Exploring the Role of E6 and E7 Oncoproteins in Cervical Oncogenesis through MBD2/3-NuRD Complex Chromatin Remodeling.

BACKGROUND: Cervical cancer is among the highest-ranking types of cancer worldwide, with human papillomavirus (HPV) as the agent driving the malignant process. One aspect of the infection's evolution is given by epigenetic modifications, mainly DNA methylation and chromatin alteration. These processes are guided by several chromatin remodeling complexes, including NuRD. The purpose of this study was to evaluate the genome-wide binding patterns of the NuRD complex components (MBD2 and MBD3) in the presence of active HPV16 E6 and E7 oncogenes and to determine the potential of identified genes through an experimental model to differentiate between cervical precursor lesions, with the aim of establishing their utility as biomarkers. METHODS: The experimental model was built using the CaSki cell line and shRNA for E6 and E7 HPV16 silencing, ChIP-seq, qRT-PCR, and Western blot analyses. Selected genes' expression was also assessed in patients. RESULTS: Several genes have been identified to exhibit altered transcriptional activity due to the influence of HPV16 E6/E7 viral oncogenes acting through the MBD2/MBD3 NuRD complex, linking them to viral infection and cervical oncogenesis. CONCLUSIONS: The impacted genes primarily play roles in governing gene transcription, mRNA processing, and regulation of translation. Understanding these mechanisms offers valuable insights into the process of HPV-induced oncogenesis.

Exploring Microbiota Diversity in Cervical Lesion Progression and HPV Infection through 16S rRNA Gene Metagenomic Sequencing.

(1) Background: Cervical cancer is a significant health concern, with the main cause being persistent infection with high-risk Human Papillomavirus (hrHPV). There is still no evidence for why viral persistence occurs in some women, but recent studies have revealed the interplay between cervical microbiota and hrHPV. This research aimed to characterize the cervicovaginal microbiota in cervical lesion progression and HPV infection status. (2) Methods: This study included 85 cervical specimens from women from the north-eastern region of Romania. DNA was isolated from cervical secretion for HPV genotyping and 16S ribosomal RNA gene NGS sequencing. (3) Results: Our study revealed a distinct pattern within the studied group when considering Lactobacillus species, which differs from findings reported in other populations. Specifically, the presence of Lactobacillus iners coupled with the absence of Lactobacillus crispatus alongside Atopobium spp., Prevotella spp., and Gardnerella spp. could serve as defining factors for severe cervical lesions. The results also showed a significant association between microbiota diversity, HPV infection, and cervical lesion progression. (4) Conclusions: As the microbiota profile seems to vary among different populations and individuals, a deeper comprehension of its composition has the potential to develop personalized detection and treatment approaches for cervical dysplasia and cancer.

Update on placenta accreta spectrum disorders by considering epidemiological factors, ultrasound diagnosis and pathological exam - literature review and authors' experience.

The aim of this paper was to correlate the circumstances that could lead to an abnormal invasion of placenta with the updated requirements to perform screening by ultrasound for all pregnant women prone to develop this pathology. To screen in the middle trimester of gestation for placenta accreta spectrum (PAS) disorders sets up an in-time referral opportunity for pregnant women prenatally detected with this pathology to a medical center with elevated level of expertise in the management of PAS disorders, able to act permanently by a multidisciplinary team (MDT) and to have access at medical resources including blood bank available. The literature review reveals especially useful data for clinical practice as regards novel explanations related to the etiology and physiopathology of PAS disorders, the composition of the MDT and the relevance of an indispensable pathologist physician at the time of Cesarean hysterectomy involved in the selection of best samples with the purpose of avoiding the possibility of losing undiagnosed cases with litigation implications. Conclusions show that the prenatal diagnosis of PAS disorders is possible so decreasing the risk of mortality and morbidity of pregnant women. Screening in the second trimester of pregnancy for PAS disorders becomes mandatory as the number of births by Cesarean section is expected to rise past three-fold until 2030. The professional expertise of the pathologist physician could be enriched by immunohistochemical staining in all suspected cases of placental invasion in myometrium wall.

Predicting Placenta Accreta Spectrum Disorders in a Cohort of Pregnant Patients in the North-East Region of Romania-Diagnostic Accuracy of Ultrasound and Magnetic Resonance Imaging.

Background: Placenta accreta spectrum (PAS) disorders are associated with high mortality and morbidity due to postpartum hemorrhage, hysterectomy, and organ injury, and a multidisciplinary team is required for an individualized case management. In this study, we assessed the diagnostic and prognostic accuracy of the most important ultrasonographic (US) and magnetic resonance imagining (MRI) markers for PAS disorders. Material and Methods: The study included 39 adult pregnant patients with at least one previous cesarean delivery and both US and MRI investigations for placenta previa evaluated at the tertiary maternity hospital 'Cuza Voda', Iasi, between 2019 and 2021. The following US signs were evaluated: intra-placental lacunae, loss of the retroplacental hypoechoic zone, myometrial thinning < 1 mm, bladder wall interruption, placental bulging, bridging vessels, and the hypervascularity of the uterovesical or retroplacental space. The MRI signs that were evaluated were intra-placental dark T2 bands, placental bulging, loss of the retroplacental hypointense line on T2 images, myometrial thinning, bladder wall interruption, focal exophytic placental mass, and abnormal vascularization of the placental bed. Results: The US and MRI signs analyzed in our study presented adequate sensitivities and specificities for PAS, but no sign proved to be a useful predictor by itself. The presence of three or more US markers for accretion was associated with a sensitivity of 84.6.6% and a specificity of 92.3% (p < 0.001). The presence of three or more MRI signs supplemented these results and were associated with a sensitivity of 92.3% and a specificity of 61.5% for predicting PAS (p < 0.001). Moreover, US and MRI findings were correlated with FIGO grading and severity of PAS. Conclusions: Even though no US or MRI finding alone can predict PAS with high sensitivity and specificity, our study proves that the presence of three or more imagistic signs could significantly increase the diagnostic accuracy of this condition. Furthermore, US and MRI could be useful tools for evaluating prognostic and perinatal planning.

Physicians' Opinion and Practice of Vitamin K Administration at Birth in Romania.

(1) Background. Vitamin K is recommended worldwide as a standard of care for the prophylaxis of vitamin-K-deficiency bleeding (VKDB). This is also a standard practice in Romania, but due to the rising number of refusals by parents of basic interventions in the neonatal period, we aimed to assess the Romanian neonatologists' opinions and current practice regarding vitamin K administration at birth. (2) Methods. We designed and conducted an electronic survey addressed to 110 physicians working in Romanian hospitals. (3) Results. Half of respondents are accustomed to receiving refusals for vitamin K administration once or twice a year. When parents refuse vitamin K administration, they usually refuse other neonatal interventions, according to 90.9% of the responding physicians, and this situation has occurred more frequently during the last two years. The number of refusals and especially their increase are more frequent in level III hospitals (p = 0.0304, p = 0.0036, respectively). Only 22.7% of the physicians responded that they would recommend an oral preparation of vitamin K in the absence of intramuscular prophylaxis. (4) Conclusion. Efforts should be made to address parents' concerns and to have available alternatives to the intramuscular administration of vitamin K.

Risk Factors for Urological Complications Associated with Caesarean Section-A Case-Control Study.

Background and Objectives: Acute urologic complications, including bladder and/or ureteric injury, are rare but known events occurring at the time of caesarean section (CS). Delayed or inadequate management is associated with increased morbidity and poor long-term outcomes. We conducted this study to identify the risk factors for urologic injuries at CS in order to inform obstetricians and patients of the risks and allow management planning to mitigate these risks. Materials and Methods: We reviewed all cases of urological injuries that occurred at CS surgeries in a tertiary university centre over a period of four years, from January 2016 to December 2019. To assess the risk factors of urologic injuries, a case-control study of women undergoing caesarean delivery was designed, matched 1:3 to randomly selected women who had an uncomplicated CS. Electronic medical records and operative reports were reviewed for socio-demographic and clinical information. Descriptive and univariate analyses were used to characterize the study population and identify the risk factors for urologic complications. Results: There were 36 patients with urologic complications out of 14,340 CS patients, with an incidence of 0.25%. The patients in the case group were older, had a lower gestational age at time of delivery and their newborns had a lower birth weight. Prior CS was more prevalent among the study group (88.2 vs. 66.7%), as was the incidence of placenta accreta and central praevia. In comparison with the control group, the intraoperative blood loss was higher in the case group, although there was no difference among the two groups regarding the type of surgery (emergency vs. elective), uterine rupture, or other obstetrical indications for CS. Prior CS and caesarean hysterectomy were risk factors for urologic injuries at CS. Conclusions: The major risk factor for urological injuries at the time of CS surgery is prior CS. Among patients with previous CS, those who undergo caesarean hysterectomy for placenta previa central and placenta accreta are at higher risk of surgical haemostasis and complex urologic injuries involving the bladder and the ureters.

How the SARS-CoV-2 Pandemic Period Influenced the Health Status and Determined Changes in Professional Practice among Obstetrics and Gynecology Doctors in Romania.

Background and Objectives: The beginning of the SARS-Cov-2 pandemic period has had a strong impact on patients' life, but also on doctors. The main goal of this research is to identify the difficulties related to the professional activity and personal life of obstetrics and gynecology doctors. Material and Methods: In total, 94 physicians from a single university center answered to an online questionnaire. Socio-demographic, health, family, and job-related data were collected. Data were processed using SPSS (v.25). Results: 7.4% of the doctors were confirmed infected with SARS-Cov-2 during the first 6 months of the pandemic, and 48.94% treated infected patients. Due to the large number of patients, 10.64% of the doctors have had no days-off during the last 6 months, and 22.34% of them have had new medical problems that led them to see a specialist. Seventeen to nineteen percent mentioned an increasing number of working hours and shifts per month due to the pandemic period, more than 10% used pills to cope with work-stress, and 25% of them had sleep disorders along with appetite loss. Extra-protection rules and negative consequences of wearing special equipment were identified: thermal discomfort that caused decreasing resistance and concentration during the surgery (52%), reduced mobility and accuracy of surgical or medical gestures (40%), and intraoperative visibility (47%). Doctors who were working with confirmed pregnant women preferred caesarean section. Conclusions: Working under the stress of an infection with SARS-Cov-2 is causing a lot of pressure and determines changes in personal, familial, social, and professional life. Understanding the challenges that ob-gyn doctors are facing will help institutions to better provide support.

Serological Parameters and Vascular Investigation for a Better Assessment in DVT during Pregnancy-A Systematic Review.

Pregnancy and the postpartum period represent a condition characterized by a thrombotic predisposition. The majority of pregnant women do not face acute or severe thrombotic events. In general, mild inconveniences such as leg swelling or moderately painful thrombotic events (phlebitis) are encountered. However, when pregnancy is associated with inherited or acquired deficits that affect homeostasis, the risk of acute or even life-threatening events can increase significantly. The major consequence is the loss of the fetus or the venous thromboembolism that endangers the mother's life. Venous thromboembolism is caused by deep vein thrombosis, therefore timely detection and especially the assessment of the extent of the thrombotic event are crucial. In this paper we have summarized the most important paraclinical investigations. The study emphasizes the importance of selecting the methods of investigation. The right choice allows establishing a correct diagnosis and individualizing the treatment.

Role of ProEx C immunocytochemistry in cervical high-grade squamous intraepithelial lesions detection.

BACKGROUND: Screening programs using Papanicolaou smear along with human papillomavirus (HPV) testing led to a significantly decrease of cervical cancer rates. Nevertheless, both assessments have limited specificity for revealing cervical high-grade lesions. The main problem is how to identify the real precursor of cervical squamous cell carcinomas (SCC), namely high-grade squamous intraepithelial lesions (HSIL). AIM: The aim of our study was to conclude if ProEx C might be used as a marker for high-grade cervical intraepithelial neoplasia (CIN). MATERIALS AND METHODS: In this study, we detected the immunochemical expression of anti-ProEx C antibody in liquid-based cytology (LBC) samples. We analyzed a total number of 125 cervical cytology specimens. RESULTS: In 48% of all cases, ProEx C was found to be positive. The percentage increased from 0% in negative for intraepithelial lesion or malignancy (NILM) cases to 100% in SCC cases. CONCLUSIONS: ProEx C may be utilized to improve the accuracy of cytological diagnosis on cervical smears, according to the findings of this study. This marker is also useful in detecting unrevealed high-grade lesions on atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesions (LSIL) smears, being very useful in establishing the conduct of these cases.

Morphological and histopathological changes in placentas of pregnancies with intrauterine growth restriction.

AIM: The definition of fetal growth restriction (FGR) refers to the incapability of a fetus to achieve the appropriate estimated growth, with expected fetal weight below the 10th percentile calculated for its gestational age. Placental factors and hypoxemia are considered to be essential elements with influence on intrauterine growth restriction (IUGR) and fetal death. The purpose of the present study was to investigate the macroscopic and microscopic pathological findings regarding the placentas in pregnancies complicated by influence on IUGR. PATIENTS, MATERIALS AND METHODS: Our study included 42 third-trimester pregnant patients admitted to the Cuza Voda Hospital of Obstetrics and Gynecology, Iasi, Romania, in the last three years. Soon after delivery, the 42 placentas were collected and analyzed; 32 placentas came from cases previously diagnosed with influence on IUGR and were included in our study group. Ten other placentas included in the control group were selected from uncomplicated pregnancies. Standard Hematoxylin-Eosin (HE) staining method, as well as Periodic Acid-Schiff (PAS) staining, and immunohistochemical techniques for cluster of differentiation 31 (CD31) and collagen IV were used in order to highlight the morphological features of the studied placentas. RESULTS: Our study revealed that reduced placental dimensions and eccentric umbilical cord insertion are correlated with the birthweight of the fetuses with IUGR (p<0.05). The most common histological finding in our study group was placental infarction later correlated with IUGR, but a certain causality could not be demonstrated, as this finding was also present in normal pregnancies. Other histopathological findings were also present in the influence on IUGR group, such as fibrin deposits, diffuse calcification, chronic villitis, avascular chronical villi, with no significant statistical correlations. CD31 was strongly immunoexpressed in the villous endothelial cells. Collagen IV presented a strong immunoreaction in the basement membrane and mesenchyme of the placental villi. CONCLUSIONS: Our study revealed a correlation between the dimensions of the diameters and volume of the maternal placenta and the presence of influence on IUGR. Moreover, it confirms the available data suggesting that the place of insertion of the umbilical cord is correlated with the weight of the fetus. Further studies with extended panel antibodies are needed in order to determine and complete the role of these morphological changes in the development of influence on IUGR.

Ovarian strumal carcinoid - case report.

Strumal carcinoid represents a rare form of ovarian teratoma, consisting of both thyroid tissue and carcinoid structures. The carcinoid component is a well-differentiated neuroendocrine tumor with excellent prognosis. Strumal carcinoid tumors are commonly found in peri-menopausal women who are not usually interested in preserving their fertility and who are thus open to radical surgical treatment. In this report, we present a 24-year-old, nulliparous patient with strumal carcinoid, confirmed by histopathology and a large panel of immunohistochemistry (IHC) markers, who wished to preserve her fertility. In this case, a conservative surgical treatment (salpingo-oophorectomy) served to preserve vital and reproductive prognosis, and correct tumor classification was of extreme importance. The morphological examination of strumal carcinoid showed struma ovarii with a thyroid follicle-like structure [positive for thyroid transcription factor 1 (TTF1), thyroglobulin, CD56, cytokeratin (CK) 19, and negative for Hector Battifora and mesothelioma 1 (HBME1)], and a neuroendocrine cell component with a trabecular arrangement and island growth (positive for synaptophysin, chromogranin, CD56, and CK7 negative), which were interlocked and intimately associated. Papillary thyroid carcinoma of follicular type was ruled out by CD56 positivity and HBME1 negativity. Medullary thyroid carcinoma with strumal component was excluded by calcitonin negative staining. Solid rosette-like structures with negative glial fibrillary acidic protein (GFAP) staining ruled out a neuroectodermal component. A multilocular mucinous cystadenoma was identified without other teratoma components. Strumal carcinoid requires a meticulous examination to rule out other entities with malignant behavior and poor prognosis. In this case, a conservative treatment is sufficient to remove the tumor, preserving vital and reproductive prognosis.

LINC01101 and LINC00277 expression levels as novel factors in HPV-induced cervical neoplasia.

Recently long non-coding RNAs were identified as new factors involved in gene expression regulation. To gain insight into expression pattern of these factors related to E7 HPV18 oncogene, this study uses HeLa cell culture transfected with E7-siRNA. Gene expression profile was investigated using microarray analysis. After analysing the microarray results, we identified 15,387 RNA species differentially expressed in E7-siRNA-transfected cells compared with controls (fold change >2). The expression profiles of lncRNA species highlighted 731 lncRNAs and 203 lincRNAs. We selected two lincRNAs (LINC01101 and LINC00277) and we evaluated the expression profile in HPV-induced neoplasia. Both lincRNAs investigated display a significantly reduced pattern of expression in cervical lesions and cancer, associated with clinical parameters. A connection between HPV presence and lincRNAs was noted. hrHPV-positive samples exhibit significantly reduced LINC01101 and LINC00277 expression level (P < 0.05). These results provide new insights into involvement of lncRNA in HPV-induced cervical cancer, enriching our understanding of their potential role in this pathology.

Pregnancy during Adolescence and Associated Risks: An 8-Year Hospital-Based Cohort Study (2007-2014) in Romania, the Country with the Highest Rate of Teenage Pregnancy in Europe.

Aim. To determine pregnancy and delivery outcomes among teenagers. Materials and Methods. An 8-year retrospective comparative hospital-based cohort study is analysing singleton pregnancy comorbidities and delivery parameters of a teenage group under the age of 20 compared with a young adult group 20-24 years of age in a university hospital. Results. Teenage is a risk factor for preterm birth <37 weeks (1.21 [1.08-1.35]), foetal growth restriction (1.34 [1.21-1.48]), episiotomy (1.27 [1.21-1.34]), uterine revision (1.15 [1.06-1.25]), APGAR <7 at 1 min (2.42 [1.21-1.67]), cephalopelvic disproportion (1.26 [1.07-1.48]), and postpartum haemorrhage (1.42 [1.25-1.62]); however, caesarean delivery occurs less frequently in teenagers than in adults (0.75 [0.70-0.80]). The following comorbidities are risk factors for teenage pregnancy (risk ratio [CI 95%]): anaemia (1.13 [1.10-1.17]), low urinary tract infection (1.10 [1.03-1.18]), pediculosis (2.42 [1.90-3.00]), anogenital condyloma (1.50 [1.04-2.17]), and trichomoniasis (1.74 [1.12-2.68]). The risks for hepatitis B and hepatitis C, premature rupture of membranes, and placenta praevia were lower compared with those in the young adult group, respectively, 0.43 (0.26-0.71), 0.90 (0.85-0.96), and 0.29 (0.20-0.41), while the risk for gestational diabetes and preeclampsia were the same in both groups. Conclusion. Considering the high risks for teenage pregnancy, this information should be provided to pregnant adolescent women and their caregivers.

Factors Influencing Burnout Syndrome in Obstetrics and Gynecology Physicians.

AIM: To identify the influence of environmental, personality, and alexithymia factors on burnout syndrome among obstetrics and gynecology physicians. MATERIAL AND METHODS: A total of 116 physicians (35 men and 81 women) completed questionnaires and sociodemographic data sheets. The Maslach Burnout Inventory, the Big Five Inventory, and the Toronto Alexithymia Scale were used to measure burnout, innate personality traits, and alexithymia, respectively. The t-test and Pearson correlations were used for other measurements. RESULTS: Of the 116 study physicians, 12.9% have suffered or still suffer from depression and 35.3% have had or still have problems related to insomnia. Regarding emotional exhaustion and depersonalization factors, men obtained lower scores than women (18.73 ± 13.48 versus 24.14 ± 11.71 for emotional exhaustion; 5.97 ± 5.45 versus 7.70 ± 5.29 for depersonalization). Self-reported depression was related to higher scores for all 3 domains, to higher total scores for alexithymia and neuroticism, and to lower scores for extraversion, conscientiousness, and openness. Residents and consultants had markedly different scores. CONCLUSION: The results of this study will help obstetrics and gynecology physicians cope with professional burnout and to consider personality traits, alexithymia domains, and environmental factors when finding strategies to cope with their professional stress.

Intrapartum diagnostic of Roberts syndrome - case presentation.

Roberts syndrome is a rare disease, with multiple limb and skeletal abnormalities (called "pseudothalidomide disease"). There are only around 150 cases described in literature. We present a case of Roberts syndrome, diagnosed in moment of delivery, after a pregnancy without prenatal follow-up. The stillborn baby was naturally delivered by a 17-year-old primiparous woman at 38 weeks of amenorrhea. The pregnancy was not followed due to socioeconomic and family situation, and no prenatal ultrasound was performed. The male baby has 2650 g and presented several morphological abnormalities and tight double umbilical abdominal loop. The macroscopic evaluation showed: dolichocephaly, hypoplastic inferior maxilla with micrognathia, antimongoloid palpebral slant, pterygium colli, abnormal and lower implanted ears, superior limbs phocomelia, syndactyly at lower left limb and tetradactyly in all limbs, bilateral cryptorchidism, pancreatic aplasia. Roberts syndrome is a rare genetic disease with recessive autosomal transmission generated by mutations in ESCO2 gene, located on chromosome 8. The disease should be easy to diagnose by antenatal ultrasound examination, but in our case, the lack of prenatal follow-up determined the diagnostic at term. We believe consider this case is an argument towards introducing ultrasound-screening compulsory to all pregnancies. To identify a possible genetic mutation, further investigations of the parents are in progress, but classically the disease has a recessive autosomal transmission.