Endocervical Cells Research Articles

Estrogen induces genomic instability in high-risk HPV-infected cervix and promotes the carcinogenesis of cervical adenocarcinoma

High-risk human papillomavirus (HPV) infection is the major cause of cervical cancer. However, the factors that modulate the process from infection to carcinogenesis are poorly understood. Although cervical cancer is clinically considered an estrogen-independent tumor, the role of estrogen in cervical cancer, particularly cervical adenocarcinoma, remains controversial. In this study, we showed that estrogen/GPR30 signaling induced genomic instability, which leads to carcinogenesis in high-risk HPV-infected endocervical columnar cell lines.The expression of estrogen receptors in a normal cervix was confirmed through immunohistochemical analysis which showed that G protein-coupled receptor 30 (GPR30) was predominantly expressed in endocervical glands and estrogen receptor-α (ERα) was expressed at higher levels in the squamous epithelium than in the cervical gland. E2 increased the proliferation of cervical cell lines, particularly normal endocervical columnar and adenocarcinoma cells via GPR30 rather than ERα, and increased the accumulation of DNA double-strand breaks (DSBs) in high-risk HPV-E6-expressing cells. The increase in DSBs was caused by the impairment of Rad 51 and accumulation of topoisomerase-2-DNA complexes under HPV-E6 expression. In addition, chromosomal aberrations increased in cells with E2-induced DSB accumulation. Collectively, we conclude that E2 exposure in high-risk HPV-infected cervical cells increases DSBs, leading to genomic instability and thus carcinogenesis via GPR30.

A Comparison of Conventional Pap Smear and Liquid-Based Cytology for Cervical Cancer Screening.

Early diagnosis and treatment of preinvasive lesions have made cervical cytology one of the most effective methods of cancer screening in industrialized nations, which have seen a sharp decline in the incidence and death of invasive cancer. The aim of this study is to compare liquid-based cytology (LBC) and conventional Pap on cervical smears. From July 2018 to June 2022, 600 patients were included in this cross-sectional study, which was done at the Pathology Department of a Tertiary Care Facility in Western Maharashtra. Of the 600 patients, 570 (95%) had good conventional Pap smear (CPS), whereas 30 (5%) had poor ones. Five hundred and ninety-two (98.6%) LBC smears were satisfactory, whereas 8 (1.4%) were unsatisfactory. Endocervical cells were seen in 294 (49%) CPS, whereas 360 (60%) LBC smears showed endocervical cells. The morphology of inflammatory cells was similar in both techniques. Hemorrhagic background was seen in 212 (35%) CPS and 76 (12.6%) LBC smears. Only two samples showed diathetic background, which was seen on both CPS and smear. Out of the satisfactory smears in the case of CPS, 512 (85%) cases were reported as negative for intraepithelial lesion or malignancy (NILM), whereas 58 (9.7%) cases were reported as epithelial cell abnormality. In LBC smears, 526 (87.3%) were reported as NILM, whereas 66 (11%) were reported as epithelial cell abnormality. Organisms were detected in 208 (34%) CPS and 162 (27%) LBC smears. Screening time was 5 ± 1 min for CPS, whereas it was 3 ± 1 min for LBC smear. Mortality will be decreased using LBC on a bigger scale in nations where many smears can be made and screened in a short amount of time, with the provision of doing human papillomavirus-based testing on the remaining sample.

HBXIP knockdown inhibits FHL2 to promote cycle arrest and suppress cervical cancer cell proliferation, invasion and migration.

Hepatitis B X-interacting protein (HBXIP) and four and a half LIM domain 2 (FHL2) have been reported to serve as independent biomarkers for cervical cancer. The present study evaluated the effects of HBXIP on cervical cancer in terms of its cellular malignant characteristics. Reverse transcription-quantitative PCR and western blotting were used to assess the mRNA and protein expression levels of HBXIP and FHL2 in the human endocervical epithelial End1/E6E7 cell line and the cervical cancer HeLa, CaSki, C33A and SiHa cell lines. After knocking down HBXIP expression by transfection of small interfering RNAs targeting HBXIP, cell cycle progression was assessed using flow cytometry with PI staining. Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine staining, wound healing and Transwell assays were used to assess cell proliferation, migration and invasion, respectively. Furthermore, co-immunoprecipitation assay was used to evaluate the potential binding relationship between HBXIP and FHL2. Western blotting was used for the analysis of HBXIP and FHL2, cell cycle-associated proteins, including cyclin D1 and cyclin D2, metastasis-associated proteins, including MMP2 and MMP9, and Wnt/β-catenin signaling-associated proteins, including β-catenin and c-Myc. Both HBXIP and FHL2 were found to be highly expressed in cervical cancer cells compared with that in the human endocervical epithelial cell line. HBXIP knockdown suppressed the proliferation, invasion and migration of HeLa cells, but promoted cell cycle arrest at the G0/G1 phase. HBXIP was demonstrated to interact with FHL2, and HBXIP knockdown also inhibited FHL2 mRNA and protein expression. By contrast, FHL2 overexpression reversed the inhibitory effects of HBXIP knockdown on the malignant characteristics of cervical cancer cells. Furthermore, HBXIP knockdown blocked the Wnt/β-catenin signaling pathway in HeLa cells, which was also partially reversed by FHL2 overexpression; the decreased β-catenin and c-Myc expression caused by HBXIP knockdown was increased again after FHL2 was overexpressed. In conclusion, these results suggest that HBXIP knockdown suppressed the malignant characteristics of cervical cancer cells through the downregulation of FHL2 expression, indicating a promising insight into the therapeutic target of cervical cancer.

Hormonal regulation of non-cystic fibrosis transmembrane conductance regulator ion channels in the endocervix

To characterize ion channel expression and localization in the endocervix under different hormonal conditions using a nonhuman primate primary endocervical epithelial cell model. Experimental. University-based, translational science laboratory. We cultured and treated conditionally reprogrammed primary rhesus macaque endocervix cells with estradiol and progesterone and measured gene expression changes for several known ion channel and ion channel regulators of mucus secreting epithelia. Using both rhesus macaque endocervical samples and human samples, we localized channels in the endocervix using immunohistochemistry. The relative abundance of transcripts was evaluated using real-time polymerase chain reaction. Immunostaining results were evaluated qualitatively. Compared with controls, we found that estradiol increased gene expression for ANO6, NKCC1, CLCA1, and PDE4D. Progesterone down-regulated gene expression for ANO6, SCNN1A, SCNN1B, NKCC1, and PDE4D (P≤.05). Immunohistochemistry confirmed endocervical cell membrane localization of ANO1, ANO6, KCNN4, LRR8CA, and NKCC1. We found several ion channels and ion channel regulators that are hormonally sensitive in the endocervix. These channels, therefore, may play a role in the cyclic fertility changes in the endocervix and could be further investigated as targets for future fertility and contraceptive studies.

Chlamydia trachomatis induces the transcriptional activity of host YAP in a Hippo-independent fashion.

The obligate intracellular pathogen Chlamydia trachomatis is the causative agent of the most common bacterial sexually transmitted disease worldwide. While the host response to infection by this pathogen has been well characterized, it remains unclear to what extent host gene expression during infection is the product of Chlamydia-directed modulation of host transcription factors. To identify transcription factors potentially modulated by Chlamydia during infection, we infected immortalized endocervical epithelial cells (End1/E6E7) with the anogenital C. trachomatis serovar L2, harvesting polyadenylated RNA for bulk RNA-sequencing. Subsequent experiments elucidating the mechanism of infection-mediated YAP activation assayed YAP target gene expression via qRT-PCR, YAP nuclear translocation via quantitative immunofluorescence, and YAP phosphorylation via Western blotting. RNA sequencing of Chlamydia-infected endocervical epithelial cells revealed gene expression consistent with activity of YAP, a transcriptional coactivator implicated in cell proliferation, wound healing, and fibrosis. After confirming induction of YAP target genes during infection, we observed an infection-dependent increase in YAP nuclear translocation sensitive to inhibition of bacterial protein synthesis. While Hippo-mediated phosphoinhibition of YAP at S127 was unaffected by C. trachomatis infection, Hippo-independent phosphorylation at Y357 was increased. Infection did not enhance nuclear translocation of Y357F mutant YAP, illustrating a requirement for phosphorylation at this residue. Pharmacological inhibition of host Src-family kinase activity attenuated YAP Y357 phosphorylation, but not nuclear translocation - which was instead sensitive to inhibition of Abl. Our results define a transcriptome-altering mechanism of pathogen-directed YAP activation that bypasses canonical inhibition by the Hippo kinase cascade, with a potential link to chlamydial fibrosis and other advanced disease sequelae. Additional study is required to determine the specific role of infection-associated Y357 phosphorylation and Abl activity in chlamydial induction of YAP.

Soluble E-cadherin: A marker of genital epithelial disruption.

The genital epithelial barrier is a crucial first line of defence against HIV, and epithelial disruption may enhance HIV susceptibility. Assessment of genital epithelial integrity requires biopsies, but their collection is not practical in many research settings. A validated biomarker of genital epithelial barrier integrity would therefore be useful. The purpose of this study was to evaluate soluble E-cadherin (sE-cad) as a marker of genital epithelial disruption. Using in vitro models of endocervical and foreskin epithelial cells, we assessed changes in sE-cad, IL-6, IL-1β, and IL-1α levels following mechanical disruption. We also assessed changes in sE-cad levels in vivo in cervicovaginal secretions after epithelial disruption by endocervical cytobrush sampling in Canadian women, and assessed the relationship between levels of sE-cad in coronal sulcus swabs to membrane-bound E-cadherin in the overlying foreskin tissue in Ugandan men. sE-cad levels immediately increased after in vitro epithelial physical disruption with the degree of elevation dependent on the extent of disruption, as did levels of IL-1β and IL-1α; this was followed by a delayed increase in IL-6 levels. In vivo results confirmed that sE-cad levels in cervicovaginal secretions were elevated 6h after cytobrush sampling when compared to baseline. Furthermore, levels of sE-cad in the prepuce were inversely correlated with the amount of membrane-bound E-cadherin of overlying tissue. Our results validate the use of sE-cad as a marker of epithelial disruption and demonstrate that the processes of physical disruption and inflammation in the genital tract are strongly intertwined.

Comparative Analysis of Conventional Cytology and Liquid-Based Cytology in the Detection of Carcinoma Cervix and its Precursor Lesions.

The conventional smears (CS) and Liquid based cytology (LBC) are important tools to detect carcinoma cervix and its precursor lesions. The present study was done to compare the cytomorphological features of cervical lesions using both techniques and compare with the histopathological diagnosis. This was a prospective observational study over a period of 1.5 years at a tertiary care hospital. A total of 969 women in the age group of 21-65 years presenting with either routine screening or complaints of vaginal bleeding, discharge, or pelvic pain were enrolled for the study. Both the CS and LBC smears were analyzed and compared with the corresponding histopathology diagnosis. The data was analyzed using Statistical Package for the Social Sciences (SPSS) software and P values <0.05 were considered significant. There were 8.57% unsatisfactory smears in CS as compared to 0.5% in LBC smears. Liquid-based cytology was superior to conventional preparations in terms of smear adequacy, lesser hemorrhagic and inflammatory background, and presence of more endocervical cells. Liquid-based cytology showed a better yield in detecting all the types of epithelial cell lesions with a concordance rate of 73.9% between the two techniques. On histopathology correlation of these lesions, LBC had a higher sensitivity (96.67%) and diagnostic accuracy (99.08%) as compared to CS (73.33% and 92.66%, respectively). Liquid-based cytology is superior to conventional cytology for the detection of epithelial cell lesions. Reduction in the unsatisfactory smears, a cleaner background, and better representation of the sample are more significantly appreciated on LBC in contrast to CS.

Xenobiotic metabolites modify immune output of the cervicovaginal epithelium: potential mechanisms underlying barrier disruption

Xenobiotic metabolites are exogenous biochemicals that can adversely impact reproductive health. We previously identified xenobiotics in cervicovaginal (CV) fluid during pregnancy in association with spontaneous preterm birth (sPTB). Suggesting bioactive potential, in vitro data reveal that these xenobiotics can modify CV epithelial barrier function. We hypothesize that xenobiotics dysregulate epithelial immune output as a mechanism to disrupt the CV barrier. Vaginal (VK2), ectocervical (Ecto), and endocervical (Endo) cell lines were treated with xenobiotics diethanolamine (DEA, 2.5mM), ethyl glucoside (EG, 5mM), tartrate (TA, 2.5mM) for 24 hours. A 41-plex cytokine Luminex panel and 5-plex matrix metalloproteinase (MMP) Luminex panel were run on cell media (n=3 per condition). Data were analyzed with a five-parameter logistic curve. One-way ANOVAs with Dunnett’s multiple comparisons tests were performed. DEA induces inflammatory cytokines while EG and TA generally exert anti-inflammatory effects across all cells (Fig. 1). DEA upregulates IL-6, IL-8, IP-10, MCP-1, GRO, fractalkine, MMP-1, MMP-9, and MMP-10 (p< 0.05 for all), factors involved in acute inflammation and recruitment of monocytes, neutrophils, and lymphocytes. EG and TA downregulate multiple cytokines, including RANTES and MCP-1 (p< 0.05 for all), which serve as chemotactic factors. VK2 cells exhibit higher inflammatory tone compared to cervical cells, with approximately twice the number of differentially detected cytokines after xenobiotic treatment. Cervical cells demonstrate similar immunologic responses to xenobiotics compared to VK2 cells. Xenobiotic metabolites present in the CV space during pregnancy modify epithelial immune output, unveiling potential immunologic pathways through which environmental exposures may contribute to the pathogenesis of cervical remodeling preceding sPTB. Future work identifying exogenous sources of these xenobiotics is warranted and offers potential to modify aspects of the environment to improve pregnancy outcomes. SMFM/AAOGF (KG), 5R01NR014784 (ME)

The level of inadequate Pap smears collected by the Gynecology Service of Fier Regional Hospital in Albania

This study aimed to determine the level of smears inadequate for Pap test in a population of women in Fier Prefecture in Albania. Material and methods: This is a cross-sectional study including 1254 women who showed up for a Pap test at the gynecology service at the Fier Regional Hospital during 2013 and 2014. The women presented to our service either on their own initiative to perform the Pap test or recommended by the family doctor for this examination. Standard criteria were applied to classify a cervical smear as inadequate for interpretation. Reasons for the inadequate taking of Pap smears are discussed. Results: A total of 234 patients (96.2% females) were included. The average age of women in our study was 39.8 years ± 10.7 (34.2% were 31-40 years old and 27.1% were 41-50 years old). Overall, 9.6% of the cervical smears they were taken inadequately (there was a need to repeat the obtaining of the smear). Almost all materials were obtained from the exocervix and endocervix (in 98.1% of cases for which there is information on this variable). The absence of endocervical cells was the most frequent reason for classifying a smear as unsuitable for interpretation. Insufficient equipment, scarce material for cytological analysis and poor technical preparation can lead to the non-detection of abnormalities and errors in the microscopic analysis of smears. Conclusions: Each member of the team is responsible for the accuracy of the results, but also for the adequacy of the sample. Inadequate smears could lead to the need for repeat tests and increased health care costs. Regular internal control, work quality monitoring as well as continuous training of all experts involved in obtaining, storing and interpreting cervical smears could reduce the level of inadequate Pap smears. Keywords: Albania, adequacy, cervical cancer, cervical smear, Pap test.

Never stop wondering: When cells become PAP-art on the slides.

The project named Victoria's cells was created to train health care personnel, especially in low-income countries. This innovative approach is designed to associate benign and malignant cellular images and/or patterns with a range of shapes and color shades to evoke animals, common objects, and colorful aquariums. The project makes use of familiar images to capture the viewer's interest as an aid for cytological interpretation. Cervicovaginal cytology is processed with conventional and liquid-based cytology. The images are visually compelling to highlight the importance of studying cells and their diagnostic significance. Infectious diseases as well as malignant cells are thereby easily recognized. The pictures are organized into different sections, including Victoria's zoo, Victoria's fantasy, and malignant mockery. Branching mycelia resemble a starfish; squamous metaplasia recalls a sea turtle's shell. Among others, different patterns of endometrial, endocervical, and squamous cells can resemble fish tanks populated by cells with the shapes of pufferfish, anglerfish, whales, scorpions, and garfish. The sea transitions to the earth, with a sly cat, a little elephant, a dog, and a koala. Other cellular preparations resemble a gymnast, a geisha, and a plunging diver as well as hummingbirds, a heron, a water lily, and a peony. The malignant mockery section is composed of squamous intraepithelial lesion patterns that resemble monsters, eyes, a foul tongue, eagles, and feathers. In conclusion, the recognition of visual images can make the study of cytology simpler and more enjoyable and serve the final objectives of prevention and cure.

Cytopathologic features of human papillomavirus–independent, gastric-type endocervical adenocarcinoma

BackgroundGastric-type endocervical adenocarcinoma (GEA) is unrelated to human papillomavirus (HPV) infection and is clinically aggressive compared with HPV-associated usual-type endocervical adenocarcinoma (UEA). The cytological diagnosis falls short of a definitive diagnosis of GEA and is often categorized as atypical glandular cells (AGCs). To improve cytologic recognition, cytological findings of HPV-independent GEA were analyzed and the results compared with HPV-associated UEA.MethodsCervical Papanicolaou (Pap) smears from eight patients with a histopathologic diagnosis of GEA and 12 control cases of UEA were reviewed. All slides were conventionally prepared and/or liquid-based prepared (ThinPrep) and stained following the Pap method. A mucinous background, architectural, nuclear, and cytoplasmic features were analyzed and compared with UEA.ResultsPreoperative cytologic diagnoses of the eight GEA cases were AGCs, favor neoplastic in three cases, adenocarcinoma in situ in one case, and adenocarcinoma in four cases. Cytologically, monolayered honeycomb-like sheets (p = .002) of atypical endocervical cells with vacuolar granular cytoplasm (p = .001) were extensive in GEA, and three-dimensional clusters (p = .010) were extensive in UEA. Although the differences were not statistically significant, background mucin (p = .058), vesicular nuclei (p = .057), and golden-brown intracytoplasmic mucin (p = .089) were also discriminatory findings for GEA versus UEA.ConclusionsAlthough GEA is difficult to diagnose on cytologic screening, GEA can be recognized based on cytologic features of monolayered honeycomb sheets of atypical endocervical cells with abundant vacuolar cytoplasm and some golden-brown intracytoplasmic mucin. UEA cases are characterized by three-dimensional clusters.

Exosomes from Ureaplasma parvum-infected ectocervical epithelial cells promote feto-maternal interface inflammation but are insufficient to cause preterm delivery.

This study determined if exosomes from ectocervical epithelial (ECTO) cells infected with Ureaplasma parvum (U. parvum) can carry bacterial antigens and cause inflammation at the feto-maternal interface using two organ-on-chip devices, one representing the vagina-cervix-decidua and another one mimicking the feto-maternal interface, and whether such inflammation can lead to preterm birth (PTB). Exosomes from U. parvum-infected ECTO cells were characterized using cryo-electron microscopy, nanoparticle tracking analysis, Western blot, and Exoview analysis. The antigenicity of the exosomes from U. parvum-infected ECTO cells was also tested using THP-1 cells and our newly developed vagina-cervix-decidua organ-on-a-chip (VCD-OOC) having six microchannel-interconnected cell culture chambers containing cells from the vagina, ectocervical, endocervical, transformation zone epithelia, cervical stroma, and decidua. The VCD-OOC was linked to the maternal side of our previously developed feto-maternal interface organ-on-a-chip (FMi-OOC). Cell culture media were collected after 48 h to determine the cytokine levels from each cell line via ELISA. For physiological validation of our in vitro data, high-dose exosomes from U. parvum-infected ECTO cells were delivered to the vagina of pregnant CD-1 mice on E15. Mice were monitored for preterm birth (PTB, < E18.5 days). Exosomes from ECTO cells infected with U. parvum (UP ECTO) showed significant downregulation of exosome markers CD9, CD63, and CD81, but contained multiple banded antigen (MBA), a U. parvum virulence factor. Monoculture experiments showed that exosomes from UP ECTO cells delivered MBA from the host cell to uninfected endocervical epithelial cells (ENDO). Moreover, exposure of THP-1 cells to exosomes from UP ECTO cells resulted in increased IL-8 and TNFα and reduced IL-10. The OOC experiments showed that low and high doses of exosomes from UP ECTO cells produced a cell type-specific inflammatory response in the VCD-OOC and FMi-OOC. Specifically, exosomes from UP ECTO cells increased pro-inflammatory cytokines such as GM-CSF, IL-6, and IL-8 in cervical, decidual, chorion trophoblast, and amnion mesenchymal cells. The results from our OOC models were validated in our in vivo mice model. The inflammatory response was insufficient to promote PTB. These results showed the potential use of the VCD-OOC and FMi-OOC in simulating the pathophysiological processes in vivo.

Histopathological findings in hysterectomy for cervical stenosis in postmenopausal women: A retrospective case series.

Objective:To analyze the histopathological findings in postmenopausal women who underwent hysterectomy for postsurgical cervical stenosis, evaluating the incidental findings of preinvasive or invasive uterine and cervical disease.Methods:Retrospective case series of postmenopausal women who underwent hysterectomy for postsurgical cervical stenosis at Gynecological Oncology Unit of Istituto di Ricovero e Cura a Carattere Scientifico Centro di Riferimento Oncologico Aviano—National Cancer Institute from January 2014 to January 2021.Results:During the study period, 36 women underwent hysterectomy for postsurgical cervical stenosis at our institution. Cervical stenosis occurred 10.2 ± 5.6 years from the onset of menopause. In particular, 26 (72.2%) patients underwent a single loop electrosurgical excision procedure or carbon dioxide (CO2)-laser conization before the onset of stenosis. The remaining 10 (27.8%) women had multiple surgical excision before the onset of stenosis. At the final histopathological analysis, 17 (47.2%) patients had a preinvasive or invasive gynecological disease. In particular, 9 cases of cervical disease (including 1 case of endocervical squamous cell carcinoma pT1a) and 6 cases of endometrial hyperplasia emerged. Also, 2 cases of tubo-ovarian diseases were found.Conclusions:Postsurgical cervical stenosis is a challenging clinical condition, especially in women treated for cervical intraepithelial neoplasia or microinvasive cervical cancer. As shown, cervical stenosis can prevent an adequate gynecological follow-up and a prompt diagnosis of malignancies. Therefore, postmenopausal women with cervical stenosis should be carefully counseled, and hysterectomy could be a reasonable option, especially in those cases in which a conservative approach is not feasible, failed, or is not accepted by the patient.

Immune parameters of HIV susceptibility in the female genital tract before and after penile-vaginal sex

BackgroundIn women, most HIV infections are acquired through penile-vaginal sex. Inflammation in the female genital tract (FGT) increases the risk of HIV acquisition and transmission, likely through recruitment of HIV target cells and disruption of epithelial barrier integrity. Although sex may have important immune and epithelial effects, the impact of receptive penile-vaginal sex on the immune correlates of HIV susceptibility in the female genital tract is not well described.MethodsSTI-free heterosexual couples were recruited to the Sex, Couples and Science (SECS) Study, with the serial collection of cervical secretions (CVS), endocervical cytobrushes, blood and semen before and up to 72 h after either condomless (n = 29) or condom-protected (n = 8) penile-vaginal sex. Immune cells were characterized by flow cytometry, and immune factors including cytokines and soluble E-cadherin (sE-cad; a marker of epithelial disruption) were quantified by multiplex immunoassay. Co-primary endpoints were defined as levels of IP-10 and IL-1α, cytokines previously associated with increased HIV susceptibility.ResultsHere we show that cervicovaginal levels of vaginal IP-10, sE-cad and several other cytokines increase rapidly after sex, regardless of condom use. The proportion of endocervical HIV target cells, including Th17 cells, activated T cells, and activated or mature dendritic cells (DCs) also increase, particularly after condomless sex. Although most of these immune changes resolve within 72 h, increases in activated cervical CD4 + T cells and Tcm persist beyond this time.ConclusionsPenile-vaginal sex induces multiple genital immune changes that may enhance HIV susceptibility during the 72 h post-sex window that is critical for virus acquisition. This has important implications for the mucosal immunopathogenesis of HIV transmission.

SPECTRUM OF CERVICAL LESIONS IN LIQUID BASED CYTOLOGY SMEAR IN A TERTIARY CARE CENTRE IN NORTH INDIA: A 3 YEAR RETROSPECTIVE STUDY

Cervical lesions are one of the most common cause of female morbidity, cervical cancer being the second most common malignancy only after breast in women in India as well as the world. The aim of this study was to analyse the spectrum of cervical lesions in Liquid based cytology and categorizing them according to the Bethesda System for Reporting Cervical cytology. This is a 3 year retrospective study . Total number of cases collected were 1228, of which majority were in the age group of 21-30 years comprising of 450 cases (36.6%) . The most common complain was whitish discharge followed by pain lower abdomen. Out of 1228 cases ,1131 (92.10%) were NILM, 11(0.89%) LSIL,10(0.80%) each of HSIL &amp;ASCUS,5(0.42%) Atypical endocervical cells(NOS), 4(0.35%) SCC and 2(0.16%) Atypical glandular cells(NOS) .55 cases (4.48%) were inadequate. Out of 1131cases of Negative for intraepithelial lesion or Malignancy, 1048 were non -neoplastic and 83 were associated with specic infection. Premalignant and malignant lesions of the cervix are very common, therefore proper screening and categorization of cervical lesion is necessary for treatment and follow up,thereby reducing the mortality and morbidity.

Functional evaluation of the cystic fibrosis transmembrane conductance regulator in the endocervix†.

The cystic fibrosis transmembrane conductance regulator (CFTR) is an apical membrane chloride/bicarbonate ion channel in epithelial cells. Mutations in CFTR cause cystic fibrosis, a disease characterized by thickened mucus secretions and is associated with subfertility and infertility. CFTR function has been well characterized in vitro and in vivo in airway and other epithelia studies. However, little is known about CFTR function in the cervix in health and its contribution to cyclic regulation of fertility from endocervical mucus changes. Contributing to this research gap is the lack of information on the effect of sex steroid hormones on CFTR expression in cervical epithelial cells across the menstrual cycle. Herein, we demonstrate the hormonal regulation of CFTR expression in endocervical cells both in vitro and in vivo, and that conditionally reprogrammed endocervical epithelial cells can be used to interrogate CFTR ion channel function. CFTR activity was demonstrated in vitro using electrophysiological methods and functionally inhibited by the CFTR-specific inhibitors inh-172 and GlyH-101. We also report that CFTR expression is increased by estradiol in the macaque cervix both in vitro and in vivo in Rhesus macaques treated with artificial menstrual cycles. Estrogen upregulation of CFTR is blocked in vivo by cotreatment with progesterone. Our findings provide the most comprehensive evidence to date that steroid hormones drive changes in CFTR expression. These data are integral to understanding the role of CFTR as a fertility regulator in the endocervix.

Primary fallopian tube epithelial cells are resistant to Chlamydia trachomatis infection due to high expression of ido1.

Abstract Chlamydia trachomatis (Ct) is the most prevalent sexually transmitted bacterial infection in the world. In ~50% of cisgender women, Ct ascends from the cervix to infect the upper genital tract and increased ascension is linked to hormonal therapies. Oviduct infection induces inflammation which can lead to tubal scarring and sequelae of infertility and ectopic pregnancy. Fortunately, these complications occur in a minority of infected women, suggesting that intrinsic factors may protect the fallopian tubes from infection or tissue damage. In vitro inoculation of primary fallopian tube epithelial (FTE) cells with Ct serovar E resulted in a 10-fold reduction in infected cells compared to transformed endocervical HeLa cells. Indoleamine 2,3-dioxygenase (IDO) in the cytoplasm of epithelial cells acts as a key mediator of chlamydial host defense by metabolizing tryptophan, an essential Ct nutrient. We determined that uninfected primary FTE cells expressed on average 9 times higher levels of ido1 transcripts by qPCR compared to HeLa cells, and that in vitro infectivity of FTE cells was rescued by addition of an IDO-inhibitor (9-fold), tryptophan (21-fold), or indole (a metabolite that bypasses tryptophan starvation; 16-fold). Induction of IDO has mostly been attributed to IFNγ, but it can also be induced by Type I IFNs. Detection of high constitutive ido1 transcription in primary FTE cells indicates their ability to produce IDO independently of IFNγ. Ongoing studies are examining the role of Type I IFNs, including IFNɛ, which is constitutively expressed by reproductive tract epithelium and regulated by hormones. This work has implications for defining factors that contribute to protection of women from Ct-induced disease. The study was funded through a Translational Team Science Award (UNC School of Medicine and NC TRaCS) and also funded by NIAID NIH R01 AI067678 to U.M.N., NIAID NIH R01 AI119164 to T.D., and NIH grant DK065988 and Cystic Fibrosis Foundation grant BOUCHE15R0 to S.H.R.

Atypical glandular cells (AGC): Cytology of glandular lesions of the uterine cervix.

The Pap smear is a well-known screening tool for squamous lesions of the uterine cervix. However, its screening role in glandular lesions is less effective. The incidence of squamous cell carcinoma of the cervix has dramatically decreased with the advent of Pap smear and recent understanding related to HPV carcinogenesis of cervical cancers including the advent of HPV vaccines. However, in recent years, the incidence of glandular abnormalities, diagnosed on Pap smears, has increased with greater sensitivity and precision.The incidence of atypical glandular cells (AGC) is approximately 0.18–0.74% of all cervical smears with a reported prevalence of 2.5% among all Pap smears. A high degree of suspicion, good clinical history, and the presence of diagnostic cytomorphological findings are essential for the proper interpretation of glandular cell abnormalities. A methodical approach to evaluate Pap smear greatly helps interpretation and avoids the diagnostic pitfalls. The Bethesda System for reporting cervical cytology has categorized glandular cell abnormalities into various categories as follows: Endocervical adenocarcinoma in situ (AIS)Atypical glandular cells (AGCs) Endocervical cells: a1 NOS or specify in comments; a2 Favor neoplasticEndometrial cells: NOS or specify in comments Adenocarcinoma (AdCa) EndocervicalEndometrialExtrauterineNOS Subtle differences in quantitative and qualitative cytologic features are essential for distinguishing one category from another. In this chapter, we highlight an organized approach for the interpretation of glandular abnormalities in Pap smear for our readers. This is an overview of the Bethesda categories, the reason for classification, and differential diagnosis with key characteristic features. An approach to the methodical evaluation of hyperchromatic crowded groups is discussed with key cytomorphologic differences. An algorithmic approach is suggested to facilitate the interpretation of various AGC categories.

Mucus secretions from a conditionally reprogrammed primary endocervical cell culture

To determine if the secretions collected from a conditionally reprogrammed primary endocervical cell culture are suitable surrogates for mucus studies. Experimental. University research center. Female rhesus macaque (n = 2). None. Quantitative proteomic analysis using tandem mass tag mass spectrometry liquid chromatography/tandem mass spectrometry. We identified 3,047 proteins, common proteins present in both primary endocervical cell cultures and the mucus of rhesus macaques. We found a 71% overlap in the top 500 most prevalent proteins in the samples. Cell culture secretions contained many essential mucus proteins, including MUC5B, the primary mucin of the endocervix. Similarities in secreted proteins suggest that conditionally reprogrammed primary endocervical cells could be used to study mucus secretion invitro.

Nonneoplastic Cervical Cytology.

Diagnostic cytology of cervix can be made strong if normal cytology is known thoroughly. Cervical lining comprises three layers of squamous cells, the basal, intermediate, and superficial cells. Knowing the dimensions of these cells, especially the intermediate cells, helps to diagnose the squamous intraepithelial lesions accurately. Furthermore, recognizing the parabasal cells in the menopausal smears, either singly or as syncytial aggregates, is important to avoid overdiagnosis of squamous intraepithelial lesions. The other cell type in the cervical lining is the endocervical glandular epithelium. Exfoliated endocervical cells may at times resemble endometrial glandular cells. The morphology and differences between these two cell types have been highlighted. It is essential to recognize and report endometrial cells in women of 40 years and above according to the recent Bethesda System for Reporting Cervical Cytology. The squamous epithelium of cervix and vagina is highly sensitive to estrogen and progesterone hormones. Hence, the Pap smears, if desired, can help in evaluating the hormonal status of the woman. The ratio of parabasal, intermediate, and superficial squamous cells can help in calculating various maturation indices.