CA 125 in peritoneal fluid from patients with endometriosis.

This study was performed to evaluate CA 125 in peritoneal fluid as an indicator of endometriosis. Peritoneal fluid from patients with mostly minimal and mild endometriosis (n = 43) and normal controls (n = 17) was collected at laparoscopy or laparotomy. The median concentration of CA 125 in peritoneal fluid did not differ significantly between patients and controls (79 IU/ml versus 76 IU/ml). In patients with endometriosis, a significantly increasing concentration of CA 125 in peritoneal fluid was seen from the early follicular to the late luteal phase; a similar change was not observed in the controls. In 14 patients, peritoneal fluid was sampled again after treatment with danazol and a significant reduction in median CA 125 concentration (76.5 IU/ml versus 57 IU/ml), peritoneal fluid volume (17.5 ml versus 10.5 ml) as well as reduced endometriosis scores (4 versus 2) were found. In controls, the concentration of CA 125 was about 10 times higher in peritoneal fluid than in serum. As the peritoneal levels of CA 125 did not differ significantly between patients with endometriosis and controls and as the reduction seen after danazol treatment did not correlate with the decrease of endometriotic implants, it is concluded that the monitoring of CA 125 in peritoneal fluid will not be useful in the diagnosis or control of endometriosis.     

Effect of peritoneal fluid supplemented with exogenous progesterone on sperm motility in vitro

Since progesterone has been claimed to induce acrosomal reactionand hyperactivated motility of human spermatozoa, the presentstudy was undertaken to determine if its presence at concentrationssimilar to those of peri-ovulatory follicular fluid could influencethe effect of peritoneal fluid on spermmotility in vitro. Tothis end, 11 sperm samples were incubated at 37°C with fiveperitoneal fluids with/without exogenous progesterone, and spermmotility was assessed using a computer-assisted analyser attime (t) = 0, 2.5, 5 and 24 h. Overall there was no observableconstant trend for enhancement or inhibition of sperm motility.Progesterone generally induced a negative effect on those spermsamples with high velocities in the native peritoneal fluidsand a positive effect on those sperm samples demonstrating lowmotility in the native peritoneal fluids. The incorporationof progesterone into the incubation medium seemed to resultin a ‘tuning’ of sperm velocity to around 30–50µm/s. However, a given sperm sample reacted differentlywhen incubated with various peritoneal fluids and, reciprocally,different semen samples incubated with the same peritoneal fluidshowed very variable motility patterns. The greater variabilityof the effects exerted by progesterone on sperm motility couldarise from the fact that each sperm sample may contain subpopulationsof gametes with different sensitivity to progesterone.     

Comparative analysis of motility characteristics of Percoll-selected spermatozoa populations from fresh and cryopreserved semen

The proportion and quality of motility of spermatozoa in normozoospermicejaculates were assessed using computerassisted semen analysis.The ejaculate was split and the motility re-assessed followingseparation on a Percoll gradient with or without cryomediumand cryopreservation. Cryopreservation caused a significantdecrease in the proportion of motile spermatozoa and in theirvelocity and amplitude of lateral head displacement. The initialdecrease in the proportion of motile spermatozoa was found tobe in part an effect of the cryomedium. The use of Percoll gradientseparation did not initially change these effects but after4 h incubation differences in velocity and amplitude of lateralhead displacement between samples were no longer evident. Percoll-selected,cryopreserved spermatozoa had both a stable proportion of motilespermatozoa and a stable velocity for at least 48 h, whereasin fresh spermatozoa populations, similarly separated usingPercoll, the proportion of motile spermatozoa had decreasedby 24 h and the velocity was lower at 48 h. Percoll preparationis an effective method for the selection of motile spermatozoafrom cryopreserved semen which, after a short incubation, havesimilar motility characteristics to fresh spermatozoa.     

Endometriosis: Effect of peritoneal fluid from infertile women with endometriosis on ionophore-stimulated acrosome loss

The effect of peritoneal fluid (PF) from endometriosis patientswas studied in spontaneous and stimulus-induced (Ca-ionophore;A23187) acrosome reactions. PF samples were obtained from 21infertile women with endometriosis and five normal women (controls).Sperm acrosomes were examined by staining with Pisum sativumagglutinin labelled with fluorescein isothiocyanate. The incidenceof spontaneous acrosome reaction after 1 and 6 h of incubation(6.7 ± 1.6 and 6.9 ± 1.4 respectively) was significantly(P < 0.001) lower when the incubation was performed withPF from endometriosis patients in comparison with spermatozoaincubated in PF from the control group (12.8 ± 1.1 and12.8 ± 0.8). Similarly, the incidence of A23187-inducedacrosome reaction after 1 and 6 h of incubation (19.8 ±2.7 and 20.0 ± 2.4) was significantly (P < 0.001)lower when spermatozoa were incubated with PF from endometriosispatients in comparison with spermatozoa incubated with PF fromthe control group (34.6 ± 9.8 and 34.4 ± 1.1).The incidence of A23187-inducible acrosome reaction was alsosignificantly (P < 0.001) lower when the incubation was performedwith PF from endometriosis patients (13.1 ± 2.8 and 13.1± 2.4) when compared with that from the control group(21.8 ± 2.6 and 21.6 ± 1.5). No relationship wasfound between the stage of endometriosis and the incidence ofacrosome loss. In conclusion, the PF from endometriosis patientsdecreased both spontaneous and stimulus-induced acrosome reaction.This may represent a mechanism for the detrimental effect ofthe PF from endometriosis patients on the spermatozoa-oocyteinteraction and partially explain the aetiology of infertilityin patients with endometriosis.     

Decreased amounts of antibodies to 22 and 18 kDa antigens in the peritoneal fluid of patients with endometriosis

Accumulated evidence implicates immunological alterations inendometriosis. The purpose of this study was to look for variationsin antibodies to distinct antigens in peritoneal fluid of womenwith and without endometriosis. Peritoneal fluid was aspiratedfrom 17 women undergoing laparoscopy for tubal ligation and37 patients complaining of symptoms of pain and/or infertility.Peritoneal fluid antibodies to a standard preparation of peritonealfluid antigens were detected by Western blot analysis usingperoxidase-labelled anti-human immunoglobulin G antibodies specificto the Fc region. Antibodies to distinct antigens were quantifiedby estimating the ratio of the relative optical density betweensamples and a standard amount of antibodies. Marked changeswere found in the antibody detection to two antigens havingapparent molecular weights of 22 and 18 kDa. The intensity ofthe antibody signal was significantly weaker in the peritonealfluid from endometriosis patients (0.36 ± 0.06 and 0.46± 0.06) compared with that in women without endometriosis(0.62 ± 0.08 and 0.75 ± 0.06). It was also weakerin patients without endometriosis presenting with infertility(036 ± 0.07 and 0.47 ± 0.08), but only the 18kDa antigen result was significant After adjusting for infertility,the P values for the 18 and 22 kDa bands were 0.03 and 0.28(not significant) respectively in the group of endometriosispatients. These changes were not related to the phase of themenstrual cycle. These data suggest an alteration in the immuneresponse to two distinct antigens in the peritoneal fluid fromwomen with endometriosis and infertility. Further evaluationof these two antigens and their antibodies would be of interestto help understand endometriosis and its associated infertility.     

Increased levels of interleukin-15 in the peritoneal fluid of women with endometriosis: inverse correlation with stage and depth of invasion

BACKGROUND: Interleukin (IL)-15 is a novel cytokine with immunoregulatory and angiogenic properties. We compared IL-15 levels in the peritoneal fluid (PF) of women with and without endometriosis. METHODS: PF samples were obtained from 55 women with endometriosis (23 with superficial peritoneal implants, 19 with deep endometriotic implants and 13 with ovarian endometriomas). Eighteen women with normal pelvic anatomy undergoing tubal sterilization served as controls. RESULTS: PF IL-15 concentrations were increased in women with endometriosis (2.7 +/- 0.5 pg/ml) versus controls (2.1 +/- 0.3 pg/ml; P < 0.001). However, IL-15 levels were higher in women with superficial peritoneal implants (2.9 +/- 0.5 pg/ml) than women with deep endometriotic implants (2.6 +/- 0.4 pg/ml; P = 0.01) or ovarian endometriomas (2.2 +/- 0.4 pg/ml; P < 0.001). IL-15 was also higher in women with deep implants than in those with endometriomas (P < 0.05). PF IL-15 correlated inversely with both depth of invasion (r = -0.52) and the stage of endometriosis (r = -0.42). PF IL-15 levels demonstrated little variation during the menstrual cycle, and did not discriminate between women with infertility or pelvic pain. CONCLUSION: PF IL-15 levels are increased in women with endometriosis. However, IL-15 levels are inversely correlated with the depth of invasion and disease stage, suggesting a possible role for this cytokine in the early pathogenesis of endometriosis.     

Effects of peritoneal fluid from endometriosis patients on the release of vascular endothelial growth factor by neutrophils and monocytes

BACKGROUND: An increase in the level of the vascular endothelial growth factor (VEGF) production has been reported in the peritoneal fluid (PF) of endometriosis patients. This suggests that changes in the vascular permeability and angiogenesis play an important role in the pathophysiology of this disease. This study examined the effects of the PF obtained from endometriosis patients on the release of VEGF by neutrophils and monocytes. METHODS: Neutrophils and monocytes were obtained from young healthy volunteers and cultured with the PF obtained from either endometriosis patients (EPF) (n=18) or a control group (CPF) (n=4). A human monocyte/macrophage cell line, THP-1, was cultured with either 10% EPF or 10% CPF. The PF and culture supernatants were assayed for VEGF using ELISA. Real-time PCR and Western blotting were used to measure the VEGF mRNA and protein expression level, respectively. RESULTS: The VEGF levels were higher in the EPF than in the CPF (591+/-75 versus 185+/-31 pg/ml, P<0.05). However, the level of VEGF released by THP-1 cells in CPF and EPF was similar. The EPF induced the release of VEGF by neutrophils, but no VEGF was released by monocytes. The VEGF mRNA expression levels in the neutrophils were higher in the EPF, which was abrogated by cycloheximide, suggesting that the EPF induces the production of VEGF in neutrophils. Neutralizing antibodies against IL-8 and TNF-alpha did not completely prevent the EPF-induced release of VEGF by the neutrophils, even though these growth factors stimulated the release of VEGF by neutrophils. There was a positive correlation between the VEGF and IL-10 concentrations in the EPF (correlation coefficient=0.549, P=0.012, n=18), but the neutralizing antibody of IL-10 did not affect the release of VEGF by the EPF-treated neutrophils. CONCLUSION: The EPF induced the production and release of VEGF by neutrophils, suggesting that neutrophils may be a source of peritoneal VEGF. In addition, neutrophil-derived VEGF might be a marker for diagnosing endometriosis.     

Adverse effects of hydrosalpinx fluid on sperm motility and survival

The negative impact of hydrosalpinx on IVF outcome is well recognized but some reports indicate that tubal infertility with hydrosalpinx is a heterogeneous entity and may have different effects on the outcome. The embryotoxic effects of hydrosalpinx fluid (HF) have been documented in mouse but not human embryos. This study examined the effects of HF on sperm motility and survival after various periods of incubation. Fifteen infertile patients with hydrosalpinx shown on ultrasound monitoring during ovarian stimulation underwent aspiration of HF after egg collection. Electrolytes, glucose and pyruvate concentrations were within the physiological ranges found in normal human tubal fluid. Sperm motility and velocities remained unchanged after 5 h of incubation with various concentrations of HF but the percentage of motile spermatozoa was significantly reduced after 24 h of incubation. Both 50 and 100% HF were potentially cytotoxic (survival indices <85%). The detrimental effect seemed to be dependent on the concentrations of HF. Low osmolarity, low lactate concentrations or the protein content may be responsible for the loss of sperm motility. A human sperm survival test using HF may be useful in selecting appropriate treatment options for patients with hydrosalpinx undergoing IVF treatment or tubal surgery.     

Modulation of neutrophil apoptosis by plasma and peritoneal fluid from patients with advanced endometriosis

BACKGROUND: The increased production of pro-inflammatory chemoattractant cytokines for neutrophils in endometriosis suggests that changes in the immune system play an important role in the pathophysiology of endometriosis. The effects of plasma and peritoneal fluid from patients with advanced endometriosis on the apoptosis of neutrophils were investigated. METHODS: Apoptotic changes of neutrophils were evaluated by morphological changes using Giemsa staining. Apoptosis was confirmed by DNA electrophoretic analysis. RESULTS: Compared with the plasma (n = 20) and peritoneal fluid (n = 5) of healthy controls, the addition of 10% plasma (n = 20) and peritoneal fluid (n = 10) from patients with endometriosis to an in-vitro culture of neutrophils from healthy subjects reduced the percentage of apoptotic cells from 65.3 +/- 6.6 to 27.2 +/- 4.6% (P < 0.001) and from 45.3 +/- 4.8 to 10.5 +/- 4.3% (P < 0.001) respectively. Neutralizing interleukin-8 antibody abrogated the delay of neutrophil apoptosis induced by peritoneal fluid, but not in the plasma of endometriosis patients. CONCLUSIONS: These findings show that interleukin-8 is one of the neutrophil survival factors in the peritoneal fluid of endometriosis patients and that an unidentified survival factor is also present in the plasma of patients with endometriosis.     

Hepatocyte growth factor concentrations are elevated in peritoneal fluid of women with endometriosis.

The concentrations of hepatocyte growth factor (HGF) in peritoneal fluid (PF) from women with endometriosis (n = 36) and without endometriosis (n = 40) were measured. All of the PF samples examined contained detectable concentrations of HGF. The HGF concentrations in PF from women with stage III/IV endometriosis (0.906 ng/ml, 0. 561-1.185; median, interquartile range) were significantly higher (P < 0.0001) than those from women without endometriosis (0.315 ng/ml, 0.251-0.472). The HGF concentrations from women with stage I/II endometriosis (0.417 ng/ml, 0.310-1.023) appeared to be intermediate. There were no apparent variations detected among the HGF concentrations in women in the follicular or luteal phases regardless of the presence of endometriosis. Interestingly, HGF concentrations in PF from women on gonadotrophin releasing hormone analogues, independent of the presence of endometriosis, were comparable with those from untreated women. Given the known mitogenic property of HGF in human endometrial cells, these results suggest that HGF might play a role in the progression of endometriosis.     

Embryotoxicity of peritoneal fluid in women with endometriosis. Its relation with cytokines and lymphocyte populations

BACKGROUND: The goals of the present work were to study the embryotoxic effects of peritoneal fluid (PF) in women with or without endometriosis, and to relate any embryotoxicity to the severity of endometriosis, infertility or achievement of pregnancy, cytokine concentrations and lymphocyte populations. METHODS: Sixty-six consecutive women of reproductive age, 54 with endometriosis (21 infertile) and 12 infertile without endometriosis, and another 12 fertile women as control group, were included in this study. They all underwent laparoscopy or laparotomy in the second half of the cycle, and PF was collected from the pouch of Douglas. The embryotoxicity of the PF was assessed by means of a mouse embryo assay, and expressed as the number of embryos that did not reach blastocyst stage. Cytokines and lymphocyte populations present in PF were also studied and correlated with embryotoxicity. RESULTS: PF embryotoxicity was increased in women with endometriosis, but there was little correlation with the severity of the disease. However, although a clear relationship to the presence of infertility was not found, embryotoxicity appeared to be lower in those infertile patients with endometriosis who later became pregnant. We found a significant increase in embryotoxicity in the presence of high cytokine concentrations, especially with interleukin-6, and less so with interleukin-8 (P < 0.05). No good correlation was observed with lymphocyte populations, but CD56 (NK) cells were significantly increased in the PF of women with endometriosis. In general, the correlations for embryotoxicity were better when PF was diluted at 20% (91.4 +/- 17 versus 68.1 +/- 31, P < 0.01). CONCLUSIONS: These results suggest that alteration in the production of cytokines in the PF, especially IL-6, besides contributing to the endometriosis and its evolution, probably increases embryotoxicity. However, no correlation was found between the latter and associated infertility.     

Interleukin-10 gene promoter polymorphisms and their protein production in peritoneal fluid in patients with endometriosis

Although associations of interleukin-10 (IL-10) gene promoter polymorphisms and their protein production with endometriosis risk have been reported, the correlations remain controversial. The objective of this study was to determine IL-10 gene promoter polymorphisms at -1082, -819 and -592 sites and their protein production in peritoneal fluid (PF) in patients with and without endometriosis. IL-10 gene promoter polymorphisms at -1082 site were detected by amplification refractory mutation system (ARMS)-PCR and that at -819 and -592 sites was genotyped by restriction fragment length polymorphism (RFLP)-PCR. Protein levels of IL-10 in PF were measured by enzyme-linked immunosorbent assay (ELISA). There were no significant differences in the genotype and allele frequencies of IL-10 gene promoter polymorphisms at position -1082 between the endometriosis and the control groups. However, the frequency of -819 or -592 C alleles was significantly increased in patients with endometriosis compared with controls. The protein levels of IL-10 in PF were statistically higher in the endometriosis group than in the control group. Moreover, the polymorphisms at -1082, -819 and -592 sites were associated with protein levels of IL-10 in PF in the endometriosis group while in the control group only the polymorphisms at position -1082 correlated with protein levels. Increased frequency of -819 or -592 C allele and increased protein production of IL-10 in PF in patients with endometriosis compared with controls and correlations of polymorphisms at -819 and -592 sites with protein levels of IL-10 in PF in patients with endometriosis may suggest that polymorphisms at -819 and -592 sites and their protein production are associated with endometriosis risk.     

Endometriosis: Insulin-like growth factor binding proteins in peritoneal fluid of women with minimal and mild endometriosis

This prospective cohort study was carried out in a university-basedInfertility clinic to determine the profile of insulin-likegrowth factor binding proteins (IGFBPs) in patients with mildendometriosis and no obvious mechan ical factor contributingto infertility. A total of 26 patients with minimal and mildendometriosis and 10 controls contributed peritoneal fluid atsurgery. The variety, expression and levels of IGFBPs were determinedby radio inununoassay and Western ligand blots (WLBs) with quantitationby laser densitometer. A 27 kDa species was significantly lowerand a 31 kDa species tended to be lower in patients with endometriosisas determined by quantitative laser densitometer. The levelsof IGFBP-3 detected by radioimmunoassay and by WLB were correlatedin the control group and in the patients with endometriosisin the follicular phase but not in patients with endometriosisin the luteal phase. The level of 27 kDa species seen on WLBsdid not appear to correspond to IGFBP-1 determined by radiomimunoassayand IGFBP-3 levels in luteal phase endometriosis patients alsodeparted from values determined by radioimmunoassay. These discrepancies suggest a complex system to control levels of IGFin the peritoneum involving multiple binding proteins and proteases.The IGFBPs of patients with endometriosis may contribute toreproductive dysfunction and be able to serve as markers.     

Vascular endothelial growth factor (VEGF) concentrations are elevated in peritoneal fluid of women with endometriosis

Active endometriosis is characterized by hypervascularizationboth within and surrounding the implant; therefore the presenceof angiogenic factors in the peritoneal environment would beof great importance. Vascular endothelial growth factor (VEGF)is a potent angiogenic factor involved in both physiologicaland pathological angiogenesis. We sought to determine if VEGFwas present in the peritoneal fluid of women with and withoutendometriosis, and to establish if differences exist betweenthese groups. VEGF was present in all patients sampled. Thefluid from patients with endometriosis contained significantlygreater amounts of VEGF than controls. Cyclic variations inVEGF concentration were seen in fluid from patients with endometriosis,the VEGF concentration in proliferative phase being significantlyhigher than in the secretory phase. The concentration of VEGFin this fluid was also significantly higher than that foundin the proliferative and secretory phases of women without endometriosis.No cyclic variations in VEGF were seen in the control group.We suggest that elevated levels of VEGF in the peritoneal fluidof patients with endometriosis may be critical in the pathogenesisof endometriosis.     

Andrology: The predictive value of computer-assisted semen analysis in the context of a donor insemination programme

In this paper we examine the value of both conventional andcomputer-assisted semen analysis (CASA) using the Hamilton-ThornHTM-S 2030 in predicting the in-vivo fertility of cryopreserveddonor semen. Semen samples were examined prospectively and dataon the conventional criteria of semen quality, sperm morphometryand movement were collected. Of 61 ejaculates identified, 33achieved pregnancies (‘successful’) and 28 failedto do so (‘unsuccessful’), despite inseminationinto at least four different normal female recipients. Whenthe post-thaw semen profiles were compared, no differences wereobserved between the two groups in respect of the conventionalcriteria of semen quality determined by conventional laboratorytechniques; however, there were differences in respect of bothmorphometry and movement characteristics determined by the HTM-S.When multiple logistic regression was used to examine the abilityof the variables measured to predict the achievement of pregnancy,the conventional criteria of semen quality were of no value(X² = 6.67, P = 0.353). However, the CASA assessment successfullypredicted outcome in 86.9% of cases (X² = 44.3, P = 0.0021).It was concluded that CASA assessment is of significant valuein predicting the ability of an ejaculate to achieve pregnancy.     

A combination of interleukin-6 and its soluble receptor impairs sperm motility: implications in infertility associated with endometriosis

BACKGROUND: We previously reported that the level of interleukin (IL)-6 is increased in the peritoneal fluid of women with endometriosis. This study was undertaken to assess the effects of IL-6 and soluble IL-6 receptor (sIL-6R) on in vitro sperm motility. METHODS: Sperm (n = 20) were cultured with IL-6 or sIL-6R, or with a combination of both. After 24 h cultures, sperm motility was evaluated using a computer-assisted semen analysis system. Gene and protein expressions of IL-6, IL-6 receptor (IL-6R), and glycoprotein 130 (gp130) were examined in sperm by RT-PCR analysis and western blot analysis. RESULTS: Addition of IL-6 or sIL-6R individually to the culture media had no affect on sperm motion. However, adding a combination of IL-6 and sIL-6R dose-dependently reduced the percentage of motile and rapidly moving sperm. Adding anti-IL-6R antibody abolished these adverse effects. Sperm expressed the gp130 gene and protein, but not IL-6 or IL-6R. CONCLUSIONS: A combination of IL-6 and sIL-6R may be associated with gp130 expressed in the sperm and reduce sperm motility. IL-6 and sIL-6R may contribute to the pathogenesis of endometriosis-associated infertility.     

The effect of intracytoplasmic sperm injection and semen parameters on blastocyst development in vitro

The present study compares the development and quality of blastocysts derived from conventional oocyte insemination with those derived from intracytoplasmic sperm injection (ICSI). Oocytes were collected from patients undergoing ovarian stimulation with human menopausal gonadotrophins for IVF. Patients with normal semen were assigned to conventional oocyte insemination while those with progressive motility <20% and/or normal sperm morphology < or =4% were assigned to ICSI. Resulting embryos were cultured for up to 6 days. The mean number and percentage of embryos reaching the blastocyst stage and the mean number and percentage of blastocysts of high quality on days 5-6 were assessed for both treatment groups and compared. The influence of paternal factors (sperm concentration, motility, progressive motility, morphology) on blastocyst development and quality were assessed by regression analyses. Significantly more ICSI-derived embryos arrested at the 5- to 8-cell stage (P = 0.024) concomitant with the activation of the paternal genome than those derived from conventional oocyte insemination. Significantly fewer ICSI-derived embryos reached the blastocyst stage on days 5-6 (P<0.001) and significantly fewer ICSI-derived embryos were of high quality (P = 0.002) compared with conventional oocyte insemination. When treatment groups were combined and evaluated by regression analysis, progressive motility and sperm morphology were significantly correlated with diminished blastocyst development and quality (P < 0.05). From these data, we conclude that paternal factors and/or performing ICSI in cases of severe male factor infertility may have a detrimental effect on blastocyst development and their quality.     

Andrology: Evaluation of a computer-aided semen analysis system with sperm tail detection

The aim of this study was to evaluate the Stroemberg-Mika cellmotion analyser (SM-CMA) which uses tail detection in orderto discriminate between immotile spermatozoa and other particles.Analysis of the spermatozoa by the SM-CMA can easily be checkedon a video monitor. The semen samples were from donors and patientsvisiting the fertility unit of the University Hospital, Hanzeplein,The Netherlands. Both fresh semen samples and purified spermsuspensions were used to estimate sperm counts and motilitycharacteristics. We tested the use of the x10 objective insteadof the x20 and we assessed the ways in which motility characteristcswere influenced by temperature. We found a considerable discrepancybetween sperm concentrations measured manually and by computerizedanalysis, both in semen samples and in purified sperm suspensions.The SM-CMA correctly recognizes motile spermatozoa, but underestimatesimmotile ones. Although temperature affects motility characteristics,in routine measurements the influence of short cooling periods,which are unavoidable, was nil. We found that using the x10objective can be useful, especially at low sperm concentrations.In our opinion, the SM-CMA system is, despite some shortcomingsin its user-interface, a useful and versatile instrument forexamination of human semen samples, with desirable features.     

Tumour necrosis factor alpha concentrations in the peritoneal fluid of infertile women with minimal or mild endometriosis are lower in patients with red lesions only than in patients without red lesions

Tumour necrosis factor alpha (TNFalpha) of peritoneal fluid is believed to have important pro-inflammatory and angiogenic activities in the complex mechanisms of development of peritoneal endometriotic lesions. We have evaluated the concentrations of TNFalpha and macrophages in peritoneal fluid of infertile women with minimal or mild endometriosis and related them to the presence of peritoneal red lesions alone (red lesions only group; n = 11) or their absence (non-red lesions group; n = 36). A group of 39 infertile normo-ovulatory patients with normal pelvic anatomy was used as controls. TNFalpha concentrations did not differ between controls and either group of patients. Patients with red lesions only had significantly lower concentrations of TNFalpha in peritoneal fluid (P < 0.05) and had a higher proportion of samples with undetectable concentrations (P < 0.05) than patients without red lesions. The significant difference in TNFalpha concentrations was present when comparing the groups of patients in the proliferative phase but not in the secretory phase. Macrophage concentrations were not different in the groups. Our findings are compatible with an impairment of macrophage function and therefore lend support to the theory that an inappropriate immunological response of the peritoneal environment to regurgitated endometrium may play a part in the initial phases of endometriotic implants.     

The effect of endometriosis, cycle stage, lymphocyte suppression and pregnancy on CA-125 levels in peritoneal fluid and serum in baboons

BACKGROUND: Serum CA-125 during the mid-follicular phase has been reported to be a clinically useful and reproducible marker in the diagnosis of advanced endometriosis in women. This study was undertaken to document the effect of the menstrual cycle, pregnancy and lymphocyte suppression on CA-125 levels in peritoneal fluid (PF) and serum in baboons with a normal pelvis and baboons with endometriosis. METHODS: CA-125 levels were measured in 264 serum samples that were serially obtained during one menstrual cycle from 10 animals with and without endometriosis. In addition, CA-125 levels were determined in 204 archived samples (serum, n = 112 and PF, n = 92) obtained from 32 female baboons with or without endometriosis. The CA-125 assays were performed by radioimmunoassay using kits from Centocor (Malvern, PA, USA). RESULTS: Serum CA-125 levels were at their highest during menstruation and decreased progressively during the follicular and luteal phase. PF CA-125 levels were increased during the follicular phase in baboons with a normal pelvis, but no cyclic changes were observed in animals with endometriosis. Serum CA-125 levels were unaffected by induction, lymphocyte suppression or pregnancy. Induction of endometriosis resulted in increased PF CA-125 levels, whereas lymphocyte suppression or pregnancy had no effect. CONCLUSION: In baboons, serum CA-125 originates mainly from eutopic endometrium whereas the main source of PF CA-125 seems to be the peritoneum or ectopic endometrium. The baboon appears to be a valid model to further study the relationship between endometriosis and CA-125.