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Biomedical subjects

C L Barratt

Publications and source records attributed to C L Barratt.

At least 55 records · Page 3Linked to original sources
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

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↗
PubMed1995

A time course study of capacitation and the acrosome reaction in human spermatozoa using a revised chlortetracycline pattern classification.

OBJECTIVE: To study the time course of capacitation, spontaneous, and A23187-induced acrosome reaction of human spermatozoa during 8 hours incubation in vitro using the chlortetracycline (CTC) assay with a revised fluorescent pattern classification. DESIGN: Fertile donor spermatozoa were isolated by direct swim-up and incubated in Earle's balanced salt solution for up to 8 hours. At hourly intervals, spermatozoa were stained with CTC before and after the addition of A23187 to induce the acrosome reaction. SETTING: The University Clinic, Jessop Hospital for Women, Sheffield, United Kingdom. PATIENTS: Donors participating in the Donor Insemination Program. MAIN OUTCOME MEASURES: Eight fluorescent patterns identified by the CTC assay and acrosome-reacted spermatozoa detected by indirect immunofluorescence using 18.6 monoclonal antibody. RESULTS: Using a statistical model defined by analysis of deviance allowed rationalization of the CTC pattern classification by grouping together patterns that showed a similar and significant change over time. In addition, spontaneous and A23187-induced acrosome-reacted spermatozoa identified by the CTC assay were shown to be correlated significantly to those identified by indirect immunofluorescence. CONCLUSION: The CTC assay using a revised pattern classification offers a more precise description of human spermatozoa capacitation in vitro. Also, CTC-identified acrosome reaction (both spontaneous and A23187 induced) was confirmed independently by indirect immunofluorescence.

Acrosome↗
PubMed1994

The more accurate timing of insemination with regard to ovulation does not create a significant improvement in pregnancy rates in a donor insemination program.

OBJECTIVE: To determine if the accurate prediction of ovulation before artificial insemination of frozen donor semen was justified by increased pregnancy rates. DESIGN: A retrospective study of inseminations, over a 2-year period, administered to women who were enrolled on a donor insemination program and who were monitored using one of four ovulation timing regimens. Also, a study of the significance of follicle status at the time of insemination with relevance to pregnancy was carried out. SETTING: A university teaching hospital donor insemination program. PATIENTS: Patients entered in this study were undergoing insemination with thawed, cryopreserved, donor semen. MAIN OUTCOME MEASURES: Pregnancy and follicle status at the time of insemination. RESULTS: There was no statistically significant difference in pregnancy rates between the four methods of ovulation detection most commonly used for artificial insemination. Inseminations several hours either side of ovulation did not result in a significantly different pregnancy rate than inseminations carried out closer to ovulation. CONCLUSION: More accurate and expensive methods of detecting changes in ovulation result in no significant increase in fecundity than other less expensive timing techniques.

Female↗
PubMed1994

The sequential effects of human cervical mucus, oviductal fluid, and follicular fluid on sperm function.

OBJECTIVE: To compare the sequential effects of human cervical mucus (CM), oviductal fluid, and follicular fluid (FF) on sperm motility, hyperactivation, and the acrosome reaction during 9 hours of incubation in vitro. DESIGN: Spermatozoa from a fertile donor were allowed to penetrate human CM in vitro and then were recovered and incubated in either 30% human oviductal fluid or a simple salt solution for 3 hours before challenge with 20% FF. Thus, the sequential effects of the fluids collected from the female reproductive tract on sperm hyperactivation and acrosome reaction were examined. SETTING: The donor insemination program at the University Clinic within the Jessop Hospital for Women, Sheffield, United Kingdom. PATIENTS: All donors used in this study were involved in the donor insemination program. MAIN OUTCOME MEASURES: Sperm motility, hyperactivation, and percentage of acrosome-reacted spermatozoa were measured. RESULTS: This study used an in vitro technique to mimic sperm transport in vivo. Spermatozoa sequentially incubated with female reproductive tract secretions in vitro showed a maintenance in the level of FF-induced hyperactivated motility and a decreased response to the FF-induced acrosome reaction when compared with sperm challenged with FF in the absence of prior incubation with human oviductal fluid. CONCLUSION: It is suggested that human oviductal fluid may serve to delay the FF-induced acrosome reaction.

Acrosome↗
PubMed1994

Computer-assisted measurement of hyperactivation in human spermatozoa: differences between European and American versions of the Hamilton-Thorn motility analyser.

Spermatozoa from healthy donors of proven fertility were analysed for percentage hyperactivation, mean curvilinear velocity (VCL), mean progressive velocity (VSL), amplitude of lateral head displacement (ALH) and percentage linearity (LIN) using two versions of the Hamilton-Thorn motility analyser (HTMA) running at different frame rates. The sort fraction criteria developed using a machine utilizing 30 Hz gave comparable figures for the percentage spermatozoa displaying motility patterns concomitant with hyperactivation when compared to that developed for a machine using a frame rate of 25 Hz. Statistically significant differences were observed when criteria developed using 30 Hz machines were transferred to those using 25 Hz. The frame rate dependency of VSL and VCL are thought to account for these differences; values for ALH and LIN were largely unaffected by frame rate. These results have implications for workers using the European version of the HTMA (at 25 Hz) with sort criteria, containing frame rate-dependent variables, which have been developed by American colleagues using the machines of a higher frame rate. More appropriate sort criteria for workers using European (25 Hz) machines are suggested.

Computers↗
PubMed1994

Human oviductal fluid prolongs sperm survival.

OBJECTIVE: To compare the effect of human oviductal fluid on sperm motility and hyperactivation during 9 hours' incubation in vitro with follicular fluid (FF) and medium controls. DESIGN: Fertile donor spermatozoa were allowed to penetrate human cervical mucus in vitro and then recovered and incubated in either 30% human oviductal fluid, 20% FF, or medium for up to 9 hours. Sperm motion characteristics were measured using a sperm motility analyzer. SETTING: The donor insemination program at the University Clinic within the Jessop Hospital for Women, Sheffield, United Kingdom. PATIENTS: All donors used in this study were involved in the donor insemination program. MAIN OUTCOME MEASURES: Sperm motility, hyperactivation, curvilinear velocity, progressive, lateral head displacement, and linearity were measured using a sperm motility analyzer. RESULTS: After 9 hours' incubation, spermatozoa in human oviductal fluid had a significantly higher percentage motility than sperm incubated in FF or the control medium. A more linear sperm motion was consistently observed in the spermatozoa incubated in human oviductal fluid: significantly different from FF and media at 3 hours and 6 hours. The effect of human oviductal fluid on maintaining sperm motility was not affected by the addition of P. CONCLUSION: Human oviductal fluid can maintain sperm motility in a mechanism that is not mediated by the low concentration of P. We suggest that human oviductal fluid is a favorable environment for sperm survival.

Body Fluids↗
PubMed1993

The effects of clomiphene citrate and cyclofenil on cervical mucus volume and receptivity over the periovulatory period.

OBJECTIVE: To determine the effects of clomiphene citrate (CC) and cyclofenil on cervical mucus (CM) volume and receptivity sampled serially over the periovulatory period. DESIGN: Using prospective luteinizing hormone (LH) timing CM volume and receptivity were compared in standard CC and cyclofenil-stimulated cycles using normal ovulatory cycles as controls. LOCATION: The Donor Insemination Unit at the University Research Clinic, Sheffield, United Kingdom. PATIENTS: Twenty anovulatory patients and 10 normally ovulating patients, all of whom were participating in a treatment cycle of donor insemination. INTERVENTIONS: The 20 anovulatory patients were allocated at random into two groups: group 1 was administered 50 mg of CC on days 2 to 6 of the menstrual cycle; group 2 was administered 400 mg of cyclofenil on days 3 to 12 of the menstrual cycle. All the patients were given a single treatment of donor insemination 24 to 36 hours after the onset of the LH surge. RESULTS: Clomiphene citrate and cyclofenil were shown to exert differential impacts on CM quantity and quality. In terms of quantity, the CC patients produced significantly lower volumes of CM than the cyclofenil patients and controls. In terms of quality, the CC patients and controls produced CM of similar receptivity, whereas the cyclofenil patients produced CM that was significantly more receptive to sperm than both the CC patients and controls. CONCLUSIONS: Neither CC nor cyclofenil exerted a detrimental impact on CM quality throughout the periovulatory period.

Adult↗
PubMed1993

The contribution of a hidden male factor to unexplained infertility.

OBJECTIVE: To assess sperm function in patients with unexplained infertility in comparison with normal fertile men. DESIGN: Prospective study. SETTING: Infertility outpatient clinic and donor insemination program, University Department of Obstetrics and Gynecology at the Jessop Hospital for Women, Sheffield, United Kingdom. PATIENTS: Nineteen patients with unexplained infertility and nineteen normal fertile men from a donor insemination program. MAIN OUTCOME MEASURES: Monitoring of reactive oxygen species generation; computerized assessment of the hyperactivated motility pattern of the spermatozoa under capacitating conditions, with and without the addition of follicular fluid (FF); evaluation of sperm-zona pellucida (ZP) binding. RESULTS: Reactive oxygen species generation was not significantly different between the study and control group. Follicular fluid-induced hyperactivation (HA) was significantly lower in patients with unexplained infertility than in the donors, but no difference was found comparing spontaneous HA (without FF) between the two groups. Follicular fluid-induced HA was completely absent in 39% of infertile men and only one fertile man. A low sperm-ZP binding ratio (< or = 0.3) was observed in 28% (5/18) of patients with unexplained infertility. In all, nine infertile men (47%) had 0% FF-induced HA and/or low sperm-ZP binding ratio. CONCLUSIONS: Assessing FF-induced HA and sperm-ZP binding may have clinical significance in distinguishing a subgroup of men with unexplained infertility with sperm function abnormalities in which the fertilizing capacity of the spermatozoa might be reduced. These patients cannot be identified by the conventional semen analysis. Both tests may also be useful in planning the appropriate treatment for couples with unexplained infertility.

Female↗
PubMed1993

Prospective study of leukocytes and leukocyte subpopulations in semen suggests they are not a cause of male infertility.

OBJECTIVE: To determine the effects of leukocytes in semen on sperm quality and the ability to achieve conception. DESIGN: A prospective analysis of 512 couples attending a regional infertility clinic. Leukocyte subsets were quantified using a monoclonal antibody-based staining procedure. In addition to basic seminal parameters (density, motility, morphology, and antisperm antibodies), reactive oxygen species and immature germ cells were also quantified in the semen of each patient. The presence or absence of a treatment-independent conception was determined 22 months after the start of the study. Semen parameters were then related to the ability to conceive. SETTING: University-based center for reproductive medicine. PARTICIPANTS: Success or failure to conceive was recorded from 512 couples. Couples were then selected to minimize the influence of any pathology of the female on outcome. A final study group of 229 couples, in which the women had regular menstrual cycles, ovulatory midluteal serum P levels of > 18 nmol/L, and patent fallopian tubes was finally selected for analysis. MAIN OUTCOME MEASURE: Pregnancy. RESULTS: Leukocyte concentration (total or individual subsets) was not associated with either reduced semen quality or conception rates. Similarly, neither reactive oxygen species or antisperm antibody (immunobead) concentration had any bearing on the outcome. Of all semen parameters measured, only the level of immature germ cells was found to be negatively associated with the rate of conception. CONCLUSION: Measurement of seminal leukocytes in routine semen analysis appears to be of little prognostic value with regard to male fertilizing potential. As reactive oxygen species and antisperm measurement were of similar predictive value, the term "immunologic male infertility" should be redefined.

Autoantibodies↗
PubMed1993

Sperm numbers and distribution within the human fallopian tube around ovulation.

This study aimed to determine the number and distribution of spermatozoa within the human Fallopian tubes around ovulation. Parous women, undergoing total abdominal hysterectomy for menorrhagia, were inseminated with either partner's semen (3/10) or donor semen (7/10). Approximately 18 h later both Fallopian tubes were ligatured into ampullary, isthmic and intramural regions. These were removed and assessed for sperm content by flushing, scanning electron microscopy (SEM) or homogenization. A median of 251 spermatozoa were recovered (range, 79-1386). The number of spermatozoa within each tube was not significantly different. The ovulatory ampulla contained a significantly (P < or = 0.01) larger percentage of spermatozoa than the non-ovulatory ampulla. The number of motile spermatozoa inseminated was not significantly correlated to the number of spermatozoa recovered, but a trend was identified. The time between the onset of the luteinizing hormone surge and hysterectomy was significantly correlated (P < or = 0.01) to the number of spermatozoa within the intramural regions, but not to the tubal sperm distribution. Spermatozoa were not observed, by SEM, bound to the tubal epithelium. These data suggest that, after artificial insemination at least, sperm access to the human Fallopian tube may be controlled, but that ovulation does not affect the redistribution of spermatozoa between tubal regions and that the isthmus does not appear to act specifically as a sperm reservoir.

Adult↗
PubMed1993

Uterine flushing: a method to recover spermatozoa and leukocytes.

The process of sperm transport from the cervix, where a leukocytic reaction is initiated, through the uterus to gain access to the site of fertilization is very poorly understood. This preliminary study was designed to utilize a uterine flushing technique to determine firstly, the number of spermatozoa that can be recovered from the uterine cavity at 4 h post-insemination, around the time of ovulation, and secondly, to establish whether the spermatozoa initiate a leukocytic response while present. Uterine flushing was carried out in 10 potentially fertile women at 4 h post-insemination with donor semen, 24-36 h after the onset of the luteinizing hormone (LH) surge. The flush fluid was analysed for the numbers of spermatozoa and leukocytes present. In 8/10 women spermatozoa were retrieved from the uterus, in consistently low numbers (median 46, range 3-415). In 5/5 women leukocytes were recovered (median, 2.75 x 10(8)/l, range 2.0 x 10(8)-12.7 x 10(8)/l) from an origin other than peripheral blood contamination. These results suggest firstly that the flushing technique was a consistent method for retrieving spermatozoa and leukocytes from the uterine cavity, secondly that only low numbers of spermatozoa can be retrieved on flushing and thirdly that the leukocytic reaction to spermatozoa extends to the uterine cavity.

Female↗
PubMed1993

Prognostic significance of computerized motility analysis for in vivo fertility.

OBJECTIVE: To determine the predictive value of quantitative motility characteristics produced by the Hamilton Thorn Motility (HTM) Analyzer (Hamilton Thorn Research, Beverley, MA) for in vivo conception. DESIGN: A prospective analysis of 222 couples attending a regional infertility clinic. The measurements were made on a semen sample, and the presence or absence of a treatment-independent conception up to 22 months later was determined. The semen variables were then correlated to conception. SETTING: University based center for reproductive medicine. PATIENTS, PARTICIPANTS: The presence or absence of an in vivo conception was recorded in 222 couples in whom the influence on fertility of the female partner was minimized, i.e., normal in terms of history and examination, a regular menstrual cycle, ovulatory (midluteal serum P > 18 nmol/L [5.6 ng/mL]), and the outcome of the hysterosalpingogram was normal. The median follow-up time was 13 months (range, 5 to 22 months). INTERVENTIONS: None. MAIN OUTCOME MEASURE: Pregnancy. RESULTS: A number of variables were significantly related to time to conception. When a forward stepwise analysis was performed, the total number of spermatozoa was selected on the first step, and average path velocity was selected on the second step. No other variables were selected. The final variables consisted of the total number of spermatozoa and average path velocity. CONCLUSION: The measurement of quantitative motility and sperm number using a HTM Analyser is of clinical value.

Electronic Data Processing↗
PubMed1992

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↗