Biomedical subjects

I D Cooke

Publications and source records attributed to I D Cooke.

At least 55 records · Page 3Linked to original sources

PubMed1996

Changes in nuclear morphology in the human endometrial glandular epithelium in women with unexplained infertility.

In the light and electron microscopical study reported here, we documented the structure of the nucleus, nucleolus and nuclear channel system (NCS) in the uterine glandular epithelium in both fertile and infertile women during the early luteal phase. Nuclear volume was found to be larger in the infertile group at day 5 after the luteinizing hormone surge (LH+5) compared to day-matched fertile subjects. A two-way analysis of variance performed on nucleolar volume data from fertile and infertile women biopsied on days LH+5, +5, and +6 revealed a significant effect of condition but no effect of day or interaction. Nucleolar volume decreased from day LH+2 to day LH+6 in the fertile group, the sharpest decrease occurring between day LH+3 and day LH+4. The largest mean volume of the NCS was found at day LH+5 in the fertile group and day LH+6 in the infertile group. The results suggest a delay in the development of this organelle in the infertile women. The present study has documented alterations in the nuclei of uterine glandular cells from infertile patients. In these infertile women, there is also a delayed elaboration of the secretory apparatus and this delay correlates well with the delayed/reduced expression of a luteal-specific glyco-protein.

Analysis of Variance↗

PubMed1996

Direct assessment of cryopreservation of human spermatozoa using a cryomicroscope and computer-aided sperm analysis.

Use of a cryostage has enabled direct observation of human spermatozoa as they are cryopreserved and thawed. Crystallization and recrystallization events are readily observed. In combination with computer-aided semen analysis (CASA) equipment it was possible to determine the consequence of altering the cooling, freezing and thawing rates of a temperature-rate profile on sperm motility. Increasing the cooling rate to 50 degrees C/min resulted in significantly lower pre-freeze to post-thaw ratios for average path velocity (VAP, 13%), mean straight line velocity (VSL, 35%), mean linearity (LIN, 28%) and straightness (STR, 24%), while the ratio of the number of cells crossing the field of view (NCF) significantly increased (30%) compared to a standard freeze-thaw temperature rate profile. The NCF pre-freeze to post-thaw ratio was associated with the percentage of cell recovery after cryopreservation. Faster thaw rates resulted in better survival of the cells, perhaps due to the shorter time during which recrystallization occurred. The NCF ratios were significantly higher (33 and 30% for thaw rates of 50 and 100 degrees C/min respectively) than for the standard profile samples. Previous studies on cell survival have shown a link between the cooling and thaw rates. The cryostage should prove invaluable in future studies to identify the causes of cryodamage to spermatozoa. When used in combination with CASA, changes to sperm function during cryopreservation can be accurately measured.

Computers↗

PubMed1996

Comparative study of the effect of human cervical mucus and a cervical mucus substitute, Healonid, on capacitation and the acrosome reaction of human spermatozoa in vitro.

The present study was designed to investigate the effect of human cervical mucus on capacitation and the acrosome reaction of human spermatozoa and compare its effect to that of a cervical mucus substitute, sodium hyaluronate (Healonid). Spermatozoa from donors of proven fertility were isolated from semen using cervical mucus, Healonid or a direct swim-up (acting as the control). Sperm capacitation and the acrosome reaction were monitored by the chlortetracycline assay. In the mucus-treated group, there was a significantly higher percentage of capacitated spermatozoa, but a low incidence of spontaneous and A23187-induced acrosome reactions compared to the control. The use of Healonid during sperm isolation mimicked the effect of mucus relatively successfully. Since mucus and Healonid show very little chemical similarity, this finding would imply that cervical mucus exerts a physical effect during its interaction with spermatozoa, although a chemical effect cannot be completely dismissed. In conclusion, this study confirms early reports describing the ability of cervical mucus to capacitate spermatozoa but at the same time conserve sperm function. The finding that Healonid exerts an almost identical effect on spermatozoa would lend support to its use as a cervical mucus substitute during in-vitro fertility assessments and research studies.

Acrosome↗

PubMed1996

Soluble CD23 protein in the peritoneal fluid of patients with endometriosis.

Activated B cells are known to produce soluble CD23 protein (sCD23) from their membranes. We recently showed a higher concentration of sCD23 in the serum of patients with endometriosis. As the commonest site of endometriosis is the peritoneal cavity, we sought to evaluate the concentration of sCD23 in the peritoneal fluid of 47 fertile symptomatic women with endometriosis and 35 fertile women without endometriosis. Endometriosis was diagnosed by laparoscopy and confirmed by histopathology. There was a statistically significant difference between the sCD23 concentrations in the endometriosis and control groups (P < 0.05). When endometriosis was defined based on the revised American Fertility Society (AFS) Score, patients with mild (AFS 1 and II) but not severe (AFS III and IV) endometriosis showed a higher and significant difference in the concentration of sCD23 when compared with the controls (P < 0.05). There was no significant correlation between the peritoneal fluid concentration of sCD23 and the phase of the menstrual cycle. We conclude that the concentration of sCD23 is higher in the peritoneal fluid of patients with endometriosis when compared with the controls, suggestive of B cell activation in patients with endometriosis. Furthermore, mild endometriosis is immunologically more active than severe endometriosis as defined by the current classification.

Ascitic Fluid↗

PubMed1996

The human ovarian granulosa cell: a stereological approach.

The human ovarian granulosa cell is perhaps the most widely studied endocrine cell, but little quantitative structural information exists for this cell. In the present study new and traditional stereological probes have been employed to provide quantitative structural information on these functionally important cells. Granulosa cells were obtained from follicular aspirations from 10 women during in vitro fertilisation procedures. Initially 2 methods were used to estimate the mean nuclear volume of these cells: the mean number weighted nuclear volume was estimated by the Selector and the mean volume weighted nuclear volume by the point sampled intercept method. It was found that the difference between the 2 volume estimates was only 8.5%. The volume weighted mean nuclear volume was used as an estimate of nuclear volume. This was subsequently corrected (taking the percentage difference as the empirical bias) and combined with fractional cell volumes (Vv) to produce estimates of cell, mitochondrial, lipid and nucleolar volume. The proportion of the cell occupied by the nucleus had a remarkably low interindividual variation (CV = 7.6%). The proportion of the nucleus occupied by euchromatin also had a striking low variation (CV < 6%). All other cellular parameters had CVs of less than 35%. The lipid composition of these cells showed the greatest interindividual variability, with a CV of 42% for relative and 54% for absolute volume. The present study outlines a simple protocol for the quantitation of granulosa cell structure using new unbiased stereological probes and providing baseline structural information.

Cell Nucleus↗

PubMed1995

Endocervical mucus pH is inversely related to serum androgen levels and waist to hip ratio.

OBJECTIVE: To determine whether an inter-relationship exists between endocervical mucus pH, serum androgen levels, and waist to hip (W:H) ratio. SETTING: Donor insemination clinic, University of Sheffield, Jessop Hospital, Sheffield, United Kingdom. PATIENTS: One hundred patients receiving donor insemination. INTERVENTIONS: Waist, hip, height, and weight measurements were recorded for 100 patients receiving donor insemination. At the onset of the LH surge when patients attended for their first insemination, a sample of endocervical mucus and blood was taken. The Insler score of the mucus was recorded and the pH was measured with multirange pH paper. Of the 100 patients studied, 10 cases had an endocervical mucus pH < 6 despite Insler scores of > 10. Serum T, DHEAS, and androstenedione levels were measured in the 10 patients with pH < 6 and in 26 patients selected at random from the 90 patients with pH > or = 6. RESULTS: The W:H ratio was significantly higher in the patients with pH < 6 than pH > or = 6. No significant difference was seen in body mass index between the latter groups. All serum androgen levels were significantly higher in the patients with pH < 6 than pH > or = 7. CONCLUSION: A potential link (serum androgen levels) between a powerful predictive factor of fertility (W:H ratio) and a potential mechanism of subfertility (low endocervical mucus pH) has been demonstrated. This observation warrants further detailed evaluation particularly as it has been shown that a low endocervical mucus pH may be corrected by simple inexpensive treatment.

Androgens↗

PubMed1995

A double-blind randomized placebo cross-over controlled trial using the antioxidant vitamin E to treat reactive oxygen species associated male infertility.

OBJECTIVE: To determine the effectiveness of the in vivo administration of vitamin E as treatment for reactive oxygen species-associated male infertility. SETTING: University-based center for reproductive medicine. DESIGN: Double-blind randomized placebo cross-over controlled trial. PATIENTS, PARTICIPANTS: Thirty healthy men with high levels of reactive oxygen species generation in semen and a normal female partner. INTERVENTIONS: Patients were allocated to two groups according to the blinded randomization. Each patient received either 600 mg/d of vitamin E (Ephynal, 300 mg tablets; F. Hoffman-La Roche Ltd., Basle, Switzerland) (order A) or identical placebo tablets (order B) for 3 months. Then after a 1-month wash-out period the patients were crossed-over to the other treatment. MAIN OUTCOME MEASURES: Improvement in the in vitro function of the spermatozoa measured by conventional semen analysis, computerized motility assessment, determination of reactive oxygen species generation, binding to the zona pellucida of the unfertilized human oocyte in a competitive zona binding assay, development of hyperactivated motility (both spontaneous and in the presence of 20% of the natural agonist, human follicular fluid) and pregnancy. RESULTS: Rise in the blood serum vitamin E levels after treatment accompanied by improvement in one of the sperm function tests: the zona binding assay. The zona binding ratio for order A improved from 0.2 (range 0 to 0.5) before treatment to 0.5 (range 0.1 to 1.0) after treatment, the corresponding values for order B were 0.2 (range 0 to 1.0) before treatment and 0.3 (range 0.1 to 0.7) after treatment. CONCLUSION: Oral administration of vitamin E significantly improves the in vitro function of human spermatozoa as assessed by the zona binding test.

Adult↗

PubMed1995

Oral steroid therapy for subfertile males with antisperm antibodies in the semen: prediction of the responders.

This study was performed to examine the effectiveness of steroid therapy in subfertile men with antisperm antibodies and infertility lasting > 1 year, to predict those who would respond positively, and to evaluate the effect of the therapy on semen parameters and antisperm antibodies. The patients included 48 subfertile couples in whom the male partner had > or = 20% motile spermatozoa with bound antibodies of immunoglobulin (Ig)G, IgA or a combination of both, and were treated with prednisolone, 40 mg a day, for the first 10 days, then 5 mg on days 11 and 12 of the partner's cycle for 9 months. Twelve couples became pregnant; a cumulative conception rate of 30.2% was achieved at 9 months. The pregnant group started with significantly higher concentrations of IgG (tail) and grade I motility (P = 0.03 and P = 0.02 respectively). Multi-covariate discrete logistic regression analysis on the initial screening semen samples predicted a higher chance of conception for those with high levels of IgG (tail) (P = 0.006, sensitivity = 33%, specificity = 93%, correct = 75%, false positive = 33% and false negative = 24%). In the pregnant group, prednisolone caused a significant increase in grade I motility (P = 0.03). In the non-pregnant group, there was a significant increase in grade I motility (P = 0.0002), amplitude of lateral head displacement (P = 0.03), curvilinear velocity (P = 0.02) and decrease in grade IV motility (P = 0.03) following prednisolone treatment. In both groups there was suppression of the total antisperm antibody concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

PubMed1995

Inter-observer variability at laparoscopic assessment of pelvic adhesions.

A total of 25 women undergoing videolaparoscopy and a dye test to investigate pelvic adhesions and infertility were evaluated to determine whether applying a score system to pelvic adhesions was reproducible, and its place in determining prognosis or management. The surgeon performing the laparoscopies scored each case using the American Fertility Society (AFS) system, gave an estimate of the likely prognosis from microsurgery and recorded a recommended management [surgery or in-vitro fertilization (IVF)]. The video images and histories were independently reviewed by two other surgeons experienced in tubal surgery, who also scored each case and recorded prognosis from surgery and recommended management. Several differences in adhesion scores for each case were noted. Agreement was present between surgeons regarding estimated prognosis for microsurgery and also in recommending management, particularly when prognosis was considered poor and IVF was recommended. Despite some reproducibility, the AFS score system did not predict outcome or management decisions. However, the surgeons' fundamental impression of the state of the pelvis and recommendation for either surgery or IVF were more consistent and reproducible. Any future proposed system for adhesion classification will need to account for, and be predictive of, the decreasing number of patients for whom surgery (rather than IVF) is the preferred management.

Adnexal Diseases↗

PubMed1995

Hyperactivation may assist human spermatozoa to detach from intimate association with the endosalpinx.

The behaviour of human spermatozoa was observed during incubation with epithelial cells isolated from the isthmic and ampullary sections of human uterine (Fallopian) tubes. During incubation, spermatozoa were observed to bind to the epithelial cells of the tube (the endosalpinx), and individual spermatozoa attached and detached at intervals. The kinematic characteristics of spermatozoa during these behaviour patterns were determined. The results showed that detached spermatozoa typically had an increased curvilinear velocity and amplitude of lateral head displacement, accompanied by a decrease in their linearity. Significantly (P < 0.01) more of the detaching spermatozoa were hyperactivated than were spermatozoa prior to attachment for both isthmic (35.3 +/- 5.5 versus 4.0 +/- 3.3%; mean +/- SEM) and ampullary (26.0 +/- 7.0 versus 2.0 +/- 1.4%) regions. Incubation with epithelial cells from either region produced no differences in any category of sperm behaviour. Furthermore, there was no significant difference between regions in the amount of time spermatozoa spent bound (33.6 +/- 12.9 and 20.6 +/- 3.0 s for isthmic and ampullary tissue respectively). These results support the hypothesis that hyperactivation may assist spermatozoa in breaking connections with epithelial cells.

Cell Adhesion↗

PubMed1995

An analysis of the outcome of microsurgical and laparoscopic adhesiolysis for infertility.

We evaluated 81 women with adnexal adhesions and no male factor who underwent microsurgical (n = 59) and laparoscopic (n = 22) adhesiolysis for infertility. The cumulative conception rates for all 81 patients at 12 and 24 months were 41 and 44% respectively. The impact of the following variables on cumulative conception rates for all patients was examined: age, duration of infertility, type of infertility, ovulatory status, presence and stage of endometriosis, adhesion grade, adnexal status (bilateral or unilateral disease, unilateral tubal absence), history of previous surgery, history of pelvic inflammatory disease and treatment modality (microsurgical versus laparoscopic). The results of independent comparisons of subgroups within each of these variables may be biased because of the interrelationships between the variables. To overcome this problem, a stepwise Cox's proportional hazards regression analysis was employed. Our analysis showed that the single most significant variable influencing the cumulative conception rates was the duration of infertility (P < 0.005). For every additional year of infertility, the probability of pregnancy after adhesiolysis (microsurgical or laparoscopic) was reduced by approximately 20%. Cumulative conception rates at 12 and 24 months after microsurgical adhesiolysis were 36 and 40% respectively, while after laparoscopic adhesiolysis they were 57% at 12 and 24 months. When imbalances were adjusted between the two treatment groups, there was no statistically significant difference between the cumulative conception rates for microsurgical and laparoscopic adhesiolysis.

Adnexal Diseases↗

PubMed1995

An analysis of the outcome of microsurgical and laparoscopic adhesiolysis for chronic pelvic pain.

We evaluated the outcome of microsurgical (n = 72) and laparoscopic (n = 51) adhesiolysis in women who complained of chronic pelvic pain. Adhesion severity was not significantly different between the two treatment groups. The influences of the following variables on the outcome of all 123 cases of adhesiolysis were examined: (i) surgical modality (microsurgical or laparoscopic), (ii) history of infertility, (iii) associated dyspareunia and (iv) aetiological factors of adhesive disease (endometriosis, pelvic inflammatory disease and previous laparotomy). To adjust for differences in follow-up intervals, overall and subgroup cumulative rates of pain persistence/recurrence were calculated and compared. To adjust for interrelationships between variables and to correct fo differences between the treatment groups, a proportional hazards regression analysis was employed. This analysis showed that the cumulative rate of pain persistance/recurrence at 24 months was not significantly different after microsurgical (44%) and laparoscopic (53%) adhesiolysis. From all the variables which were examined, the only one which appeared to influence the impact of surgical adhesiolysis for chronic pelvic pain was a history of previous laparotomy. A history of previous laparotomy was associated with approximately three times higher rates of pain persistence or recurrence. This effect did not depend on whether previous laparotomy was carried out for pain or for other indications. The most likely explanation for the failure of these patients who had a previous laparotomy to respond to surgery is that they intrinsically have a higher rate of adhesion formation and reformation. This can only be confirmed with a prospective study where all patients will undergo a second-look laparoscopy.

Adolescent↗

PubMed1995

The interaction in vitro of human spermatozoa with epithelial cells from the human uterine (fallopian) tube.

The physical interaction between human spermatozoa and the epithelium of the human uterine (Fallopian) tube was investigated in vitro using a variety of techniques. The 'live' observation of human spermatozoa incubated with 1 day old cultures of tubal epithelium demonstrated that spermatozoa can show a strong physical interaction with epithelial cells; contact with the epithelium appeared to be random and there was no evidence of any taxis toward epithelial cells. The physical interaction (or 'binding') was resistant to gentle washing and was maintained following the addition of glutaraldehyde fixative. The intimate nature of the interaction was confirmed ultrastructurally where both spermatozoa and epithelial membranes were observed to be in close apposition. These results are the first descriptions of sperm-epithelial 'binding' in the human. They are similar to other observations made in a variety of non-human mammalian species. It is suggested that this interaction may be an important feature of normal sperm transport in the human uterine tube in vivo.

Cell Communication↗

PubMed1995

Clinical value of sperm morphology for in-vivo fertility: comparison between World Health Organization criteria of 1987 and 1992.

The World Health Organization (WHO, 1992) has suggested new criteria for scoring sperm morphology. This study compares the clinical value of the new criteria, i.e. classification of a man as fertile or infertile, to those previously established by the WHO (1987). Papanicolaou-stained semen smears from 166 men attending our infertility clinic, whose fertility status was known, were scored using both methods. Using logistic discriminant analysis for compositional data, no difference between these two sets of criteria with respect to predicting pregnancy outcome was observed. The categorization of the abnormalities (head, midpiece, tail) provides no extra clarification. The WHO (1992) cut-off point of 30% for normal forms is not appropriate, as approximately half of the men in the fertile group had a normal sperm morphology below this limit. In conclusion, the present WHO (1992) classification of sperm morphology is of no additional clinical value. Studies on sperm morphology should concentrate on obtaining biological data on, and measurements of, spermatozoa which are functionally active. Only then can the definition of normal be achieved and clinically useful criteria be adopted.

Discriminant Analysis↗

PubMed1995

Serum-soluble CD23 in patients with endometriosis and the effect of treatment with danazol and leuprolide acetate depot injection.

Activated B cells have recently been shown to produce soluble CD23 from their membranes. The serum-soluble CD23 concentration in 21 patients with pelvic pain diagnosed as having endometriosis and confirmed by histology, and in 18 patients without pelvic pain, who had a normal pelvis during laparoscopic sterilization, was studied by chemiluminescent enzyme-linked immunosorbent assay. The endometriosis patients were randomized to 3 months of either danazol or leuprolide acetate injection. Serum was taken before and after 3 months of therapy. The serum-soluble CD23 concentration was significantly elevated in patients with endometriosis when compared with the controls (P < 0.0001). There was no correlation between soluble CD23 concentrations and the severity of endometriosis (r = 0.48, P > 0.05). The serum concentration of soluble CD23 decreased significantly on treatment with danazol but not leuprolide acetate (P < 0.05). We conclude that the elevation of soluble CD23 in patients with endometriosis suggests that there is activation of B cells, which respond to danazol but not leuprolide acetate injection.

B-Lymphocytes↗

PubMed1995

The prevalence of endometrial immunoglobulin G antibodies in patients with endometriosis.

The purpose of this study was to investigate the frequency of antibodies in the serum of patients with endometriosis reacting with endometrial, ovarian, thyroid and skeletal muscle antigens. A total of 55 fertile patients with pelvic pain who had endometriosis diagnosed by laparoscopy and confirmed by histology formed the study group, while 43 fertile patients without pelvic pain and undergoing tubal sterilization formed the control group. Eutopic endometrial membrane antigen was prepared from biopsies taken from patients or controls by an ultracentrifugation technique and used in an enzyme-linked immunosorbent assay (ELISA) for immunoglobulin (Ig) G, IgA and IgM endometrial antibodies. Ovarian, thyroid and skeletal muscle antigens were prepared similarly and used in an ELISA. There was a significant difference in the prevalence of IgG antibodies to eutopic endometrial antigens derived from either control or endometriosis patients (P < 0.05). IgA or IgM antibodies against endometrial antigens were not detected. Endometrial antibodies were detectable in both the luteal and the follicular phases of the menstrual cycle. Immunoblot analysis demonstrated the presence of endometrial antigens with molecular weights of 60 and 66 kDa in the eutopic endometrium of patients with endometriosis. Antibodies to thyroid, ovary or skeletal muscle were not detected in these patients. These findings indicate that endometrial antibodies of the IgG class can be detected in approximately 50% of patients with endometriosis but it remains unclear whether these represent a pathologically distinct subgroup of such patients.

Adult↗

PubMed1995

A light microscopical morphometric study of the luminal epithelium of human endometrium during the peri-implantation period.

Different aspects of the histology of luminal epithelial cells were examined quantitatively at the light microscopical level. For the first time, using a variety of morphometric indices, an attempt was made to quantify and compare the cellular events in a group of well defined fertile controls and infertile subjects during the peri-implantation period. The most striking observation was that the majority of the features examined, including all nuclear parameters and most cell dimensions, were similar and did not differ significantly between the fertile controls and the infertile subjects over the period of study. The coefficients of variation of most of the parameters investigated were similar and within a narrow range in both the fertile and infertile population, indicating little variation either between individuals at a particular stage or between subjects on different days. This low inter-subject variability may suggest that the cellular events which we have been able to quantify at the light microscope level in the uterine surface epithelium around the anticipated time of implantation are precisely regulated. Such precise regulation does not mean that changes do not occur in luminal epithelial cells during this time, but that those which do occur are well controlled and co-ordinated. Possible changes in biochemistry, physiology and ultrastructure of these cells remain to be determined.

Adult↗

PubMed1995

Implementing comprehensive quality control in the andrology laboratory.

Comprehensive quality control procedures were integrated into the routine semen analysis workload of a large university-based andrology laboratory. Methods were chosen to match as far as possible those which have been used successfully for many years in disciplines such as clinical chemistry. Levey-Jennings and cusum charts were plotted in order to monitor the immunobead-binding test for antisperm antibodies and a video-taped control sample for computerized semen analysis. A cryopreserved semen control was also charted. Daily manual sperm counts were plotted against the corresponding computer-assisted semen analysis (CASA) value. Multiple readings of 30 slides were used to monitor morphology assessments. Monthly means for morphology were also calculated regularly. Coefficients of variation were calculated for all variables and were found to be more appropriate for some aspects, such as CASA, than for others, such as morphology, when difference from the previous reading of the same slide was found to be more useful. These integrated quality control procedures had a direct influence on the production of results from the laboratory. Together with a high standard of technician training, comprehensive routine quality control based on repeated analyses of control samples is an effective way of assuring the validity of semen analysis results.

Antibodies↗