Peripheral leucocyte molecular indicators of inflammation and oxidative stress are altered in dairy cows with embryonic loss.

Objective of experiment was to determine whether oxidative stress (OS) and inflammation altered embryonic loss in dairy cows. Blood samples were collected at days 0, 16, 32 and 60 after timed (AI) from 200 Holstein cows to determine embryonic loss based on interferon-stimulated gene-15 (ISG15) mRNA expression (day 16) and ultrasound at day 32 and day 60. Leucocyte expressions of mRNA TLR2, TLR4, TNF-α, IL1B, IL10, STAT3 (inflammation), PTGS2, PTGES (prostaglandin synthesis), and PLA2G4A and ALOX5AP (eicosanoid metabolism) at days 0 and 16 were determined. Plasma redox status for antioxidant enzymatic activities of glutathione peroxidase (GPX), superoxide dismutase (SOD), total antioxidant capacity (TAC), and concentrations of malondialdehyde (MDA) were determined at days 0, 16, 32 and 60. All antioxidant-redox responses were beneficially significant in pregnant cows diagnosed pregnant at day16 and sustained pregnancy to day 60 compared to non-pregnant cows at day16 or pregnant at day16 and lost embryos by days 32 or 60. The leucocyte mRNA expressions of TLR2, TLR4, STAT 3, IL1B, PTGS2, PLA2G4A and ALOX5AP were greater and PTGES was lower at day16 in pregnant cows that lost embryos early (P < 0.05). In conclusion peripheral leucocyte molecular indicators of inflammation and plasma indicators of OS were altered in pregnant cows undergoing embryonic losses compared to cows with a sustained pregnancy.

Coagulation parameters predictive of repeated implantation failure in Chinese women: A retrospective study.

Repeated implantation failure (RIF) greatly influences pregnancy rate after assisted reproductive technologies (ART) with elusive causes. Our study aimed to explore coagulation parameters in association with RIF and establish a model to predict the risk of RIF in Chinese women.Coagulation parameters, including prothrombin time (PT), thrombin time (TT), activated partial prothrombin time (APTT), D-dimer (DD), fibrin degradation products (FDP), fibrinogen (FG), and platelet aggregation induced by arachidonic acid (AA) and adenosine diphosphate (ADP) were measured in RIF patients and controls. A logistic regression model was built by using the purposeful selection to select important factors for the prediction of RIF.Between 92 RIF patients and 47 controls, we found a statistically significant difference in all of the coagulation parameters except APTT, FDP and platelet aggregation induced by ADP. The purposeful selection method selected PT (odds ratio [OR] = 0.28, 95% CI: 0.12-0.66, P = .003), APPT (odds ratio [OR] = 0.76, 95% CI: 0.63-0.91, P = .004), TT (odds ratio [OR] = 0.75, 95% CI: 0.53-1.08, P = .124), and platelet aggregation induced by AA (odds ratio [OR] = 1.27, 95% CI: 1.11-1.44, P = .0003) as important predictors of RIF risk. ROC curve analysis indicated that the area under ROC curve (AUC) of the model was 0.85 with an optimal cut-off point of the predicted probability being P = .65, leading to a sensitivity of 0.83 and a specificity 0.75.We found that coagulation parameters including PT, APTT, TT and platelet aggregation induced by AA are predictive of RIF in Chinese women. Our results highlight the potential of anti-coagulation therapies to lower the risk of RIF.

Decreased ANGPTL4 impairs endometrial angiogenesis during peri-implantation period in patients with recurrent implantation failure.

Insufficient endometrial angiogenesis during peri-implantation impairs endometrial receptivity (ER), which contributes to recurrent implantation failure (RIF) during in vitro fertilization and embryo transfer (IVF-ET). Angiopoietin-like protein 4 (ANGPTL4) acts as a multifunctional secretory protein and is involved in the regulation of lipid metabolism and angiogenesis in various tissues including the endometrium. Herein, we found decreased ANGPTL4 expression in endometrial tissue and serum during peri-implantation period in 18 RIF-affected women with elevated uterine arterial impedance (UAI) compared with the pregnancy controls. ANGPTL4 and peroxisome proliferator-activated receptor gamma (PPARγ) expression were up-regulated upon decidualization on human endometrial stromal cells (HESCs). Rosiglitazone promoted the expression of ANGPTL4 in HESCs and human umbilical vein endothelial cells (HUVECs) via PPARγ. ANGPTL4 promoted the proliferation, migration and angiogenesis of HUVECs in vitro. Our results suggest that decreased abundance of ANGPTL4 in endometrial tissues impairs the endometrial receptivity via restraining endometrial angiogenesis during decidualization; while rosiglitazone-induced ANGPTL4 up-regulation in hESCs and HUVECs through PPARγ. Therefore, ANGPTL4 could be a potential therapeutic approach for some RIF-affected women with elevated UAI.

NK and trophoblast cells interaction: cytotoxic activity on recurrent pregnancy loss.

The trial objective was to determine the peripheral blood NK cells cytotoxic activity effect on trophoblast cells at recurrent pregnancy loss (RPL). The investigation involved non-pregnant women with PRL in proliferating and secretory menstrual cycle phases (PMCPh and SMCPh, respectively); women of 6-7 weeks pregnancy with RPL in past medical history; healthy fertile non-pregnant women in PMCPh and SMCPh, women of 6-7 weeks physiological pregnancy, nulliparity healthy women with regular menstrual function in PMCPh and SMCPh. NK cells cytotoxic activity was determined using peripheral blood mononuclear cells. The target cells were JEG-3 line trophoblasts. It has been established that NK cells cytotoxic activity effect on trophoblasts is lower in SMCPh than in PMCPh in non-pregnant fertile women. The NK cells cytotoxic activity was higher in SMCPh than in PMCPh in non-pregnant women with PRL and also higher than the same value in SMCPh in non-pregnant fertile women. The increased NK cells cytotoxic activity values in SMCPh in women with RPL may be the reason for miscarriage.

Longitudinal profiles of relaxin and progestagens during pregnancy, pregnancy loss and false pregnancy in the killer whale (Orcinus orca).

The circulating pattern of immunoreactive relaxin and progestagens based on monthly and gestational stage (early, mid, late) profiles were determined during pregnancies that resulted in live calves (LIVE, n = 30), stillbirths (STILLB, n = 3), abortions (ABORT, n = 5) and presumptive false pregnancies (FALSE, n = 8), and during the follicular (n = 34) and luteal phase (n = 58). Monthly LIVE relaxin concentrations steadily increased during gestation, but values did not significantly exceed those of the luteal phase until 9 months prior to parturition, peaking during the final month at 2356 ng/ml. Relaxin surged (P < 0.05) during the final week of gestation (36,397 ng/ml), undergoing a 3 and 9-fold increase compared with concentrations in the preceding two weeks, respectively. Monthly relaxin production did not differ among each reproductive state with the exception of months-13-16 where concentrations were higher (P < 0.001) for STILLB than LIVE. Relaxin concentration was reduced (P < 0.0001) by 849% in placental versus maternal serum collected within 1 day of labor. Mid- and late-pregnancy progestagen concentrations were lower for FALSE (P < 0.001) compared with STILLB and LIVE. Late pregnancy progestagen concentrations were reduced for FALSE (P < 0.05) and ABORT (P < 0.02) compared with LIVE and STILLB. Monthly progestagen production in ABORT tended to be lower than LIVE across a range of gestational months (Months 2, 7, 8, 11) but this difference only became significant during months 14 and 15. Results indicate that relaxin is primarily produced by the CL during pregnancy, and that concentrations could not be used to differentiate from non-pregnant females until the final 6 months of gestation. In addition, as would be expected from a primarily CL product, relaxin cannot be used to detect abnormal pregnancies. Conversely, progestagens, which are produced by both the placenta and CL can be used to differentiate FALSE from normal pregnancy and may be useful indicators of fetal health in the killer whale.

Embryotoxic cytokines-Potential roles in embryo loss and fetal programming.

Cytokines in the reproductive tract environment at conception mediate a dialogue between the embryo and maternal tissues to profoundly influence embryo development and implantation success. Through effects on gene expression and the cell stress response, cytokines elicit an epigenetic impact with consequences for placental development and fetal growth, which in turn affect metabolic phenotype and long-term health of offspring. There is substantial evidence demonstrating that pro-survival cytokines, such as GM-CSF, CSF1, LIF, HB-EGF and IGFII, support embryos to develop optimally. Less attention has been paid to cytokines that adversely impact embryo development, including the pro-inflammatory cytokines TNF, TRAIL and IFNG. These agents elicit cell stress, impair cell survival and retard blastocyst development, and at sufficiently high concentrations, can cause embryo demise. Experiments in mice suggest these so-called 'embryotoxic' cytokines can harm embryos through pro-apoptotic and adverse programming effects, as well as indirectly suppressing uterine receptivity through the maternal immune response. Embryotrophic factors may mitigate against and protect from these adverse effects. Thus, the balance between embryotrophic and embryotoxic cytokines can impart effects on embryo development and implantation, and has the potential to contribute to endometrial 'biosensor' function to mediate embryo selection. Embryotoxic cytokines can be elevated in plasma and reproductive tract tissues in inflammatory conditions including infection, diabetes, obesity, PCOS and endometriosis. Studies are therefore warranted to investigate whether excessive embryotoxic cytokines contribute to infertility and recurrent implantation failure in women, and compromised reproductive performance in livestock animals.

Maternal serum amyloid A level as a novel marker of primary unexplained recurrent early pregnancy loss.

OBJECTIVE: To assess maternal serum amyloid A (SAA) levels among women with primary unexplained recurrent early pregnancy loss (REPL). METHODS: A prospective study was conducted among women with missed spontaneous abortion in the first trimester at Ain Shams University Maternity Hospital, Cairo, Egypt, between January 21 and December 25, 2014. Women with at least two consecutive primary unexplained REPLs and no previous live births were enrolled. A control group was formed of women with no history of REPL who had at least one previous uneventful pregnancy with no adverse outcomes. Serum samples were collected to measure SAA levels. The main outcome was the association between SAA and primary unexplained REPL. RESULTS: Each group contained 96 participants. Median SAA level was significantly higher among women with REPL (50.0 µg/mL, interquartile range 26.0-69.0) than among women in the control group (11.6 µg/mL, interquartile range 6.2-15.5; P<0.001). The SAA level was an independent indicator of primary unexplained REPL, after adjusting for maternal age and gestational age (odds ratio 1.12, 95% confidence interval 1.06-1.19; P<0.001). CONCLUSION: Elevated SAA levels found among women with primary unexplained REPL could represent a novel biomarker for this complication of pregnancy.

Monitoring of embryonic and fetal losses in different breeds of goats using real-time B-mode ultrasonography.

Compared to cattle and sheep, few studies had been undertaken to evaluate the incidence of embryonic and fetal losses (EFL) in goats. The objectives of the present study were to characterize the timing of EFL and to identify the factors that are associated with EFL in goats such as breed, age, parity, method of estrous synchronization, and breeding. Moreover, this study aimed to ensure whether a relationship existed between serum progesterone (P4) and EFL. Goats (n = 151) of different breeds (70 Zaraiebi, 42 Damascus, and 39 Cross goats [Baladi × Damascus]) were evaluated by ultrasonography to monitor EFL during different stages of gestation (D20-23, D26-29, D33-36, D40-45, and D47-54 after breeding). Blood samples were collected at D7, D20, and at each ultrasonographic scanning to clarify changes of serum P4 levels concurrently with EFL. Results revealed that 45 of 109 goats (41.28%) were exposed to EFL. A higher EFL % was observed between D20 to 23 and D47 to 54 (19.61%) compared with D47 to 54 to birth (11.76%). Moreover, a higher EFL % was observed in Zaraiebi goats compared with others. Age and goat parity had no significant effect on the EFL % in all goats. A high EFL % were observed in goats synchronized by P4 sponge, as well as artificially inseminated goats compared with goats with spontaneous estrus, and bred by natural mating, respectively. Serum P4 at D7 or D20 after breeding showed nonsignificant difference between normal pregnant goats and goats that experienced EFL. Unlike goats that experienced partial EFL, goats that experienced total EFL between D20 to 23 and D26 to 29 showed an abrupt P4 reduction (85.06%; P < 0.01) suggesting the probability of endocrine disruption of the CL. However, goats that were exposed to total EFL between D26 and 29 to D33 to 36 showed a low P4 reduction (24.90%; P < 0.05), which might be considered as an effect rather than a cause of EFL. In conclusion, different factors such as breed, estrous synchronization, breeding, and stage of pregnancy may be involved in EFL in goats. Therefore, improvement of the goat management in the early stage of pregnancy is important to decrease EFL % in goats. Although the P4 did not show any significant difference between normal pregnancy goats and goats that experienced EFL, CL disruption should be taken into the consideration, at least, in goats exposed to total embryonic losses.

The pro-inflammatory and anti-inflammatory cytokine profile in peripheral blood of women with recurrent implantation failure.

Limited information is available on the balance state of pro- and anti-inflammatory cytokines in patients with recurrent implantation failure (RIF). This study assessed the pro- and anti-inflammatory cytokines in plasma of 34 patients with RIF, compared with those of 25 women with a successful pregnancy in the first IVF/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycle. The IFN-γ, IL-1ß, IL-6 and IL-4 concentrations were higher, whereas the TGF-ß1 concentration was lower in the RIF group compared with the control group. Furthermore, the ratios of pro-inflammatory and anti-inflammatory cytokines IFN-γ/IL-4, IFN-γ/IL-10, IFN-γ/TGF-ß1, IL-6/IL-10, IL-6/TGF-ß1, IL-1ß/TGF-ß1 and TNF-α/TGF-ß1 were higher in the RIF group (all P < 0.01). The results suggested a shift toward a pro-inflammatory state in peripheral blood of the patients with RIF.

Low levels of naturally occurring regulatory T lymphocytes in blood of mares with early pregnancy loss.

Early pregnancy loss is a major reason for low reproductive efficiency in the horse. In humans and mice, low numbers of regulatory T cells (Treg cells) are linked to miscarriage. The percentage of Treg cells in oestrous mares at the start of the breeding season was evaluated in relation to the outcome of subsequent pregnancy. For identification and quantification of Treg cells, a highly sensitive and specific qPCR assay targeting the Treg-specific demethylated region in the equine forkhead box transcription factor (FOXP3) gene was established. In a total of 108 mares, pregnancy was followed until detection of early pregnancy loss (n=17), abortion without identification of an infectious or apparent cause (n=9) or birth of a viable foal (n=82). Measured Treg-cell levels did not significantly differ between mares that conceived (82%; 1.50±0.04%) or did not get pregnant (18%; 1.45±0.10%). The Treg-cell percentage at oestrus before breeding was significantly different (P<0.05) between mares that either underwent early pregnancy loss up to Day 40 of pregnancy (1.29±0.07%) and mares that aborted (1.61±0.15%) or gave birth to a live foal (1.52±0.05%). These results suggest that low levels of Treg cells in mares can contribute to pregnancy loss up to Day 40 after ovulation.

[Reproductive problems in women with PCOS, the impact of PAL-1 CARRIERS OF 4G PAI -1 polymorphism and BMI].

Approximately 7-12% of women in reproductive age are affected by PCOS[2] and 40 to 70 percent of them are overweight contributing to the clinical picture of PCOS and increased reproductive and metabolic disorder. In order to investigate the role of PAl-1 as a possible risk factor for the development of PCOS a group of 67 women with polycystic ovarian disease and 70 healthy controls were investigated for levels of PAI-1 and carriage of the promoter polymorphism 675 4G/5G in gene of PAl-1. The results of the DNA analysis showed a high carriage of polymorphism 675 4G/4G in promoter of PAI-1 gene in women with PCOS but not as significant (OR = 1.6645, p = 0.141). Serum levels of PAI-1 were significantly higher in total group of patients compared to controls. The levels of PAI-1 is correlated with carriage of 675 4G/5G polymorphism in the gene for PAI-1 (R = 0.534, p = 0.03) as well as wih BMI, like correlation coefficients were higher in the group with PCOS (0.572, p = 0.04). Data from the disease history showed a higher percentage of women with reproductive problems: 61.5% (early pregnancy loss and infertility) significantly higher in the group with PCOS (70.1% compared to 54.1%). The carriers of polymorphism 4G are at greater risk for early pregnancy loss than those with 5G (61.45% as compared to 36.8%), which confirms that carriage of the polymorphism 4G/5G 675 gene PAI-1 has a specific in multifactorial pathogenesis and expression of PCOS.

Relationship of thyroid hormone levels and thyroid autoantibodies with early pregnancy loss and infertility.

Prevalence of abnormalities in thyroid hormones and thyroid autoantibodies in patients with a history of early pregnancy loss (EPL) (n = 17) and unexplained infertility (UI) (n = 25), were compared with that of 45 control patients. TSH, antithyroid peroxidase (anti-TPO) and antithyroglobulin (anti-TG) antibody levels in UI and EPL groups were similar to that of the control group. TSH was normal in 39, 22 and 13 of control, UI and EPL patients, respectively. Among patients with a normal TSH, fT4 was higher (p < 0.001) and fT3 was lower (p < 0.001) in infertile patients when compared with the control group. Thyroid function tests seem to be associated with infertility but their association with EPL is weaker. Infertility seems to be associated with a high fT4 and low fT3 status. Thyroid autoantibodies do not seem to be associated with either infertility or EPL.

Progesterone is essential for protecting against LPS-induced pregnancy loss. LIF as a potential mediator of the anti-inflammatory effect of progesterone.

Lipopolysaccharide (LPS) administration to mice on day 7 of gestation led to 100% embryonic resorption after 24 h. In this model, nitric oxide is fundamental for the resorption process. Progesterone may be responsible, at least in part, for a Th2 switch in the feto-maternal interface, inducing active immune tolerance against fetal antigens. Th2 cells promote the development of T cells, producing leukemia inhibitory factor (LIF), which seems to be important due to its immunomodulatory action during early pregnancy. Our aim was to evaluate the involvement of progesterone in the mechanism of LPS-induced embryonic resorption, and whether LIF can mediate hormonal action. Using in vivo and in vitro models, we provide evidence that circulating progesterone is an important component of the process by which infection causes embryonic resorption in mice. Also, LIF seems to be a mediator of the progesterone effect under inflammatory conditions. We found that serum progesterone fell to very low levels after 24 h of LPS exposure. Moreover, progesterone supplementation prevented embryonic resorption and LPS-induced increase of uterine nitric oxide levels in vivo. Results show that LPS diminished the expression of the nuclear progesterone receptor in the uterus after 6 and 12 h of treatment. We investigated the expression of LIF in uterine tissue from pregnant mice and found that progesterone up-regulates LIF mRNA expression in vitro. We observed that LIF was able to modulate the levels of nitric oxide induced by LPS in vitro, suggesting that it could be a potential mediator of the inflammatory action of progesterone. Our observations support the view that progesterone plays a critical role in a successful pregnancy as an anti-inflammatory agent, and that it could have possible therapeutic applications in the prevention of early reproductive failure associated with inflammatory disorders.

First trimester assessment of pentraxin-3 levels in women with primary unexplained recurrent pregnancy loss.

OBJECTIVE: To evaluate the potential role of measuring first-trimester maternal Pentraxin-3 levels in patients with primary unexplained recurrent pregnancy loss. STUDY DESIGN: A case control study was conducted in Ain Shams University Maternity Hospital. Cases included 45 women with primary unexplained recurrent pregnancy loss and early pregnancy failure admitted for medical or surgical termination of pregnancy. Controls (45 women) included a matched group of apparently healthy pregnant women who had at least one previous uneventful pregnancy with no previous obstetric history of adverse pregnancy outcomes. Maternal venous blood samples were collected for assay of Pentraxin-3 using enzyme-linked immunosorbent assay. The main outcome measure was the pregnancy outcome in women with elevated Pentraxin-3 levels. RESULTS: 90 participants were statistically analyzed. In the patient group, the mean Pentraxin-3 level was 12.00 ± 4.07 ng/ml, while in the control group it was 1.69 ± 0.91 ng/ml. The difference was statistically significant (p<0.001). In the patient group, Pentraxin-3 showed a significant positive correlation with the number of previous miscarriages (p=0.038). CONCLUSION: Abnormally elevated Pentraxin-3 levels indicate the presence of an abnormally exaggerated intrauterine inflammatory or innate immune response that may cause pregnancy failure in women with primary unexplained recurrent pregnancy loss.

Predictive value of serum human chorionic gonadotropin ratio, progesterone and inhibin A for expectant management of early pregnancies of unknown location.

OBJECTIVE: To evaluate serum human chorionic gonadotropin (hCG) ratio, progesterone and inhibin A as single parameters and in combination for the prediction of spontaneous resolution of pregnancies of unknown location (PUL). STUDY DESIGN: Prospective observational study of 105 consecutive patients with a diagnosis of PUL. Serum levels of hCG, progesterone and inhibin A were determined at the first visit and after 2 days. Patients were followed clinically until a final diagnosis of spontaneously resolving PUL, viable or non-viable intrauterine pregnancy, or ectopic pregnancy with need of laparoscopic intervention had been reached. Different combinations of hCG ratio (hCG at 48 h/hCG at 0 h), s-progesterone and s-inhibin A were investigated to find the best predictor for successful expectant management. RESULTS: The final pregnancy outcomes were: 52 spontaneously resolving PUL (49.5%), 37 viable intrauterine pregnancies (35.2%), 8 non-viable intrauterine pregnancies (7.6%), 7 ectopic pregnancies (6.7%), and one molar pregnancy (1.0%). An hCG ratio<0.80 predicted spontaneously resolving PUL with positive and negative predictive values (PPV and NPV), sensitivity, and specificity of 0.98, 0.78, 0.72, and 0.99, respectively. In patients with hCG ratio ≥ 0.80, a combination of s-progesterone < 20 nmol/l and s-inhibin A < 30 pg/ml predicted spontaneously resolving PUL with PPV, NPV, sensitivity and specificity of 0.92, 0.96, 0.85, and 0.98 respectively. CONCLUSION: Our results suggest that patients with PUL and hCG ratio < 0.80 display a high probability of spontaneously resolving PUL with minimum need of follow-up. In cases of hCG ratio ≥ 0.80, a combination of s-progesterone < 20 nmol/l and s-inhibin A < 30 pg/ml, may be a reliable predictor of spontaneously resolving PUL. The safety of this approach should be tested in large prospective studies.

Anticardiolipin antibodies and recurrent early pregnancy loss: a century of equivocal evidence.

In 1987, Nigel Harris cautioned against over-diagnosing the antiphospholipid syndrome (APS). In what was a rather prophetic editorial titled 'The Syndrome of the Black Swan', he suggested that while patients with APS do indeed exist, they are probably much more unusual than many medical professionals might like to believe. He expressed concern that the value of studying antiphospholipid antibodies (aPLs) as interesting non-organ specific autoantibodies, would become lost in a 'sea of over-interpreted and over-reported laboratory and clinical findings'. It is our contention that 25 years later, this prediction has come to pass, particularly with respect to one type of aPL and its relation to a clinical event, namely anticardiolipin antibodies and early recurrent pregnancy loss. In this commentary, we trace the evolution of the current dogma and propose that reevaluation of available data from an alternative perspective results in quite a different understanding, the acceptance of which would necessitate not only a revision of the classification criteria for APS but also the subsequent revision of many diagnoses.

Candidate biomarkers for acute rejection of pregnancy after in-utero cell-based therapy in pre-immune embryos via ultrasound-guided celocentesis.

PROBLEM: A maternal serum biomarker profile analysis was performed to determine potential indicators of acute rejection of pregnancy following in-utero cell-based treatments in pre-immune embryos. METHOD OF STUDY: We used an established non-human primate model for in-utero stem cell therapy at 38-42 days from fertilization. The maternal serum concentrations of nine candidate biomarkers for acute rejection of pregnancy were determined before and after the injection of different cocktails of human umbilical cord blood stem cells into the gestational sac. All animals were then followed until delivery. RESULTS: Twenty-four hours after celocentesis, two of the animals aborted. These two animals received a cocktail of haemopoietic stem cells with the highest concentration of human CD3(+) cells and showed a twofold increase in maternal serum IL-6 and a threefold increase in prolactin after the procedure. The remaining six animals delivered at term live and normal newborns and only demonstrated an increase in prolactin after the celocentesis procedure. CONCLUSION: IL-6 and prolactin are master immunoregulators with pleiotropic biological functions that have different maternal serum concentrations depending on pregnancy outcome. These findings suggest that increases in maternal serum prolactin and IL-6 concentration may be associated with acute rejection of pregnancy after in-utero stem cell therapy.

Antiphospholipid syndrome: laboratory testing and diagnostic strategies.

The antiphospholipid syndrome (APS) is diagnosed in patients with recurrent thromboembolic events and/or pregnancy loss in the presence of persistent laboratory evidence for antiphospholipid antibodies. Diagnostic tests for the detection of antiphospholipid antibodies include laboratory assays that detect anticardiolipin antibodies, lupus anticoagulants, and anti-ß(2)-glycoprotein I antibodies. These assays have their origins beginning >60 years ago, with the identification of the biologic false positive test for syphilis, the observation of "circulating anticoagulants" in certain patients with systemic lupus erythematosus, the identification of cardiolipin as a key component in the serologic test for syphilis, and the recognition and characterization of a "cofactor" for antibody binding to phospholipids. Although these assays have been used clinically for many years, there are still problems with the accurate diagnosis of patients with this syndrome. For example, lupus anticoagulant testing can be difficult to interpret in patients receiving anticoagulant therapy, but most patients with a thromboembolic event will already be anticoagulated before the decision to perform the tests has been made. In addition to understanding limitations of the assays, clinicians also need to be aware of which patients should be tested and not obtain testing on patients unlikely to have APS. New tests and diagnostic strategies are in various stages of development and should help improve our ability to accurately diagnose this important clinical disorder.

Quantitative and functional changes in peripheral natural killer cells in women with reproductive failure after artificial insemination with donor sperm.

Considerable evidence indicates that quantitative and/or functional abnormalities of natural killer (NK) cells are associated with reproductive failure. The objective of this study was to determine if peripheral NK percentage and activity are altered in women with reproductive failure after artificial insemination by donor (AID), and which parameters can best discriminate women with AID failure and normal controls. A case-control study of 20 fertile controls, 25 patients undergoing more than four treatment cycles without success (Group I), 25 experiencing biochemical pregnancy loss (Group II), and 25 undergoing embryo growth arrest or miscarriage (Group III) was performed. The peripheral NK percentage and cytotoxicity was evaluated by flow cytometry. Our results showed that a significantly increased percentage of NK cell cytotoxicity (NKCC) was detected in all study groups compared with the controls. The percentage of CD56(+) NK cells was elevated in Groups II and III compared with the controls. The percentage of CD56(+)CD16(+) NK cells in the study groups was higher than that in the controls. By using receiver operating characteristic curve and logistic regression analysis, we found that NKCC combined with CD56(+)CD16(+) NK percentage had good predictive power. The overall predicted percentage correct was 0.89. These results suggest that enhanced NKCC and an elevated cytotoxic NK percentage may be associated with AID failure and can be biomarkers for predicting AID-induced failure.

Acquired activated protein C resistance, thrombophilia and adverse pregnancy outcomes: a study performed in an Irish cohort of pregnant women.

The combination of thrombophilia and pregnancy increases the risk of thrombosis and the potential for adverse outcomes during pregnancy. The most significant common inherited risk factor for thrombophilia is activated protein C resistance (APCR), a poor anticoagulant response of APC in haemostasis, which is mainly caused by an inherited single-nucleotide polymorphism (SNP), factor V G1691A (FV Leiden) (FVL), referred as inherited APCR. Changes in the levels of coagulation factors: FV, FVIII, and FIX, and anticoagulant factors: protein S (PS) and protein C (PC) can alter APC function causing acquired APCR. Prothrombin G20210A and methylenetetrahydrofolate reductase (MTHFR) C677T are prothrombotic SNPs which in association with APCR can also increase the risk of thrombosis amongst Caucasians. In this study, a correlation between an acquired APCR phenotype and increased levels of factors V, VIII, and IX was demonstrated. Thrombophilic mutations amongst our acquired APCR pregnant women cohort are relatively common but do not appear to exert a severe undue adverse effect on pregnancy.