Serum luteinzing hormone pulsatility and intratesticular testosterone and oestradiol concentrations in idiopathic infertile men with high and normal fofficle stimulating hormone serum concentrations

The purpose of the study was to evaluate pulsatile luteinizinghormone (L release and intratesticular concentrations of testosteroneand oestradlol in infertile men, to determine if alterationsin gonadotrophin secretion are associated with changes in thetesticular concentrations of steroids. Patients with idiopathicoligo/azoospermia were divided into a high follicle stimulatinghormone (FSH) group (n=5) and a normal FSH group (n = 6). Bloodsamples were taken every 15 mm for 6 h to determine LH, FSH,testosterone, oestradiol, sex hormone binding globulin, bioactiveLH and bioavailable testosterone. The patients underwent a bilateraltesticular biopsy for histological assessment and to determinetestosterone and oestradiol concentrations. Serum measure mentswere compared with those of seven fertile men. The high FSHgroup had a higher concentration of serum UI and oestradiolthan normal men (P < 0.01) and showed a lower frequency ofLII pulses than the normal FSH group and control men (P <0.01). Intratesticular oestradiol was higher in the high FSHgroup (P < 0.001), with a lower testosterone/oestradlol ratio(P < 0.01). Patients showed a negative correlation betweenthe serum testosterone/LH ratio and FSH (r = -–0.75; P< 0.01) and a positive correlation between the testicularoestradiol concentration and serum FSH (r=0.86; P<0.01).The histopathological examination only showed a smaller tubediameter in the high FSll group (P < 0.05). These data seemto indicate that a higher intratesticular concentration of oestradiolwith a lower testosterone/oestradiol ratio in the high FSH groupcould have a deleterious effect on spermatogenesis.     

Evaluation of {beta}-endorphin and interleukin-6 in seminal plasma of patients with certain andrological diseases

Human semen contains large amounts of opioid peptides and cytokines.We have measured the concentrations of interleukin (IL)-6 in140 semen samples and of -endorphinin 77 semen samples. Themedian concentration of endorphinin seminal plasma from normozoospermicmen(n = 23) was 154.7 pg/ml (10th—90th percentiles, 42.0—774.6),and there was no significant difference in the -endorphin concentrationamong normozoospermic, oligozoospermic (n= 28), asthenozoospermic(n= 15), azoospermic(n= 4) and post-vasectomy (n= 7) samples.There was no correlation between -endorphin concentration andsperm characteristics, nor with blood hormones. Endorphinconcentration was lower in cases with immunelogical infertility,as revealed by a positive direct mixedantiglobulin reactiontest (n = 12) ( > 0.01), than inmatched controls. The medianconcentration of IL-6 insamples with normal sperm concentration,motility andmorphology with or without white blood cells (n=39) was 26.1 pg/ml (10th–90th percentiles, 7.3–172.3),and there was no significant difference in the IL-6 concentrationamong normozoospermic, oligozoospermic (n= 46),asthenozoospermic(n= 32), azoospermic (n= 13) and post-vasectomy (n= 10) samples.The IL-6 concentration was significantly higher in cases ofvaricocele (n= 22)without white blood cells in semen (P <0.001) than in matched controls without varicocele (n= 23).In addition, the IL-6 concentration was elevated (P < 0.0001)in cases with accessory sex gland inflammation (n= 40). IL-6concentration was positively correlated with white blood cellsin semen (n= 60, r = 0.59, P < 0.0001), but there was nocorrelation with -endorphin concentration. The IL-6 concentrationchosen to differentiate between cases with and without accessorygland inflammation was 45.3 pg/ml, with a specificity of 80.6%and a sensitivity of 92.5%. It is concluded that -endorphinin seminal plasma playsan immune suppressive role, and thatincreased IL-6 concentration may be related to testicular dysfunctionincases with varicocele. Furthermore, IL-6 is an accurate markerof accessory sex gland inflammation.     

Relationships between motility parameters, morphology and acrosomal status of human spermatozoa

A total of 130 semen samples were examined for motility (bycomputer-assisted sperm analysis), morphology and acrosomalstatus. A high positive correlation was found between percentagesof normal forms and progressive motility in the whole semen(r = 0.539, P < 0.0001) as well as in the Percoll fraction(r = 0.702, P < 0.0001). Among the specific abnormalities,acrosome defects were most highly correlated with progressivemotility (r = –0.492, P < 0.0001, in the Percoll fraction).The percentage of total spontaneously acrosome-reacted spermatozoain the Percoll fraction was negatively correlated with the progressivemotility (r = –0.499, P < 0.0001) and with the percentageof normal forms (r = –0.430, P < 0.0001). Surprisingly,the percentage of total spontaneously acrosome-reacted spermatozoawas poorly linked with head abnormalities but displayed significantpositive correlations with the percentages of bent tails (r= 0359, P < 0.0001) and of coiled tails (r = 0371, P <0.0001). These data suggest that sperm defects are often linkedtogether, reflecting spermiogenesis and/or epididymal dysfunctions.     

Andrology: Insulin-like growth factor-I and {alpha}2-macroglobulin in seminal plasma correlate with semen quality

Insulin-like growth factor-I (IGF-I) and ₂-macroglobulin (₂-M)are believed to be involved in the development of germ cells.IGF-I is mainly controlled by concentrations of human growthhormone (HGH), influences cell proliferation and differentiationand its action is mediated by insulin-like growth factor-bindingproteins (IGFBP), placental protein 14 (PP14) and prostate-specificantigen (PSA). ₂-M acts as a broad spectrum proteinase inhibitorand a binding protein for many cytokines and hormones, e.g.inhibin and activin. This study was designed to identify concentrationsof these molecules in seminal plasma in normal semen samplesof healthy men, correlations with semen quality, the relationshipof IGF-I and ₂-M with factors affecting male fertility, andwhether vasectomy influences the concentrations of these molecules.Concentrations of IGF-I and₂-M in human seminal plasma wererelated to semen quality, basal concentrations of HGH, testosterone,IGFBP-3, soluble fibronectin receptor (sFNR), PSA and PP14 inseminal plasma and to serum concentrations of luteinizing hormone(LH) and follicle stimulating hormone (FSH). Commercially availableassays were used to analyse 69 semen samples of various qualityand 11 post-vasectomy samples. IGF-I concentrations in seminalplasma were significantly correlated with the percentage ofmorphologically normal spermatozoa (r = 0.748, P = 0.00001)and sperm concentration (r = 0301, P = 0.011), but negativelycorrelated with serum FSH (r = –0.506, P = 0.00006) andPSA in seminal plasma (r = –0388, P = 0.0009). Total ₂-Mwas significantly correlated with sperm count (r = 0.423, P= 0.0005), percentage of progressively motile spermatozoa (r= 0.444, P = 0.00019), quality of motility (sperm motile efficiency,r = 802, P = 0.00001) and straight line velocity (r = 0.411,P = 0.0013). Correlation between the sperm concentration andHGH in seminal plasma was weak (r = 0.287, P = 0.015). Vasectomyreduced the concentration of total ₂-M (P = 0.00008) and HGH(P = 0.0068) in the seminal plasma; IGF-I was also reduced aftervasectomy when the total ejaculate amount was considered. ThusIGF-I and ₂-M are significant for the germ cell development:IGF-I hi the maturation of spermatozoa and ₂-M in progressivemotility.     

Andrology: The presence of Mullerian inhibiting substance in human seminal plasma

Müllerian inhibiting substance (MIS), produced by testicu-IarSertoli cells, is present in adult male serum. The first aimof this study was to determine if MIS is present in seminalplasma. Using an enzyme-linked immunosorbent assay (ELISA),we measured MIS concentrations in seminal plasma from 23 donorsexhibiting normal (WHO criteria) sperm qualities, and 169 patientswith subnormal sperm parameters. The second aim of this studywas to examine a potential relationship between MIS and spermmotility. MIS concentrations in seminal plasma ranged from 0.5to 3.6 ng/ml in donors and from 0.5 to 17.8 ng/ml in patients.Motility index (MI, mean ± SEM) for all patient sampleswas lower compared with donors (113.3 ± 3.2 and 1983± 13.5, P < 0.00001), while mean MIS concentration(± SEM) was higher (4.2 ± 03 and 1.4 ±0.2, P < 0.0003). When the patients were stratified intoGroups I (motility < 50%, n = 42) and D (motility >50%,n = 127), the MI (mean ± SEM) values were 623 ±3.8 and 130.2 ± 2.7 respectively (P < 0.0001 for bothcompared with donors) and mean MIS concentrations (±SEM) were 5.4 ± 0.6 and 3.9 ± 03, respectively(P < 0.0001 and P < 0.001 compared with donors). The inverserelationship between MIS concentration in seminal plasma andmotility index suggests that MIS may have a function in modulatingmotility.     

Andrology: Relationship between resazurin reduction test, reactive oxygen species generation, and {gamma}-glutamyltransferase

The production of reactive oxygen species (ROS) from spermatozoahas been measured by chemiluminescence in the two fractionsof a Percoll gradient column (47 and 90%). Chemiluminescentsignals were recorded in each fraction after addition of luminoland horse-radish peroxidase (basal state), and after stimulationwith formylmethionyl-leucyl-phenyl-alanine (FMLP) and phorbolester (PMA). We found an inverse correlation between the reducingcapacity of the semen as estimated by the resazurin test, andthe production of ROS by the spermatozoa, especially after stimulationwith PMA (r = 0.51, P <0.001). -glutamyltransferase activityof seminal plasma was inversely correlated with ROS (r = –0.47,P < 0.01) and the resazurin test result (r = –0.43,P < 0.001) suggesting a possible role of prostatic secretionsin the protection of spermatozoa against oxygen radicals. Theresazurin test has a positive predictive value of 92.5% at acriterion value of colour scale 4 to discriminate between normaland excessive ROS production by spermatozoa, with sensitivity79% and specificity 74%. In addition, at criterion value ofcolour scale 3, the resazurin test could distinguish betweensamples with normal or low activity of -glutamyltransferase,with sensitivity 57% and specificity 93%. It is concluded thatthe result of the resazurin test can be influenced by the oxidativestress of spermatozoa and by prostatic function.     

Ultrasonic texture and volume of testicles in infertile men

Ultrasound scanning of the testes and surgical biopsy were performedin 95 infertile men to evaluate the use of ultrasound In maleinfertility. Ultrasonic testicular volume was calculated usingthree measurements and the formula of an ellipsoid, and theultrasonic texture was evaluated and given a score from 1 to5, indicating increasing degrees of irregularity. The medianscore was 3 (range 1–5), which was higher than previouslyfound in nonnal men (median score 2; range 1–5; P <0.0001). The ultrasonic texture score was lower in testes witha uniform pattern of 100% spermatogenic tubules compared withthe rest, both for the right (P < 0.001) and for the left(P < 0.0005) testis. Texture score was correlated with thenumber of obliterated tubules for both testes (P <0.001).The mean ultrasonic testicular volume of the right testis was10.30 ml, and that of the left 10.26 ml. Both were smaller comparedwith the findings in normal men (P <0.0001). Ultrasonic testicularvolume was negatively correlated with texture score (P <0.001). A positive correlation between ultrasonic volume andsperm count was seen (P < 0.001). Sperm count was negativelycorrelated with texture score if calculated together with datafrom 119 men from the general population (P < 0.001). Thestudy shows that ultrasonic volume and texture are valuableparameters in the evaluation of infertile men.     

Sequential clomiphene citrate and human menopausal gonadotrophin for ovulation induction: comparison to clomiphene citrate alone and human menopausal gonadotrophin alone

The need for frequent injections and monitoring, the possibilityof multiple gestations, and the higher cost compared to clomiphenecitrate, prevents many clinicians from using human menopausalgonadotrophin (HMG) for ovulation induction. A sequential medicationregimen, in which HMG is taken after clomiphene, overcomes theseproblems. We retrospectively compared per cycle fecundity andbirth rates in 119 cycles of clomiphene—HMG, 524 cyclesof clomiphene alone, 57 cycles of HMG alone, and 79 cycles ofconcurrent HMG and clomiphene in patients receiving intra-uterineinsemination (IUI), who were free of endometriosis or tubaldisease. Per cycle fecundity for clomiphene—HMG was 22%[95% confidence interval (CI) 12–34%], double that ofclomiphene alone (11%) (95% CI 8–14%) (P < 0.01), andequal to HMG alone (18%) (95% CI 7–29%) or HMG and clomiphenetogether (19%) (95% CI 10–28%). The multiple birth ratefor clomiphene—HMG (7/21) equalled that for HMG alone(3/12) and HMG and clomiphene together (3/8). The average numberof ampoules of HMG required [follicle stimulating hormone (FSH)75 mIU, luteinizing hormone (LH) 75 mIU] was decreased by 65%from 24.5 ± 1.0 for HMG or HMG and clomiphene togetherto 8.6 ± 0.3 for clomiphene—HMG (P < 0.001).Per cycle fecundity was identical when one, two or three ampoulesof HMG per day were administered after clomiphene. We concludethat ovulation induction with sequential clomiphene—HMGresults in fecundity double that of clomiphene alone and equalto HMG alone or concurrent with clomiphene, thereby reducingthe requirement for HMG.     

Physiology: Enhanced sensitivity of the extrinsic coagulation system during ovarian stimulation for in-vitro fertilization

The tissue factor activity in blood monocytes was investigatedduring ovarian stimulation for in-vitro fertilization (IVF)in 13 women. Blood samples were taken prior to hormonal stimulation(days 2–3 of the menstrual cycle, median serum oestradiolconcentration 70 pmol/1) and the day after ovulation inductionwith human chorionic gonadotrophin (days 11–13, medianserum oestradiol concentration 6270 pmol/l). The tissue factoractivity in unstimulated monocytes and factor VII concentrationwere unchanged during the treatment. However, the tissue factoractivity in lipopolysaccharide-stimulated monocytes was on averagemore than twice as high after stimulation (P < 0.02). A positivecorrelation was found between the tissue factor activity andthe serum concentration of oestradiol (r = 0.514, P < 0.02).The tumour necrosis factor (TNF)- increased during ovarian stimulation(P = 0.05), and there was a positive correlation between thechange in TNF- and the change in tissue factor activity (r =0.663, P < 0.05). Our results indicate an enhanced sensitivityof the extrinsic coagulation system during IVF treatment sincemore tissue factor is available upon stimulation. It is suggestedthat this may be important in thrombotic situations. Furtherstudies are necessary to elucidate the mechanism behind thisresponse.     

Physiology: Growth rates and antrum formation of mouse ovarian follicles in vitro in response to follicle-stimulating hormone, relaxin, cyclic AMP and hypoxanthine

The culture of individual intact follicles in vitro, from smallpre-antral to pre-ovulatory stages, will improve our abilityto perform controlled experiments studying follicle growth andfemale gamete development. This study was undertaken to characterizefurther the conditions required for physiological follicle growthin vitro. We cultured a total of 398 pre-antral follicles ofimmature mice (aged 26–28 days) with an initial diameterof 140–340 µm for 4 days in vitro, using individualmicro-cultures under paraffin oil. Summarizing the results ofall groups, 50 follicles were damaged (12.6%) and, of thoseremaining intact (n = 348), 60 (17.2%) became atretic, 195 (56.0%)became antral and 26 (7.5%) ovulated. The most advanced folliclesgrew to 400–500 µm diameter. The presence of follicle-stimulatinghormone (FSH) in the medium significantly stimulated folliclegrowth in vitro (P < 0.03), in a manner proportional to theinitial diameter over the range of 140–250 µm initialdiameter, with larger follicles being refractory. FSH also significantlyincreased the proportion of follicles forming antra (P <0.001) and their likelihood of ovulating in vitro (P < 0.01),and reduced the frequency of atresia (P < 0.01). Dibutyryl-cyclicAMP mimicked FSH, significantly stimulating growth of largefollicles (P < 0.05) and antrum formation (P < 0.01).Hypoxanthine also stimulated antrum formation (P < 0.01)but did not significantly affect follicle growth. Porcine relaxinhad no significant effect on mouse follicle growth or antrumformation. The optimal conditions for mouse follicle growthin vitro have not yet been defined, but selection of folliclesof < 250 µm diameter and inclusion of FSH or dibutyryl-cyclicAMP in the culture medium are recommended.     

Proinflammatory cytokine polymorphisms and the risk of preterm birth and low birthweight in a Japanese population

Pregnancy and parturition involve a complex and poorly understoodmolecular and biological interplay between mother and fetus.Inflammatory cytokines have been reported to be associated withfetal growth and parturition. The aim of this study was to examinewhether common proinflammatory cytokine polymorphisms are associatedwith preterm birth (PTB), low birthweight or intrauterine growthrestriction in a Japanese population. We assessed a consecutiveseries of 414 women who had singleton deliveries in Sapporo,Japan between 2001 and 2005. Genotyping of IL1A –889C/T,+4845G/T (A114S), IL1B –511C/T, –31C/T, IL2 –384T/Gand IL6 –634C/G polymorphisms was determined by an allelicdiscrimination assay. The risk of PTB significantly increasedin women carrying the IL1A –889T allele (CC genotype [reference];CT genotype, odds ratios (OR): 2.5; 95% confidence intervals(95% CI): 1.4–4.8; CT+TT genotypes [dominant genotypemodel], OR: 2.5, 95% CI: 1.3–4.6). Similarly, the riskof PTB significantly increased in women carrying the IL1A +4845Tallele (GG genotype [reference]; GT genotype, OR: 2.4, 95% CI:1.3–4.4; GT+TT genotypes [dominant genotype model], OR:2.3, 95% CI: 1.2–4.2). The frequency of the IL1A TT haplotypein mothers with PTB was significantly higher than in motherswho had a term birth (P < 0.001), whereas the frequency ofthe IL1A CG haplotype in mothers who had a PTB was significantlylower (P < 0.001). Our findings suggest that the polymorphismsand haplotypes in the IL1A gene are associated with PTB in Japanesewomen.     

Relationship of biochemical pregnancy to pre-ovulatory endometrial thickness and pattern in patients undergoing ovulation induction

In order to assess the relationship between pre-ovulatory endometrialthickness and pattern and biochemical pregnancy, the pregnancyoutcome was retrospectively analysed in 81 patients undergoingovulation induction evaluated by vaginal ultrasound on the dayof human chorionic gonadotrophin (HCG) administration or luteinizinghormone (LH) surge. Biochemical pregnancies occurred in 7/32(21.9%) pregnancies when endometrial thickness was <9 mm,compared to 0/49 when endometrial thickness was 9 mm on theday of HCG administration or LH surge (P < 0.0025). Clinicalabortions occurred in 5/32 (15.6%) pregnancies when endometrialthickness was 6–8 mm, compared to 6/49 (12.2%) when endometrialthickness was 6–8 mm (NS). Endometrial thickness was relatedto the cycle day of HCG or LH surge (r = 0.37, P < 0.001)but was unrelated to oestradiol level on the day of HCG administrationor LH surge (r = 0.12). Biochemical pregnancies were relatedto endometrial pattern (r = – 0.22, P = 0.02) but wereunrelated to maternal age or previous abortions. Clinical abortionswere related to age (r = 0.26, P = 0.01) and to previous abortion(r = 0.25, P = 0.013) but were unrelated to endometrial pattern.Neither biochemical pregnancy nor clinical abortion was relatedto oestradiol or LH levels on the day of HCG administrationor LH surge. These findings suggest that the majority of biochemicalpregnancies do not result from karyotypically abnormal embryos,as do clinical abortions.     

Lead, magnesium, selenium and zinc in human seminal fluid: comparison with semen parameters and fertility

The concentrations of lead, magnesium, selenium and zinc inseminal fluid from men with variable semen quality (sperm morphology,density and motility) and fertility were determined by atomicabsorption spectrometer without or with Zeeman background correction.The mean (?SD) concentration of selenium in the samples (n =142) was 28.8 ? 9.5 µg/l, which was about a third of thecorresponding serum value (77.8 ? 13.3 µg/l, n = 140).The serum selenium level was significantly (P < 0.001) higherin infertile than in fertile men, but the seminal fluid didnot show such a difference. No correlation was obtained betweenselenium values in seminal plasma and sperm density or motility.The levels of lead in seminal fluid were very low with no correlationto the levels of magnesium, selenium and zinc or the semen qualities.The seminal fluid lead concentration was significantly (P <0.001) higher in infertile (3.6 ? 3.2 µg/l, n = 79) thanin fertile men (1.7 ? 1.0 µg/l, n = 39). Magnesium (103.5? 49.2 mg/l, n = 90) and zinc (141.1 + 71.7 mg/l, n = 157) concentrationsin seminal fluid were comparable with previous reports. Bothminerals showed a positive correlation to the seminal fluidselenium, while only zinc displayed a borderline correlationwith sperm density. The present findings indicate that the determinationof seminal fluid selenium may not offer any advantages overzinc and magnesium measurement in the fertility assessment andits role in human semen remains obscure. The low lead concentrationsin the present material is a clear indication of low industrialexposure.     

Endocrinology: Follicular fluid insulin-like growth factor-I and insulin-like growth factor-II concentrations vary as a function of day 3 serum follicle stimulating hormone

We determined follicular fluid concentrations of insulin-likegrowth factor (IGF)-I, IGF-II and inhibin as a function of day3 serum follicle stimulating hormone (FSH) in 16 women undergoingfollicular fluid aspiration in preparation for in-vitro fertilizationand embryo transfer. Follicular fluid concentrations of IGF-Iand IGF-II were significantly less in the ‘low’FSH group as compared to the ‘high’ FSH group. Themean IGF-I concentration was 67.6 ng/ml [confidence intervals(CI) 51.6–92.5] in the ‘low’ FSH group comparedto 87.1 ng/ml (CI 72.8–104.2; P < 0.025) in the ‘high’FSH group. Mean IGF-II concentrations were 354.8 ng/ml (CI 297.8–422.9)in the ‘low’ FSH group compared to 489.8 ng/ml (CI384.6–624.5; P < 0.05) in the ‘high’ FSHgroup. Follicular fluid inhibin concentrations did not differbetween groups. These differences in follicular fluid IGF asa function of day 3 FSH may raise questions regarding the rolegrowth factors play in the physiological processes of the ageingfollicle.     

Andrology: Seminal volume and total sperm number trends in men attending subfertllity clinics in the Greater Athens area during the period 1977-1993

The trends for such Important parameters of male fertility asseminal volume and total sperm number were assessed in men livingpermanently in the Greater Athens area over a prolonged periodof time. To this end, the records of three andrological laboratoriesemploying the same method for semen evaluation were analysedretrospectively. Out of 23 850 men examined from 1977 to 1993(17 years) for couple subfertillty, a total of 2385 (10%) wereselected for evaluation by a randomization procedure. Analysisof the data included (i) estimation of mean seminal volume andtotal sperm number per year, (ii) assessment of percentage frequencydistribution for each seminal parameter and (iii) evaluationof seminal volume and total sperm number changes in relationto the year of observation and age of the subjects. A significantdecrease (P < 0.01) of total sperm number was observed overthe years with a mean (± SEM) of 154.3 ± 19.2x 10⁶ at the beginning (1977), dropping to 130.1 ± 13.3x 10⁶ in the final year (1993). Mean seminal volume was lowerin the final year of observation, but its difference from theinitial year value was not significant Frequency distributionanalysis showed a marked decline in the 240–400 x 10⁶sub-set of the range of sperm number values from 16.9 ±4.5% (1977) to 10.6 ± 1.6% in the final year (P <0.01). Multiple regression analysis of seminal volume, totalsperm number, age and year of assessment revealed a significantdecline of the two seminal parameters along the years of observation(P < 0.05 and P < 0.0001 respectively). Over the sameperiod, a marked deterioration of some air pollution indiceswas observed in that area. It is concluded that in this raciallyand ethnically homogeneous sample of men, living under the sameenvironmental conditions, a significant decline in seminal volumeand total sperm number occurred over the 17 years of observation.     

Pregnancy: Activation of protein kinase C is required for oxytocin-induced contractility in human pregnant myometrium

Intracellular mediators regulating the initiation of parturitionare not fully understood. This study was designed to determinethe possible mechanism of oxytocin-induced uterine contractilityduring labour. In-vitro isometric contraction studies were performedwith longitudinal strips of human pregnant myometrium in thepresence and absence of the protein kinase C inhibitors, staurosporineand RO 31–8220, and the tyrosine kinase inhibitor, genistein.Phospholipase D activity was measured by employing the transphosphatidylationreaction. Staurosporine significantly reduced oxytocin-stimulatedcontractile activity with mean activity reduced by >50% followingthe addition of 10^–6M staurosporine (P < 0.01), whileaddition of 10^–5 M resulted in a measured mean contractileactivity of –10% of the control (P < 0.001, n = 5).Similarly, uterine activity was minimal with oxytocin applicationfollowing incubation with RO 31–8220, mean contractileactivity being reduced by -40% by the addition of 10^–7M RO 31–8220 (P < 0.05) and by –87% by the additionof either 10^–6 or 10^–5 M (P < 0.01, n = 3). Conversely,addition of genistein (10^–7 and 10^–6 M) had littleeffect on oxytocin-induced contractions, although at a higherconcentration (10^–5 M) a significant reduction in oxytocin-inducedcontractile activity was observed (P < 0.01). Oxytocin evokedphospholipase D activation in a concentration- and time-dependentmanner in cultured human pregnant myometrial cells (n = 4).These results indicate that activation of protein kinase C andtyrosine kinase are involved in the regulation of oxytocin-mediatedmyometrial contractile activity and that a coupled phospholipaseD/phosphatidate phosphohydrolase pathway may play a role inthe sustained stimulation of myometrial activity during labour.     

The diagnosis of male infertility--prospective time-specific study of conception rates related to seminal analysis and post-coital sperm--mucus penetration and survival in otherwise unexplained infertility

Infertile women without any inherent female infertility factorsand able to secrete normal cervical mucus were studied prospectivelyin relation to post-coital sperm—mucus penetration (PCT)and their partner's seminal analysis, excluding men with azoospermia.Time-specific cumulative conception rates calculated as forlife-table analysis were related to each measured seminal variableon routine analysis of 2–3 samples (volume, density, proportionwith progressive motility, and proportion with normal morphology);to various derivatives from combinations of these variables;to seminal findings after vital staining; and to the PCT results.The best seminal predictor of fertility was the motile normalsperm density (MNSD), the 18 month conception rates being 57.4%+ 4.6 (SE) and 30.2% + 5.9 (ratio 1.9, P < 0.001) above andbelow a derived threshold value of 4 x 10⁶/ml. The PCT led torates of 55.6% ± 4.3 and 14.9% ± 5.1 (ratio 3.73,P < 0.001) for positive and negative results, respectively.The PCT also gave rise to a significantly distinct intermediatepoor-psitive sub-group (conception rate 30.6% ± 9.0).Seminal analysis (the MNSD) did not affect the conception rateassociated with a positive PCT but helped to discriminate furtherwith a negative PCT (conception rates 22.5% ± 8.7 withan MNSD above 4 x 10⁶/ml versus 5.6% ± 4.8 below, P <0.05). The PCT was the single best predictor of fertility butseminal analysis (the MNSD) was of additional value after anegative PCT.     

Seminal plasma can be a predictive factor for male infertility

Seminal plasma from ejaculates of 10 healthy, fertile volunteersand 63 infertile males was analysed for superoxide dismutase(SOD) and xanthine oxidase (XO) activities using a chemiluminometer.There was no statistically significant difference in the activityof either enzyme between control and infertile populations (113±74 IU/ml for SOD and 1.17 ± 0.52 IU/l for XO) in samplesfrom normozoospermic ejaculates. Sperm progressive motilitywas positively correlated with SOD activity in seminal plasmaof corresponding ejaculates (P < 0.05) and negatively withXO activity (P < 0.001). An ‘oxido-sensitive’index was defined as the SOD/XO ratio and was found to be inverselyrelated to sperm progressive motility samples (P < 0.01).Analysing this index among all tested samples of semen includingthose with pathological spermiograms, as well as normospermic(N) samples we found statistically significant (elevated) differencesin oligoasthenoteratospermia (OAT) in comparison with N (P <0.05); OAT samples were also significantly different from oligospermic(O) and oligoteratospermic (OT) samples (P < 0.05). Thissuggests that the ‘oxido-sensitive’ index of seminalplasma may be a simple diagnostic factor, useful in the determinationof male infertility.     

Placental and ovarian hormones in anembryonic pregnancy

The circulating levels of human chorionic gonadotrophin (HCG),pregnancy-associated plasma protein-A (PAPP-A), Schwangerschaftprotein 1 (SP-1), oestradiol and progesterone were measuredin 81 pregnant patients between 4 and 11 weeks gestation, followingin-vitro fertilization and embryo transfer. The patients weredivided as follows: singleton anembryonic pregnancies, n = 22;singleton pregnancies which spontaneously aborted followingthe demonstration of fetal heart activity, n = 7; and normalsingleton pregnancies, n = 52. The levels of all substancesmeasured were significantly reduced in women with anembryoniccompared to those with singleton pregnancies which proceededto term. The serum levels of SP-1, weeks 6–8 (P < 0.01);HCG, weeks 6–8 (P < 0.05); oestradiol, weeks 5–8(P < 0.05) and progesterone, weeks 6–8 (P < 0.05),were lower in anembryonic pregnancies than in those of pregnancieswhich spontaneously aborted. These differences may be a reflectionof the fact that miscarriage, after the demonstration of fetalheart activity, represents fetal demise at a later stage inpregnancy. In anembryonic pregnancies, significant associationswere found between HCG and both oestradiol and progesteronelevels from weeks 6 and 8, suggesting that in the absence ofan embryo, HCG is the prime determinant of steroid synthesisby the corpus luteum.     

The effect of age on the cyclical patterns of plasma LH, FSH, oestradiol and progesterone in women with regular menstrual cycles

Luteinizing hormone (LH), follicle-stimulating hormone (FSH),oestradiol and progesterone concentrations in plasma were obtaineddaily throughout the menstrual cycles of 94 regularly cyclingwomen, aged between 24 and 50 years. Although mean LH concentrationschanged little with advancing age, mean FSH concentrations weresignificantly (P< 0.001) elevated from the age of 39 years.FSH concentrations in the oldest women studied (48–50years) were 3-fold greater than in the younger controls (womenaged 23–35 years). LH concentrations rose slightly (P< 0.05) during the last 5 years only. The increase in FSHconcentration was not, however, uniform across the cycle, butwas confined predominantly to the mid-follicular and post-ovula-toryphases (i.e. those times in the normal menstrual cycle whencirculating inhibin concentrations appear to be minimal). Despitethe clear increases in FSH concentration, there was little alterationin the mean steroid profiles which remained within the normalfertile range throughout the last decade of reproductive life.The only exception to this was a small, transient, but significant(P< 0.05) decrease in pre-ovulatory oestradiol concentrationbetween the ages of 36 and 38 years, which was followed by atransient increase (P< 0.01) in oestradiol concentrationbetween 39 and 44 years. However, no corresponding significantchanges in mean progesterone concentrations were observed.