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Biomedical subjects

I D Cooke

Publications and source records attributed to I D Cooke.

At least 19 recordsLinked to original sources

The endometrial biopsy as a predictive factor of pregnancy rate in women with unexplained infertility.

To investigate whether or not there is any relation between endometrial morphology and subsequent pregnancy rate, 47 couples with unexplained infertility were followed up for 3 years after they had had an endometrial biopsy. Each woman had an LH-timed endometrial biopsy performed in the luteal phase of the cycle. The biopsy was dated chronologically according to the luteinizing hormone (LH) surge and histologically using morphometric criteria. None of the women received any form of treatment in the endometrial biopsy cycle. Of 47 women with unexplained infertility, 36 (76.6%) (Group I) had 'in phase' endometrial development and 11 (23.4%) (Group II) had retarded endometrium. Women with normal endometrial development had a higher pregnancy rate than women with retarded endometrial development (50% vs. 9%; P less than 0.02). The fecundability during treatment cycles was higher in Group I than in Group II (0.051 vs. 0.008; P less than 0.05). In women with 'in phase' endometrium, treatment increased the monthly probability of conception (0.051 vs. 0.006; P less than 0.001). A precisely timed endometrial biopsy should be considered as part of the investigations for women with unexplained reproductive failure, to help determine prognosis.

Adult

The impact of infertility on psychological functioning.

To explore the impact of infertility on psychological functioning 130 couples presenting with primary infertility were assessed at their initial visit to an infertility clinic. Of these, 116 couples were assessed on a second occasion some 7-9 months later when in most cases the medical tests were complete. Measures of personality, psychopathology, perceived social support, sex role identity and marital state were obtained from both partners. The set was subsequently divided into five subgroups on the basis of the diagnosis made or the outcome (female cause, male cause, female and male cause, unexplained and pregnant). The results show little evidence of psychopathology in the sample, depression scores remained low throughout the period of investigation. The results also indicated stable marital relationships. Scores on tests of anxiety and psychiatric morbidity declined between the first and second assessment except in the case of men who were diagnosed with a fertility problem. The implications of these findings are discussed in the increased use of donor insemination which circumvents rather than treats the problem of male infertility.

Adaptation, Psychological

Endometrial lymphoid tissue in the timed endometrial biopsy: morphometric and immunohistochemical aspects.

OBJECTIVES: The purpose of this study was to provide a morphometric profile of endometrial granulated lymphocytes and to investigate qualitative and quantitative differences in leukocyte subsets in precisely timed luteal phase endometrial biopsies. STUDY DESIGN: Endometrial biopsies were obtained from 24 normal fertile women at 4, 7, 10, and 13 days after the luteinizing hormone surge. Endometrial granulated lymphocytes were assessed morphometrically in 2 microns resin sections. Eleven monoclonal antibodies were used to characterize leukocytes in frozen sections. Semiquantitation was performed with a Quantimet 970 image analyzer. Data were analyzed with one-way analysis of variance. RESULTS: CD8+ (T suppressor-cytotoxic) cells increased significantly from 4 to 7 days after the luteinizing hormone surge, whereas CD68+ macrophages increased from days 10 to 13. Lymphocytes with an unusual phenotype (CD56+, CD38+, CD2+) increased dramatically after 7 days. The volume fraction of endometrium occupied by the nuclei of endometrial granulated lymphocytes did not alter, but their mean nuclear diameter and axial ratio decreased from days 7 to 13. CONCLUSION: The morphometric findings indicate in situ proliferation of endometrial granulated lymphocytes rather than migration from the peripheral circulation. T lymphocytes, macrophages, and endometrial granulated lymphocytes increase significantly between certain stages of the luteal phase.

Adult

An analysis of the variation of plasma concentrations of placental protein 14 in artificial cycles.

OBJECTIVE: To analyze the factors affecting the variation of plasma concentration of placental protein 14 (PP14) in artificial cycles. DESIGN: The effects of different hormone replacement therapy (HRT) regimens were examined in a crossover design. SETTING: Jessop Hospital for Women, Sheffield, United Kingdom. PATIENTS: Eighteen women with premature ovarian failure: 6 associated with Turner's syndrome and 12 with idiopathic premature ovarian failure. INTERVENTIONS: Four different HRT regimens; 36 study cycles. MAIN OUTCOME MEASURES: Plasma PP14 concentrations on days 1, 15, 19, and 29 of the artificial cycles. RESULTS: In cycles treated with a standard HRT, the levels were similar to those of the natural cycle. Subjects with Turner's syndrome did not have elevated PP14 levels, whereas the majority (9/12 [75%]) of those with idiopathic premature ovarian failure had elevated levels on day 29 of the cycle. Levels of PP14 were reduced when either the doses of estradiol valerate were reduced to 1/3 or the doses of progesterone (P) were reduced to 1/5 of the standard HRT. CONCLUSIONS: Plasma levels of PP14 are dependent not only on P stimulation but also on adequate estrogen priming.

Adult

Round cells and sperm fertilizing capacity: the presence of immature germ cells but not seminal leukocytes are associated with reduced success of in vitro fertilization.

Fifty semen samples produced for IVF by patients diagnosed as having unexplained infertility were screened for leukocytes, leukocyte subsets, and immature germ cells using a mAB-based staining procedure. Nonfertilizing ejaculates were found to contain significantly larger numbers of immature germ cells, although no significant differences in leukocyte content were observed between groups. Neither sperm density or progressive motility were significantly different between fertilizing and nonfertilizing groups. We conclude that seminal leukocytes have little if any influence on the fertilizing capacity of the spermatozoa in patients undergoing IVF for unexplained infertility, but the presence of large numbers of germinal elements is associated with reduced fertilizing capacity and may be indicative of an immature sperm population.

Fertilization in Vitro

In vitro fertilization in the natural cycle.

In vitro fertilization in the natural or spontaneous reproductive cycle was first described by Edwards and his colleagues in 1980 following the birth of their first natural cycle IVF baby 2 years earlier. Many groups attempted to follow their lead but it was almost ten years later that the next publication of success appeared (Foulot et al, 1989). The concept of IVF in the natural cycle is particularly attractive and so in 1987 our group also started evaluating the technique. Initial success in the patients with tubal lesions was not translated to patients with other infertility indications. Unfortunately the technique as initially developed was relatively inefficient with significant procedural losses at each stage. Over the succeeding four years a number of changes have been introduced and the efficiency considerably improved. Although these changes have improved take-home baby rates, overall pregnancy rates per embryo have not altered and are still lower than spontaneous in vivo pregnancy rates. It is likely that in the future, with current developments in culture techniques and greater understanding of gamete biology, this situation will change significantly.

Female

How well does ultrasonographic measurement of endometrial thickness predict the results of histological dating?

In this study, transabdominal ultrasonographic measurement of endometrial thickness was performed prior to sampling of the endometrium for histological examination in regularly cycling women. In the natural cycles, the results of ultrasonography and histological dating were significantly correlated (r = 0.76, P less than 0.0001, n = 63). Endometrial histology was likely to be proliferative if the thickness was less than 8 mm, and likely to be secretory if the thickness was greater than 9 mm. However, for a given endometrial thickness, the stage of endometrial development appeared to vary widely, suggesting that ultrasonographic measurement of endometrial thickness cannot accurately predict the results of histological dating. In the second part of the study, endometrial thickness at a defined stage of histological development (results of histological dating between days LH + 4 and LH + 6) was compared in three types of cycles: natural (n = 13), clomiphene-stimulated (n = 12) and artificial (n = 22) cycles. The endometrial thickness (mean +/- standard deviation) in artificial cycles (9.9 +/- 1.3 mm) was significantly smaller than that in natural (11.3 +/- 1.4 mm) or clomiphene-stimulated (11.2 +/- 1.3 mm) cycles, but there was no difference in endometrial thickness between the latter two types of cycle. Our observations suggest that it is unlikely that ultrasonographic measurement of endometrial thickness can replace histological examination of the endometrium in the evaluation of the luteal phase.

Adult

The poor prognostic value of low to moderate levels of sperm surface-bound antibodies.

The clinical significance of antisperm antibodies for fertility remains controversial. In this study, we determined whether the presence, isotype, region and/or amount of sperm-bound antibody was of any predictive value for future fertility in 534 men using Cox's proportional hazards model. Significant correlations between the presence of antibodies and semen parameters were recorded, such as sperm mucus penetration and sperm motility. However, low (less than 10%) negative binding and moderate (less than 50%) binding had no significant effect on the probability of conception or the time to conception. This study confirms in-vitro data suggesting that sperm function is not impaired unless the degree of antibody binding to spermatozoa is very high.

Antibodies

Effect of human cervical mucus on human sperm motion and hyperactivation in vitro.

The aim of our experiment was to examine the effect of exposure to human cervical mucus on quantitative sperm motility with specific reference to hyperactivated sperm motility. Human spermatozoa were allowed to penetrate cervical mucus for 20 min before swimming into Earle's balanced salt solution tissue culture medium for 25 min. The sperm motion characteristics were compared to those which had been obtained from a direct swim-up for 45 min. Spermatozoa treated with mucus were more 'active' than the control group. Multivariate statistical analysis indicated that cervical mucus promotes hyperactivated motility and that sperm sub-populations exposed to cervical mucus are very heterogeneous, as indicated by the numbers and motility characteristics of spermatozoa.

Cervix Mucus

Recovery of artificially inseminated spermatozoa from the fallopian tubes of a woman undergoing total abdominal hysterectomy.

Investigation of the process of sperm transport within the human female reproductive tract must involve the recovery of spermatozoa from the oviducts of women post-coitum or insemination. This case report describes the insemination of a 40-year-old woman with her partner's semen and the subsequent recovery of spermatozoa from her Fallopian tubes, removed at hysterectomy, 2 days post-ovulation. The number of spermatozoa present within the tubes and their location was ascertained using a flushing technique and scanning electron microscopy (SEM). The total number of spermatozoa present within the flushed tube was calculated to be 142, whilst the total number of spermatozoa calculated to be present within the scanned tube was 673. The proportional distribution of spermatozoa within the tubes was virtually identical by the two techniques used. The largest population of spermatozoa resided within the ampulla (66.1% SEM, 68.3% flush), and the smallest population resided within the intramural region (14.9% SEM, 13.4% flush).

Adult

The removal of morphologically abnormal sperm forms by phagocytes: a positive role for seminal leukocytes?

A preliminary investigation was undertaken further to determine the function of the leukocytic cells found in semen. We performed semen analysis and quantified leukocyte subsets using immunocytochemical staining techniques in ejaculates of 351 patients. Leukocyte profiles were examined in relation to sperm morphological data for evidence of a sperm removal/selection process. Three types of seminal phagocytic cell were found to contain spermatozoa: small polymorphonuclear leukocytes (approximately 10-12 microns), monocytes of similar size and much larger (30-40 microns) macrophages capable of engulfing multiple sperm heads. The total leukocyte count (P less than 0.01), the numbers of phagocytic cells i.e. polymorphonuclear leukocytes (P less than 0.05), monocyte/macrophages (P less than 0.01) and HLA-DR positive cells (P less than 0.01), were significantly higher in those samples with greater than 50% ideal sperm forms. Significantly fewer of these same cell types were observed in samples with greater than 50% head defects. There was no difference in the number of tail or midpiece defects between leukocytospermic (greater than 10(6)/ml) and non-leukocytospermic semen samples. Oligozoospermic samples contained significantly fewer leukocytes (P less than 0.005), although above a concentration of 5 x 10(6)/ml, the sperm number was not correlated with leukocyte number. These data, along with repeated observation of spermatozoa or sperm fragments within phagocytic cells, support the hypothesis that leukocytes have a role in the removal of abnormal spermatozoa from the ejaculate.

Alkaline Phosphatase

Prognostic value of two putative sperm function tests: hypo-osmotic swelling and bovine sperm mucus penetration test (Penetrak).

The clinical value of two inexpensive and easy to perform sperm function tests (hypo-osmotic swelling and bovine sperm mucus penetration tests) were examined in a prospective study of in-vivo conception rates in 325 couples. Both tests were of no significant value alone or in combination with traditional semen characteristics in predicting pregnancy outcome. The two most predictive factors of conception were the length of infertility and percentage of progressively motile spermatozoa. Our data support the further investigation of sperm function using more sophisticated quantitative tests, specifically the examination of sperm motility.

Adult

A prospective, randomised, cross-over study comparing the effects of clomiphene citrate and cyclofenil on endometrial morphology in the luteal phase of normal, fertile women.

OBJECTIVE: To examine the effect of two anti-oestrogens, clomiphene citrate and cyclofenil, on endometrial morphology in the luteal phase. DESIGN: A prospective, randomised, cross-over study. SETTING: Jessop Hospital for Women, Sheffield, UK. SUBJECTS: 10 women who were previously fertile and regularly cycling. INTERVENTION: The administration of clomiphene citrate or cyclofenil in the treatment cycles. A LH timed endometrial biopsy was taken on day LH + 6. MAIN OUTCOME MEASURES: Histological dating and morphometric analysis of the endometrial sample. RESULTS: Only one out of 10 subjects had abnormal endometrium. There was no difference in the results between cycles treated with clomiphene citrate and cyclofenil. CONCLUSIONS: Clomiphene citrate or cyclofenil does not have a major adverse effect on endometrial morphology in the luteal phase of normal fertile subjects. The possible adverse effects of anti-oestrogens on endometrium may have been previously overestimated.

Adult

Endometrial responses in artificial cycles: a prospective, randomized study comparing three different progesterone dosages.

OBJECTIVE: To examine the endometrial response to three different regimens of progesterone. DESIGN: A prospective, randomized study. SETTING: The Jessop Hospital for Women, Sheffield. SUBJECTS: 14 women with premature ovarian failure divided into two equal groups. INTERVENTIONS: Three different regimens of hormone replacement therapy containing standard, high and low progesterone dosages. One group received the standard regimen in one cycle and high dosage regimen in another cycle. The second group received the standard regimen in one cycle and low dosage regimen in another cycle. The order of the two dosage regimens was randomized by lottery. MAIN OUTCOME MEASURES: Ultrasonographic measurement of endometrial thickness and morphological study of endometrial biopsy specimens taken on day 19 of the cycle using the traditional dating criteria and morphometric techniques. RESULTS: Compared with standard regimen cycles, the endometrial response in cycles treated with the low dosage regimen showed significant retardation of overall endometrial development and changes in a number of morphometric measurements. The response of endometrial glands to the high dosage regimen was similar to that of the standard regimen, although the stromal cell diameter was increased. CONCLUSIONS: In women receiving hormone replacement therapy normal endometrial development depends on an adequate dosage of progesterone, but increased dosage above our standard regimen does not produce any further change in the glandular component of the endometrium.

Adult

Endometrial responses in artificial cycles: a prospective study comparing four different oestrogen dosages.

OBJECTIVE: To examine the endometrial response to four different regimens of oestrogen. DESIGN: A prospective, randomized cross-over study. SETTING: Jessop Hospital for Women, Sheffield. SUBJECTS: Twenty one women with premature ovarian failure divided into three equal groups. INTERVENTIONS: Four different regimens of hormone replacement therapy: variable, fixed 1 mg, fixed 2 mg and fixed 4 mg oestrogen dosages. Each woman received the variable dosage regimen in one cycle and crossed over to receive one of the three fixed dose regimens (1 mg, Group 1; 2 mg, Group 2; 4 mg Group 3) in another cycle. MAIN OUTCOME MEASURE: Ultrasonographic measurement of endometrial thickness and outpatient endometrial biopsy on day 19 of the artificial cycle; analysis of endometrial specimens by three separate methods: traditional histological criteria, morphometry and immunohistochemistry. RESULTS: The endometrial response was similar in those treated with the variable and the fixed 2 mg or 4 mg dosage regimens. The response was suboptimal in those treated with the fixed 1 mg dosage regimen. CONCLUSIONS: Normal endometrial development requires adequate priming of the endometrium by oestrogen, which may be administered in a sequential, variable dosage fashion, or simply by a fixed daily dosage regimen. However, the minimum daily dose required is likely to be 2 mg of oestradiol valerate. No adverse effect on the endometrium was observed at a daily dose of 4 mg oestradiol valerate, which produced plasma levels of oestradiol above the reference ranges of the natural cycle.

Adult

Outpatient falloposcopy: intra-luminal imaging of the fallopian tube by trans-uterine fibre-optic endoscopy as an outpatient procedure.

OBJECTIVE: To develop trans-uterine falloposcopy using the linear eversion technique in outpatients. DESIGN: A prospective, descriptive study of the introduction and application of this new technique. SETTING: A tertiary referral University Research Clinic independent of operating theatres and inpatient admission. SUBJECTS: Twenty-one infertile women with previous evidence of fallopian tube disease. MAIN OUTCOME MEASURES: Rate of successful tubal cannulation and procedure completion, tubal section visualized, condition of the tubal epithelium, subsequent treatment plan. RESULTS: A bilateral examination was attempted in 19/21 patients and completed in 18. Two early falloposcopies with inadequate analgesia were aborted due to pain after only one tube had been examined without an attempt to identify or cannulate the contralateral tubal ostium. In one patient, neither ostium was identified or cannulated. After ostial localization, 37/37 tubes were cannulated. If the ostium was not localized but 'blind' cannulation attempted, 1/3 were cannulated successfully. Epithelium characteristic of the ampulla was seen in 31/38 (82%) tubes, and of the proximal tube only in 3/38 (8%). Successful bilateral assessment noted complete bilateral proximal obstruction in 2/18 patients, non-obstructive bilateral endotubal damage in 7/18, unilateral damage in 3/18 and bilateral healthy tubes in 6/18. Unilateral examination in two patients found minimal evidence of damage in one and severe intra-tubal adhesions in the other. After falloposcopy, 11/21 patients elected for IVF, 9/21 tubal surgery and 1/21 for IVF whilst awaiting surgery. CONCLUSION: Falloposcopy is a technique which can be performed in an outpatient clinic. It is well tolerated, and high rates of luminal cannulation and visualization are possible. Atraumatic access to the tube and visualization of its lumen offer exciting opportunities for diagnosis and treatment of tubal conditions.

Ambulatory Care

The leukocytic reaction of the human uterine cervix.

PROBLEM: Sperm interaction with the immune system of the human cervix is poorly understood. METHOD: The leukocytic response of the human cervix to sperm was examined in a closely monitored patient population (N = 10), using monoclonal antibody cell identification techniques. Baseline data were collected from both cervical mucus and smears sampled before treatment by donor insemination. Donor insemination was timed to coincide with ovulation by monitoring plasma LH concentrations twice daily. Following insemination the numbers of leukocytes were recorded in cervical mucus and smear samples taken over a 24-h period relative to the time of treatment. Controls treated with "pure sperm," seminal plasma, cryopreservative, and cervical smearing alone were also included in the study. RESULTS: Only those women treated with sperm cells exhibited substantial elevations in leukocyte numbers following inseminations. Additionally, serial cervical smearing induced an inflammatory response of the cervix. In all the women, the neutrophil was the predominant leukocyte of the cervix both during the baseline and treatment periods (median values ranged from 77 to 86%). Macrophages, T-helper lymphocytes, and T-suppressor lymphocytes were also detected, but only in low numbers (2-10.6%). Two patients and one control ("pure sperm") became pregnant during their study cycle. CONCLUSIONS: We conclude that the leukocytic reaction is a physiological response of the cervix to sperm, the function of which remains to be established.

Cell Movement

Origin of reactive oxygen species in human semen: spermatozoa or leucocytes?

Peroxidative damage induced by reactive oxygen species (ROS) has been proposed as one of the major causes of defective sperm function. In previous studies of the production of ROS in semen, the contribution of contaminating leucocytes was not assessed. We determined the levels of ROS in 60 semen samples from men attending our infertility clinic and demonstrated by performing extraction experiments with antibody-coated magnetic beads that, within this unselected population of patients, leucocytes were the major source of ROS in the low-density Percoll fraction. Of the sperm motion parameters examined using computerized semen analysis, beat-cross frequency was the only one significantly affected by the ROS in semen.

Cell Separation