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Papers by Kelly Tilleman

Reproductive BioMedicine Online

T cell reactivity against citrullinated proteins does not differ between healthy persons and pati... more T cell reactivity against citrullinated proteins does not differ between healthy persons and patients with inflammatory arthritis Objectives: Rheumatoid Arthritis (RA) and Spondyloarthropathy (SpA) are two inflammatory auto-immune diseases. They can be distinguished by clinical presentation and the presence of antibodies to citrullinated proteins (ACPA), which is specific for RA. Production of these serological markers indicates a predominant role for B-cells in the pathology of RA. However, little is known about the effect of citrullinated proteins on T-cell reactivity. Methods: PBMC were isolated from healthy volunteers (n=6), RA-patients (CCP+ (n=5) and CCP-(n=5)) and SpA-patients (n=6). The IFN-production was evaluated by ELISpot analysis. PBMC (500000/100µL) were stimulated with in vitro citrullinated (cit) and non-citrullinated (non-cit) human cell extract, each at a concentration of 200µg/ml and 20U/ml IL2 was added. In order to identify the cells that were crucial in the citrulline-induced T cell reactivity, depletion experiments for CD4, CD8 and HLA-DR were performed according to the manufacturer's protocol (Miltenyi Biotec). In parallel, 10^6 PBMC were cultured for 7 days in the presence of cit (80µg/ml) or non-cit (80µg/ml). Supernatants were collected and the secretion of cytokines was evaluated by multiple ELISAs (detecting IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17a, IFN , TNF , G-CSF and TGF). Results: In the cultures of PBMCs of healthy persons, the RA and the SpA patients, the number of spot forming counts (SFC) when stimulated with cit was higher than when stimulated with non cit (significant difference= SFC (cit)-SFC (non cit) > (SD (cit)+ SD (non cit)). A significant difference was seen in 5 of the 6 healthy volunteers and in all the SpA patients. Unexpectedly this phenomenon was less distinct in RA (2/5 CCP+ and 4/5 CCP-), while patients with inflammatory arthritis contain high amounts of citrullinated proteins in their joints. After depletion of HLA DR positive cells a major drop in reactivity was observed, which illustrates the crucial role of an antigen presenting cell in the citrulline-specific T cell reactivity. Depletion experiments for CD4 and CD8 positive cells showed that both cell types were involved. The cytokine profile, determined from the multiple ELISAs showed a great resemblance between healthy, RA and SpA patients. For IL2, IL10, IL17 and IL6 an overall greater response to cit compared to non cit was detected. On the contrary, for TGF , a greater response to non cit, compared to cit, was seen. Conclusion: These data show that citrulline-reactive IFN producing T-cells are present in the repertoire of RA patients, SpA patients and healthy people. This implicates that T cell stimulation with citrullinated proteins is a universal mechanism and that ACPA production in RA is not due to the presence of a unique set of citrulline-reactive T cells in the periphery of RA patients.

Journal of Assisted Reproduction and Genetics, Jul 20, 2019

Purpose The importance of the surrounding ovarian stromal cells and extracellular matrix in the d... more Purpose The importance of the surrounding ovarian stromal cells and extracellular matrix in the development and maturation of follicles has recently gained attention. An aberrant extracellular matrix has been described in ovaries of patients with polycystic ovary syndrome where a more rigid structural environment, possibly induced by endogenous testosterone, impairs normal folliculogenesis. In this context, we describe the textural parameters of the ovarian cortex of transgender men after prolonged testosterone administration compared to the textural parameters of the non-exposed ovarian cortex originating from female oncological patients. Methods Texture profile analysis (TPA) was performed on ovarian cortex (5 × 5 mm) of oncological and transgender patients in order to measure stiffness, hardness, cohesiveness, and springiness of the ovarian cortex (LRXplus universal testing system). Statistical analysis was performed using repeated measurements mixed models and the Spearman rank order correlation test (IBM SPSS Statistics 23). Results A total of 36 frozen-thawed cortical strips (5 × 5 mm) were subjected to TPA. The superficial part of cortex fragments originating from transgender persons (fragments < 1.4 mm; N = 10) appeared to be significantly stiffer compared to cortex derived from oncology patients (fragments < 1.4 mm; N = 7) (6.78 ± 1.38 N/mm versus 5.41 ± 0.9 N/mm respectively, p = 0.036). Conclusions This is the first application of TPA in ovarian cortex to study the physical properties. Comparing the physical properties, we objectively describe an increased cortical stiffness in the most outer part of the ovarian cortex following prolonged testosterone administration in transgender men compared to the ovarian cortex of oncological patients. This preliminary and novel approach could be the start of future research to understand the physical properties of ovarian tissue.

Although auto-antibodies against citrullinated proteins are well-known for diagnosis of rheumatoi... more Although auto-antibodies against citrullinated proteins are well-known for diagnosis of rheumatoid arthritis, the exact position of citrulline in these proteins, important to evaluate the conformation of the epitopes, remains elusive. The purpose of this study was to specifically modify citrullinated peptides to make them discernable from non-citrullinated peptides and to identify them by liquid chromatography coupled to mass spectrometry (LC-MS). Methods Citrullinated and non-citrullinated synthetic peptides were modified at 37°C with 2,3butanedione (BD) in TFA and were analysed by nanoLC-MS and by infusion tandem MS. Next, these peptides were spiked in a Cytochrome C-digest, after which the complete mixture was modified, followed by LC-MS analysis. Peptides were also purified from mixtures by immunoprecipitation (IP) with anticitrulline antibody bound to protein A-agarose beads and subsequently modified and identified. Results Modification with BD was specific for citrullinated peptides and produced a mass shift of 50Da. Complete modification was established after 16h with 50mM BD/30 µl TFA. Based on MSMS-spectra, the butanedione-adduct was proved to be covalently bound to the citrulline residue of the peptide and the amino acid sequence of the modified peptide could still be established at a concentration of 25pmol/µl. When synthetic citrullinated peptides were spiked into a cytochrome C-digest, complete modification of the citrullinated peptide remained possible in a 1/100 dilution (160fmol citrullinated peptide/16pmol cytochrome C-digest), even though the reaction did result in significant reduction in ionization capacity (p<0.01). Modified citrullinated peptides also showed a significant shift in retention time (p<0.0001). Non-citrullinated peptides of the cytochrome C-digest were not affected. In order to obtain comparatively higher ion counts of the modified citrullinated peptides, an IP experiment was conducted. 2 nmol of citrullinated peptide could be purified from a mixture with its non-citrullinated counterpart by IP with an anticitrulline antibody. Afterwards, modification of the purified citrullinated peptides resulted in the identification of the modified peptide by the specific additional mass shift and RT shift. Moreover a gain in ion count was observed; the proportion of citrullinated peptide as opposed to non-citrullinated peptide detected increased fivefold after IP (p< 0.05). Conclusion Specific modification of citrullinated proteins with 2,3-butanedione causes a 50Damass shift and a significantly longer retention of the modified citrullinated peptides on nanoLC-MS. Non-citrullinated peptides remained unaffected in mass and retention time. Also, peptides could be selectively purified from mixtures by immunoprecipitation. This opens possibilities for the identification of citrullinated peptides in complex mixtures and to identify the in vivo citrullinated status of proteins in the inflamed joint.

International Journal of Transgender Health

Background: Transgender healthcare is a rapidly evolving interdisciplinary field. In the last dec... more Background: Transgender healthcare is a rapidly evolving interdisciplinary field. In the last decade, there has been an unprecedented increase in the number and visibility of transgender and gender diverse (TGD) people seeking support and gender-affirming medical treatment in parallel with a significant rise in the scientific literature in this area. The World Professional Association for Transgender Health (WPATH) is an international, multidisciplinary, professional association whose mission is to promote evidence-based care, education, research, public policy, and respect in transgender health. One of the main functions of WPATH is to promote the highest standards of health care for TGD people through the Standards of Care (SOC). The SOC was initially developed in 1979 and the last version (SOC-7) was published in 2012. In view of the increasing scientific evidence, WPATH commissioned a new version of the Standards of Care, the SOC-8. Aim: The overall goal of SOC-8 is to provide health care professionals (HCPs) with clinical guidance to assist TGD people in accessing safe and effective pathways to achieving lasting personal comfort with their gendered selves with the aim of optimizing their overall physical health, psychological well-being, and self-fulfillment. Methods: The SOC-8 is based on the best available science and expert professional consensus in transgender health. International professionals and stakeholders were selected to serve on the SOC-8 committee. Recommendation statements were developed based on data derived from independent systematic literature reviews, where available, background reviews and expert opinions. Grading of recommendations was based on the available evidence supporting interventions, a discussion of risks and harms, as well as the feasibility and acceptability within different contexts and country settings. Results: A total of 18 chapters were developed as part of the SOC-8. They contain recommendations for health care professionals who provide care and treatment for TGD people. Each of the recommendations is followed by explanatory text with relevant references. General areas related to transgender health are covered in the chapters Terminology, Global Applicability, Population Estimates, and Education. The chapters developed for the diverse population of TGD people include Assessment of Adults, Adolescents, Children, Nonbinary, Eunuchs, and Intersex Individuals, and people living in Institutional Environments. Finally, the chapters related to gender-affirming treatment are Hormone Therapy, Surgery and Postoperative Care, Voice and Communication, Primary Care, Reproductive Health, Sexual Health, and Mental Health. Conclusions: The SOC-8 guidelines are intended to be flexible to meet the diverse health care needs of TGD people globally. While adaptable, they offer standards for promoting optimal health care and guidance for the treatment of people experiencing gender incongruence. As in all previous versions of the SOC, the criteria set forth in this document for gender-affirming medical interventions are clinical guidelines; individual health care professionals and programs may modify these in consultation with the TGD person.

International Journal of Transgender Health

Reproductive BioMedicine Online, 2021

RESEARCH QUESTION Lack of guidance on the frequency of Chlamydia trachomatis screening in non-par... more RESEARCH QUESTION Lack of guidance on the frequency of Chlamydia trachomatis screening in non-partner donors has led to heterogeneous testing protocols. C. trachomatis was checked in sperm donations unscreened for C. trachomatis to determine the risk for C. trachomatis infection in recipients using historic sperm donations unscreened for C. trachomatis. A C. trachomatis screening protocol is proposed to harmonize C. trachomatis screening, for which a cost evaluation is provided. DESIGN Retrospective study of sperm donations carried out between 2009 and 2019 from healthy non-partner donors for whom at least one straw was available. A straw was selected from the still available donations that had not been tested for C. trachomatis in urine at the time of donation. These sperm samples were screened for C. trachomatis by nucleic acid amplification (NAT). RESULTS Forty donors were included in the analysis. The 210 analysed straws tested negative for C. trachomatis. A C. trachomatis screening protocol following the European Centre for Disease Prevention and Control (ECDC) protocol for other sexually transmitted diseases (STD), i.e. NAT C. trachomatis screening of donor eligibility and first and last donation, provided these occur within 90 days, is cost advantageous compared with screening of all samples (approximately 75% reduction). CONCLUSION A negligible risk for C. trachomatis infection was found in recipients when using historical sperm samples stored at the sperm bank. C. trachomatis screening following the ECDC protocol for other STDs is supported as it significantly reduces workload and cost compared with screening all samples.

Manual of Embryo Culture in Human Assisted Reproduction, 2021

59th ESPE Annual Meeting (ESPE 2021 Online), 2021

Fertility and Sterility, 2022

To assess the best-performing machine learning (ML) model and features to predict euploidy in hum... more To assess the best-performing machine learning (ML) model and features to predict euploidy in human embryos. Retrospective cohort analysis. Department for reproductive medicine in a university hospital. One hundred twenty-eight infertile couples treated between January 2016 and December 2019. Demographic and clinical data and embryonic developmental and morphokinetic data from 539 embryos (45% euploid, 55% aneuploid) were analyzed. Random forest classifier (RFC), scikit-learn gradient boosting classifier, support vector machine, multivariate logistic regression, and naïve Bayes ML models were trained and used in 9 databases containing either 26 morphokinetic features (as absolute [A1] or interim [A2] times or combined [A3]) alone or plus 19 standard development features [B1, B2, and B3] with and without 40 demographic and clinical characteristics [C1, C2, and C3]. Feature selection and model retraining were executed for the best-performing combination of model and dataset. The main outcome measures were overall accuracy, precision, recall or sensitivity, F1 score (the weighted average of precision and recall), and area under the receiver operating characteristic curve (AUC) of ML models for each dataset. The secondary outcome measure was ranking of feature importance for the best-performing combination of model and dataset. The RFC model had the highest accuracy (71%) and AUC (0.75) when trained and used on dataset C1. The precision, recall or sensitivity, F1 score, and AUC were 66%, 86%, 75%, and 0.75, respectively. The accuracy, recall or sensitivity, and F1 score increased to 72%, 88%, and 76%, respectively, after feature selection and retraining. Morphokinetic features had the highest relative predictive weight. The RFC model can predict euploidy with an acceptable accuracy (>70%) using a dataset including embryos' morphokinetics and standard embryonic development and subjects' demographic and clinical features.

European Urology Open Science, 2020

Endocrine and reproductive outcome of men born with various degrees of hypospadias

Clinical Cancer Research, 2020

Liquid biopsies offer a minimally invasive alternative to tissue biopsies for both diagnosis and ... more Liquid biopsies offer a minimally invasive alternative to tissue biopsies for both diagnosis and monitoring of treatment response. Extracellular RNAs present in biofluids have emerged as potential biomarkers in health and disease, including cancer. While current studies typically focus on plasma or serum, other biofluids may contain more informative RNA molecules for particular disease types. Here, we present an unprecedented atlas of messenger, circular, and small RNA transcriptomes of a comprehensive collection of 20 different human biofluids (amniotic fluid, aqueous humor, ascites, bile, bronchial lavage fluid, breast milk, cerebrospinal fluid, colostrum, gastric fluid, pancreatic cyst fluid, plasma, saliva, seminal fluid, serum, sputum, stool, synovial fluid, sweat, tear fluid, and urine). By means of 173 synthetic RNA spike-in controls, we compared RNA content across biofluids, revealing a more than 10,000-fold difference in RNA concentration. Of interest, the circular RNA frac...

Reproductive BioMedicine Online, 2020

This study suggests that, in women with endometriosis, an impaired implantation factor contribute... more This study suggests that, in women with endometriosis, an impaired implantation factor contributes to reduced pregnancy outcomes after fertility treatment as well as impairing ovarian function. This relationship should be researched in future prospective studies, including the role of uterine receptivity in pregnancy outcomes.

Extracellular RNAs present in biofluids have emerged as potential biomarkers for disease. Where m... more Extracellular RNAs present in biofluids have emerged as potential biomarkers for disease. Where most studies focus on plasma or serum, other biofluids may contain more informative RNA molecules, depending on the type of disease. Here, we present an unprecedented atlas of messenger, circular and small RNA transcriptomes of a comprehensive collection of 20 different human biofluids. By means of synthetic spike-in controls, we compared RNA content across biofluids, revealing a more than 10 000-fold difference in RNA concentration. The circular RNA fraction is increased in nearly all biofluids compared to tissues. Each biofluid transcriptome is enriched for RNA molecules derived from specific tissues and cell types. In addition, a subset of biofluids, including stool, sweat, saliva and sputum, contains high levels of bacterial RNAs. Our atlas enables a more informed selection of the most relevant biofluid to monitor particular diseases. To verify the biomarker potential in these bioflui...

Fertility and Sterility, 2019

Objective: To develop a random forest model (RFM) to predict implantation potential of a transfer... more Objective: To develop a random forest model (RFM) to predict implantation potential of a transferred embryo and compare it with a multivariate logistic regression model (MvLRM), based on data from a large cohort including in vitro fertilization (IVF) patients treated with the use of single-embryo transfer (SET) of blastocyst-stage embryos. Design: Retrospective study of a 2-year single-center cohort of women undergoing IVF or intracytoplasmatic sperm injection (ICSI). Setting: Academic hospital. Patient(s): Data from 1,052 women who underwent fresh SET in IVF or ICSI cycles were included. Intervention(s): None. Main Outcome Measure(s): The performance of both RFM and MvLRM to predict pregnancy was quantified in terms of the area under the receiver operating characteristic (ROC) curve (AUC), classification accuracy, specificity, and sensitivity. Result(s): ROC analysis resulted in an AUC of 0.74 AE 0.03 for the proposed RFM and 0.66 AE 0.05 for the MvLRM for the prediction of ongoing pregnancies of R11 weeks. This RFM approach and the MvLRM yielded, respectively, sensitivities of 0.84 AE 0.07 and 0.66 AE 0.08 and specificities of 0.48 AE 0.07 and 0.58 AE 0.08. Conclusion(s): The performance to predict ongoing implantation will significantly improve with the use of an RFM approach compared with MvLRM. (Fertil Steril Ò 2019;111:318-26. Ó2018 by American Society for Reproductive Medicine.) El resumen está disponible en Español al final del artículo.

Reproductive BioMedicine Online, 2017

Chloë De Roo concluded her medical studies at the University of Ghent in 2012. She is combining h... more Chloë De Roo concluded her medical studies at the University of Ghent in 2012. She is combining her specialization in gynaecology and obstetrics with fundamental and applied scientific research at the Centre for Reproductive Medicine at the Ghent University Hospital in order to obtain a PhD degree in fertility preservation. KEY MESSAGE This study describes a surprisingly normal distribution of cortical follicles in the ovaries of trans men after more than a year of testosterone treatment. This work confirms the presence and in-vitro maturation potential of cumulus-oocyte complexes obtained during the tissue processing of ovaries procured in trans men.

Quality Management in ART Clinics, 2012

The high demands and expectations of our patients are placed on our standards of work—and quite r... more The high demands and expectations of our patients are placed on our standards of work—and quite rightly too, because we would have to account for the consequences of any mistakes we might make. Although countless couples become pregnant and give birth to healthy children, the number of unsuccessful treatments still outweighs the successful ones. The ART clinics are therefore under constant pressure to improve the quality of their services and increase the percentage of successful treatment cycles. A quality control system is the tool with which this expectation is met. It is a concept adapted from the industry sector. The aim of this is to establish procedures (standard work methods), which ensure that these established levels of quality are reached and maintained. Furthermore, the result must be visible in order to prove that standards are being adhered to and met.

Reproductive BioMedicine Online

T cell reactivity against citrullinated proteins does not differ between healthy persons and pati... more T cell reactivity against citrullinated proteins does not differ between healthy persons and patients with inflammatory arthritis Objectives: Rheumatoid Arthritis (RA) and Spondyloarthropathy (SpA) are two inflammatory auto-immune diseases. They can be distinguished by clinical presentation and the presence of antibodies to citrullinated proteins (ACPA), which is specific for RA. Production of these serological markers indicates a predominant role for B-cells in the pathology of RA. However, little is known about the effect of citrullinated proteins on T-cell reactivity. Methods: PBMC were isolated from healthy volunteers (n=6), RA-patients (CCP+ (n=5) and CCP-(n=5)) and SpA-patients (n=6). The IFN-production was evaluated by ELISpot analysis. PBMC (500000/100µL) were stimulated with in vitro citrullinated (cit) and non-citrullinated (non-cit) human cell extract, each at a concentration of 200µg/ml and 20U/ml IL2 was added. In order to identify the cells that were crucial in the citrulline-induced T cell reactivity, depletion experiments for CD4, CD8 and HLA-DR were performed according to the manufacturer's protocol (Miltenyi Biotec). In parallel, 10^6 PBMC were cultured for 7 days in the presence of cit (80µg/ml) or non-cit (80µg/ml). Supernatants were collected and the secretion of cytokines was evaluated by multiple ELISAs (detecting IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17a, IFN , TNF , G-CSF and TGF). Results: In the cultures of PBMCs of healthy persons, the RA and the SpA patients, the number of spot forming counts (SFC) when stimulated with cit was higher than when stimulated with non cit (significant difference= SFC (cit)-SFC (non cit) > (SD (cit)+ SD (non cit)). A significant difference was seen in 5 of the 6 healthy volunteers and in all the SpA patients. Unexpectedly this phenomenon was less distinct in RA (2/5 CCP+ and 4/5 CCP-), while patients with inflammatory arthritis contain high amounts of citrullinated proteins in their joints. After depletion of HLA DR positive cells a major drop in reactivity was observed, which illustrates the crucial role of an antigen presenting cell in the citrulline-specific T cell reactivity. Depletion experiments for CD4 and CD8 positive cells showed that both cell types were involved. The cytokine profile, determined from the multiple ELISAs showed a great resemblance between healthy, RA and SpA patients. For IL2, IL10, IL17 and IL6 an overall greater response to cit compared to non cit was detected. On the contrary, for TGF , a greater response to non cit, compared to cit, was seen. Conclusion: These data show that citrulline-reactive IFN producing T-cells are present in the repertoire of RA patients, SpA patients and healthy people. This implicates that T cell stimulation with citrullinated proteins is a universal mechanism and that ACPA production in RA is not due to the presence of a unique set of citrulline-reactive T cells in the periphery of RA patients.

Journal of Assisted Reproduction and Genetics, Jul 20, 2019

Purpose The importance of the surrounding ovarian stromal cells and extracellular matrix in the d... more Purpose The importance of the surrounding ovarian stromal cells and extracellular matrix in the development and maturation of follicles has recently gained attention. An aberrant extracellular matrix has been described in ovaries of patients with polycystic ovary syndrome where a more rigid structural environment, possibly induced by endogenous testosterone, impairs normal folliculogenesis. In this context, we describe the textural parameters of the ovarian cortex of transgender men after prolonged testosterone administration compared to the textural parameters of the non-exposed ovarian cortex originating from female oncological patients. Methods Texture profile analysis (TPA) was performed on ovarian cortex (5 × 5 mm) of oncological and transgender patients in order to measure stiffness, hardness, cohesiveness, and springiness of the ovarian cortex (LRXplus universal testing system). Statistical analysis was performed using repeated measurements mixed models and the Spearman rank order correlation test (IBM SPSS Statistics 23). Results A total of 36 frozen-thawed cortical strips (5 × 5 mm) were subjected to TPA. The superficial part of cortex fragments originating from transgender persons (fragments < 1.4 mm; N = 10) appeared to be significantly stiffer compared to cortex derived from oncology patients (fragments < 1.4 mm; N = 7) (6.78 ± 1.38 N/mm versus 5.41 ± 0.9 N/mm respectively, p = 0.036). Conclusions This is the first application of TPA in ovarian cortex to study the physical properties. Comparing the physical properties, we objectively describe an increased cortical stiffness in the most outer part of the ovarian cortex following prolonged testosterone administration in transgender men compared to the ovarian cortex of oncological patients. This preliminary and novel approach could be the start of future research to understand the physical properties of ovarian tissue.

Although auto-antibodies against citrullinated proteins are well-known for diagnosis of rheumatoi... more Although auto-antibodies against citrullinated proteins are well-known for diagnosis of rheumatoid arthritis, the exact position of citrulline in these proteins, important to evaluate the conformation of the epitopes, remains elusive. The purpose of this study was to specifically modify citrullinated peptides to make them discernable from non-citrullinated peptides and to identify them by liquid chromatography coupled to mass spectrometry (LC-MS). Methods Citrullinated and non-citrullinated synthetic peptides were modified at 37°C with 2,3butanedione (BD) in TFA and were analysed by nanoLC-MS and by infusion tandem MS. Next, these peptides were spiked in a Cytochrome C-digest, after which the complete mixture was modified, followed by LC-MS analysis. Peptides were also purified from mixtures by immunoprecipitation (IP) with anticitrulline antibody bound to protein A-agarose beads and subsequently modified and identified. Results Modification with BD was specific for citrullinated peptides and produced a mass shift of 50Da. Complete modification was established after 16h with 50mM BD/30 µl TFA. Based on MSMS-spectra, the butanedione-adduct was proved to be covalently bound to the citrulline residue of the peptide and the amino acid sequence of the modified peptide could still be established at a concentration of 25pmol/µl. When synthetic citrullinated peptides were spiked into a cytochrome C-digest, complete modification of the citrullinated peptide remained possible in a 1/100 dilution (160fmol citrullinated peptide/16pmol cytochrome C-digest), even though the reaction did result in significant reduction in ionization capacity (p<0.01). Modified citrullinated peptides also showed a significant shift in retention time (p<0.0001). Non-citrullinated peptides of the cytochrome C-digest were not affected. In order to obtain comparatively higher ion counts of the modified citrullinated peptides, an IP experiment was conducted. 2 nmol of citrullinated peptide could be purified from a mixture with its non-citrullinated counterpart by IP with an anticitrulline antibody. Afterwards, modification of the purified citrullinated peptides resulted in the identification of the modified peptide by the specific additional mass shift and RT shift. Moreover a gain in ion count was observed; the proportion of citrullinated peptide as opposed to non-citrullinated peptide detected increased fivefold after IP (p< 0.05). Conclusion Specific modification of citrullinated proteins with 2,3-butanedione causes a 50Damass shift and a significantly longer retention of the modified citrullinated peptides on nanoLC-MS. Non-citrullinated peptides remained unaffected in mass and retention time. Also, peptides could be selectively purified from mixtures by immunoprecipitation. This opens possibilities for the identification of citrullinated peptides in complex mixtures and to identify the in vivo citrullinated status of proteins in the inflamed joint.

International Journal of Transgender Health

Background: Transgender healthcare is a rapidly evolving interdisciplinary field. In the last dec... more Background: Transgender healthcare is a rapidly evolving interdisciplinary field. In the last decade, there has been an unprecedented increase in the number and visibility of transgender and gender diverse (TGD) people seeking support and gender-affirming medical treatment in parallel with a significant rise in the scientific literature in this area. The World Professional Association for Transgender Health (WPATH) is an international, multidisciplinary, professional association whose mission is to promote evidence-based care, education, research, public policy, and respect in transgender health. One of the main functions of WPATH is to promote the highest standards of health care for TGD people through the Standards of Care (SOC). The SOC was initially developed in 1979 and the last version (SOC-7) was published in 2012. In view of the increasing scientific evidence, WPATH commissioned a new version of the Standards of Care, the SOC-8. Aim: The overall goal of SOC-8 is to provide health care professionals (HCPs) with clinical guidance to assist TGD people in accessing safe and effective pathways to achieving lasting personal comfort with their gendered selves with the aim of optimizing their overall physical health, psychological well-being, and self-fulfillment. Methods: The SOC-8 is based on the best available science and expert professional consensus in transgender health. International professionals and stakeholders were selected to serve on the SOC-8 committee. Recommendation statements were developed based on data derived from independent systematic literature reviews, where available, background reviews and expert opinions. Grading of recommendations was based on the available evidence supporting interventions, a discussion of risks and harms, as well as the feasibility and acceptability within different contexts and country settings. Results: A total of 18 chapters were developed as part of the SOC-8. They contain recommendations for health care professionals who provide care and treatment for TGD people. Each of the recommendations is followed by explanatory text with relevant references. General areas related to transgender health are covered in the chapters Terminology, Global Applicability, Population Estimates, and Education. The chapters developed for the diverse population of TGD people include Assessment of Adults, Adolescents, Children, Nonbinary, Eunuchs, and Intersex Individuals, and people living in Institutional Environments. Finally, the chapters related to gender-affirming treatment are Hormone Therapy, Surgery and Postoperative Care, Voice and Communication, Primary Care, Reproductive Health, Sexual Health, and Mental Health. Conclusions: The SOC-8 guidelines are intended to be flexible to meet the diverse health care needs of TGD people globally. While adaptable, they offer standards for promoting optimal health care and guidance for the treatment of people experiencing gender incongruence. As in all previous versions of the SOC, the criteria set forth in this document for gender-affirming medical interventions are clinical guidelines; individual health care professionals and programs may modify these in consultation with the TGD person.

International Journal of Transgender Health

Reproductive BioMedicine Online, 2021

RESEARCH QUESTION Lack of guidance on the frequency of Chlamydia trachomatis screening in non-par... more RESEARCH QUESTION Lack of guidance on the frequency of Chlamydia trachomatis screening in non-partner donors has led to heterogeneous testing protocols. C. trachomatis was checked in sperm donations unscreened for C. trachomatis to determine the risk for C. trachomatis infection in recipients using historic sperm donations unscreened for C. trachomatis. A C. trachomatis screening protocol is proposed to harmonize C. trachomatis screening, for which a cost evaluation is provided. DESIGN Retrospective study of sperm donations carried out between 2009 and 2019 from healthy non-partner donors for whom at least one straw was available. A straw was selected from the still available donations that had not been tested for C. trachomatis in urine at the time of donation. These sperm samples were screened for C. trachomatis by nucleic acid amplification (NAT). RESULTS Forty donors were included in the analysis. The 210 analysed straws tested negative for C. trachomatis. A C. trachomatis screening protocol following the European Centre for Disease Prevention and Control (ECDC) protocol for other sexually transmitted diseases (STD), i.e. NAT C. trachomatis screening of donor eligibility and first and last donation, provided these occur within 90 days, is cost advantageous compared with screening of all samples (approximately 75% reduction). CONCLUSION A negligible risk for C. trachomatis infection was found in recipients when using historical sperm samples stored at the sperm bank. C. trachomatis screening following the ECDC protocol for other STDs is supported as it significantly reduces workload and cost compared with screening all samples.

Manual of Embryo Culture in Human Assisted Reproduction, 2021

59th ESPE Annual Meeting (ESPE 2021 Online), 2021

Fertility and Sterility, 2022

To assess the best-performing machine learning (ML) model and features to predict euploidy in hum... more To assess the best-performing machine learning (ML) model and features to predict euploidy in human embryos. Retrospective cohort analysis. Department for reproductive medicine in a university hospital. One hundred twenty-eight infertile couples treated between January 2016 and December 2019. Demographic and clinical data and embryonic developmental and morphokinetic data from 539 embryos (45% euploid, 55% aneuploid) were analyzed. Random forest classifier (RFC), scikit-learn gradient boosting classifier, support vector machine, multivariate logistic regression, and naïve Bayes ML models were trained and used in 9 databases containing either 26 morphokinetic features (as absolute [A1] or interim [A2] times or combined [A3]) alone or plus 19 standard development features [B1, B2, and B3] with and without 40 demographic and clinical characteristics [C1, C2, and C3]. Feature selection and model retraining were executed for the best-performing combination of model and dataset. The main outcome measures were overall accuracy, precision, recall or sensitivity, F1 score (the weighted average of precision and recall), and area under the receiver operating characteristic curve (AUC) of ML models for each dataset. The secondary outcome measure was ranking of feature importance for the best-performing combination of model and dataset. The RFC model had the highest accuracy (71%) and AUC (0.75) when trained and used on dataset C1. The precision, recall or sensitivity, F1 score, and AUC were 66%, 86%, 75%, and 0.75, respectively. The accuracy, recall or sensitivity, and F1 score increased to 72%, 88%, and 76%, respectively, after feature selection and retraining. Morphokinetic features had the highest relative predictive weight. The RFC model can predict euploidy with an acceptable accuracy (>70%) using a dataset including embryos' morphokinetics and standard embryonic development and subjects' demographic and clinical features.

European Urology Open Science, 2020

Endocrine and reproductive outcome of men born with various degrees of hypospadias

Clinical Cancer Research, 2020

Liquid biopsies offer a minimally invasive alternative to tissue biopsies for both diagnosis and ... more Liquid biopsies offer a minimally invasive alternative to tissue biopsies for both diagnosis and monitoring of treatment response. Extracellular RNAs present in biofluids have emerged as potential biomarkers in health and disease, including cancer. While current studies typically focus on plasma or serum, other biofluids may contain more informative RNA molecules for particular disease types. Here, we present an unprecedented atlas of messenger, circular, and small RNA transcriptomes of a comprehensive collection of 20 different human biofluids (amniotic fluid, aqueous humor, ascites, bile, bronchial lavage fluid, breast milk, cerebrospinal fluid, colostrum, gastric fluid, pancreatic cyst fluid, plasma, saliva, seminal fluid, serum, sputum, stool, synovial fluid, sweat, tear fluid, and urine). By means of 173 synthetic RNA spike-in controls, we compared RNA content across biofluids, revealing a more than 10,000-fold difference in RNA concentration. Of interest, the circular RNA frac...

Reproductive BioMedicine Online, 2020

This study suggests that, in women with endometriosis, an impaired implantation factor contribute... more This study suggests that, in women with endometriosis, an impaired implantation factor contributes to reduced pregnancy outcomes after fertility treatment as well as impairing ovarian function. This relationship should be researched in future prospective studies, including the role of uterine receptivity in pregnancy outcomes.

Extracellular RNAs present in biofluids have emerged as potential biomarkers for disease. Where m... more Extracellular RNAs present in biofluids have emerged as potential biomarkers for disease. Where most studies focus on plasma or serum, other biofluids may contain more informative RNA molecules, depending on the type of disease. Here, we present an unprecedented atlas of messenger, circular and small RNA transcriptomes of a comprehensive collection of 20 different human biofluids. By means of synthetic spike-in controls, we compared RNA content across biofluids, revealing a more than 10 000-fold difference in RNA concentration. The circular RNA fraction is increased in nearly all biofluids compared to tissues. Each biofluid transcriptome is enriched for RNA molecules derived from specific tissues and cell types. In addition, a subset of biofluids, including stool, sweat, saliva and sputum, contains high levels of bacterial RNAs. Our atlas enables a more informed selection of the most relevant biofluid to monitor particular diseases. To verify the biomarker potential in these bioflui...

Fertility and Sterility, 2019

Objective: To develop a random forest model (RFM) to predict implantation potential of a transfer... more Objective: To develop a random forest model (RFM) to predict implantation potential of a transferred embryo and compare it with a multivariate logistic regression model (MvLRM), based on data from a large cohort including in vitro fertilization (IVF) patients treated with the use of single-embryo transfer (SET) of blastocyst-stage embryos. Design: Retrospective study of a 2-year single-center cohort of women undergoing IVF or intracytoplasmatic sperm injection (ICSI). Setting: Academic hospital. Patient(s): Data from 1,052 women who underwent fresh SET in IVF or ICSI cycles were included. Intervention(s): None. Main Outcome Measure(s): The performance of both RFM and MvLRM to predict pregnancy was quantified in terms of the area under the receiver operating characteristic (ROC) curve (AUC), classification accuracy, specificity, and sensitivity. Result(s): ROC analysis resulted in an AUC of 0.74 AE 0.03 for the proposed RFM and 0.66 AE 0.05 for the MvLRM for the prediction of ongoing pregnancies of R11 weeks. This RFM approach and the MvLRM yielded, respectively, sensitivities of 0.84 AE 0.07 and 0.66 AE 0.08 and specificities of 0.48 AE 0.07 and 0.58 AE 0.08. Conclusion(s): The performance to predict ongoing implantation will significantly improve with the use of an RFM approach compared with MvLRM. (Fertil Steril Ò 2019;111:318-26. Ó2018 by American Society for Reproductive Medicine.) El resumen está disponible en Español al final del artículo.

Reproductive BioMedicine Online, 2017

Chloë De Roo concluded her medical studies at the University of Ghent in 2012. She is combining h... more Chloë De Roo concluded her medical studies at the University of Ghent in 2012. She is combining her specialization in gynaecology and obstetrics with fundamental and applied scientific research at the Centre for Reproductive Medicine at the Ghent University Hospital in order to obtain a PhD degree in fertility preservation. KEY MESSAGE This study describes a surprisingly normal distribution of cortical follicles in the ovaries of trans men after more than a year of testosterone treatment. This work confirms the presence and in-vitro maturation potential of cumulus-oocyte complexes obtained during the tissue processing of ovaries procured in trans men.

Quality Management in ART Clinics, 2012

The high demands and expectations of our patients are placed on our standards of work—and quite r... more The high demands and expectations of our patients are placed on our standards of work—and quite rightly too, because we would have to account for the consequences of any mistakes we might make. Although countless couples become pregnant and give birth to healthy children, the number of unsuccessful treatments still outweighs the successful ones. The ART clinics are therefore under constant pressure to improve the quality of their services and increase the percentage of successful treatment cycles. A quality control system is the tool with which this expectation is met. It is a concept adapted from the industry sector. The aim of this is to establish procedures (standard work methods), which ensure that these established levels of quality are reached and maintained. Furthermore, the result must be visible in order to prove that standards are being adhered to and met.