association of these cytokines with the outcome of in vitro fertilization (IVF). STUDY DESIGN: EGF and LIF levels determined by enzyme-linked immunosorbent assay in 60 preovulatory follicular fluids were compared with 25 IVF outcomes. RESULTS: Immunoreactive EGF and LIF could be detected in human follicular fluid. Levels of these cytokines were similar in FF obtained from follicles that resulted in fertilized oocytes and those that did not. EGF levels were significantly lower in patients establishing a pregnancy as compared to patients achieving no pregnancy (P < .007). LIF levels were similar in both groups of patients. CONCLUSION: EGF appears to be associated with IVF outcome.

Adult↗

PubMed1998

Increased recovery of viable spermatozoa through oscillating centrifugation.

OBJECTIVE: To compare the number and quality of spermatozoa recovered following the standard centrifugation method with those recovered following oscillation of the relative centrifugal field during the centrifugation method. DESIGN: Prospectively controlled in vitro study. SETTING: Private andrology laboratory. PATIENT(S): Seventeen healthy sperm donors. INTERVENTION(S): Equal halves of the same semen sample were evaluated following the standard centrifugation or following oscillation of the relative centrifugal field during centrifugation. MAIN OUTCOME MEASURE(S): Number of sperm, sperm motility, progressive sperm motility at 0 hours and after 3-hour of incubation at 5 degrees C in TES and Tris (buffer) (TEST)-yolk. RESULT(S): The number of sperm, sperm motility, progressive sperm motility at 0 hours and after 3-hour of incubation at 5 degrees C in TEST-yolk were significantly higher for the sperm recovered following oscillating centrifugation compared with standard centrifugation. CONCLUSION(S): Oscillating the relative centrifugal field during centrifugation yields higher number of viable spermatozoa.

Cell Survival↗

PubMed1998

Centrifugal pulsing increases the recovery of viable spermatozoa.

PURPOSE: Our purpose was to compare the number and quality of spermatozoa recover following the standard centrifugation method with those after pulsing the relative centrifugal field during centrifugation. METHODS: A prospectively controlled in vitro study using ejaculates from 10 healthy sperm donors was undertaken. Number of sperm and sperm motility at 0, 3, and 24 hr post-incubation at 5 degrees C in TEST-yolk results determined the efficacy of the centrifugation techniques. RESULTS: The number of sperm and sperm motility at 0, 3, and 24 hr post-incubation at 5 degrees C in TEST-yolk were significantly higher for the sperm recovered following pulse centrifugation compared with those after standard centrifugation. CONCLUSIONS: Pulsing the relative centrifugal field during centrifugation yields a higher number of viable spermatozoa.

Adult↗

PubMed1997

Strategies for ovulation induction and oocyte retrieval in the lowland gorilla.

PURPOSE: Ovulation induction and oocyte retrieval were performed in a lowland gorilla in an attempt to propagate and potentially cryopreserve embryos from an infertile animal and to advance techniques to help preserve this endangered species. RESULTS: Following 34 days of leuprolide acetate suppression, human menopausal gonadotropins were administered for 14-days in a 32-year-old wild-born lowland gorilla. Ten oocytes were retrieved by transrectal ultrasound-guided aspiration. Other approaches to oocyte recovery were not feasible in this case. A serum estradiol concentration of 4700 pg/ml at the time of human chorionic gonadotropin administration did not induce ovarian hyperstimulation. Mature oocytes were recovered from follicles measuring 14 to 24 mm in diameter, with a corresponding average serum estradiol concentration of approximately 300 pg/ml for each mature follicle. Cryopreservation of a gorilla embryo was effected from cryopreserved gorilla spermatozoa. CONCLUSIONS: Parameters for monitoring ovulation induction in the gorilla appear to be similar to those for humans. The results indicate that the use of a gonadotropin releasing hormone agonist and higher doses of gonadotropins than previously used in gorillas appear to improve oocyte recovery.

Animals↗

PubMed1997

Interferon-gamma production by the human preimplantation embryo.

PROBLEM: This study demonstrated that the human embryo produces interferon-gamma (IFNgamma). It is important to know whether IFNgamma can be produced before implantation. Therefore the aim of this study was to evaluate the profile of IFNgamma production between days 2 and 5 after in vitro fertilization. METHOD: Twenty embryos were cultured from day 2 to 5 after fertilization. The embryo stages were checked each day and the media refreshed. IFNgamma levels were estimated by an enzyme-linked immunosorbent assay. RESULTS: All embryos produced measurable IFNgamma at least for 1 day. Yields of IFNgamma were: 0.46 +/- 0.45 (n = 4) on day 2, 0.69 +/- 0.52 (n = 19) on day 3, 0.73 +/- 0.52 (n = 15) on day 4, 0.55 +/- 0.32 (n = 11) IU/ml on day 5, respectively. There was no significant difference in the IFNgamma production between in vitro culture days or between the developmental stages of embryos. CONCLUSION: IFNgamma is produced by all the embryos and seems to peak between days 3 and 4, which is just before implantation.

Blastocyst↗

PubMed1996

Glass wool column filtration versus mini-Percoll gradient for processing poor quality semen samples.

OBJECTIVE: To compare the quality and number of spermatozoa recovered from laboratory-induced severe oligozoospermic specimens processed by mini-Percoll gradient and glass wool column filtration. DESIGN: Both sperm-processing procedures were compared in similar sperm samples adjusted to contain equal low numbers of motile spermatozoa using either dilution (oligozoospermia) or with the addition of killed sperm (oligoasthenozoospermia). The spermatozoa processed by both procedures samples were evaluated for motility, response to hypo-osmotic swelling test, and the hemizona assay. PATIENTS: Five healthy fertile sperm donors. SETTING: Private Andrology Laboratory and University Hospital. MAIN OUTCOME MEASURE: Sperm motility, hypo-osmotic swelling test, and hemizona assay results determined the efficacy of the sperm-processing procedures. RESULTS: The concentration of sperm recovered after both procedures was not affected by either preparation or processing methods. Glass wool-processed sperm had higher motility in oligoasthenozoospermic samples, bound tightly to hemizonae in higher mean numbers, and demonstrated a higher percentage of membrane-intact spermatozoa in oligozoospermic samples. CONCLUSION: Laboratory-prepared oligozoospermic samples subjected to glass wool filtration yielded more functionally intact spermatozoa than mini-Percoll gradient processing.

Cell Separation↗

PubMed1996

Cystic fibrosis mutation screening in healthy men with reduced sperm quality.

The majority of men with cystic fibrosis (CF) are infertile due to a bilateral congenital absence of the vas deferens (CBAVD). However, clinically affected CF patients present a spectrum of genital phenotypes ranging from normal fertility to severely impaired spermatogenesis and CBAVD. Recently, it has become apparent that CF can manifest itself as isolated CBAVD in the absence of other clinical symptoms. The present study was undertaken to test the possible involvement of the CF gene in the aetiology of male infertility other than CBAVD. Semen specimens from 127 unrelated healthy males with various diagnoses of reduced sperm quality were screened for a panel of 13 mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Fourteen of 80 (17.5%) healthy men with infertility due to reduced sperm quality and 3 of 21 (14.3%) men with azoospermia had at least one CF mutation (one azoospermic male was a compound heterozygote). The frequency of mutations in our sample of infertile males was significantly higher than the expected CF carrier frequency in the local population (P = 0.00139). No mutations were found in a control group of 26 individuals with normal semen parameters. This increased frequency of CF mutations in healthy men with reduced sperm quality and in men with azoospermia without CBAVD suggests that the CFTR protein may be involved in the process of spermatogenesis or sperm maturation apart from playing a critical role in the development of the epididymal glands and the vas deferens.

Case-Control Studies↗

PubMed1995

Estrogenic and antiestrogenic activities of anordiol: a comparison of uterine and vaginal responses with those of clomiphene citrate.

Anordiol (2 alpha,17 alpha-diethynyl-A-nor-5 alpha-androstane-2 beta,17 beta-diol) has been variously characterized as an estrogen and as an antiestrogen. To more completely understand the pharmacological properties of this contraceptive steroid, simultaneous responses were studied in uterine, vaginal, and hepatic tissues. Rats received 4 daily sc injections with either anordiol, clomiphene citrate (CC), or the vehicle alone (C+) starting on the first day of pseudopregnancy. Uteri were traumatized on day 4 of pseudopregnancy, and rats were sacrificed 5 days later. A pseudopregnant group without uterine trauma served as a negative control (C-). Mean uterine weights per animal and cytosolic estrogen (EcR) and progesterone (PcR) receptor activities per g of DNA were all 5- to 7-fold greater in the C+ group than in the other groups (all p < 0.05). However, anordiol and CC suppressed uterine weight without suppressing the stromal proliferative response; the DNA content of the uteri of anordiol- and CC-treated rats was similar to that of C+ rats. Vaginal tissue exhibited estrogenic responses to anordiol and CC with an increase in epithelial stratification compared to the C+ and C- groups even though no difference in levels of EcR/g of DNA were expressed 5 days after the last antiestrogen dose. Binding affinities and serum E2 and progesterone (P) concentrations were not statistically different among the groups. In conclusion, anordiol produced responses in the uterus and vagina of the pseudopregnant rat which were indistinguishable from those of CC, and, therefore, we conclude that anordiol acts on these tissues as an antiestrogen.

Animals↗

PubMed1995

Milk versus TEST-yolk--preincubated sperm; in vitro fertilization outcome.

OBJECTIVE: Preincubation of sperm in TEST-yolk medium enhances in vitro fertilization (IVF) outcome. However preincubation of sperm in milk at 5 degrees C enhances the results of sperm penetration assay and hemizona assay. This study was therefore performed to determine the influence of milk on in vitro fertilization rate of human oocytes, as compared with TEST-yolk medium. STUDY DESIGN: Forty-one consecutive couples undergoing an IVF procedure were randomized. Of these 20 couples were admitted for the milk study (group 1) and 21 couples for the TEST-yolk study (group 2). Each ejaculate was Percoll-processed and the sperm pellet was resuspended in 0.5 ml of culture medium. An equal volume of heat-inactivated homogenized cow's milk (95 degrees C, 10 min) was added to sperm suspension from group 1 and an equal volume of TEST-yolk medium was added to sperm suspension from group 2. After 2 h of incubation at 5 degrees C and washing with culture medium at 37 degrees C, oocytes were inseminated. Oocytes from group 1 couples were inseminated with milk-treated spermatozoa and those from group 2 couples with yolk-treated spermatozoa. Oocytes were evaluated for fertilization after 18 h. RESULTS: Sperm preincubated in milk fertilized 75 out of 100 mature oocytes (75%). TEST-yolk-treated sperm fertilized 45 out of 64 mature oocytes (70%). The difference was not statistically significant. CONCLUSIONS: Preincubation of spermatozoa in milk, as compared with preincubation in TEST-yolk medium yields a similar IVF outcome, so milk may be a suitable alternative medium for preincubation of spermatozoa to enhance its fertilizing potential.

Animals↗

PubMed1995

Sperm nuclear maturity in spinal cord-injured men: evaluation by acidic aniline blue stain.

For many of the spinal cord-injured (SCI) men who are able to produce an ejaculate, infertility because of poor semen quality is a concern. Impaired spermatogenesis has been attributed as a possible reason for the poor semen quality. If so, events that occur during spermatogenesis may be used as a marker to evaluate the extent of spermatogenic alteration. During spermatogenesis, when the sperm nuclear condensation occurs, lysine-rich somatic histone are replaced by arginine-rich protamines in the DNA. Acidic aniline blue preferentially stains the immature sperm nucleus blue by binding to the lysine. Hence, each sperm can be individually evaluated for nuclear maturity. To test this concept, the nuclear maturity of sperm from 12 SCI men obtained by vibratory stimulation was compared with sperm samples obtained by self-masturbation from 104 non-SCI men. Sperm smears stained with acidic aniline blue were evaluated for nuclear maturity. The percent of unstained spermatozoa for non-SCI men (mean +/- SEM; 83.4 +/- 1.1%) was not statistically different from that of the SCI men (79.7 +/- 4.8%). However, the sperm motility (70.5 +/- 1.2%) and the percentage of normal sperm morphology (50.8 +/- 0.7%) of non-SCI men were significantly (P < .01) different from those of the SCI men (36.5 +/- 6.8% and 44.0 +/- 2.4%). It seems that the poor semen quality observed in SCI men is probably not caused by inadequate nuclear maturity of the spermatozoa.

Cell Nucleus↗

PubMed1995

Semen extenders to salvage ejaculate in a retrograde ejaculate environment: a potential use in spinal cord-injured men.

Infertility is a well-recognized complication in spinal cord-injured men. Vibratory stimulation and electroejaculation are being used to overcome the inability to ejaculate, but poor sperm quality remains a significant problem. Retrograde ejaculation is known to occur with both sperm retrieval techniques, and may contribute to the poor semen quality. In addition, the amount of retrograde ejaculate produced is generally of sufficient quantity worthy of salvaging. We investigated the effectiveness of various semen media in maintaining the viability of healthy sperm placed in a retrograde ejaculate environment. Six semen extenders, media used to process spermatozoa for artificial insemination and to maintain viability during cryopreservation, were studied. We found that Ham's F-10 with 3.5gm % bovine serum albumin seems to be an effective medium to maintain sperm viability over a wide range of urine concentrations (up to 60% to 80% urine), and potentially, may be useful by its infusion into the bladder before vibratory or electrostimulation to enhance the recovery of greater numbers of viable sperm from the retrograde environment.

Cell Survival↗

PubMed1995

TEST-yolk media and sperm quality.

Media prepared with egg yolk and two buffers, TES and Tris, called TEST-yolk have been shown to have beneficial effects on the survivability, fertilizing capacity and storage potential of human spermatozoa. Egg yolk lipoproteins are the critical compounds for the beneficial effects, with a synergistic effect due to the TES-Tris buffer component. TEST-yolk media have been used in the sperm penetration assay, the hemizona assay, sperm preparation for clinical in-vitro fertilization, artificial insemination with homologous spermatozoa, cryopreservation, sperm samples with a positive antisperm antibody test, and preparation for techniques requiring capacitated spermatozoa. Few harmful consequences of TEST-yolk have been reported, although controlled trials are required to evaluate therapeutic effects in the treatment of male infertility.

Cryopreservation↗