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BACKGROUND & AIMS\ud \ud Gestational diabetes mellitus (GDM) compromises the level of docosahexaenoic acid (DHA) in phospholipids of maternal and fetal red blood cells and fetal plasma. This is of some concern because of the importance of DHA for fetal neuro-visual development. We have investigated whether this abnormality could be rectified by supplementation with DHA-enriched formula.\ud \ud METHODS\ud \ud Women with GDM (n = 138) recruited from Newham University Hospital, London received two capsules of DHA-enriched formula (active-group) or high oleic acid sunflower seed oil (placebo-group) from diagnosis until delivery. Maternal (baseline and delivery) and fetal (cord blood) red blood cell and plasma phospholipid fatty acid composition, and neonatal anthropometry were assessed.\ud \ud RESULTS\ud \ud One hundred and fourteen women (58 active, 56 placebo) completed the trial. The active-group compared with the placebo-group had significantly enhanced level of DHA in plasma phosphatidylcholine (4.5% vs 3.8%, P = 0.011), red blood cell phosphatidylcholine (2.7% vs 2.2%, P = 0.022) and phosphatidylethoanolamine (9.5% vs 7.6%, P = 0.002). There was no difference in cord plasma and red blood cell phospholipid DHA between the two groups. The neonates of the two groups of women had comparable anthropometric measurements at birth.\ud \ud CONCLUSION\ud \ud Daily supplementation of 600 mg DHA enhances maternal but not fetal DHA status in pregnancy complicated by GDM. The inefficacy of the supplement to improve fetal status suggests that the transfer of DHA across the placenta maybe impaired in women with the condition. Regardless of the mechanisms responsible for the impairment of the transfer, the finding has implications for the management of neonates of women with GDM because they are born with a reduced level of DHA and the condition is thought to be associated with a risk of neuro-developmental deficits. We suggest that babies of women with GDM, particularly those not suckling, similar to the babies born prematurely require formula milk fortified with a higher level of DHA.

Preterm neonates are more susceptible to infection than term neonates. Arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3) are biologically active components of cell membrane phospholipids. Arachidonic acid is a substrate for the synthesis of eicosanoids, potent regulators of immune function. Preterm babies may have a deficiency of arachidonic acid and docosahexaenoic acid, but the impact of this deficit on maturation of the immune system is unknown. To address this we explored links between placental provision of fatty acids to cord blood mononuclear cell (CBMC) membranes using gas chromatography (GC), and maturation of the immune response with gestational age by analysing lymphocyte subsets by flow cytometry. This is the first study to examine the lipid profile of the phosphatidylcholine (PC) and phosphatidylethanolamine (PE) fractions of CBMC membranes from preterm neonates. The long chain polyunsaturated fatty acid (LCPUFA) composition of CBMC membranes was dominated by arachidonic acid in both PE (34%) and PC (15%) fractions in healthy term neonates (≥37 weeks, n=9), whilst in healthy preterm neonates

Effective tubal transport of ova, sperm and embryos is a prerequisite for successful spontaneous pregnancy. Although there is much yet to be discovered about the mechanisms involved, it is evident that tubal transit is a far more complicated process than initially thought. Propulsion of gametes and embryos is achieved by complex interaction between muscle contractions, ciliary activity and the flow of tubal secretions. Evidence is accumulating of the important and possibly pre-eminent role of ciliary motion in this process; and this review describes current knowledge about ciliary activity and its physiological regulation. There is also a description of the effects on ciliary function of cigarette smoking and various pathological states, including endometriosis and microbial infection, with consideration given as to how altered ciliary activity may impact upon fertility.

Objective To present our experience with the combined use of laparoscopy and hysteroscopy as the primary investigation for tubal and uterine factors in infertility. Design This is a prospective study of patients investigated for infertility using hysterolaparoscopy for evaluation of infertility. Setting Assisted Conception and Infertility Unit at Newham General Hospital, London. Subjects These were 117 consecutive patients, of whom 53 (45.3%) had primary and 64 (54.7%) had secondary infertility. Results Laparoscopic examination revealed at least one abnormal finding in 77 (66.3%) patients and 29 (26.6%) patients had abnormal findings at hysteroscopy. Concurrent laparoscopy and hysteroscopy were completed in 108 patients in whom eight (7.4%) had abnormal hysteroscopic findings but normal laparoscopic findings. Five out of six (83.3%) patients who had intrauterine adhesions had a history of previous uterine surgery. Out of 67 patients with abnormal laparoscopy, 21 (31.3%) had abnormal findings at hysteroscopy compared with eight out of 41 (20.0%) with normal laparoscopy (χ2 = 1.678; P > 0.05). There was no significant difference in the prevalence of intrauterine abnormality detected hysteroscopically between the patients with primary and secondary infertility (χ2 = 2.899; P > 0.05). Conclusions The findings of this study suggest that routine use of hysteroscopy adds little information to the management of infertile patients and its concomitant use should be based on the history or physical examination findings. However, certain intrauterine abnormalities do not have warning signs. Until the significance of these intrauterine abnormalities in infertility is proven, routine use of hysterolaparoscopy for infertility investigation will remain a controversial issue.

To test if docosahexaenoic acid-enriched fish oil supplementation rectifies red cell membrane lipid anomaly in pregnant women with Type 2 diabetes and their neonates, and alters fetal body composition.Women with Type 2 diabetes (n = 88; 41 fish oil, 47 placebo) and healthy women (n = 85; 45 fish oil, 40 placebo) were supplemented from the first trimester until delivery. Blood fatty acid composition, fetal biometric and neonatal anthropometric measurements were assessed.A total of 117 women completed the trial. The women with Type 2 diabetes who took fish oil compared with those who received placebo had higher percentage of docosahexaenoic acid in red cell phosphatidylethanolamine in the third trimester (12.0% vs. 8.9%, P = 0.000) and at delivery (10.7% vs. 7.4%, P = 0.001). Similarly, the neonates of the women with Type 2 diabetes supplemented with the fish oil had increased docosahexaenoic acid in the red cell phosphatidylethanolamine (9.2% vs. 7.7%, P = 0.027) and plasma phosphatidylcholine (6.1% vs. 4.7%, P = 0.020). Docosahexaenoic acid-rich fish oil had no effect on the body composition of the fetus and neonates of the women with Type 2 diabetes.A daily dose of 600 mg of docosahexaenoic acid was effective in ameliorating red cell membrane docosahexaenoic acid anomaly in pregnant women with Type 2 diabetes and neonates, and in preventing the decline of maternal docosahexaenoic acid during pregnancy. We suggest that the provision of docosahexaenoic acid supplement should be integrated in the antenatal care of pregnant women with Type 2 diabetes.

Objective: To investigate the mechanisms involved in the stimulatory effect of fallopian tube epithelial cell coculture on sperm movement characteristics. Design: Human spermatozoa were cultured with human fallopian tube epithelial cell monolayers. A microporous membrane was used to prevent sperm-to-epithelial cell contact. Sperm movement characteristics were measured at 4 and 24 hours. Setting: University hospital and fertility center. Patient(s): Voluntary donors. Intervention(s): None. Main Outcome Measure(s): Movement characteristics of human spermatozoa. Result(s): Fallopian tube epithelial cell coculture increased sperm motility, curvilinear velocity, amplitude of lateral head displacement, and hyperactivated motility, mainly at 24 hours, compared with controls. These stimulatory effects were inhibited when a microporous membrane prevented cell-to-cell contact between sperm and fallopian tube epithelial cells. Conclusion(s): Physical contact between sperm and epithelial cells in coculture systems seems to be the main factor in stimulating sperm movement characteristics, and this could be the main mechanism of in vivo sperm capacitation.

In humans 6-sulphatoxy melatonin (SaMT) is the principal metabolite of endogenous and exogenous melatonin. 5-sulphatoxy N-acetyl-serotonin (SNAS) is a minor metabolite of exogenous melatonin, but it has not been established whether the levels of endogenous SNAS in plasma derives principally from endogenous melatonin. We have developed the first radioimmunoassay (RIA) for SNAS and used it (together with RIAs for melatonin and SaMT) to determine whether endogenous SNAS derives from endogenous melatonin or from platelet serotonin. Our results show a) the values of endogenous SNAS, unlike endogenous SaMT, increased with blood collection procedures that increased the values of serotonin, b) the values of endogenous SNAS in urine or in platelet-poor plasma were approximately the same as those of endogenous SaMT, but, unlike SaMT, did not show a diurnal rhythm, and c) we confirmed that SNAS was a minor metabolite of orally ingested melatonin. Thus, our conclusion is that SNAS is a minor metabolite of exogenous melatonin, but is not a significant metabolite of endogenous melatonin. In all probability, endogenous SNAS is principally the metabolite of platelet serotonin.

Summary The oviduct provides a specialized environment for the survival and transport of the mammalian gametes to the site of fertilization within well defined time limits. The oviduct environment facilitates the process of fertilization and development of the preembryo. Furthermore, the spermatozoa and the pre-embryo, which differ antigenically from the mother, are not attacked by the immune system, which differentiates them from pathogens. The mechanisms by which all these complex processes are controlled are not fully understood. The disadvantage of cell culture systems for physiological study is that the original histological appearance is lost along with some endocrine and paracrine activities. Cells cultured on glass or plastic surfaces lose the polarization normally present in intact tissues. Premature senescence and de-differentiation may occur. It has been shown that extracellular matrix may stimulate proliferation and prevent dedifferentiation. The use of polarized cell cultures also enable the study of cellular orientation and protein secretion. Culture systems for polarized and non-polarized Fallopian tube epithelial cells have been developed in our laboratory. The subsequent experimental studies have shown that the presence of oviduct epithelial cells has a specific and significant stimulatory effect on sperm capacitation, reversing effects on the blockage of blastocyst formation. The exact mechanism of how the co-culture system achieves this is yet to be elucidated and in this regard, the unique oviduct proteins are possible candidates. Further studies of the epithelial cell cultures directed at the nature of the secretory behavior of these cells, are necessary in order to determine how these cells behave in vitro .

Elevated nocturnal melatonin is found in women with idiopathic hypogonadotropic hypogonadism (IHH), but it is not known whether this is implicated in the etiology of their GnRH deficiency. It is unlikely that nocturnal melatonin can be implicated in the etiology of the GnRH deficiency of Kallmann's syndrome (KS), because this condition is caused by defective neuronal migration in embryonic life. We therefore measured nocturnal melatonin in women with IHH and KS to determine whether it was elevated in one or both conditions and thereby to determine whether it was implicated as cause or consequence of GnRH deficiency. Four women with IHH, 3 women with KS, and 7 individually matched (age and body size) controls were recruited. Frequent day- and nighttime samples were taken for LH pulsatility studies. All patients showed absent or diminished LH pulsatility, compared with their respective controls. Samples were also taken over 24 h for melatonin and 6-sulphatoxymelatonin (the principle metabolite of melatonin and an independent marker of its secretion). Melatonin and 6-sulphatoxymelatonin levels were elevated in 6 of 7 patients (compared with their matched controls) and were significantly elevated in the KS group (compared with their controls). The finding of elevated nocturnal melatonin (and its metabolite) in GnRH-deficient women with KS (as well as IHH) suggests that nocturnal melatonin is elevated as a consequence of GnRH deficiency, irrespective of its etiology.

Objective To review the experience of Down's syndrome screening in an inner city health district. Design In this retrospective study the names of the women were obtained from the Regional Cytogenetic Laboratory and District Down's Syndrome Register. Antenatal notes of the mothers were studied. Maternal age during the affected pregnancy, screening test performed if any, gestational age at booking and at screening, screening test results, and pregnancy outcome were reviewed. Setting An inner city health district. Population Down's syndrome cases diagnosed prenatally and postnatally. Interventions None. Results Antenatal diagnosis of Down's syndrome was made in 15/45 women (33.3%) ; Edwards' syndrome was diagnosed antenatally in 2/5 women. Screening was not performed in eight women (17.8%) whose infants were affected by Down's Syndrome ; it was also not performed in one woman whose baby was affected by Edwards' syndrome due to late presentation to the antenatal clinic or loss of the blood sample. Nineteen women (42.2%) with a Down's syndrome pregnancy and two women with a Edwards' syndrome pregnancy had had a negative screening test. Three women had had a positive screening test but declined amniocentesis. Among those who had a screening test, the overall detection rate of screening was 48.6% (18/37) for Down's syndrome and 50% (2/4) for Edwards' syndrome. The detection rates in different ethnic groups did not show significant difference. Four women chose not to have termination following diagnosis of Down's syndrome, giving a total reduction rate of 21%. Conclusion Screening programmes for Down's syndrome have not yet resulted in a substantial reduction in the number of affected babies. In addition to trials at developing screening programmes with better detection rates, efforts should be made to improve the provision of the screening service and the quality of antenatal care in general with emphasis on early presentation and optimal understanding of the implications of the tests.

This study investigates whether physiological variations in ovarian hormones during the ovarian cycle (OC) are associated with changes in auditory function. Sixteen women with normal hearing underwent auditory tests and simultaneous measurements of the hormone levels four times during OC. The auditory tests included recording of otoacoustic emissions (OAEs), the medial olivocochlear (MOC) suppression and auditory brainstem responses (ABRs). The OC was defined by oestradiol and progesterone serum levels and menstrual cycle dating. A significant spontaneous OAE frequency shift [F(3,114.6)=15.8, p0.001], with the greatest shift in the late follicular phase (highest oestrogen levels), was observed. Transient evoked OAE levels showed a consistent tendency in an increase in all frequency bands in the late follicular/early luteal stage and a decrease in the late follicular stage; TEOAE inter-session comparison indicated very small statistical differences. The MOC suppression changed significantly during OC [F(3,33.8)=3.2, p=0.036], with significant inter-session difference, lower in session 2 than in session 1 (p=0.019) and lower in session 4 than in session 1 (p=0.007). The ABR wave V absolute latency changed significantly during OC [F(3,33)=3.3, p=0.03], longer in the late follicular phase. There was also a significant positive correlation of TEOAEs and ABR (wave V latency and III-V interval) and significant negative correlation of MOC suppression with oestradiol levels in the follicular phase. The results of this study reflect very small changes in auditory function during OC, and they are suggestive of an increased hearing sensitivity around the time of ovulation.

In previous studies, we reported that neonates of women with gestational diabetes mellitus (GDM) have reduced blood levels of arachidonic acid (AA) and docosahexaenoic acid (DHA) that were unrelated to maternal status. Since both AA and DHA are selectively transferred from maternal to fetal circulation by the placenta, we have investigated whether the FA composition of the placenta is altered by GDM. Thirty-six women, 11 with and 25 without GDM, were recruited from Newham General Hospital, London. The women with GDM had higher levels of di-homo-γ-linolenic (P

The coupled movement of ions and water across epithelia determines the composition and volume of fluid present in the lumen of organs. The second messenger cAMP is important in effecting electrolyte and water transport in many transporting epithelia; however, its role in Fallopian tube transport is uncertain. We have conducted electrophysiological studies on Fallopian tube epithelial cell monolayers in Ussing chambers and have demonstrated that exogenously added cAMP and agents that generate its intracellular production results in an increase in short-circuit current consistent with the transport of net electrical charge from a basal to mucosal direction. In contrast to the known effects of ATP in this tissue, the increase in short-circuit current was not explicable in terms of electrogenic chloride secretion as it was not affected by the chloride channel inhibitors, 4-acetamido-4'-isothiocyanostilbene-2,2-disulphonic acid 1 mmol/l (SITS) and frusemide. Instead the current was reduced by the sodium channel inhibitor, amiloride, and was therefore, in part, explicable in terms of electrogenic Na + absorption. These findings will enhance our understanding of the physiological mechanisms responsible for human Fallopian tubal fluid formation and composition.

This study set out to compare the growth patterns and morphological characteristics of human fallopian tube epithelial cells isolated: (1) mechanically; and (2) enzymatically. Cells were cultured in medium supplemented with fetal bovine serum and antibiotics and their epithelial nature was established by immunocytochemistry for cytokeratins. Primary cultures were polygonal in shape with centrally located nuclei, irrespective of the isolation method. Cells isolated enzymatically exhibited a higher growth rate, but the survival rate was poor after more than 2-3 passages. Mechanical isolation gave a lower yield of cells, but had a higher survival rate when sub-cultured, even beyond 8 passages. Thus, mechanically isolated cells might be useful for longer term cultures, whereas enzymatically isolated cells are best only for short-term work.

We describe a nonextraction procedure, and two extraction procedures, for RIA of melatonin in human plasma. All procedures showed a diurnal rhythm of melatonin in human subjects, with interindividual differences greater than interprocedure differences. However, further investigations demonstrated considerable variability of recovery in the nonextraction procedure, suggesting a variability of binding proteins between samples. Combining recovery and dialysis experiments in the extraction procedures, we demonstrated that chloroform was unable to extract albumin-bound melatonin from a human serum albumin solution but, paradoxically, was able to extract bound and free melatonin from a plasma sample. The methanol extraction procedure extracted free and bound melatonin from all sources. These results indicate that albumin binding can substantially affect the RIA procedures. We conclude that assays should be validated against free and bound melatonin and that the two forms should be independently investigated when assessing bioactivity.

From evidence based on the use of specific receptor subtype antagonists, it has generally been assumed that human uterine tissue contains only type 2 (AT2) angiotensin II (AII) receptor subtype. Using a monoclonal antibody, 6313/G2, directed against a specific sequence in the extracellular domain of the type 1 AII receptor (AT1), in immunocytochemical studies, we show here that AT1 receptor is expressed in human endometrium. In particular, positive staining was seen in the endometrial glandular epithelium, and in the vascular endothelium, while the myometrium and endometrial stroma were negative. The most intense staining was observed during the late proliferative phase and less in the luteal phase. The ligand binding assay, using [125I]-angiotensin II, revealed high concentrations of AII receptors both in the endometrium and in the myometrium. Competition studies using losartan (AT1 specific) and CGP42112B (AT2 specific) showed that both AT1 and AT2 receptor subtypes were present in the endometrium, though only the AT2 receptor subtype was detected in the myometrium. Immunoblotting confirmed that the antibody 6313/G2 detected a single protein with a molecular weight of approximately 60 kDa. These data clearly demonstrate the presence of endometrial AT1 receptors whose expression appears to be under hormonal control.

The fallopian tube has many active roles in the process of reproduction, which include ovum pick-up, transportation of both sets of gametes to the site of fertilisation within well-defined time limits, providing an environment in which final gamete maturation and fertilisation take place, and maintenance of the gametes and the pre-embryo, which is delivered to the uterine cavity at a time when conditions are optimal for nidation. Furthermore, the sperm and pre-embryo, which differ antigenically from the mother, are not attacked by the immune system, which differentiates between them and pathogens. The mechanisms by which all these complex processes are controlled are not all well understood, and currently the principial means of assessing tubal function is to observe tubal patency and normal morphological characteristics at endoscopy (Maguiness et al. 1992a).

BACKGROUND AND OBJECTIVE Idiopathic hypogonadotrophic hypogonadism (IHH) is a condition of gonadotrophin releasing hormone (GnRH) deficiency. IHH associated with anosmia is Kallmann’s syndrome. A variant has been described by Bauman where a patient with Kallmann's syndrome apparently regained normal hypothalamo-pituitary function 2 years after the initial diagnosis. GnRH secretory activity can be assesed by measuring LH pulsatility. Our objective was to define the pattern of LH pulsatility in men with IHH and Kallmann’s syndrome compared with those of normal controls, and to determine whether there is evidence for a Bauman variant of Kallmann's syndrome. DESIGN Patients with IHH and Kallmann’s syndrome were recruited from the endocrine clinic. Long-term hormone replacement therapy was discontinued. LH pulsatility was determined. PATIENTS Three men with IHH, 3 men with classical Kallmann’s syndrome and 5 normal male volunteers. MEASUREMENTS Baseline serum FSH, LH and testosterone. Intensive blood sampling every 10 minutes for serum LH from 1000 to 1600 h during the day and 2200 to 0400 h during the night to measure LH pulsatility. RESULTS The volunteer group showed normal LH pulsatility. In the patient group, LH secretion was apulsatile in one, showed significantly diminished amplitude in four, and there was normal pulsatility in one patient which remained normal 5 months later. CONCLUSION Three patients with idiopathic hypo-gonadotrophic hypogonadism and 2 with Kallmann's syndrome had variable degrees of GnRH deficiency. One patient with Kallmann's syndrome had apparently normal GnRH activity, which remained normal 5 months later. This patient appears to have the Bauman variant of Kallmann's syndrome.

The physiological factors which induce and maintain mammalian sperm maturation and motility generally remain unclear, although several agents are known to be involved. We describe here the application of immunocytochemical and immunoblotting methods to identify the angiotensin II type 1 (AT1) receptor in the tails of ejaculated rat and human sperm. Motility data on stimulated and unstimulated sperm from volunteers and patients attending fertility clinics showed that angiotensin II may increase both the percentage of motile sperm and their linear velocity, while the specific AT1 receptor antagonist DuP753 inhibited the action of angiotensin II on the percentage of motile sperm. In rat seminiferous tubules, AT1 receptors were present in primary spermatogonia and in spermatid tails, but immunoreactivity was not seen in sperm contained in caput or cauda epididymis, showing that AT1 receptor function is regulated during transit through the reproductive tract. Since local tissue reninangiotensin systems are present in both male and female tracts, the data suggest that angiotensin II has a role in the maintenance of sperm function and fertility. Journal of Endocrinology (1995) 144, 369–

Objective To investigate the influence of cultured human oviductal epithelial cells on the movement characteristics of human spermatozoa. Design Human spermatozoa were cultured with monolayers of human epithelial or Vero cells or conditioned media derived from these cell types. The viability and movement characteristics of the cells was subsequently analyzed at 4, 24, and 48hours. Setting University hospital and Medical Research Council laboratories. Patients, Participants Volunteer donors. Main Outcome Measures Movement characteristics of human spermatozoa. Results The presence of both Vero and oviductal epithelial cells, but not conditioned media, had a general promoting effect on sperm survival, significantly enhancing sperm viability and motility for up to 48hours of culture. In addition, the presence of oviductal epithelial cells had a specific, significant stimulatory effect on sperm capacitation, enhancing the incidence of hyperactivated motility after 4, 24, and 48hours of culture. Significantly, this effect was not observed with cocultures containing Vero cells. Conclusions The coincubation of human spermatozoa with human oviductal epithelial cells provides a convenient system for the induction and analysis of sperm capacitation.

Summary Fallopian tubes were obtained from 25 women undergoing abdominal hysterectomy. Pieces of fallopian tube mucosa were placed in culture flasks containing minimum essential medium in Earle's salts supplemented with fetal bovine serum. First passage was carried out after 7–10 days and subcultures in 4–5 days. For polarised cell culture, epithelial cells were seeded onto an extracellular matrix system. New epithelial cells were seen on day 2–3 of the primary culture and epithelial patches on day 7–10. Cells reached confluence in 4–5 days in subcultures. The cells could be subcultured for 7–11 passages with a life span of 42–60 days. Epithelial origins of the cells were confirmed by immunofluorescence staining with anti -cytokeratin antibody. Polarised cells showed a columnar pattern, microvilli on their apical surface and basally located nucleus whereas non-polarised cells were flat. It was concluded that the human fallopian tube epithelial cells can be cultured in vitro to create non-polarised and polarised cell layers by using a simple and reproducible technique and this system can be a potential model to study function of the fallopian tube.

Endometriosis and infertility are known to be associated, but it is unclear whether endometriosis causes infertility. We used contrast analogue enhancement to study the effect of peritoneal fluid from women with early stage endometriosis on the ciliary beat frequency of human fallopian tube epithelium. We obtained peritoneal fluid from six women with early stage endometriosis and from six fertile women with no evidence of endometriosis to use as controls. Fallopian tubes from hysterectomy specimens were collected from 17 women. The difference in ciliary beat frequency between fallopian tubes exposed to peritoneal fluids of women with and without endometriosis increased with the duration of incubation (mean difference at 24 h 1.35 Hz, 95% CI 0.94-1.75, p=0.01). At 24 h, ciliary beat frequency was significantly lower in the incubations with peritoneal fluid from women with endometriosis than controls (4.29 [0.15] vs 5.64 Hz [0.15], respectively, p=0.001). Impairment of ciliary action in women with endometriosis might reduce fertility.

Summary The oviduct provides a specialized environment for the survival and transport of the mammalian gametes to the site of fertilization within well defined time limits. The oviduct environment facilitates the process of fertilization and development of the preembryo. Furthermore, the spermatozoa and the pre-embryo, which differ antigenically from the mother, are not attacked by the immune system, which differentiates them from pathogens. The mechanisms by which all these complex processes are controlled are not fully understood. The disadvantage of cell culture systems for physiological study is that the original histological appearance is lost along with some endocrine and paracrine activities. Cells cultured on glass or plastic surfaces lose the polarization normally present in intact tissues. Premature senescence and de-differentiation may occur. It has been shown that extracellular matrix may stimulate proliferation and prevent dedifferentiation. The use of polarized cell cultures also enable the study of cellular orientation and protein secretion. Culture systems for polarized and non-polarized Fallopian tube epithelial cells have been developed in our laboratory. The subsequent experimental studies have shown that the presence of oviduct epithelial cells has a specific and significant stimulatory effect on sperm capacitation, reversing effects on the blockage of blastocyst formation. The exact mechanism of how the co-culture system achieves this is yet to be elucidated and in this regard, the unique oviduct proteins are possible candidates. Further studies of the epithelial cell cultures directed at the nature of the secretory behavior of these cells, are necessary in order to determine how these cells behave in vitro .

This study is based upon an analysis of 27 severely oligozoospermic men, in whom defects in sperm motility and morphologic defects were also evident, and 35 control subjects of proven fertility. Semen samples recovered from these men were assessed by conventional criteria, time-exposure photomicrography, and the zona-free hamster egg penetration test. Evidence of severely defective sperm function was obtained for the oligozoospermic group, which gave a mean ± standard error (SE) fertilization rate of 2.8% ± 1.5, compared with 44.0% ± 3.4 for the normal fertile control subjects. Multivariate discriminant analysis was used for selection of those parameters of semen quality that could most accurately identify the 70% of oligozoospermic samples that failed to exhibit any fertilizing potential in the hamster egg assay. This analysis correctly identified 89% of such samples, on the basis of seven discriminating variables, of which the most significant were percentage of motile sperm, the log of sperm density, and a log transformation of the concentration of progressive spermatozoa.

A detailed analysis of semen quality was carried out in 85 couples with unexplained infertility by the use of conventional criteria of semen analysis, time-exposure photomicrography, and the zona-free hamster egg penetration test. According to the latter, 34.1% of the male partners exhibited evidence of defective sperm function, although only 4% of these patients were devoid of any demonstrable fertilizing capacity. Of the conventional parameters of semen analysis examined, the most revealing was the morphologic character of the sperm, which was significantly poorer (P less than 0.001) in the group with unexplained infertility than in the normal fertile control group and also showed a significant relationship (P less than 0.001) with the presence of subnormal fertilizing capacity in the hamster egg assay. A majority of movement characteristics measured by time-exposure photomicrography were significantly depressed in the group with unexplained infertility, compared with the normal fertile control group. In addition, certain of the movement characteristics investigated were significantly related to the fertilizing capacity of the spermatozoa; and, in this respect, a progressive swimming speed (greater than 25 micrometers/sec), a straight swimming mode of progression, and a small amplitude of lateral head displacement (Ah) all appeared to be important qualities.

In 31 infertile patients, ovulation was induced with clomiphene citrate and oocytes were captured by laparoscopy 36 hours after injection of chorionic gonadotropin (hCG). The oocyte was incubated in tissue culture medium for 6 hours, sperm was added, and culture was continued for 1 further hour. The oocyte was then transferred directly into the uterus in 20 μl of fluid containing 10 000-20 000 sperm. In 14 patients (45%) β-hCG levels in urine indicated trophoblastic activity, and 2 patients have continuing intrauterine pregnancies.

An investigation has been carried out in normal fertile men on the correlates of fertilizing capacity as defined by the zona-free hamster egg penetration test. The results demonstrate that the apparent fertility of these men is compatible with a wide range of intrinsic sperm quality as reflected by penetration rates ranging from 14% to 90% and differences in the minimum concentration of motile spermatozoa required to initiate penetration. These differences in sperm function could not be correlated with any of the conventional parameters of semen analysis or any of a large number of sperm movement characteristics analyzed by time-exposure photomicrography, with two exceptions: (1) the percentage of progressively (greater than 25 micron/sec) motile spermatozoa exhibiting an amplitude of lateral head displacement (Ah) of less than 10 micron, which was positively correlated with penetrating capacity (P less than 0.01), and (2) the percentage of progressively (less than 25 micron/sec) motile spermatozoa exhibiting an Ah of greater than 10 micron, which was negatively correlated with penetration rate (P less than 0.05). The information obtained in this study should provide a useful basis against which to compare the properties of spermatozoa in cases of suspected infertility.

The aim of this study was to determine whether the high concentration of arachidonic acid (AA) in term placentae accumulates during pregnancy or is an inherent characteristic of placental lipids. We investigated the lipid content and fatty acid composition of the human placental phospholipids at 2 gestational periods, early in pregnancy (8 -14 wk, n 48) and at term (38 - 41 wk of gestation, n 19). The subjects were healthy, normotensive, and free of medical and obstetric complications. The lipid concentration of placentae increased from 0.8% in early gestation to 1.4% at term (P 0.0001). The mean proportions of AA were lower in the choline (P 0.05), inositol (P 0.0001), and ethanolamine (P 0.0001) phosphoglycerides of the term compared with the early placenta. In contrast, the proportions of the immediate precursor of AA, dihomo--linolenic acid (DGLA), were higher in the term placenta, particularly in the inositol and serine phosphoglycerides (P 0.0001). In sphingomyelin, the percentage of lignoceric acid was increased and that of nervonic acid was reduced at term (P 0.01). The dominance of AA, particularly in the early placenta, suggests that it has an important role for placental develop- ment, i.e., organogenesis and vascularization. There was no evidence of an accumulation of AA in the placenta toward term, which might be a trigger for parturition. In contrast, the increased proportion of DGLA (precursor of the vasorelaxant and anticoagulant prostaglandin E1) at term is more consistent with a profile favoring optimal blood flow to nourish the fetal growth spurt. J. Nutr. 135: 2566 -2571, 2005.