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Investigating selected adhesion molecules as urinary biomarkers for diagnosing endometriosis
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Investigating selected adhesion molecules as urinary biomarkers for diagnosing endometriosis Katharina Proestling , Rene´ Wenzl , Iveta Yotova , Christoph Hauser , Heinrich Husslein , Lorenz Kuessel PII: DOI: Reference:
S1472-6483(20)30067-5 https://doi.org/10.1016/j.rbmo.2020.01.014 RBMO 2337
To appear in:
Reproductive BioMedicine Online
Received date: Accepted date:
21 December 2019 21 January 2020
Please cite this article as: Katharina Proestling , Rene´ Wenzl , Iveta Yotova , Christoph Hauser , Heinrich Husslein , Lorenz Kuessel , Investigating selected adhesion molecules as urinary biomarkers for diagnosing endometriosis, Reproductive BioMedicine Online (2020), doi: https://doi.org/10.1016/j.rbmo.2020.01.014
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo editing, typesetting, and review of the resulting proof before it is published in its final form. Please note that during this process changes will be made and errors may be discovered which could affect the content. Correspondence or other submissions concerning this article should await its publication online as a corrected proof or following inclusion in an issue of the journal. © 2020 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.
Investigating selected adhesion molecules as urinary biomarkers for diagnosing endometriosis Katharina Proestlinga, René Wenzla, Iveta Yotovaa, Christoph Hausera, Heinrich Hussleina*, Lorenz Kuessela a
Department of Gynecology and Obstetrics, Medical University of Vienna, Waehringer Guertel 18-20, 1090
Vienna, Austria
*Corresponding author. E-mail address: [email protected]
Abstract Research Question: Are selected cell adhesion molecules useful as urinary biomarkers for diagnosing endometriosis? Design: Prospective, longitudinal study (the Endometriosis Marker Austria) in patients who underwent laparoscopic surgery for benign gynecological pathologies. A total of 149 patients not receiving hormonal treatment for at least 3 months prior to recruitment were included and preoperative urine protein levels of soluble VCAM-1 (sVCAM-1), ICAM-1 (sICAM-1), E-selectin, and P-selectin were measured using a magnetic bead-based multiplex assay, normalized to creatinine levels of each sample. Levels were correlated with endometriosis status, menstrual cycle phase, body mass index, cigarette smoking and severity and entity of the lesions. Results: Urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin did not differ between women with (n=84) and without (n=65) endometriosis and among subgroups. Accordingly, ROC analysis to examine the value of using sVCAM-1, sICAM-1, E-selectin and P-selectin levels and sVCAM/sICAM ratio to diagnose endometriosis were not significant. In addition we examined whether the serum sVCAM-1 levels correlate with the urine levels of the protein in the same women, which revealed no significant correlations for sVCAM or sICAM. Conclusion: Although our previous study suggests that serum sVCAM is a promising biomarker for diagnosing endometriosis, we found no significant differences in urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin between women with and without endometriosis. Other markers should be studied in an effort to establish a truly non-invasive, urinary test for diagnosing endometriosis.
Keywords: endometriosis; biomarker; cell adhesion molecules; VCAM-1; ICAM-1
Introduction About 10% of reproductive-age women suffer from endometriosis, an estrogen dependent, chronic, disease causing pelvic pain and subfertility (Vigano et al., 2004). Laparoscopy is currently the gold standard for establishing the diagnosis of endometriosis, but it is expensive and carries surgical risks. At present a non-invasive test is not available in clinical practice, although the ability to accurately diagnose patients with endometriosis using a reliable, non–invasive approach would be of great value (Fassbender et al., 2015). Diagnostic markers have been evaluated in various body fluids, including urine, which is increasingly favored as a fluid for biological testing in biomarker discovery (Liu et al., 2015). The pathogenesis of endometriosis is not fully understood; A widely accepted hypothesis is that endometriosis originates from endometrial cells that adhere and implant on peritoneal surfaces via retrograde menstrual reflux. Cell adhesion molecules mediate cell adherence to extracellular matrix of the host tissue, enable cancer related biological processes and are related to the regulation of inflammatory and immune responses (Ohene-Abuakwa and Pignatelli, 2000). Contributing to a growing body of evidence regarding the role of cell adhesion molecules in endometriosis, our group demonstrated a significant overexpression of Vascular cell adhesion molecule-1 mRNA and elevated serum levels of soluble vascular cell adhesion molecule-1 (sVCAM1) in women with endometriosis, suggesting that sVCAM-1 is a promising diagnostic serum biomarker for endometriosis (Kuessel et al., 2017). Here we aimed to examine the value of selected cell adhesion molecules as urinary biomarkers for diagnosing endometriosis.
Materials & Methods Women were included in the EMMA study and samples and subgroups were collected and classified as described previously (Kuessel et al., 2017). Urine samples of women not on hormonal treatments for at least 3 months prior to recruitment were included.
Urine samples were obtained prior to laparoscopy, centrifuged within one hour of collection and stored in aliquots at -80°C. The urine concentrations of sVCAM-1, sICAM-1, E-selectin and P-selectin were measured in duplicate on the Luminex Bio-Plex 200 System (BioRad, California, USA) using Multiplex Luminex kits from RnD Systems (LKT007, RnD Systems, MN, USA) in accordance with the manufacturer’s protocol. The results below detection limit were set to 0pg/ml for the statistical analysis. Urine concentrations of each sample were normalized to the urine creatinine concentration for each sample expressed as pg/mg. Creatinine in urine was measured in duplicate using the Urinary Creatinine Assay Kit (STA-378; Cell Biolabs, San Diego, CA, USA). All statistical tests were performed using SPSS version 17.0. Non-parametric Mann‒Whitney U test and Kruskal‒Wallis test were used for comparison between groups. For paired statistics, the Wilcoxon signed-ranks test was used. Receiver operating characteristics (ROC) analysis was used to examine the diagnostic value. Spearman correlation analyses were used for correlation analysis between serum and urine levels of sVCAM-1 and sICAM-1. Differences with a p-value <0.05 were considered statistically significant.
Results In our present study, 84 (56.4%) women had and 65 (43.6%) did not have endometriosis. Women’s characteristics are shown in Table 1. Measured urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin did not differ significantly among groups (Table1). Accordingly, ROC analysis examining the value of using sVCAM-1, sICAM1, E-selectin and P-selectin levels and sVCAM/sICAM ratio to diagnose endometriosis were not significant (p=0.472, p=0.265, p=0.323, p=0.161, and p=0.545 for VCAM-1, ICAM-1, E-selectin and P-selectin levels and sVCAM/sICAM ratio, AUC<0.6 for all).
Exploratory subgroup analysis revealed no significant differences in urine levels between controls and woman with mild or severe endometriosis. Levels were not dependent on BMI, menstrual cycle phase, or cigarette smoking status (Table1).
Finally, by analyzing the experimental data from our current and previous study (Kuessel et al., 2017), we examined whether the serum sVCAM-1 and sICAM-1 levels correlate with the urine levels of the protein in the same women (n=59). Our analysis revealed no significant correlations (sVCAM-1: r=0.061; p=0.645 and sICAM-1: r=-0.197 p=0.135).
Discussion The field of endometriosis has an urgent need for an efficient, sensitive, non-invasive tool for diagnosing endometriosis. Researchers have searched for a suitable biomarker for several decades, primarily focusing on a multitude of different candidate markers in serum. Although urine is easier to access, shows a significantly narrower dynamic range of proteins and a less complex fluid composition compared to blood, only few endometriosis biomarker studies were urinebased (Fassbender et al., 2015). A systematic review aiming to evaluate the utility of urinary biomarkers found that most of the included studies were of retrospective design, evaluated either a specific phase of menstrual cycle or ignored cycle phase, and included only a limited number of samples; Furthermore, none of the potential biomarkers has been validated in separate patient cohorts (Fassbender et al., 2015; Liu et al., 2015). Our present study also has clear limitations, in particular the small sample size in the analyzed subgroups. However, in this study, we used a well-characterized, prospective study cohort and our robust design enabled us to perform exploratory subgroup analysis for lesion entities, disease stage, and menstrual cycle. Examining the utility of selected cell adhesion molecules as urinary biomarkers and investigating the correlation between serum and urine levels of sVCAM-1 and sICAM-1 in the same women, we found no significant differences between women with and without endometriosis, no correlation of urine levels with disease severity or patient characteristics, and no correlation between serum and urine levels of sVCAM-1 or sICAM-1. This lack of correlation between serum and urine levels of sVCAM-1 points to differences in the protein regulation in these biofluids. Published data from others and our own experience have shown that analyzing a small group of proven or suspected factors in the pathogenesis of the disease is unable to cover all aspects of the complex regulation of proteins and hypotheses can’t easily be transferred from one biofluid to another.
In conclusion, although our previous study suggests that serum sVCAM is a promising biomarker for diagnosing endometriosis, we found no significant differences between urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin between women with and without endometriosis. We believe that reporting information regarding well desingned negative trials can significantly contribute to focusing research on well-selected, better diagnostic targets.
Acknowledgements This study was co-funded by the Ingrid Flick Foundation (Grant no. FA751C0801), which played no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors declare no competing interests.
References Fassbender, A., Burney, R.O., O, D.F., D'Hooghe, T., and Giudice, L. , 2015. Update on Biomarkers for the Detection of Endometriosis. Biomed Res Int 2015, 130854. Kuessel, L., Wenzl, R., Proestling, K., Balendran, S., Pateisky, P., Yotova, s., Yerlikaya, G., Streubel, B., and Husslein, H. , 2017. Soluble VCAM-1/soluble ICAM-1 ratio is a promising biomarker for diagnosing endometriosis. Hum Reprod 32, 770-779. Liu, E., Nisenblat, V., Farquhar, C., Fraser, I., Bossuyt, P.M., Johnson, N., and Hull, M.L. , 2015. Urinary biomarkers for the non-invasive diagnosis of endometriosis. Cochrane Database Syst Rev, CD012019. Ohene-Abuakwa, Y., and Pignatelli, M. , 2000. Adhesion Molecules as Diagnostic Tools in Tumor Pathology. Int J Surg Pathol 8, 191-200. Vigano, P., Parazzini, F., Somigliana, E., and Vercellini, P. , 2004. Endometriosis: epidemiology and aetiological factors. Best Pract Res Clin Obstet Gynaecol 18, 177-200.
Katharina Proestling received her Ph.D. in Molecular Biology of Malignant Diseases from the Medical University of Vienna in 2009. After several years in cancer research she focused on endometriosis research. She is currently a PostDoc in the Department of Obstetrics and Gynecology at the Medical University of Vienna, Austria.
Table 1. Median Urine Levels of Adhesion Molecules in indicated Subgroups Endometriosisa No (n=65) Yes (n=84) Menstrual Cycle Phasea Proliferative (n=70) Secretory (n=75) rAFS scoreb Mild (1 and 2; n=43) Severe (3 and 4; n=41) BMIb Underweight (n=13) Normal weight (n=81) Overweight (n=31) Obesity (n=24) Cigarette Smokinga No (n=106) Yes (n=43) Entity of the lesionb Peritoneal (n=19) Ovarian (n=13) DIE (n=0) Combined (n=44) Others (n=6)
sICAM-1 pg/mg p-value 0.472 5575 (59057) 6153 (264217) 0.719 6267 (26020) 5575 (262883) 0.076 7707 (262883) 5296 (17733) 0.377 5212 (15892) 6184 (264217) 5125 (11719) 7652 (13063) 0.592 6040 (26020) 5575 (264069) 0.926 7192 (262883) 8200 (23863) 6040 (17733) 5641 (9765)
sVCAM-1 pg/mg p-value 0.262 370 (20717) 779 (4400305) 0.117 906 (52144) 381 (4400306) 0.533 673 (4400306) 862 (4325) 0.561 592 (4241) 539 (4400306) 966 (5713) 438 (20717) 0.632 542 (52144) 673 (4400306) 0.459 409 (4400306) 1368 (4077) 828 (4325) 164 (4254)
E-Selectin pg/mg p-value 0.232 292 (21318) 412 (149769) 0.459 475 (17631) 275 (149783) 0.258 309 (149769) 466 (17631) 0.921 264 (4998) 372 (149774) 484 (6487) 404 (9907) 0.603 406 (21318) 285 (149770) 0.429 298 (149769) 525 (6266)
P-Selectin pg/mg p-value 0.161 485 (20621) 551 (56417) 0.937 544 (7386) 478 (56417) 0.324 530 (56417) 680 (11287) 0.903 616 (2503) 530 (56417) 551 (4090) 442 (6010) 0.630 533 (20640) 530 (56417) 0.337 531 (56417) 643 (3440)
417 (17631) 80 (4405)
648 (11287) 251 (7337)
Data are reported as median values (pg) per mg creatinine; ranges (i.e. the difference between the largest and smallest values) are shown in parentheses; aThe non-parametric Mann‒Whitney U test and bKruskal‒Wallis test were used for comparison between groups; BMI, body mass index; rAFS, revised American Fertility Society score
Investigating selected adhesion molecules as urinary biomarkers for diagnosing endometriosis Katharina Proestling , Rene´ Wenzl , Iveta Yotova , Christoph Hauser , Heinrich Husslein , Lorenz Kuessel PII: DOI: Reference:
S1472-6483(20)30067-5 https://doi.org/10.1016/j.rbmo.2020.01.014 RBMO 2337
To appear in:
Reproductive BioMedicine Online
Received date: Accepted date:
21 December 2019 21 January 2020
Please cite this article as: Katharina Proestling , Rene´ Wenzl , Iveta Yotova , Christoph Hauser , Heinrich Husslein , Lorenz Kuessel , Investigating selected adhesion molecules as urinary biomarkers for diagnosing endometriosis, Reproductive BioMedicine Online (2020), doi: https://doi.org/10.1016/j.rbmo.2020.01.014
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo editing, typesetting, and review of the resulting proof before it is published in its final form. Please note that during this process changes will be made and errors may be discovered which could affect the content. Correspondence or other submissions concerning this article should await its publication online as a corrected proof or following inclusion in an issue of the journal. © 2020 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.
Investigating selected adhesion molecules as urinary biomarkers for diagnosing endometriosis Katharina Proestlinga, René Wenzla, Iveta Yotovaa, Christoph Hausera, Heinrich Hussleina*, Lorenz Kuessela a
Department of Gynecology and Obstetrics, Medical University of Vienna, Waehringer Guertel 18-20, 1090
Vienna, Austria
*Corresponding author. E-mail address: [email protected]
Abstract Research Question: Are selected cell adhesion molecules useful as urinary biomarkers for diagnosing endometriosis? Design: Prospective, longitudinal study (the Endometriosis Marker Austria) in patients who underwent laparoscopic surgery for benign gynecological pathologies. A total of 149 patients not receiving hormonal treatment for at least 3 months prior to recruitment were included and preoperative urine protein levels of soluble VCAM-1 (sVCAM-1), ICAM-1 (sICAM-1), E-selectin, and P-selectin were measured using a magnetic bead-based multiplex assay, normalized to creatinine levels of each sample. Levels were correlated with endometriosis status, menstrual cycle phase, body mass index, cigarette smoking and severity and entity of the lesions. Results: Urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin did not differ between women with (n=84) and without (n=65) endometriosis and among subgroups. Accordingly, ROC analysis to examine the value of using sVCAM-1, sICAM-1, E-selectin and P-selectin levels and sVCAM/sICAM ratio to diagnose endometriosis were not significant. In addition we examined whether the serum sVCAM-1 levels correlate with the urine levels of the protein in the same women, which revealed no significant correlations for sVCAM or sICAM. Conclusion: Although our previous study suggests that serum sVCAM is a promising biomarker for diagnosing endometriosis, we found no significant differences in urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin between women with and without endometriosis. Other markers should be studied in an effort to establish a truly non-invasive, urinary test for diagnosing endometriosis.
Keywords: endometriosis; biomarker; cell adhesion molecules; VCAM-1; ICAM-1
Introduction About 10% of reproductive-age women suffer from endometriosis, an estrogen dependent, chronic, disease causing pelvic pain and subfertility (Vigano et al., 2004). Laparoscopy is currently the gold standard for establishing the diagnosis of endometriosis, but it is expensive and carries surgical risks. At present a non-invasive test is not available in clinical practice, although the ability to accurately diagnose patients with endometriosis using a reliable, non–invasive approach would be of great value (Fassbender et al., 2015). Diagnostic markers have been evaluated in various body fluids, including urine, which is increasingly favored as a fluid for biological testing in biomarker discovery (Liu et al., 2015). The pathogenesis of endometriosis is not fully understood; A widely accepted hypothesis is that endometriosis originates from endometrial cells that adhere and implant on peritoneal surfaces via retrograde menstrual reflux. Cell adhesion molecules mediate cell adherence to extracellular matrix of the host tissue, enable cancer related biological processes and are related to the regulation of inflammatory and immune responses (Ohene-Abuakwa and Pignatelli, 2000). Contributing to a growing body of evidence regarding the role of cell adhesion molecules in endometriosis, our group demonstrated a significant overexpression of Vascular cell adhesion molecule-1 mRNA and elevated serum levels of soluble vascular cell adhesion molecule-1 (sVCAM1) in women with endometriosis, suggesting that sVCAM-1 is a promising diagnostic serum biomarker for endometriosis (Kuessel et al., 2017). Here we aimed to examine the value of selected cell adhesion molecules as urinary biomarkers for diagnosing endometriosis.
Materials & Methods Women were included in the EMMA study and samples and subgroups were collected and classified as described previously (Kuessel et al., 2017). Urine samples of women not on hormonal treatments for at least 3 months prior to recruitment were included.
Urine samples were obtained prior to laparoscopy, centrifuged within one hour of collection and stored in aliquots at -80°C. The urine concentrations of sVCAM-1, sICAM-1, E-selectin and P-selectin were measured in duplicate on the Luminex Bio-Plex 200 System (BioRad, California, USA) using Multiplex Luminex kits from RnD Systems (LKT007, RnD Systems, MN, USA) in accordance with the manufacturer’s protocol. The results below detection limit were set to 0pg/ml for the statistical analysis. Urine concentrations of each sample were normalized to the urine creatinine concentration for each sample expressed as pg/mg. Creatinine in urine was measured in duplicate using the Urinary Creatinine Assay Kit (STA-378; Cell Biolabs, San Diego, CA, USA). All statistical tests were performed using SPSS version 17.0. Non-parametric Mann‒Whitney U test and Kruskal‒Wallis test were used for comparison between groups. For paired statistics, the Wilcoxon signed-ranks test was used. Receiver operating characteristics (ROC) analysis was used to examine the diagnostic value. Spearman correlation analyses were used for correlation analysis between serum and urine levels of sVCAM-1 and sICAM-1. Differences with a p-value <0.05 were considered statistically significant.
Results In our present study, 84 (56.4%) women had and 65 (43.6%) did not have endometriosis. Women’s characteristics are shown in Table 1. Measured urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin did not differ significantly among groups (Table1). Accordingly, ROC analysis examining the value of using sVCAM-1, sICAM1, E-selectin and P-selectin levels and sVCAM/sICAM ratio to diagnose endometriosis were not significant (p=0.472, p=0.265, p=0.323, p=0.161, and p=0.545 for VCAM-1, ICAM-1, E-selectin and P-selectin levels and sVCAM/sICAM ratio, AUC<0.6 for all).
Exploratory subgroup analysis revealed no significant differences in urine levels between controls and woman with mild or severe endometriosis. Levels were not dependent on BMI, menstrual cycle phase, or cigarette smoking status (Table1).
Finally, by analyzing the experimental data from our current and previous study (Kuessel et al., 2017), we examined whether the serum sVCAM-1 and sICAM-1 levels correlate with the urine levels of the protein in the same women (n=59). Our analysis revealed no significant correlations (sVCAM-1: r=0.061; p=0.645 and sICAM-1: r=-0.197 p=0.135).
Discussion The field of endometriosis has an urgent need for an efficient, sensitive, non-invasive tool for diagnosing endometriosis. Researchers have searched for a suitable biomarker for several decades, primarily focusing on a multitude of different candidate markers in serum. Although urine is easier to access, shows a significantly narrower dynamic range of proteins and a less complex fluid composition compared to blood, only few endometriosis biomarker studies were urinebased (Fassbender et al., 2015). A systematic review aiming to evaluate the utility of urinary biomarkers found that most of the included studies were of retrospective design, evaluated either a specific phase of menstrual cycle or ignored cycle phase, and included only a limited number of samples; Furthermore, none of the potential biomarkers has been validated in separate patient cohorts (Fassbender et al., 2015; Liu et al., 2015). Our present study also has clear limitations, in particular the small sample size in the analyzed subgroups. However, in this study, we used a well-characterized, prospective study cohort and our robust design enabled us to perform exploratory subgroup analysis for lesion entities, disease stage, and menstrual cycle. Examining the utility of selected cell adhesion molecules as urinary biomarkers and investigating the correlation between serum and urine levels of sVCAM-1 and sICAM-1 in the same women, we found no significant differences between women with and without endometriosis, no correlation of urine levels with disease severity or patient characteristics, and no correlation between serum and urine levels of sVCAM-1 or sICAM-1. This lack of correlation between serum and urine levels of sVCAM-1 points to differences in the protein regulation in these biofluids. Published data from others and our own experience have shown that analyzing a small group of proven or suspected factors in the pathogenesis of the disease is unable to cover all aspects of the complex regulation of proteins and hypotheses can’t easily be transferred from one biofluid to another.
In conclusion, although our previous study suggests that serum sVCAM is a promising biomarker for diagnosing endometriosis, we found no significant differences between urine levels of sVCAM-1, sICAM-1, E-selectin and P-selectin between women with and without endometriosis. We believe that reporting information regarding well desingned negative trials can significantly contribute to focusing research on well-selected, better diagnostic targets.
Acknowledgements This study was co-funded by the Ingrid Flick Foundation (Grant no. FA751C0801), which played no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors declare no competing interests.
References Fassbender, A., Burney, R.O., O, D.F., D'Hooghe, T., and Giudice, L. , 2015. Update on Biomarkers for the Detection of Endometriosis. Biomed Res Int 2015, 130854. Kuessel, L., Wenzl, R., Proestling, K., Balendran, S., Pateisky, P., Yotova, s., Yerlikaya, G., Streubel, B., and Husslein, H. , 2017. Soluble VCAM-1/soluble ICAM-1 ratio is a promising biomarker for diagnosing endometriosis. Hum Reprod 32, 770-779. Liu, E., Nisenblat, V., Farquhar, C., Fraser, I., Bossuyt, P.M., Johnson, N., and Hull, M.L. , 2015. Urinary biomarkers for the non-invasive diagnosis of endometriosis. Cochrane Database Syst Rev, CD012019. Ohene-Abuakwa, Y., and Pignatelli, M. , 2000. Adhesion Molecules as Diagnostic Tools in Tumor Pathology. Int J Surg Pathol 8, 191-200. Vigano, P., Parazzini, F., Somigliana, E., and Vercellini, P. , 2004. Endometriosis: epidemiology and aetiological factors. Best Pract Res Clin Obstet Gynaecol 18, 177-200.
Katharina Proestling received her Ph.D. in Molecular Biology of Malignant Diseases from the Medical University of Vienna in 2009. After several years in cancer research she focused on endometriosis research. She is currently a PostDoc in the Department of Obstetrics and Gynecology at the Medical University of Vienna, Austria.
Table 1. Median Urine Levels of Adhesion Molecules in indicated Subgroups Endometriosisa No (n=65) Yes (n=84) Menstrual Cycle Phasea Proliferative (n=70) Secretory (n=75) rAFS scoreb Mild (1 and 2; n=43) Severe (3 and 4; n=41) BMIb Underweight (n=13) Normal weight (n=81) Overweight (n=31) Obesity (n=24) Cigarette Smokinga No (n=106) Yes (n=43) Entity of the lesionb Peritoneal (n=19) Ovarian (n=13) DIE (n=0) Combined (n=44) Others (n=6)
sICAM-1 pg/mg p-value 0.472 5575 (59057) 6153 (264217) 0.719 6267 (26020) 5575 (262883) 0.076 7707 (262883) 5296 (17733) 0.377 5212 (15892) 6184 (264217) 5125 (11719) 7652 (13063) 0.592 6040 (26020) 5575 (264069) 0.926 7192 (262883) 8200 (23863) 6040 (17733) 5641 (9765)
sVCAM-1 pg/mg p-value 0.262 370 (20717) 779 (4400305) 0.117 906 (52144) 381 (4400306) 0.533 673 (4400306) 862 (4325) 0.561 592 (4241) 539 (4400306) 966 (5713) 438 (20717) 0.632 542 (52144) 673 (4400306) 0.459 409 (4400306) 1368 (4077) 828 (4325) 164 (4254)
E-Selectin pg/mg p-value 0.232 292 (21318) 412 (149769) 0.459 475 (17631) 275 (149783) 0.258 309 (149769) 466 (17631) 0.921 264 (4998) 372 (149774) 484 (6487) 404 (9907) 0.603 406 (21318) 285 (149770) 0.429 298 (149769) 525 (6266)
P-Selectin pg/mg p-value 0.161 485 (20621) 551 (56417) 0.937 544 (7386) 478 (56417) 0.324 530 (56417) 680 (11287) 0.903 616 (2503) 530 (56417) 551 (4090) 442 (6010) 0.630 533 (20640) 530 (56417) 0.337 531 (56417) 643 (3440)
417 (17631) 80 (4405)
648 (11287) 251 (7337)
Data are reported as median values (pg) per mg creatinine; ranges (i.e. the difference between the largest and smallest values) are shown in parentheses; aThe non-parametric Mann‒Whitney U test and bKruskal‒Wallis test were used for comparison between groups; BMI, body mass index; rAFS, revised American Fertility Society score