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Biomedical subjects

S Fishel

Publications and source records attributed to S Fishel.

At least 19 recordsLinked to original sources

'Curing' empty follicle syndrome.

We report a novel method of rescuing empty follicle syndrome (EFS) and provide evidence that it is a drug-related problem rather than a clinical dysfunction. In a preliminary study we established that in EFS the serum beta-human chorionic gonadotrophin (beta-HCG) concentrations 36 h after HCG administration never exceeded 10 mIU/ ml. beta-HCG concentrations were thus used to confirm EFS when oocytes were not retrieved from one ovary after controlled ovarian hyperstimulation. The procedure was suspended leaving intact all follicles in the second, ovary. After confirmation of EFS, a second HCG from a different batch was administered and 36 h later mature oocytes were retrieved from the intact ovary, suggesting a fault with the HCG previously administered. Three patients have been treated in this way. In the first case, four out of five mature eggs were fertilized after intracytoplasmic sperm injection (ICSI) resulting in the transfer of three top grade (grade 1) embryos. In the second case all seven mature oocytes fertilized after in-vitro fertilization (IVF) and three grade 1 embryos were transferred resulting in a twin pregnancy, now delivered. In the third case, five out of nine oocytes were fertilized after ICSI and one out of the three treated with high insemination concentration IVF fertilized, resulting in the transfer of three ICSI embryos.

Chorionic Gonadotropin

Human fertilization with round and elongated spermatids.

Human spermatids from ejaculate and testicular tissue have been utilized for evaluating human fertilization by intracytoplasmic sperm injection (ICSI) and, where possible, compared with spermatozoa utilizing sibling oocytes. Round and elongated spermatids obtained from ejaculates were either prepared through Percoll gradients or isolated and washed individually using subzonal insemination needles (SUZI; 10-14 microm internal diameter). Seminiferous tubules obtained after biopsy were placed into HEPES-buffered Earle's medium and dissected using 21-gauge needles. Spermatogenic cells and spermatozoa were isolated and washed individually using SUZI needles. Spermatozoa were subsequently injected into the ooplasm using 5 microm (internal diameter) ICSI needles, whereas 8-9 microm (internal diameter) needles were used for spermatid injection. Only metaphase II oocytes (n = 207) were injected: 64 with round spermatids, 92 with elongated spermatids and 51 with spermatozoa; the fertilization rate was 30, 24 and 67% respectively. There was a significant (P < 0.001) increase in the fertilization rate using spermatozoa compared with spermatids. The fertilization rate was not different between round and elongated spermatids, although the fertilization rates for round and elongated spermatids in the ejaculate were 33 and 18% respectively, compared with 22 and 38% respectively when testicular spermatids were utilized. In three patients sibling oocytes were used to compare round and elongated spermatids found in the ejaculate with spermatozoa extracted from seminiferous tubules. The fertilization rate was 24% for spermatids and 79% for testicular spermatozoa. This result suggests that, should only spermatids be available in the ejaculate, a testicular biopsy in the hope of obtaining testicular spermatozoa would be worth while.

Cell Size

Comparison of second trimester biometry in singleton and twin pregnancies conceived with assisted reproductive techniques.

The objective of this study was to investigate the size of singleton vs twin pregnancies at the time of a second trimester dating scan. The analysis included 86 infants from 63 pregnancies achieved with assisted reproductive techniques, comprising 40 singletons and 46 twins. Measurements of second trimester biparietal diameter (n = 85) and femur length (n = 74) were plotted against the precisely known gestational age. A common regression line was calculated for each parameter and the residuals for singletons and twins were compared. Gestational age and weight at birth were also analysed for each group. There was no significant difference between singletons and twins in biparietal diameter or femur length in second trimester. In contrast, twins had a lower mean birthweight, gestational age at birth, and weight-for-gestational age centile compared with singletons. Singleton babies from these pregnancies had an average birthweight centile of 49.8% (i.e. close to the median for spontaneously conceived pregnancies in our population). We concluded that the same pregnancy dating charts can be used for singletons and twins. At corresponding gestational age, twins are smaller than singletons at birth because of slower growth in the third trimester.

Anthropometry

Flow cytometric analysis of granulosa cells from follicular fluid after follicular stimulation.

Granulosa cells are not easily accessible, unless they are examined in follicular fluid after oocyte retrieval. These samples are usually contaminated with blood. We have set up a general technique for analysis of granulosa cells without physically separating them from blood cells. The sample is stained with CD45, which is a pan-leukocyte marker, and granulosa cells are consecutively selected as CD45 negative during flow cytometric analysis. Analysis of forward scatter of the granulosa cells, which is correlated to cell size, shows a wide size range throughout the whole population rather than two distinct populations as previously suggested.

Female

Endometrial thickness as a predictor of pregnancy after in-vitro fertilization but not after intracytoplasmic sperm injection.

An ultrasonographic evaluation of the endometrium was performed in 158 patients undergoing ovarian stimulation for an in-vitro assisted reproduction programme. Endometrial thickness was evaluated in 109 patients undergoing in-vitro fertilization (IVF) for female indications and in 49 patients undergoing intracytoplasmic sperm injection (ICSI) for male indications. The maximal endometrial thickness was measured on the day of human chorionic gonadotrophin (HCG) administration by longitudinal scanning of the uterus on the frozen image using electronic callipers placed at the junction of the endometrium-myometrium interface at the level of the fundus. Cases in which the endometrial thickness was >/=10 mm were included in group A; cases in which the endometrial thickness was <10 mm were assigned to group B. The age of the patients, serum 17-beta oestradiol concentrations on the day of HCG administration, the length of follicular stimulation, the number of follicles, 17-beta oestradiol concentrations per follicle on the day of HCG and the number of embryos transferred were analysed in each case. When comparing endometrial thickness and results in IVF and ICSI patients, an endometrium <10 mm predominated in IVF patients (27.5%) compared with those undergoing ICSI (16.7%) (P = 0.05); conversely an endometrium >=10 mm was more frequent in ICSI than in IVF patients. The incidence of pregnancy was higher in IVF group A patients (32/79; 41%) than in IVF group B patients (5/30; 17%) (P = 0.03), whereas no significant difference was found between ICSI group A (13/42; 31%) and ICSI group B (3/7; 43%) patients. Thus, a higher percentage of IVF patients had thin endometrium when compared with ICSI patients; thin endometrium was a prognostic indicator of pregnancy only in the case of a female indication for infertility (IVF). A thin endometrium in cases of female infertility may reflect a previous or present uterine pathology, whereas in indications of male infertility (i.e. cases using ICSI), in the absence of any associated uterine pathology, the presence of a thin endometrium is not predictive.

Cytoplasm

Predicting empty follicle syndrome.

OBJECTIVE: To determine a simple predictive test for empty follicle syndrome before oocyte retrieval. DESIGN: Retrospective correlation analysis. SETTING: Patients attending Nottingham University Research and Treatment Unit, a tertiary, university-based assisted reproductive technologies (ART) program between April 1, 1994 and March 31, 1995. PATIENT(S): Six women in whom no oocytes were retrieved after superovulation for ART (empty follicle syndrome) were compared with 11 women with successful oocyte retrieval. INTERVENTION(S): Subcutaneous buserelin acetate plus IM hMG and hCG were used for superovulation. Oocyte retrieval was transvaginal and ultrasound guided. MAIN OUTCOME MEASURE(S): Ultrasound measurement of follicular growth and serum E2 levels during superovulation. Serum beta-hCG levels before and 36 hours after hCG administration IM. Number of oocytes retrieved. RESULT(S): Before hCG administration, beta-hCG was not detectable in the serum. The serum beta-hCG 36 hours after hCG was 209 +/- 16.7 mIU/mL (conversion factor to SI units, 1.0; mean +/- SEM, range 106 to 290 mIU/mL) in women with successful oocyte retrieval and 4 +/- 1.8 mIU/mL (range 0 to 9 mIU/mL) in empty follicle syndrome. This difference was significant. CONCLUSION(S): Empty follicle syndrome is associated with very low bioavailability of beta-hCG and can be predicted by measuring serum beta-hCG level 36 hours after IM hCG administration.

Adult

Intracytoplasmic sperm injection (ICSI) versus high insemination concentration (HIC) for human conception in vitro.

The use of high insemination concentration (HIC) for in vitro fertilization (IVF) was compared with intracytoplasmic sperm injection (ICSI) in cases of male factor infertility. Sibling oocytes (n = 252) from 24 patients were used, 123 for HIC and 129 for ICSI. Although the incidence of fertilization was decreased with HIC (48% v. 61%), this treatment was nevertheless a viable option for many patients, especially when ICSI was not available. However, there was a higher incidence of cytoplasmic fragmentation of embryos after HIC compared with ICSI (36% v. 10%, P = 0.003) and the outcome was significantly affected by the severity of teratozoospermia. Using a cut-off of 5% normal forms, the incidence of fertilization with HIC for the group with < 5% normal forms was 37% compared with 72% for the group with > 5% normal forms; there was also a significant decrease in cleavage rate (P = 0.05) and the number of regular embryos (P = 0.005), and an increase in cytoplasmic fragmentation (P = 0.006) in patients with < 5% normal forms. No distinction was made between cases of teratozoospermia when ICSI was used. The present study confirms the value of HIC as a first line treatment for male infertility, as long as ICSI remains significantly more expensive and concerns on safety are mooted. However, the use of sibling oocytes for ICSI is recommended, especially in cases with < 5% normal sperm morphology.

Adult

Computer image sperm selection as a novel approach to subzonal insemination in the human.

Utilizing real-time computer image analysis, individual spermatozoa were selected using microaspiration. Selection criteria were based on potential hyperactivation motility characteristics; the amplitude of lateral head displacement > 7.5 microns, curvilinear velocity > 70 microns/s and linearity of < 30%. For this pilot study, 16 patients (eight in each group) were recruited. Using subzonal insemination (SUZI), up to five (mean = 4.4 +/- 0.3) spermatozoa selected using computer-image sperm selection (CISS) were micro-injected, or up to 15 (mean = 12.8 +/- 1.3 SD) unselected spermatozoa. In the group which utilized CISS, 28 out of 49 (57%) oocytes were fertilized compared with 13 out of 52 (25%) utilizing conventional SUZI (P < 0.04); polyspermy was 20% (n = 10) and 2% (n = 1) respectively. CISS with SUZI showed increased efficiency in achieving fertilization and is a novel approach to studying individual sperm function in a sperm egg bioassay where gamete ratios are close to unity.

Adult

A positive correlation between expression of beta 1-integrin cell adhesion molecules and fertilizing ability of human spermatozoa in vitro.

The purpose of this study was to investigate firstly whether beta 1-integrin cell adhesion molecules are expressed by human spermatozoa, and secondly whether there is any relationship between the expression of beta 1-integrin cell adhesion molecules and the fertilizing ability of human spermatozoa in vitro. A total of 50 semen samples were examined. The samples were obtained from the male partners of couples undergoing in-vitro fertilization (IVF) for either unexplained, tubal or male factor infertility. A panel of six monoclonal antibodies against beta 1-integrin cell adhesion molecules and immunohistochemical techniques were used to identify the presence of these molecules on the spermatozoa. The percentage of spermatozoa showing strong immunolabelling with each monoclonal antibody was assessed in each sample. The relationship between these results and the aetiology of infertility and incidence of fertilization was examined. beta 1-Integrins, and primarily the ones with alpha 4-, alpha 5- and alpha 6-chains, were expressed by human spermatozoa. Compared with semen samples from unexplained or male factor infertility patients, samples from tubal infertility patients had a significantly higher (P < 0.05) percentage of spermatozoa expressing adhesion molecules. There was a positive correlation between the expression of alpha 4, alpha 5 and alpha 6 adhesion molecules and the fertilizing ability of spermatozoa. The positive correlation between the presence of certain beta 1-integrin cell adhesion molecules and the fertilizing ability of human spermatozoa suggests that integrins may be putative determinants in egg-sperm recognition and interaction.

Adult

Sperm function and its manipulation for microassisted fertilization.

Comprehension of the intricate complexities of sperm function is clearly crucial to the success of attempts to manipulate it for the purposes of assisted conception. This is particularly important when considering various procedures for microassisted fertilization since these bypass critical physiological events that are mandatory for normal fertilization, to varying degrees. Methylxanthine derivatives such as pentoxifylline are useful agents for the management of oligoasthenozoospermic patients. This is particularly so for procedures such as SUZI where adequate motility of spermatozoa injected into the perivitelline space is crucial for fusion with the vitelline membrane to achieve fertilization. The generation of minute concentrations of reactive oxygen species in vitro may prove to be a valuable technique in this respect, in the light of recent evidence for their involvement in capacitation and hyperactivation. Induction of the acrosome reaction by non-invasive, non-toxic agents should markedly improve success rates for microassisted fertilization. Acrosin appears to play a central role in this and, therefore, it would seem prudent to monitor levels of acrosin activity in samples of spermatozoa used in assisted conception procedures. With respect to microassisted fertilization, the potential to select recently acrosome-reacted spermatozoa coated by activated acrosin promises to be a major improvement. Current methods employed for determination of the fertilization potential of spermatozoa are clearly inadequate (Polansky and Lamb, 1988; Aitken, 1990). In fact, the prevailing evidence suggests that no single parameter of sperm function reflects this potential (Zaneveld and Jeyendran, 1988). Therefore, we have both a scientific and a moral responsibility to investigate these processes further. Subsequently, we should be in a position to identify individual gametes with the potential for fertilization and so utilize procedures that result in maximal fertilization rates with minimal risk of polyploidy or abnormality.

Autoantibodies

Subzonal insemination and zona breaching techniques for assisting conception in vitro.

The following comments can be made with regard to zona breaching procedures (PZD) and SUZI. First and foremost, it is important to establish that fertilization cannot occur with microdrop IVF or conventional IVF before proceeding with microassisted fertilization technology. PZD increases the rate of polyspermy, and this is significantly higher than when utilizing SUZI. SUZI increases the incidence of implantation in cases exhibiting severe teratozoospermia. SUZI increases the fertilization rate per patient and per oocyte in cases where IVF has previously failed on more than one occasion. SUZI increases the incidence of fertilization when very few motile sperm are available, and there is an increase in the incidence of cytoplasmic fragmentation with SUZI, which might be alleviated by improving methods of penetrating the zona pellucida, for example by using the Sonic Sword or utilizing a thinner, sharper injection needle alone or in addition to increasing the perivitelline space by dehydrating the oocyte (Yang et al, 1988). Morphology is an important parameter in predicting the outcome of IVF and therefore in guiding patients on the relevance of microinjection technology.

Adult

Micro-assisted fertilization in patients who have failed subzonal insemination.

During the past 3 years we have used direct injection of spermatozoa into the cytoplasm of the oocyte (DISCO) in an attempt to procure conception in vitro where subzonal insemination (SUZI) has failed. Acknowledging the paucity of information about this invasive procedure in humans, it was clearly the only alternative for some patients. A total of 58 patients (cycles) who had had previously failed SUZI elected for this approach. Sibling oocytes, where possible, were used as a comparison between DISCO, SUZI, partial zona dissection (PZD) and microdrop in-vitro fertilization (IVF). Fertilization and embryo transfer was achieved by 66% (n = 38) of patients. Patients with embryos derived solely from DISCO (n = 19) produced four clinical pregnancies (21%), four patients had embryo transfer from SUZI embryos only with no pregnancies, and 15 had embryo transfer from a mixture of SUZI and DISCO embryos, with one dizygotic twin pregnancy from a combined SUZI and DISCO embryo transfer (7%). Twins, one male and one female, have been delivered, there has been one miscarriage and two pregnancies are ongoing. Of the 627 oocytes, 251 were used for DISCO and 71 (28%) were fertilized, 296 were used for SUZI and 29 (10%) were fertilized; 70 were used for PZD, two (3%) were fertilized, and none of the 10 undergoing microdrop IVF were fertilized. Of the 58 patients (cycles) 31% (n = 19) had fertilization with DISCO only, 8% (n = 4) with SUZI only, 28% (n = 15) with both SUZI and DISCO, while only one (2%) had fertilization with PZD.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult