Human Papillomavirus Infections and the Role Played by Cervical and Cervico-Vaginal Microbiota-Evidence from Next-Generation Sequencing Studies.

This comprehensive review encompasses studies examining changes in the cervical and cervico-vaginal microbiota (CM and CVM) in relation to human papillomavirus (HPV) using next-generation sequencing (NGS) technology. HPV infection remains a prominent global health concern, with a spectrum of manifestations, from benign lesions to life-threatening cervical cancers. The CM and CVM, a unique collection of microorganisms inhabiting the cervix/vagina, has emerged as a critical player in cervical health. Recent research has indicated that disruptions in the CM and CVM, characterized by a decrease in Lactobacillus and the overgrowth of other bacteria, might increase the risk of HPV persistence and the progression of cervical abnormalities. This alteration in the CM or CVM has been linked to a higher likelihood of HPV infection and cervical dysplasia. NGS technology has revolutionized the study of the cervical microbiome, providing insights into microbial diversity, dynamics, and taxonomic classifications. Bacterial 16S rRNA gene sequencing, has proven invaluable in characterizing the cervical microbiome, shedding light on its role in HPV infections and paving the way for more tailored strategies to combat cervical diseases. NGS-based studies offer personalized insights into an individual's cervical microbiome. This knowledge holds promise for the development of novel diagnostic tools, targeted therapies, and preventive interventions for cervix-related conditions, including cervical cancer.

Postgraduate Training in Pediatric and Adolescent Gynecology: Trainees' Perspectives.

OBJECTIVE: The purpose of this study was to evaluate resident trainees' perspectives on the pediatric and adolescent gynecology (PAG) training in obstetrics and gynecology training programs in Europe. STUDY DESIGN: This study was a cross-sectional survey using an online questionnaire, on the basis of the PAG training in obstetrics and gynecology section of the European Board & College of Obstetrics and Gynaecology Project of Achieving Consensus in Training curriculum. We aimed to survey the national programs in 35 European Network of Trainees in Obstetrics and Gynaecology (ENTOG) member countries. Taking part in the survey was voluntary. The questionnaire was shared on the ENTOG online platforms. RESULTS: Ninety obstetrics and gynecology trainees in 33 of 35 countries responded to our questionnaire. Of the 35 ENTOG member countries, 33 participated in the survey, and a total of 90 responses were collected, giving a response rate of 9% of all European trainees and representing 94% of the member countries. Only 27% of trainees reported having a PAG rotation during their training program, and a PAG elective was only available to 34% of the trainees. Forty-one percent reported that PAG training was not included in their curriculum (no official rotations or lectures planned). Despite the lack of formal training, 72% of trainees felt able to diagnose and manage prepubertal vaginal bleeding and adnexal masses in children and adolescents by the end of their training. Most (58%) also confirmed that they could determine indications for treatment of vulval, vaginal, perineal, and rectal conditions. However, despite scoring positively for the management and counseling of subjects that often overlap with adult patients, such as "contraception in adolescents with health problems," "acute abdominal pain," "menstrual abnormalities," and "vaginal discharge," the study revealed poorer scores when the trainees were asked about more specific PAG topics such as "premature puberty" and "developmental disorders of the genital tract." CONCLUSION: Most core training programs across Europe do not include formal PAG training, and trainees reported a need to improve the provision of core PAG training in Europe.

The implementation of the European Working Time Directive in Europe and its impact on training in obstetrics and gynaecology: A ten year follow-up.

OBJECTIVE: To reassess the compliance with the European Working Time Directive (EWTD) in the member countries of the European Network of Trainees in Obstetrics and Gynaecology (ENTOG) and to investigate the impact of the EWTD on training. STUDY DESIGN: In this observational, cross-sectional study, an online questionnaire, containing multiple-choice questions and open questions, was distributed among Obstetrics and Gynaecology trainees in 33 ENTOG member countries. The questionnaire was designed as a follow-up of a similar survey conducted by ENTOG in 2009 and assessed the overall compliance with the EWTD, the adaptations needed to achieve this compliance, the impact of the EWTD on the quality of training and the well-being of trainees. The answers were analysed using descriptive statistics in Microsoft Excel. RESULTS: 59 responses from 28/33 (84.8%) ENTOG member countries were collected. Only 5 out of 28 (17.9%) countries were found to be nationally compliant with EWTD. There were no clear differences in the compliance between different types of the hospitals (university/teaching/district), but a trend was observed towards higher rate of implementation in smaller hospitals (<1500 deliveries per year). Regarding the changes needed to become EWTD-compliant and yet maintain high-quality training, the most common suggestions were: hiring extra junior doctors, restructuring training, having less doctors on duty simultaneously, consultants performing more hands on work, dedicated training sessions, reduction of administrative tasks and simulation training for surgical skills. The majority of trainees, 7 out of 12, (58.3%) in the EWTD-compliant hospitals experienced a positive effect on their training, whereas the majority of trainees in non-compliant hospitals, 31 out of 47, (66%) were uncertain about the impact of the EWTD on the quality of training. Among the positive changes, better work-life balance and more consultants available out of the daily working hours were listed. CONCLUSIONS: Despite the introduction and implementation of the EWTD over two decades ago, the compliance rates across Europe remain low and seem not to have altered in the last ten years. In order to ensure the quality of training and, most importantly patient safety, we suggest that European nations keep striving to implement the EWTD for doctors in training. We also suggest for nations that have yet to implement this directive to use the strategies as an exemplar in countries that follow EWTD.

Free Amino Acid Alterations in Patients with Gynecological and Breast Cancer: A Review.

Gynecological and breast cancers still remain a significant health problem worldwide. Diagnostic methods are not sensitive and specific enough to detect the disease at an early stage. During carcinogenesis and tumor progression, the cellular need for DNA and protein synthesis increases leading to changes in the levels of amino acids. An important role of amino acids in many biological pathways, including biosynthesis of proteins, nucleic acids, enzymes, etc., which serve as an energy source and maintain redox balance, has been highlighted in many research articles. The aim of this review is a detailed analysis of the literature on metabolomic studies of gynecology and breast cancers with particular emphasis on alterations in free amino acid profiles. The work includes a brief overview of the metabolomic methodology and types of biological samples used in the studies. Special attention was paid to the possible role of selected amino acids in the carcinogenesis, especially proline and amino acids related to its metabolism. There is a clear need for further research and multiple external validation studies to establish the role of amino acid profiling in diagnosing gynecological and breast cancers.

The influence of the COVID-19 outbreak on European trainees in obstetrics and gynaecology: A survey of the impact on training and trainee.

OBJECTIVE: The purpose of this study is to evaluate how the obstetrics and gynaecology residency program and trainees have been affected by the Corona Virus Disease-19 (COVID-19) pandemic in Europe. STUDY DESIGN: This study is a cross-sectional explorative survey using an online questionnaire. The questionnaire comprised of 40 questions that were subdivided into 4 subjects; workload, specialist training aspects in obstetrics and gynaecology, health and safety of the trainee and women's health and maternal health issues. Inclusion criteria consisted of being a trainee in Obstetrics and Gynaecology (ObGyn) at the time of the COVID-19 pandemic in Europe or trainees who had recently finished their training during the time of the outbreak. Taking part in the survey was voluntary. The questionnaire was shared on the website of the European Network for Trainees in Obstetrics and Gynaecology (ENTOG), ENTOG social media, in the ENTOG-newsletter and through the national representatives of ENTOG. RESULTS: 110 ObGyn trainees from 25 different countries responded to the questionnaire. Almost all trainees (95 %, N = 105) reported an effect on their training due to COVID-19 pandemic. Training was interrupted in 21 % of cases (n = 23). Trainees observed a decrease in educational activities or lectures and a decrease in number of patients. The possibility of training surgical skills decreased, because 67 % (N = 74) trainees reported that surgeries were cancelled. Trainees expressed concerns about reaching the goals of their ObGyn specialist training in 60 % (n = 66) of cases. A decrease in workload was experienced during the first COVID-19 wave in Europe by 60 % (n = 66) of trainees. On average these trainees worked 33 % less hours compared to a normal workweek. Although 22 % (n = 24) were expected to be available continuously for 24 h a day and 7 days a week for unscheduled duties, 15 % (n = 16) were deployed to work on special COVID-units. Concerning preparation, 45 % of the trainees (n = 50) had not received any training for treating COVID-positive patients. Trainees claimed to have enough personal protective equipment (PPE), although problems were reported. Any form of psychosocial support was arranged for 65 % of trainees (n = 71) by the hospital or department. The results of the survey suggest that obstetric care was not affected much (92 % (n = 102) of the respondents said at least necessary care continued) while patients in need for reproductive medicine were affected the most; out of the 110 departments 58 % (n = 60) were closed and 35 % (n = 36) reduced their activities. Access to family planning and benign gynaecology were also significantly reduced; 77 % and 87 % respectively of the departments were less accessible or only open to emergency cases. CONCLUSION: COVID-19 pandemic has had a tremendous effect on the ObGyn training in Europe. Exposure to learning opportunities, surgeries and teaching has been decreased during the outbreak and may result in a decrease in quality of care provided to women in the future if impairment of training is not recovered.

Serum Free Amino Acid Profiling in Differential Diagnosis of Ovarian Tumors-A Comparative Study with Review of the Literature.

Proper preoperative ovarian cancer (OC) diagnosis remains challenging. Serum free amino acid (SFAA) profiles were investigated to identify potential novel biomarkers of OC and assess their performance in ovarian tumor differential diagnosis. Serum samples were divided based on the histopathological result: epithelial OC (n = 38), borderline ovarian tumors (n = 6), and benign ovarian tumors (BOTs) (n = 62). SFAA profiles were evaluated using aTRAQ methodology based on high-performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Levels of eleven amino acids significantly differed between OC+borderline and BOTs. The highest area under the receiver operating characteristic curve (AUC of ROC) (0.787) was obtained for histidine. Cystine and histidine were identified as best single markers for early stage OC/BOT and type I OC. For advanced stage OC, seven amino acids differed significantly between the groups and citrulline obtained the best AUC of 0.807. Between type II OC and BOTs, eight amino acids differed significantly and the highest AUC of 0.798 was achieved by histidine and citrulline (AUC of 0.778). Histidine was identified as a potential new biomarker in differential diagnosis of ovarian tumors. Adding histidine to a multimarker panel together with CA125 and HE4 improved the differential diagnosis between OC and BOTs.

MALDI-MSI-A Step Forward in Overcoming the Diagnostic Challenges in Ovarian Tumors.

This study presents the use of matrix-assisted laser desorption and ionization mass spectrometry imaging (MALDI-MSI) directly on the tissue of two ovarian tumors that often present a diagnostic challenge, a low-grade serous borderline ovarian tumor and ovarian fibrothecoma. Different spatial distribution of m/z values within the tissue samples was observed, and regiospecific peaks were identified. Among the 106 peaks in the borderline ovarian tumor five, regiospecific peaks (m/z: 2861.35; 2775.79; 3368.34; 3438.43; 4936.37) were selected using FlexImaging software. Subsequently, the distribution of those selected peaks was visualized on the fibrothecoma tissue section, which demonstrated the differences in the tissue homo-/heterogeneous structure of both tumors. The comparison with the histopathological staining of the ovarian borderline tumor tissue section, obtained during serial sectioning, showed a close correlation of the molecular map with the morphological and histopathological features of the tissue and allowed the identification of different tissue types within the sample. This study highlights the potential significance of MSI in enabling morphological characterization of ovarian tumors as well as correct diagnosis and further prognosis than thus far seen in the literature. Osteopontin, tropomyosin and orosomucoid are only a couple of the molecules investigated using MALDI-MSI in ovarian cancer research. This study, in line with the available literature, proves the potential of MALDI-MSI to overcome the current limitations of classic histopathological examination giving a more in-depth insight into the tissue structure and thus lead to the more accurate differential diagnosis of ovarian tumors, especially in the most challenging cases.

Wide spectrum targeted metabolomics identifies potential ovarian cancer biomarkers.

AIMS: Despite of almost a hundred years of research on cancer metabolism, the biological background of cancerogenesis and cancer-related reprogramming of metabolism remains not fully understood. In order to comprehensively and effectively diagnose and treat the deadliest diseases, the mechanisms underlying these diseases have to be discovered urgently. Among the gynecological malignancies, ovarian cancer is the most common cause of death. The aim of the study was to search for potential cancer-related differences in concentrations of metabolites and interactions between them in serum of women with ovarian cancer and benign ovarian tumor in comparison with healthy controls using targeted metabolomics. These metabolites might serve as biomarkers in the future. MAIN METHODS: We used wide spectrum targeted metabolomics to evaluate serum concentrations of metabolites related to ovarian cancer and compared them against benign ovarian tumors and healthy controls. The measurements were performed using high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry technique in highly-selective multiple reaction monitoring mode. KEY FINDINGS: In this study we confirmed our previous findings about the role of histidine and citrulline in ovarian cancer as well as we indicated new lipid compounds (lysoPC a C16:1, PC aa C32:2, PC aa C34:4 and PC aa C 36:6) potentially involved in cancer metabolism. SIGNIFICANCES: We indicated interesting interactions between metabolites for further in-depth research which could potentially serve as clinically useful biomarkers in future. Moreover, the presented work attempts to visualize a possible 3D-network of relationships between the molecules found to be related to ovarian malignancy.

Assessment of diagnostic utility of multivariate diagnostic models in differential diagnosis of ovarian tumors.

INTRODUCTION: Ovarian cancer (OC) diagnosis remains a clinical challenge due to lack of early symptoms and insufficient accuracy of the available diagnostic methods. The purpose of this study was to determine whether osteopontin could be useful in differential diagnosis of ovarian tumors. MATERIAL AND METHODS: Serum samples from 92 patients qualified for surgical treatment due to ovarian mass were divided into 2 groups according to the histopathological result: OC including borderline ovarian tumors (n = 39) and benign ovarian tumors (BOTs) (n = 53). CA125, HE4 and osteopontin concentrations were measured in all patients. Areas under the receiver operating characteristic curves (AUC of ROC) were used to compare the discriminative ability of the univariate and multivariate diagnostic models. RESULTS: The addition of osteopontin to ROMA significantly improved the diagnostic performance of the test in 3 of the 5 analyses: 1) in the OC vs BOT group (from AUC of 0.955 to 0.975), 2) in premenopausal women OC vs BOT (from AUC of 0.828 to 0.892) and 3) in the FIGO I-II stage OC vs BOT (from AUC of 0.865 to 0.895). It did not alter the diagnostic performance of multifactor tests in the group of postmenopausal women nor in OC FIGO III-IV stage group. Osteopontin was also the best single marker to differentiate between early stage OC and BOTs (AUC of 0.863). CONCLUSIONS: Osteopontin improves the diagnostic performance of a multimarker OC diagnostic test and could be useful in differential diagnosis of ovarian tumors, especially in pre-menopausal women and for early stage OC.

Serum angiogenesis profile in gestational trophoblastic neoplasm using multiplex immunoassay.

AIMS: Gestational trophoblastic neoplasms (GTN) exemplify a rare, mostly curable but highly aggressive disease. It is often associated with a rapid formation of distant metastases and most likely with an intense neoangiogenesis processes. The aim of the study was to analyze markers in serum of patients with GTN before chemotherapy compared to healthy pregnant women. MAIN METHODS: In this study sixteen protein angiogenesis markers were evaluated in serum of 21 patients with GTN before chemotherapy and compared with healthy pregnant women. Markers were measured using BioPlex Pro Human Cancer Biomarker Panel 1 immunoassay. t-Tests and receiver operating characteristic curves were used for statistical analysis. KEY FINDINGS: Receiver operator curve analysis identified six proteins (sTIE-2, osteopontin, sIL-6α, sVEGFR-2, sEGFR, PECAM-1) which had sufficient sensitivity and specificity (AUC > 0,70) to distinguish GTN patients before the treatment from pregnant controls. The levels of three proteins (sTIE-2, osteopontin and sIL-6α) were altered in GTN patients before the treatment as compared to healthy controls (p = 0,0112; p = 0,0442; p = 0,0488, respectively) and thereby may serve as potential disease markers. SIGNIFICANCE: Serum concentration of proteins related to angiogenesis changes in the course of GTN and may appear useful in the diagnostic process of this disease.

Understanding Ovarian Cancer: iTRAQ-Based Proteomics for Biomarker Discovery.

Despite many years of studies, ovarian cancer remains one of the top ten cancers worldwide. Its high mortality rate is mainly due to lack of sufficient diagnostic methods. For this reason, our research focused on the identification of blood markers whose appearance would precede the clinical manifestation of the disease. ITRAQ-tagging (isobaric Tags for Relative and Absolute Quantification) coupled with mass spectrometry technology was applied. Three groups of samples derived from patients with: ovarian cancer, benign ovarian tumor, and healthy controls, were examined. Mass spectrometry analysis allowed for highlighting the dysregulation of several proteins associated with ovarian cancer. Further validation of the obtained results indicated that five proteins (Serotransferrin, Amyloid A1, Hemopexin, C-reactive protein, Albumin) were differentially expressed in ovarian cancer group. Interestingly, the addition of Albumin, Serotransferrin, and Amyloid A1 to CA125 (cancer antigen 125) and HE4 (human epididymis protein4) improved the diagnostic performance of the model discriminating between benign and malignant tumors. Identified proteins shed light on the molecular signaling pathways that are associated with ovarian cancer development and should be further investigated in future studies. Our findings indicate five proteins with a strong potential to use in a multimarker test for screening and detection of ovarian cancer.

Usefulness of Amino Acid Profiling in Ovarian Cancer Screening with Special Emphasis on Their Role in Cancerogenesis.

The aim of this study was to quantitate 42 serum-free amino acids, propose the biochemical explanation of their role in tumor development, and identify new ovarian cancer (OC) biomarkers for potential use in OC screening. The additional value of this work is the schematic presentation of the interrelationship between metabolites which were identified as significant for OC development and progression. The liquid chromatography-tandem mass spectrometry technique using highly-selective multiple reaction monitoring mode and labeled internal standards for each analyzed compound was applied. Performed statistical analyses showed that amino acids are potentially useful as OC biomarkers, especially as variables in multi-marker models. For the distinguishing metabolites the following metabolic pathways involved in cancer growth and development were proposed: histidine metabolism; tryptophan metabolism; arginine biosynthesis; arginine and proline metabolism; and alanine, aspartate and glutamine metabolism. The presented research identifies histidine and citrulline as potential new OC biomarkers. Furthermore, it provides evidence that amino acids are involved in metabolic pathways related to tumor growth and play an important role in cancerogenesis.

MALDI-TOF-MS analysis in discovery and identification of serum proteomic patterns of ovarian cancer.

BACKGROUND: Due to high mortality and lack of efficient screening, new tools for ovarian cancer (OC) diagnosis are urgently needed. To broaden the knowledge on the pathological processes that occur during ovarian cancer tumorigenesis, protein-peptide profiling was proposed. METHODS: Serum proteomic patterns in samples from OC patients were obtained using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF). Eighty nine serum samples (44 ovarian cancer and 45 healthy controls) were pretreated using solid-phase extraction method. Next, a classification model with the most discriminative factors was identified using chemometric algorithms. Finally, the results were verified by external validation on an independent test set of samples. RESULTS: Main outcome of this study was an identification of potential OC biomarkers by applying liquid chromatography coupled with tandem mass spectrometry. Application of this novel strategy enabled the identification of four potential OC serum biomarkers (complement C3, kininogen-1, inter-alpha-trypsin inhibitor heavy chain H4, and transthyretin). The role of these proteins was discussed in relation to OC pathomechanism. CONCLUSIONS: The study results may contribute to the development of clinically useful multi-component diagnostic tools in OC. In addition, identifying a novel panel of discriminative proteins could provide a new insight into complex signaling and functional networks associated with this multifactorial disease.

Diagnostic Value of Serum Angiogenesis Markers in Ovarian Cancer Using Multiplex Immunoassay.

As cancer development involves pathological vessel formation, 16 angiogenesis markers were evaluated as potential ovarian cancer (OC) biomarkers. Blood samples collected from 172 patients were divided based on histopathological result: OC (n = 38), borderline ovarian tumours (n = 6), non-malignant ovarian tumours (n = 62), healthy controls (n = 50) and 16 patients were excluded. Sixteen angiogenesis markers were measured using BioPlex Pro Human Cancer Biomarker Panel 1 immunoassay. Additionally, concentrations of cancer antigen 125 (CA125) and human epididymis protein 4 (HE4) were measured in patients with adnexal masses using electrochemiluminescence immunoassay. In the comparison between OC vs. non-OC, osteopontin achieved the highest area under the curve (AUC) of 0.79 (sensitivity 69%, specificity 78%). Multimarker models based on four to six markers (basic fibroblast growth factor-FGF-basic, follistatin, hepatocyte growth factor-HGF, osteopontin, platelet-derived growth factor AB/BB-PDGF-AB/BB, leptin) demonstrated higher discriminatory ability (AUC 0.80-0.81) than a single marker (AUC 0.79). When comparing OC with benign ovarian tumours, six markers had statistically different expression (osteopontin, leptin, follistatin, PDGF-AB/BB, HGF, FGF-basic). Osteopontin was the best single angiogenesis marker (AUC 0.825, sensitivity 72%, specificity 82%). A three-marker panel consisting of osteopontin, CA125 and HE4 better discriminated the groups (AUC 0.958) than HE4 or CA125 alone (AUC 0.941 and 0.932, respectively). Osteopontin should be further investigated as a potential biomarker in OC screening and differential diagnosis of ovarian tumours. Adding osteopontin to a panel of already used biomarkers (CA125 and HE4) significantly improves differential diagnosis between malignant and benign ovarian tumours.