Copeptin is associated with the severity of endometriosis

Purpose

Endometriosis is defined as the presence of endometrial glands and stroma in ectopic locations and may be associated with local and systemic inflammatory processes. Copeptin is elevated in acute and chronic inflammation conditions. The aim of the present study was to determine whether serum copeptin levels were altered in women with endometriosis and played a role in the pathophysiology of the disease.

Methods

A total of 86 women were recruited for this case–control study. 50 patients with surgically proven endometriosis were included, while 36 patients without endometriosis comprised the control group. Patients were classified as having minimal, mild, moderate and severe disease in accordance with American Society of Reproductive Medicine revised classification. Two subgroups were formed by combining patients with minimal and mild disease and with moderate and severe disease (Stage 1–2, stage 3–4; respectively). Levels of copeptin, tumor markers (CA-125, CA-19-9, CA-15-3) and C-reactive protein in serum were measured.

Results

Serum copeptin, CA-125, CA-15-3 and CA-19-9 levels were higher in the endometriosis group (p: 0.002; 0.001; 0.017; 0.015; respectively). Copeptin and CA-19-9 levels were significantly higher in stage 3–4 group as compared to stage 1–2 group (p: 0.004; 0.036 respectively). Serum copeptin levels were positively correlated with stage of the disease and size of endometriomas. ROC analysis revealed that CA-125 had the highest AUC for predicting endometriosis (0.938; 95 % confidence interval 0.882–0.993; p: 0.001).

Conclusions

Serum copeptin levels were significantly higher in patients with endometriosis as compared to healthy controls. Moreover, severity of the disease was correlated with serum copeptin levels.     

Effects of amifostine on endometriosis, comparison with N-acetyl cysteine, and leuprolide as a new treatment alternative: a randomized controlled trial

Purpose

To assess the effects of amifostine, N-acetyl cysteine (NAC), and leuprolide as a scavenger in a rat endometriosis model.

Methods

This is a prospective randomized animal study. Setting The Animal Laboratory of Medical University. Animals 40 rats were used for transplantation of an autologous fragment of endometrial tissue onto the inner surface of the abdominal wall. After allowing 3 weeks for growth, laparotomies were performed to check the implants. Then animals were randomized into four groups: Group I amifostine (200 mg/day loading dose after 20 mg/kg/day, p.o.); Group II NAC (200 mg/day, p.o.); Group III leuprolide acetate 1 mg/kg single dose, sc; and Group IV (controls) no medication. Three weeks later, implants were evaluated morphologically. Serum and peritoneal TNF-alpha levels were evaluated. The transmission electron microscopic examination of the peritoneal samples and ovaries was also performed.

Results

Leuprolide acetate, amifostine and NAC caused significant decreases in the mean implant areas and significant decreases in serum and peritoneal TNF-alpha levels. On comparing all groups, these reductions were higher in Group II. According to the transmission electron microscopic findings, leuprolide seems to be protecting normal structure of peritoneum best when compared to the other groups.

Conclusions

Amifostine, NAC and leuprolide caused regression of endometriosis in this experimental rat model by a yet unsettled mechanism.     

Diagnostic biomarkers of pro-inflammatory immune-mediated preterm birth

Purpose

Pro-inflammatory immunity, either infectious or sterile-derived, is one of the major causes of preterm birth and associated with enhanced maternal and fetal morbidity and mortality. Diagnosing intrauterine inflammation at an early stage is tremendously important. Amniotic fluid interleukin (IL)-6 concentration is currently the most investigated diagnostic tool for detecting intrauterine inflammation.

Methods

Amniotic fluid samples were obtained from women with no signs of intrauterine infection [amniocentesis (n = 82), cesarean section (n = 110), spontaneous delivery (n = 20) and those with clinical signs of intrauterine infection or inflammation (AIS, n = 16)]. Amniotic fluid was screened by commercial ELISAs for IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-15, IL-17, growth regulated oncogene-α (gro) α, macrophage inflammatory protein (MIP) 1α, MIP1β, histone, tumor necrosis factor (TNF) α, proIL1β and interferon γ-induced protein (IP) 10.

Results

ProIL-1β, MIP1β, IL-10 and IL-8 levels were significantly elevated in the AIS group, whereas IL-4 levels were significantly lower in the AIS group. No significant differences were found regarding IL-2, IL-6, IL-12, IL-15, IL-17, GROα, MIP1α, histone, TNFα, ProIL1β and IP10.

Conclusion

MIP1β, IL-4, IL-8, IL-10 and proIL-1β might be potential singular biomarkers in diagnosing intrauterine inflammation. The combinations of elevated levels of IL-17/GROα, MIP1β/IL-15 and histone/IL-10 are new potentially advantageous biomarker combinations.     

Combination of non-invasive and semi-invasive tests for diagnosis of minimal to mild endometriosis

Objectives

Non-surgical diagnostic approach for endometriosis would be of great gain to both physicians and patients. The aim of this study was to evaluate the diagnostic value of serum measurement of IL-6 combined with the presence of nerve fibres in the functional layer of endometrium for diagnosis of minimal-mild endometriosis.

Methods

In this prospective study 114 women who underwent laparoscopy for infertility and/or pelvic pain were divided into two groups: control cases (40 cases) with no pathologic findings; and endometriosis patients (74 cases) [subdivided into stages 1–2 or minimal-mild (MM) and stages 3–4 or moderate-severe cases]. Blood was drawn one day before laparoscopy and stored for subsequent analysis of IL-6. Endometrial biopsy was obtained prior to laparoscopy and Immunohistochemistry was performed using the pan-neuronal marker protein gene product 9.5(PGP9.5). Then laparoscopic diagnosis of endometriosis confirmed by histopathology was done.

Results

Serum IL-6 with a threshold of 15.4 pg/ml was found to be able to diagnose MM endometriosis with 89.5 % sensitivity and 82.5 % specificity, but sensitivity and specificity of presence of nerve fibres in the functional layer of endometrium were 92 % and 80 % respectively. When two diagnostic modalities were combined the sensitivity and specificity were raised to 100 and 92.5 % respectively.

Conclusions

Combination of both serum IL-6 and presence of nerve fibres in the endometrium is more reliable method for diagnosis of MM endometriosis than in single test.     

Expression of human β-defensin-2 in the eutopic and ectopic endometrial tissues in patients with endometriosis

Objective

To investigate the expression of human β-defensin-2 (hBD-2) in the endometrium of patients with endometriosis (EMS) and explore the potential role of hBD-2 in the pathogenesis of EMS.

Design

Prospective controlled study.

Sample

50 women including EMS patients undergoing laparoscopic ovarian cystectomy and non-EMS patients undergoing hysterectomy for uterine fibroids.

Setting

Large university teaching hospital.

Methods

Patients were divided into EMS and non-EMS groups. The gene expressions of hBD-2, interleukin (IL)-1β and tumor necrosis factor (TNF)-α in the endometrial tissues of each group were detected with real-time quantitative polymerase chain reaction (PCR), and hBD-2 protein expression with immunohistochemical method.

Results

The gene expression levels of hBD-2, TNF-α, and IL-1β as well as the positive expression rate of hBD-2 protein in the ectopic endometrium of EMS patients were significantly higher than those in the eutopic endometrium of EMS and non-EMS patients (all P < 0.05). Correlation analysis showed that the gene expression levels of hBD-2 in the ectopic and eutopic endometrium of EMS patients were positively correlated with the gene expression levels of IL-1β and TNF-α (P < 0.01).

Conclusion

High levels of hBD-2 gene and protein expressions in the ectopic endometrium of EMS patients may be an important contributor in the pathogenesis of EMS. TNF-α and IL-1β may promote the upregulation of hBD-2 expression.     

Intrafollicular inflammatory cytokines but not steroid hormone concentrations are increased in naturally matured follicles of women with proven endometriosis

Purpose

The aim of this study was to assess whether the intrafollicular cytokine profile in naturally developed follicles is different in women with endometriosis, possibly explaining the lower reproductive outcome in endometriosis patients.

Methods

A matched case-control study was conducted at a university-based infertility and endometriosis centre. The study population included 17 patients with laparoscopically and histologically confirmed endometriosis (rAFS stages II–IV), each undergoing one natural cycle IVF (NC-IVF) treatment cycle between 2013 and 2015, and 17 age-matched NC-IVF women without diagnosed endometriosis (control group). Follicular fluid and serum was collected at the time of follicle aspiration. The concentrations of inflammatory cytokines (IL-1β, IL-6, IL-8, IL-15, IL-18, TNF-α) and hormones (testosterone, estradiol, AMH) were determined in follicular fluid and serum by single or multiplexed immunoassay and compared between both groups.

Results

In the follicular fluid, IL-1β and IL-6 showed significantly (P?<?0.001 and 0.01, respectively) higher median concentrations in the endometriosis group than in the control group and a tendency towards endometriosis severity (rAFS stage) dependence. The levels of the interleukins detectable in follicular fluid were significantly higher than those in the serum (P?<?0.01). Follicular estradiol concentration was lower in severe endometriosis patients than in the control group (P?=?0.036). Follicular fluid IL-1β and IL-6 levels were not correlated with estradiol in the same compartment in neither patient group.

Conclusions

In women with moderate and severe endometrioses, some intrafollicular inflammatory cytokines are upregulated and not correlated with intrafollicular hormone concentrations. This might be due to the inflammatory microenvironment in endometriosis women, affecting follicular function and thereby possibly contributing to the reproductive dysfunction in endometriosis.

     

The diagnostic and prognostic value of serum YKL-40 in endometrial cancer

Purpose

To evaluate the diagnostic and prognostic value of serum YKL-40 in endometrial cancer (EC).

Methods

Serum YKL-40 levels were detected and compared in 34 of the 50 cases with EC before surgery, in 22 of the 34 with EC after surgery, in 30 cases with uterine myoma, and in 30 healthy women as normal controls. Receiver operating characteristics (ROC) curves were adopted for diagnosis and calculation of area under each ROC curve in EC. The progression-free survival (PFS) and overall survival (OS) between YKL-40 positive and negative patients were compared in the follow-up.

Results

The mean pre-operative serum YKL-40 values were significantly higher than that in the uterine myoma cases and in the healthy women (P = 0.000). The mean post-operative serum YKL-40 in the 22 EC cases was significantly lower than pre-operative serum YKL-40 levels in these cases (P = 0.000). There were critical differences between the area under ROC curve for YKL-40 and CA125 (P = 0.053). The PFS and OS for the YKL-40-positive patients were significantly shorter than those for the YKL-40-negative patients.

Conclusion

Preliminary investigations have shown that serum YKL-40 level may have a definite clinical value in the diagnosis and prognosis of EC.     

Comparative study between VEGF-A and CA-125 in diagnosis and follow-up of advanced endometriosis after conservative laparoscopic surgery

Objective

To evaluate the role of serum level of VEGF-A in comparison to CA-125 in diagnosis and follow-up of patients with advanced endometriosis after conservative laparoscopic surgery.

Methods

A prospective randomized case–control study was performed on patients referred for laparoscopy complaining of unexplained primary infertility with or without chronic pelvic pain. Thirty patients with advanced endometriosis; stage III–IV were included (study group), another 30 women without endometriosis or any other medical conditions were settled as a control group. Pre-operative blood samples were collected from study and control cases. Post-operative blood samples were collected from 25 treated patients in the follicular phase of the third menstrual cycle; 5 cases were drop-outs. Serum level of cancer antigen-125 (CA-125) and vascular endothelial growth factor (VEGF-A) were assayed by using enzyme linked immunosorbent assay (ELISA) kit.

Results

There was a statistically significant difference in serum CA-125 and VEGF-A level in patients with advanced endometriosis before conservative laparoscopic surgery and those without endometriosis (p < 0.001) and after conservative laparoscopic surgery (p < 0.001). High sensitivity (93.3 %), specificity (96.7 %) and accuracy (95.0 %) of VEGF-A assay than in CA-125 distinguishing between patients with endometriosis from those without endometriosis; CA-125 has 70.0 %sensitivity, 90.0 % specificity and 85.0 % accuracy. Percentage of decrease of VEGF-A level after operation was higher than that of CA-125 (45.9 vs. 25.8 %) p < 0.001, respectively.

Conclusion

The use of VEGF-A for diagnosis of advanced endometriosis at cut-off 680 pg/ml and for follow-up is better than CA-125.     

Deep endometriosis inflicting the bladder: long-term outcomes of surgical management

Objective

To assess the efficacy and safety of laparoscopic treatment of bladder endometriosis, especially in cases of full thickness endometriotic nodules.

Design

Retrospective review of medical records.

Setting

Tertiary medical center and a referral center for endometriosis.

Population

Sixty-nine patients with bladder endometriosis that underwent surgery between January 2005 and December 2011.

Methods

The records of all patients with bladder endometriosis were reviewed and the pre-, intra- and postoperative information of patients who underwent surgery was collected.

Main outcome measures

Efficacy, safety and long-term outcome of laparoscopic treatment of bladder endometriosis.

Results

The mean age of 69 patients with bladder endometriosis was 31.3 ± 4.6 years. Preoperative urinary symptoms (such as frequency, urgency, dysuria and others) were present in 28 (40.0 %) patients. Laparoscopy was performed in all patients. Deep detrusor involvement was found in 45 (65.2 %) patients. Of these, 21 patients underwent partial cystectomy due to a full thickness lesion. Deep nodule resection without bladder invasion was performed in 24 (34.8 %) patients and bladder nodule coagulation and ablation in the remaining 24 (34.8 %) patients with superficial involvement. No intraoperative complications were noted. Postoperative follow-up results were available for all patients. After a median (range) follow-up period of 60 (4–92) months, 92.7 % of the patients were asymptomatic or reported improvement in symptoms.

Conclusions

After a long-term follow-up surgical management of bladder endometriosis is strongly recommended. During surgery, careful inspection and full excision of bladder lesions should be performed. Laparoscopic excision is a safe and efficacies approach.     

Atorvastatin exerts anti-nociceptive activity and decreases serum levels of high-sensitivity C-reactive protein and tumor necrosis factor-α in a rat endometriosis model

Purpose

The purpose of this study was to examine the effects of atorvastatin in the treatment of experimental endometriosis.

Methods

Endometriosis was induced in 24 female rats. 4 weeks after the procedure dimensions of the foci were recorded. Rats were divided into three groups: in Group 1 (n = 8), a daily dose of 10 mg/kg atorvastatin was given for 14 days. In the second group (n = 8), a single dose of 1 mg/kg leuprolide acetate was injected intraperitoneally. The rats in Group 3 (n = 8) were received 1 mg/kg i.p. 0.9 % NaCl. At the end of the treatment, laparotomy was performed, and the dimensions of the endometriotic foci were recorded. Biochemical, histopathological and immunohistochemical studies were performed and nociception was compared in groups.

Results

Atorvastatin treatment exhibited significant analgesic activity in hot plate model (P = 0.022). The serum hs-CRP and tumor necrosis TNF-α levels were similar between the Group 2 and Group 3 (P > 0.05); however atorvastatin caused significant decrease in both serum markers. The histological and immunohistochemical scores were also found to be markedly lower in Group 1 and Group 2 (P < 0.05).

Conclusion

Atorvastatin treatment may have a therapeutic potential in the treatment of endometriosis through its anti-inflammatory and anti-nociceptive properties.     

Evaluation of novel serum biomarkers and the proteomic differences of endometriosis and adenomyosis using MALDI-TOF–MS

Background

Both endometriosis and adenomyosis are common benign gynecological diseases. This study aimed to find the novel noninvasive, biochemical diagnostic markers for detection of endometriosis and adenomyosis, and evaluate the correlation of these two diseases at the protein level.

Methods

Serum samples from patients with endometriosis or adenomyosis were compared with control groups to detect specific serum biomarkers and to explore the different protein fingerprint of endometriosis and adenomyosis using MALDI-TOF–MS.

Result(s)

There were 13 protein peaks abnormally expressed in endometriosis as well as twelve in adenomyosis compared with control groups (P < 0.05). And five-peak mass was found downregulated significantly both in the women with endometriosis and adenomyosis. The common diagnostic model of endometriosis and adenomyosis we set up had a lower sensitivity and specificity than the separate diagnostic model of these two diseases.

Conclusion(s)

MALDI-TOF–MS technology plays an important role in screening the diagnostic biomarkers of endometriosis and adenomyosis. And our study found the correlation between endometriosis and adenomyosis in protein fingerprint and it is hard to separate the endometriosis from adenomyosis with the serum biomarkers.     

Circulating apelin levels are associated with cardiometabolic risk factors in women with previous gestational diabetes

Purpose

Women with previous gestational diabetes mellitus (pGDM) are at high risk for type 2 diabetes and cardiovascular disorders. In this study, we aimed to compare plasma apelin levels between women with and without pGDM, and to investigate the possible association of apelin with cardiometabolic risk factors.

Methods

Among 252 consecutive Caucasian women with GDM being included in a prospective postpartum follow-up protocol, 141 women eligible for the study protocol were enrolled. Control group consisted of 49 age- and body mass index-matched healthy women without pGDM. Circulating apelin, IL-6 and plasminogen activator inhibitor levels, and carotid intima media thickness (IMT) were measured. To evaluate carbohydrate intolerance, 75-g oral glucose tolerance test was performed. Fasting insulin and lipids were measured, and homeostasis model assessment index was calculated.

Results

Plasma apelin levels were reduced in women with pGDM (p < 0.001). In multiple regression analysis, apelin was negatively associated with fasting (r ² 0.090, β ?0.273, p = 0.001) and post-load glucose (r ² 0.061, β ?0.187, p = 0.022), serum IL-6 (r ² 0.082, β ?0.234, p = 0.002), and carotid IMT (r ² 0.057, β ?0.168, p = 0.033).

Conclusions

Our results suggested that suppressed apelin levels were associated with increased cardiovascular risk in women with pGDM.     

Association of the progesterone receptor gene polymorphism (PROGINS) with endometriosis: a meta-analysis

Background

Reported associations of progesterone receptor gene polymorphism (PROGINS) with endometriosis have been inconsistent.

Aim of the study

To evaluate the association between the PROGINS polymorphism and the risk of endometriosis.

Methodology

A meta-analysis of 12 published case–control studies with a total sample size of 3,321 (1,323 cases/1,998 controls) was performed. We estimated the risk (odds ratio [OR] 95 % confidence intervals) of endometriosis association with the PROGINS polymorphism.

Results

An association between the presence of the variant allele and risk of endometriosis was found, more in the homozygous and recessive models (OR 1.41–1.43, p = 0.15–0.17), and less in the dominant and co-dominant models (OR 1.22, p = 0.11–0.15). Reanalysis without the studies whose controls deviated from the Hardy–Weinberg Equilibrium did not materially alter the dominant and co-dominant effects (OR 1.19–1.22, p = 0.19–0.32), but exacerbated the homozygous and recessive effects (OR 1.59, p = 0.09). The subgroups based on geography showed increased risk associations, consistently significant in the European (OR 1.52–2.72, p = 0.0008–0.03) but not in the Brazilian studies, where ORs ranged from reduced (OR 0.70–0.74, p = 0.54–0.61) to increased (OR 1.11, p = 0.75) risks. Heterogeneity was confined in all comparisons to the dominant and co-dominant models (I ² = 38–70 %), except in the European subgroup, which had zero heterogeneity (I ² = 0 %) in all genetic models, as did all homozygous and recessive effects.

Conclusion

This meta-analysis provides a comprehensive profile of the role of the PROGINS polymorphism in endometriosis by exploring the magnitude of the summary effects with modifier analysis. This magnitude is expressed with modulation or exacerbation of the summary effects, as defined by the parameters of the analysis. Thus, the results showed trend towards an increased risk of the variant PROGINS allele and susceptibility for the endometriosis.     

Effect of lesion location on endometriotic adhesion and angiogenesis in SCID mice

Objective

To evaluate the effects of lesion location on adhesion and angiogenesis of transplanted endometriotic lesions in SCID mice.

Methods

Three groups of female SCID mice included intraperitoneal (i.p.) (n = 12), subcutaneous (s.c.) (n = 12), and mock surgery (control) (n = 12). At 2 weeks after ovariectomy, the mice were transplanted with eutopic endometrium from endometriosis patients either subcutaneously or sutured within the peritoneal, or underwent mock surgery. After 4 weeks, the mice were sacrificed to evaluate the adhesion and volume changes of the implanted lesions. Furthermore, semiquantitative immunohistochemical staining was performed to analyze expression of MMP-2 and TIMP-2 as adhesion makers, and vWF, VEGF, and HIF-1α as angiogenesis markers.

Results

Adhesion occurred in 9 of 12 mice in the i.p. group, 3 of 12 mice in the s.c. group, and 3 of 12 mice in the control group. Fisher’s exact test showed that the difference of adhesion occurrence between i.p. and s.c. groups was statistically significant (p < 0.05). Graft volume changes were higher in the s.c. group than those in the i.p. group. MMP-2 expression was higher in the s.c. group than that in the i.p. group (p < 0.01). There was no significant difference of TIMP-2 expression between s.c. and i.p. groups. vWF, VEGF, and HIF-1α expression was significantly higher in the s.c. group than that in the i.p. group (p < 0.01).

Conclusions

Lesion location might be involved in the pathological changes of endometriosis. The intraperitoneal location is related to endometriotic adhesion, whereas the subcutaneous location is related to the infiltration of endometriotic lesions.     

CCDC22 gene polymorphism is associated with advanced stages of endometriosis in a sample of Brazilian women

Purpose

Based on the assumption that genetic factors are involved in the etiology of endometriosis, this study aimed to investigate the possibility of rs498679 (TLR4 gene), rs1799964 (TNF-α gene), rs3024496 (IL-10 gene), and rs2294021 (CCDC22 gene) polymorphisms being associated with the occurrence of this disease in a sample of Brazilian women.

Methods

We conducted a case-control study with 100 women with histological confirmation of endometriosis (endometriosis group) and 100 women submitted to laparoscopy for benign disorders, in which the absence of endometriosis was confirmed (control group). All samples were genotyped by real-time PCR technique for rs498679, rs1799964, rs3024496, and rs2294021 polymorphisms.

Results

No significant difference was observed in genotypic or allelic frequencies between control and endometriosis groups for rs498679 (TLR4 gene), rs1799964 (TNF-α gene), rs3024496 (IL-10 gene), neither when comparing endometriosis subgroups (I–II versus III-IV). On the other hand, significant difference between stages I–II and III–IV of the disease was found in genotypic and allelic frequencies for the rs2294021 (CCDC22 gene) SNP (p = 0.048 and p = 0.017, respectively).

Conclusion

Our results suggest that the rs2294021 (CCDC22 gene) polymorphism could be associated with increased susceptibility to endometriosis in Brazilian women when the allele C is present. In order to clarify this result, further studies should be conducted on a larger population.

     

Intrafollicular interleukin-8, interleukin-12, and adrenomedullin are the promising prognostic markers of oocyte and embryo quality in women with endometriosis

Purpose

The study aimed to investigate key intrafollicular prognostic factors among various cytokines and angiogenic molecules for prediction of mature oocytes and good-quality embryos in women with endometriosis undergoing in vitro fertilization (IVF).

Methods

Paired follicular fluid and serum samples were collected from 200 women with advanced stage endometriosis and 140 normal ovulating women during oocyte retrieval. The concentrations of cytokines (pro-inflammatory: IL-1β, TNF-α, IL-2, IL-8, IL-12, IFN-γ; anti-inflammatory: IL-4, IL-6, IL-10) and angiogenic molecules (vascular endothelial growth factor (VEGF), adrenomedullin, angiogenin) were determined in follicular fluid and serum using ELISA. Expression of these molecules was subjected to multivariate analysis for the identification of major predictive markers of oocyte and embryo quality. Receiver operating characteristic (ROC) curve was applied to determine the best cutoff point for the discrimination between mature and immature oocytes in these women.

Results

Significant increases in levels of cytokines and angiogenic molecules were observed in women with endometriosis compared to controls (P?<?0.001). From the validated partial least squares-discriminant analysis (PLS-DA) model, IL-8, IL-12, and adrenomedullin were identified as the most important factors contributing to endometriosis and were negatively associated with oocyte maturity and embryo quality.

Conclusion

The levels of IL-8, IL-12, and adrenomedullin may be good indicators of embryo and oocyte quality in endometriosis patients undergoing IVF. Further studies are necessary to ascertain the potential of these markers for oocyte and embryo developmental competence which may help improve the chances of a successful IVF in endometriosis patients.