Cytopathology

Intercenter Study of Interobserver Variability With Optical and Digital Imaging in Cervicovaginal Cytology. Simulation of a Quality Control Program

ABSTRACT Introduction Participation in diagnostic intercomparison programs is a mandatory requirement for accreditation of cytology laboratories. Results described by these programs in cervico vaginal cytology (CVC) show a significant number of discrepancies. We hypothesised whether the use of IA systems, such as the ThinPrep Genius system, would improve diagnostic concordance. Methods We simulated an intercomparison program between two centres in which four observers used the same 30 cases of CVC with negative diagnoses, ASCUS, LSIL and HSIL and observed them with light optic microscopy (OM) and two Genius (G1 and G2) systems, one from each centre. Results The diagnostic agreement between OM and the two Genius was similar, with overall agreement values of 82% for G1 and 78% for G2 and OM, and similar kappa values for the three techniques. However, both Genius diagnosed glandular lesions that were not included in the selected cases and were not diagnosed with OM. Conclusion The concordance obtained in both OM and Genius is acceptable and falls within the range of that obtained in what is described in intercomparison programs. We must consider the greater affinity for diagnosing glandular lesions with Genius and the fact that there may be discrete differences between two different devices, probably due to calibration.

Keratinization in atypical glandular cell clusters as a cytological clue to endometrioid carcinoma on cervical cytology

AbstractIntroductionSpecific diagnosis of endometrial carcinomas on cervical cytology is difficult with few useful cytomorphological clues reported. This study reviews a cohort of cervical cytology to investigate the presence of keratinization in atypical glandular cells (AGC), an undescribed cytomorphological clue for identifying endometrial endometrioid carcinomas on cervical cytology.MethodsCervical cytology slides from patients with a histologic diagnosis of endometrial endometrioid carcinoma were reviewed for the presence of keratinization associated with AGCs. Corresponding histology slides were reviewed for tumour grading and degree of squamous differentiation.ResultsIn total, 42 cases of cervical cytology specimens from 41 patients were retrieved, including 7 (16.7%) with keratinization associated with AGCs seen and 35 (83.3%) without. Comparison of histologic grading did not demonstrate an association with the presence of keratinization on cytology (p = 0.565). Corresponding histology slides were available for 37 cases. Cytologic and histologic keratinization were associated statistically (p = 0.002). Frank keratinization was seen on histologic slides of five cases, with four also showing cytologic keratinization. Area of squamous differentiation, including squamous morule formation, did not correlate with keratinization on cytologic preparation (p = 0.185).ConclusionHistologic and cytologic keratinization are observed in endometrioid endometrial carcinomas. Such is reflected in cervical cytology by the presence of orangeophilic, rigid and acellular fragments within or associated with AGC clusters. Keratinization, when identified with AGCs, should be regarded as a cytologic clue suggestive of an endometroid carcinoma of endometrial origin.

Evaluation of Diagnostic Accuracy of Directly Sampled Endometrial Cytology Using ThinPrep for Endometrial Malignancies: Comparison With Existing Endometrial Liquid‐Based Cytology

ABSTRACTObjectiveThe aim of this study was to evaluate the accuracy of detecting malignancies by directly sampled endometrial cytology using globally adopted ThinPrep with a novel preparation technique.MethodsMedical records and reports of pathology and cytology from June 2019 to March 2022 were reviewed. We selected 112 endometrial cytology specimens using ThinPrep with the novel preparation technique, where the clinical course or pathological samples confirmed negative or positive results. Eight cytotechnologists or cytopathologists examined the cytology specimens and provided reports based on standardised criteria from the descriptive reporting system for endometrial cytology (the Yokohama System).ResultsThe 112 specimens were evenly smeared and well prepared, featuring high quality, with no issues hindering microscopic examination. Examiners unfamiliar with endometrial cytology using ThinPrep showed high diagnostic accuracy, demonstrating that this modality of preparation for endometrial cytology is feasible for clinical use.ConclusionsThe novel preparation method using ThinPrep successfully provided high‐quality, standardised specimens. Furthermore, employing the Yokohama System enabled high accuracy in detecting endometrial malignancies, even for examiners with minimal experience with this cytological technique. This suggests that ThinPrep‐based endometrial cytology can be globally adopted with ease, potentially contributing significantly to the early detection of endometrial cancer.

The British Association for Cytopathology's recommended code of practice for cytology laboratories participating in the United Kingdom cervical screening programmes (2022) – A new edition for a new decade

This article gives an overview of the changes recently made to format and content of the BAC code of practice for cervical screening laboratories, reflecting the implementation of HPV primary screening across most of the UK in 2018/19. There is a link to the code itself, which encompasses the most up to date national guidance and provides key principle and standards for laboratories to work within.

Effects of the menstrual cycle on cytological false‐negatives in women with persistent cervical intraepithelial neoplasia

AbstractObjectivesFalse‐negatives on cytology may be observed during follow‐ups for patients with persistent cervical intraepithelial neoplasia (CIN); however, the underlying reasons are unknown, and the relationship between the intra‐individual variability of false‐negatives and the menstrual cycle phase has not been elucidated. Therefore, this study aimed to determine whether the menstrual cycle influences cytological results to maximise the accuracy of such tests.MethodsA total of 154 liquid‐based cytological (LBC) samples were obtained during follow‐ups for 26 patients with CIN, and the relationship between cytological results and the menstrual cycle, which was classified into six phases, was analysed.ResultsAll LBC smears were satisfactory, and 20 of 154 (13.0%) specimens were negative for intraepithelial lesions or malignancy (NILM). A statistically significant association was observed between the cytological results and the phase of the menstrual cycle, χ2(2) = 19.322, P < 0.01. The association was moderately strong (Cramer's V = 0.354). The value of adjusted residuals showed a statistically significant increase in the NILM percentage as a cytological result during the early secretory phase (P < 0.01) and a statistically significant decrease in NILM during the menstrual and proliferative phases (P < 0.01).ConclusionsThe present study revealed that false‐negative cytological results were found to more likely to occur during the early secretory phase. More careful and precise microscopic observation of Pap smears collected at the early secretory phase may contribute to a reduction in the occurrence of false‐negatives and improve cytological sensitivity.

Invasive cervical cancer audit: What lessons can we learn locally and where would we stand with regard to duty of candour?

AbstractObjectiveTo identify lessons learned locally from the invasive cervical cancer audit. To estimate the impact that the application of ‘Duty of Candour’ may have upon our future service provision.MethodsRetrospective cohort study with interval analysis of all women diagnosed with cervical cancer at Sheffield Teaching Hospitals NHS Foundation Trust between 1 April 2007 to 31 December 2019. Data were collected prospectively with retrospective categorisation by screening history and invasive cervical cancer audit outcomes as satisfactory, satisfactory with learning points, and unsatisfactory.Statistical analysis was performed using the chi‐squared test and paired t‐test.ResultsCervical cancer was diagnosed in 344 women. Seventy‐eight (23%) had no record of prior cervical cytology, 108 (31%) had delayed attendance to the screening programme, 102 (30%) were detected by routine screening, and 56 (16%) were screening programme compliant. Satisfactory management was undertaken in 301 (87.5%) cases, 26 cases (7.5%) were satisfactory with learning points, and 17 cases (5%) were considered as unsatisfactory.ConclusionsSeventeen cases were applicable to the Duty of Candour process equating to 1.3 cases per year, incurring minimal impact upon future service provision. Invasive audit categorisation is subject to bias, however, with the potential for considerable intra‐ and inter‐observer variation; the authors accordingly recommend that a further study be conducted to investigate both the consistency and reproducibility of the invasive cervical cancer audit categorisation.

Clinical Performance of a Liquid Preservation Medium for Cervicovaginal Samples in DNA‐HPV Testing and Liquid‐Based Cytology for Cervical Cancer Screening

ABSTRACTIntroductionCervical cancer remains a significant global health concern, primarily associated with persistent infections by high‐risk human papillomavirus (hr‐HPV). As screening programmes evolve from traditional cytology to DNA‐HPV testing, the need for a liquid medium that maintains the integrity of cervical samples for biomolecular analysis and cytology becomes critical.MethodsThis study evaluated the performance of the candidate liquid preservation medium (PM) Cytoliq for cervical samples intended for DNA‐HPV testing and liquid‐based cytology (LBC), in comparison with the reference PM, PreservCyt‐ThinPrep. A total of 112 women aged 18–64 years underwent routine gynaecological examinations, with paired cervical samples preserved in both PM for HPV testing and genotyping (Cobas HPV test), and LBC. The study aimed for a sensitivity greater than 90% in detecting cervical intraepithelial neoplasia grade 2 or worse (CIN2+), moderate to high agreement in HPV testing results (Kappa index > 0.70) and adequate performance in LBC.ResultsThe candidate PM exhibited non‐inferior performance relative to the reference PM. DNA‐HPV testing showed a 94.5% agreement rate (Kappa = 0.88) and a sensitivity of 92.9% for CIN2+ detection. Additionally, the candidate PM performed well in LBC smear production, with no significant differences in cytological diagnoses. The agreement in LBC diagnoses was 94.0% (Kappa = 0.79) with the ThinPrep processor and 91.8% (Kappa = 0.63) with the Cytoliq processor.ConclusionThe Cytoliq PM demonstrated comparable efficacy to the reference for DNA‐HPV testing and LBC, supporting its potential as an alternative preservation medium in cervical cancer screening programmes.

Cervical cancer screening efficacy using SurePath , ThinPrep and conventional cytology: A large data set analysis from the Japan Cancer Society

Abstract Objective Over the past decade, liquid‐based cytology has replaced conventional cytology for cervical cancer screening in many countries, including Japan. We aimed to evaluate the efficacy of liquid‐based cytology using a large database and compare two major liquid‐based cytology platforms, SurePath and ThinPrep, to conventional cytology. Methods Cervical cancer screening data were collected from the Japan Cancer Society between 2015 and 2019. The efficacy of liquid‐based and conventional cytology in detecting cervical intraepithelial neoplasia (CIN) was evaluated. Detection rates and positive predictive values were compared using a Poisson regression model. Results We collected data of 3,918,149 participants, including 2,248,202 conventional cytology, 874,807 SurePath and 795,140 ThinPrep smears. The detection rate of CIN2 or more was 1.14 times higher using SurePath than that using conventional cytology (95% confidence interval [CI], 1.09–1.20; p  < 0.001). Contrastingly, the detection rate of CIN2 or more was 0.91 times lower using ThinPrep (95% CI, 0.86–0.96; p  < 0.001). The detection rates of CIN3 or more did not differ significantly between SurePath and conventional cytology (detection rate ratio, 1.04; 95% CI, 0.97–1.12; p  = 0.224). The positive predictive value ratios of CIN2 or more were 0.80 using SurePath (95% CI, 0.76–0.84; p  < 0.001) and 0.83 using ThinPrep (95% CI, 0.79–0.87; p  < 0.001) compared with conventional cytology. Conclusions Liquid‐based cytology, particularly SurePath, was useful for detecting CIN2 or higher in population‐based cervical cancer screening. Further widespread use of liquid‐based cytology methods would lead to efficient detection of cervical precancerous lesions.

Does HPV‐18 co‐infection increase the risk of cervical pathology in individuals with HPV‐16?

Abstract Objective We aimed to investigate differences between HPV‐16 mono‐ and HPV‐16/18 co‐infections in terms of cervical dysplasia and invasive cancer. Methods This multicentre, retrospective study spanned from December 2017 to December 2020, involving women who visited gynaecological oncology clinics for colposcopy with either HPV‐16 or HPV‐16/18 positivity. A total of 736 patients, 670 in Group 1 (HPV‐16 positivity) and 66 in Group 2 (HPV‐16/18 positivity), were compared for the presence of CIN2+ lesions detected by colposcopic biopsy or endocervical curettage (ECC). Exclusions included hysterectomized patients, those with prior gynaecological cancers, and patients with HPV positivity other than types 16 and 18. Results Among the included patients, 42.4% had a diagnosis of CIN2+ lesions. The cytology results demonstrated abnormal findings in 45.3% in Group 1 and 42.2% in Group 2, with no significant difference between the groups. ECC revealed CIN2+ lesion in 49 (8.7%) patients in group 1, while only 1 (1.7%) patient had CIN2+ lesion in group 2. There was no difference between 2 groups in terms of ECC result ( p  = 0.052). In group 1, 289 (43.1%) patients had CIN2+ lesion, while 23 (34.8%) patients had CIN2+ lesions in group 2. There was no difference between group 1 and 2 in terms of diagnosis of CIN2+ lesions ( p  = 0.19). Conclusion This multicentre retrospective study found no significant differences between HPV‐16 mono‐ and HPV‐16/18 co‐infections regarding cervical pathologies. Larger studies are needed to validate and further explore these findings.

Expression of palladin is associated with disease progression in metastatic high‐grade serous carcinoma

AbstractObjectiveTo analyse the expression and clinical role of the actin‐associated molecule palladin in serous effusions.MethodsPALLD mRNA expression was analysed by quantitative reverse transcription polymerase chain reaction in 83 high‐grade serous carcinoma (HGSC) effusions. Fifteen malignant mesothelioma (MM) effusions and 18 surgical HGSC specimens from the ovary were studied for comparative purposes. Palladin protein expression by immunohistochemistry was analysed in another series consisting of 261 HGSC effusions.ResultsPALLD mRNA was significantly overexpressed in HGSC compared to MM effusions (P < .001). Palladin expression by immunohistochemistry was found in HGSC cells in 106/261 (41%) effusions, most commonly focally (<5% of cells). PALLD expression was additionally higher in ovarian HGSC specimens compared to HGSC effusions (P < .001). However, immunohistochemistry showed only stromal expression of this protein in surgical specimens. PALLD mRNA expression in HGSC effusions was unrelated to clinicopathological parameters, chemotherapy response or survival. Palladin protein expression was higher in post‐chemotherapy, mainly disease recurrence, specimens compared to chemo‐naïve effusions tapped at diagnosis (P = .018), although it was unrelated to other clinicopathological parameters or survival.ConclusionPALLD mRNA is overexpressed in HGSC compared to MM effusions, and its protein product is overexpressed in post‐chemotherapy compared to pre‐chemotherapy HGSC effusions, suggesting upregulation along tumour progression. The presence of this molecule in HGSC effusions, at the mRNA or the protein level, is unrelated to disease outcome.

Cervical cytology in a woman with abdominal distension

Exfoliated tumour cells from extra‐uterine origin are rarely identified in the cervical samples, however, may represent the first manifestation of an underlying malignancy. The present case describes the distinctive cytomorphologic features of a low‐grade ovarian serous carcinoma metastasising to the cervix.

Bilateral adnexal masses in a perimenopausal female

Small cell carcinoma of the ovary is an extremely rare ovarian malignancy with an aggressive clinical course. Establishing an early and accurate clinical diagnosis is challenging, owing to the rarity and non‐specific clinical presentations. We present a case of small cell carcinoma of the ovary in a perimenopausal female, diagnosed on fine needle aspiration cytology and confirmed by immunocytochemistry.

Cytodiagnosis of bilateral abdominopelvic masses in a postmenopausal woman

Cytologic diagnosis of ovarian low-grade serous carcinoma.

Ultrasound‐guided fine needle aspiration of ovarian masses: Assessment of diagnostic accuracy and risk stratification using a categorical reporting system

AbstractIntroductionThe present study was undertaken to assess the accuracy of fine needle aspiration cytology (FNAC) and cell‐block immunocytochemistry, and to estimate the risk of malignancy, using a categorical reporting system, in the diagnosis of ovarian masses.MethodsThis was a 5‐year retrospective study of FNAs of ovarian masses. The cytological diagnoses were categorised as inadequate, non‐neoplastic, benign neoplasms, indeterminate for malignancy, suspicious for malignancy and malignant neoplasms. The cytology was correlated with the corresponding histopathology to assess the diagnostic accuracy and risk of malignancy associated with each diagnostic category.ResultsOf a total of 66 703 FNAs performed during the study period, 580 (0.9%) were performed on ovarian masses. Of these, 40 (6.9%) were reported as non‐neoplastic; 76 (13.1%) as benign neoplasms; 14 (2.4%) as indeterminate for malignancy, 48 (8.3%) as suspicious for malignancy, 337 (58.1%) as malignant neoplasms and 65 (11.2%) as inadequate for interpretation. Immunocytochemistry (ICC) was performed on 99 (17%) aspirates. Subsequent histopathology was available in 208 (35.8%) cases. On cyto‐histopathological correlation, 183 (88%) were concordant and 25 (12%) were discordant. The overall sensitivity, specificity, positive and negative predictive values and diagnostic accuracy for diagnosing ovarian malignancy were 88.4%, 85.7%, 96.8%, 60.0% and 88% respectively. Risk of malignancy for each category was 80%, 0%, 4.5%, 66.7%, 88.5% and 98.5% respectively.ConclusionsUltrasound‐guided FNAC has high specificity and diagnostic accuracy for preoperative diagnosis of ovarian malignancies and hence is a valid diagnostic procedure in certain clinical situations. Reporting using a categorical system imparts uniformity and also provides the clinicians with an associated risk of malignancy to guide further management.

Fine needle aspiration diagnosed a rare case of ovarian clear cell carcinoma metastasis to thyroid gland

Metastatic deposits from malignant melanoma, renal clear cell carcinoma, and squamous cell carcinoma involving the thyroid gland have been reported before, but there are no previous reports of ovarian clear cell carcinoma metastatic to the thyroid. Cytological features and immunocytochemistry can help distinguish between primary and metastatic thyroid malignancies.

Smear detected cervicovaginal melanoma following negative screening—a cautionary tale of rapidly developing malignancy of the lower female genital tract

Cervicovaginal melanoma is a rare and aggressive neoplasm detectable by Pap smear. Non‐HPV‐related lesions, such as melanoma, can be rapidly progressive and unsuspected. Awareness of uncommon but clinically significant lesions on Pap smear is important.

Metastatic germ cell tumour in effusion cytology

AbstractBackgroundGerm cell tumours infrequently metastasise to body cavities, where early detection on fluid samples is possible and can spearhead early treatment and survival.Materials and methodsA total of seven cases of metastatic germ cell tumours were retrieved out of 7500 effusion samples received for cytopathological examination from 2015 to 2021. Detailed cytological features of metastatic germ cell tumours in effusion samples were studied, along with a correlation between clinical, radiological, and histopathological features.ResultsA total of seven cases of metastatic germ cell tumours were analysed in effusion samples which included dysgerminoma (2), immature teratoma (2), yolk sac tumour (1), embryonal carcinoma (1), and mixed germ cell tumour (1). The smears showed predominantly discrete or loose clusters of cells. The cells with round nuclei and prominent nucleoli were helpful in detecting dysgerminoma and yolk sac tumours. Immature teratoma showed tiny groups of small cells and mature squamous cells. Serum tumour markers were raised in the majority of cases.ConclusionMetastatic germ cell tumours in effusion are uncommon, but detailed clinical history, including serum markers and characteristic cytological features, are helpful in their diagnosis.

Gastric‐type endocervical adenocarcinoma and cervical cytology: Experience at a general hospital and review of the literature

AbstractIntroductionGastric‐type endocervical adenocarcinoma (GAS) is an uncommon type of endocervical adenocarcinoma that is not associated with human papillomavirus infection. This diagnosis is relatively rare and may portend a worse prognosis than usual‐type endocervical adenocarcinoma. Subtle morphological features make it an under‐recognised diagnostic challenge. Study of the cytological features of individual cases is valuable in order to increase awareness of this entity.MethodsThe pathology database of our institution was searched for the diagnosis of GAS and all cytological and surgical specimens for each patient were reviewed. The original cytological interpretation was compared to a retrospective central review interpretation. Clinical history and follow‐up results were obtained from the electronic medical record.ResultsFour cases of GAS were identified. The findings on initial cervical cytology varied, with GAS found in both patients with negative cervical cytology and those with atypical glandular cells. Cytological findings included endocervical cells arranged in three‐dimensional clusters and honeycomb sheets with abundant vacuolar cytoplasm, and in two patients, moderate nuclear atypia with irregular nuclear membranes, coarse chromatin, hyperchromatic nuclei, and prominent nucleoli. In one patient, GAS was incidentally discovered via thorough sampling of a cystic lesion in the superior portion of the endocervical canal.ConclusionsGAS is an aggressive human papillomavirus‐independent type of endocervical adenocarcinoma with subtle morphological features and, as our study shows, varying clinical presentation. Given the aggressive nature of GAS and the difficulties in initial diagnosis, increased awareness of this entity among pathologists is crucial.

Metastatic breast cancer diagnosed on cervical cytology

An unusual case is described of a 64‐year‐old woman with a remote history of breast cancer whose first non‐nodal metastasis was discovered on a routine Pap test. The Pap test was supported by the results of cervical and endometrial biopsies.

Is micronucleus assay a suitable tumour marker in cervical carcinogenesis? Some concepts and paradigms

AbstractThis letter discusses a previously published article in Cytopathology and provides comments the author believes allow better understanding regarding validating the micronucleus assay as a tumor marker in cervical carcinogenesis.

SARS‐CoV‐2 RNA may rarely be present in a uterine cervix LBC sample at the asymptomatic early stage of COVID 19 disease

AbstractObjectiveCurrently, it is thought that uterine cervix mucosal samples present a low risk of SARS‐CoV‐2 exposure. So far, there is no evidence of SARS‐CoV‐2 detection in Papanicolaou (Pap) smears. Nevertheless, clinicians could be exposed unaware to the coronavirus while performing and handling a Pap smear. We aimed to retrospectively evaluate the presence of SARS‐CoV‐2 RNA in cervical liquid‐based cytology (LBC) samples in women who tested positive for a nasopharyngeal COVID‐19 PCR test.MethodsFrom our laboratory database, we identified patients with data on a cervical cancer screening LBC sample paired with a positive nasopharyngeal COVID‐19 PCR test. Relevant LBC samples taken within an incubation period of 14 days and post‐onset RNA shedding interval of 25 days were subsequently tested for SARS‐CoV‐2 RNA using RT‐PCR tests.ResultsThe study group consisted of 102 women. Of those, 23 LBC samples were tested. SARS‐CoV‐2 RNA was detected in one LBC sample from a 26‐year‐old asymptomatic woman taken six days before reporting headaches and knee arthralgia with a positive nasopharyngeal SARS‐CoV‐2 RT‐PCR test.ConclusionsIt is possible to detect SARS‐CoV‐2 RNA in cervical LBC samples at an early asymptomatic stage of COVID‐19. In general, this finding is infrequent in asymptomatic women who tested SARS‐CoV‐2 positive within an incubation of 14 days and a post‐onset RNA shedding period of 25 days. We fully support the current thinking that cervical LBC samples from asymptomatic women pose a low risk of SARS‐CoV‐2 exposure and can be handled in the frame of good microbiological practice and procedures.

Bulky cervical tumour showing mixed unique cell clusters in cervical cytology

The cytological features of synchronous mucinous signet‐ring cell carcinoma and squamous cell carcinoma of the cervix have not been reported before. A case is described to emphasise that these two distinct carcinoma types can occur simultaneously in a high‐risk HPV infected cervix.

Assessment of micronuclei counts as tumour marker in cervical carcinogenesis: A follow‐up study

AbstractObjectiveMicronuclei counts were performed in cervical smears with low‐grade squamous intraepithelial lesions of the cervix (LSIL) to assess its potentiality as tumour marker in cervical carcinogenesis.MethodsThe cases studied were from the ongoing rural cervical cancer screening in west Lucknow, India. Micronuclei counts were performed in the cervical smears of 100 LSIL cases, and the number of cells with micronuclei was defined as micronucleated cells (MNC) and the number of micronuclei per 1000 cells as MNC score. Human papillomavirus (HPV) DNA testing was also done in 100 LSIL cases by GeneNav qPCR test.ResultsA high MNC score was found in 20 of the 100 LSIL cases while the counts were low in the remaining 80. Persistence of LSIL was seen in 19 of the 20 LSIL cases with high MNC score while only six cases of the 80 cases with low MNC score showed persistence. The persistence of LSIL was very high in cases with high MNC score. The multiple high‐risk HPV types such as 18, 31, 33 and 35 were seen in 12 of the 100 LSIL cases and a high positivity rate was seen in women with high MNC score. The persistence of LSIL was also higher with HPV positivity.ConclusionThe study revealed correlation between high MNC score, persistence of LSIL and HPV positivity. Hence, MNC score can prove to be very useful in discriminating high‐risk LSIL cases that are less likely to regress and possibly may progress to high‐grade squamous intraepithelial lesions or carcinoma.

Cytology of an ulceroproliferative cervical mass

The article describes a case of cervical melanoma diagnosed by cervical cytology. Mucosal melanoma, especially cervical melanoma, is rare and diagnosis in cervical cytology requires awareness of this entity.

Women's preferences for communication with the cervical screening programme: A qualitative study

AbstractBackgroundIn Scotland, invitations and results for cervical screening are sent by post. We ask the question: is this an effective means of communication in the 21st century? Consideration of other ways of communicating with women may help to increase acceptability of the cervical screening programme.ObjectiveTo explore perspectives of screening‐eligible women, regarding methods for communication of invitations and results from the cervical screening programme to improve acceptability.MethodsA qualitative study design using semi‐structured face‐to‐face or telephone interviews with women aged 25‐65 years. Thirty interviews were directed using visual cues to generate discussion. Interviews were audio‐recorded and transcribed verbatim. Thematic analysis of the data was conducted using a Framework approach.ResultsThe main advantage of the postal system is its perceived formality; however, its lack of speed was a concern. Advantages of e‐communication included speed and convenience; however, concerns such as lack of confidentiality and access were mentioned. Telephone communication was deemed impractical, while face‐to‐face communication was highly regarded. Furthermore, the majority of participants felt screening appointments set at a specific date and time may improve uptake. Overall, participants believed there is no universal solution regarding the issue of communication.ConclusionAt present, the postal system may be an appropriate method for invitation and results for cervical screening; however, there may be greater scope for preference of communication to improve the acceptability of the screening programme to women.

Reappraisal of cytology‐histology correlation in cervical cytology based on the recent American Society of Cytopathology guidelines (2017) at a cancer research centre

AbstractObjectiveTo assess the impact of recently published American Society of Cytopathology (ASC) guidelines (2017) on the conduct of cervical cytology‐histology correlation (CHC).MethodsA retrospective review was conducted for cervical biopsies with their corresponding conventional cervical smears over a 7.5‐year period (January 2011‐June 2018). As per the ASC guidelines, a discrepancy assessment grid was prepared. Major cytology‐histology discordance was defined as a diagnosis of high‐grade squamous intraepithelial lesion (HSIL) or CIN2+ in one of the tests with negative result in the other. Smears and biopsies of all discordant cases were reviewed for reasons of overcall and undercall.ResultsOf the 341 cervical biopsies with corresponding Papanicolaou smear, cytology‐histology agreement was noted in 249 (73%) cases. Major discordance was observed in 22 cases (6.4%)—16 undercalls and six overcalls on cytology—while minor discrepancies were noted in 70 cases. Atypical metaplasia and repair changes were the main reasons for overcall while small HSIL cells in atrophic smear and scant HSIL cells were important causes of undercall on cytology review. Using the ASC guidelines, we could improvise upon the existing CHC methodology for categorisation of cyto‐histological pairs of cases with a cytological diagnosis of atypical glandular cells.ConclusionThe present study demonstrates, for the first time, that the recent ASC guidelines facilitate cervical CHC, especially for categorisation of cases with atypical glandular cells on cytology. Uniform application of these guidelines would standardise the conduct of cervical CHC internationally and provide scope for inter‐laboratory comparison of data as well as enhance self‐learning and peer learning.

Quality metrics in cervical cytopathology: A single institutional study

AbstractObjectivesImplementation of quality control measures ensures acceptable performance by a laboratory. This study aims to assess the quality of cervical cytopathology reporting using quality metrics like atypical squamous cells (which include both atypical squamous cells of undetermined significance and atypical squamous cell ‐cannot rule out high grade squamous intraepithelial lesion)/squamous intraepithelial lesion (ASC/SIL), cytohistological correlation (CHC) and positive predictive value (PPV) of Papanicolaou (Pap) smears for squamous lesions of cervix.MethodsA retrospective study of Pap smears from 2015 to 2020 was performed. The quality metrics analysed include diagnoses of ASCUS, ASC‐H and ASCUS/SIL ratio, CHC and PPV. Cases with cervical biopsies/hysterectomy were included for CHC, and discrepancy was defined as discordance in diagnostic category between cytology and histology in the CHC.ResultsA total of 22,695 cervical cytology smears were reported. Unsatisfactory smears (n = 290) were excluded. Squamous lesions were reported in 233 smears, and the Bethesda system of nomenclature was followed. A definitive diagnosis (SILs and SCC) was given in 74% of cases. ASCUS and ASC‐H were reported in 47 and 14 cases, respectively. The most common lesion on Pap smear was high‐grade squamous intraepithelial lesion (HSIL; n = 92), followed by low‐grade squamous intraepithelial lesion (LSIL; n = 64), and two were ungradable SIL. Squamous cell carcinoma (SCC) was reported in 14 smears. The ASC/SIL ratio was 0.38. CHC (n = 139) was 100% for ASC‐H, LSIL, SCC and 84.7% for HSIL. A review of discrepant cases suggested sampling and interpretational discrepancy in five and one cases, respectively. The PPV of Pap smear for squamous lesions was 96.4%.ConclusionIt is essential to have good quality cytopathology reports for early identification, which enables appropriate management. The most commonly used quality indicator for cytopathology is the ASCUS/SIL ratio. This study suggests the inclusion of the CHC and PPV values as quality metrics for Pap smear, since these are easily measurable and serve as a good indicator of quality in cervical cytopathology reporting.

Human telomerase RNA component (hTERC) gene expression and chromosome 7 ploidy correlate positively with histological grade of cervical intraepithelial neoplasia

AbstractObjectiveCervical cancer screening by primary human papilloma virus detection and cytology is fraught with low specificity and variable sensitivity, respectively. Cytology‐histology correlation remains modest. Biomarkers associated with early genetic events in cervical squamous carcinogenesis and detectable in cytology material are likely to be relevant. Human telomerase RNA component (hTERC) gene overexpression and aneuploidy are promising candidates in view of their reported early and consistent association with cervical squamous oncogenesis.MethodsWe analysed hTERC gene expression and chromosome 7 ploidy by fluorescent in‐situ hybridisation (FISH) in 50 women with cytological precursor squamous intraepithelial lesions and available histology outcomes. Results were expressed as percentages of cells showing ≥3 signals, mean signals/nucleus, and maximum amplitude across various cytology and histology categories. Proportions of positive cases were calculated from threshold values derived from 6 controls. Distribution of above indices with respect to ≥cervical intraepithelial neoplasia 2 (CIN2) was explored.ResultsFor both genetic aberrations, there was significant positive correlation (for all indices) between the proportion of positive cases and worsening cytological and histological outcomes (P < .05), with significant intergroup differences (P < .05). High‐grade lesions (≥CIN2) had significantly higher results compared to <CIN2 lesions (P ≤ .001). In five discordant cases with ≥CIN2 under‐ or overdiagnosed on cytology, FISH supported the histological diagnosis.ConclusionsHTERC gene amplification and chromosome 7 ploidy showed positive association with cervical squamous carcinogenesis and could be relevant in settings of discrepant cytology‐histology correlation.

Unusual uterine cervical polypoid mass presenting during pregnancy

The authors encountered a case of uterine cervical adenosarcoma with sarcomatous overgrowth during pregnancy. Cytological images of atypical stromal cells in sarcoma components were obtained in this case.

Are we managing our patients correctly following treatment for cervical glandular intraepithelial neoplasia? A review of practice at the Jessop Wing Colposcopy Unit

AbstractObjectiveWomen diagnosed with cervical glandular intraepithelial neoplasia (CGIN) remain at risk of further pre‐malignant and malignant disease and require rigorous post‐treatment follow‐up. We assess the effectiveness and safety of community cervical sampling follow‐up in women treated for CGIN.MethodsA retrospective study was conducted of women diagnosed with CGIN between April 1, 2013, and March 31, 2019, at Jessop Wing Colposcopy Unit, Sheffield, UK.ResultsOf 140 women diagnosed with CGIN, 76 had co‐existing cervical intraepithelial neoplasia (CIN). Cytologists were significantly more likely to report glandular neoplasia in the absence of co‐existing CIN, and high‐grade dyskaryosis in its presence (Ps < 0.0001). Co‐existing CIN was significantly more likely to be present with high or low‐grade compared to normal colposcopy findings (P < 0.0001). The 6‐month test of cure (TOC) was attended by 67% of women (84% within 12 months), and the 18‐month post‐treatment sampling by 52.5% of women (70% within 24 months). Colposcopy recalled 96% of women correctly for the 18‐month sampling, but 20% of women undertaking primary care samples were incorrectly recalled at 3 years instead.ConclusionsWhen CGIN is diagnosed, two dates for recall should be provided at 6 and 18 months post‐treatment to the Cervical Screening Administration Service and the centralised screening laboratory ensuring the 18‐month post‐treatment sample is correctly appointed, preventing women with HPV‐negative TOC samples being returned to 3‐year recall. Follow‐up of CGIN should be closely audited by the centralised laboratories ensuring women with CGIN are not put at additional risk.

Cytomorphological features of cervical small cell neuroendocrine carcinoma in SurePath™ liquid‐based cervical samples

AbstractSmall cell neuroendocrine carcinoma (SCNEC) of the cervix is a rare, highly aggressive tumour with poor prognosis and high propensity for distant metastases. The cytological features of SCNEC have rarely been described in cervical samples, and to the best of our knowledge, there are no previous reports using SurePath™ liquid‐based cytology. In the present report we present the cytomorphological features of histopathologically confirmed cases of cervical SCNEC in SurePath preparations. On cytological examination, all three cases demonstrated variable numbers of tumour cells, ranging from a few dispersed cells and tiny micro‐biopsies to large aggregates of small tumour cells with a high nucleus‐to‐cytoplasmic ratio, stippled chromatin, inconspicuous nucleoli, and scant cytoplasm. Immunocytochemistry for CD56 on the cervical preparation confirmed the diagnosis in one case. The presence of small tumour cells with characteristic stippled/salt‐and‐pepper type nuclear chromatin were the most consistent cytological features in these cases. Knowledge of these characteristic cytological features can help in suggesting a diagnosis of SCNEC in cervical samples which can then be confirmed by immunocytochemistry.

Cervical glandular neoplasia referrals and the diagnosis of adenocarcinoma in situ: Correlating cytology, colposcopy findings, and clinical outcomes

AbstractObjectivesTo determine the clinical outcomes of women with possible glandular neoplasia of endocervical type on cervical cytology, and review all diagnoses of cervical adenocarcinoma in situ (AIS) over a 5 year period at our institution.Study designA retrospective case‐note review was conducted of all women referred to colposcopy with possible glandular neoplasia of endocervical type on cervical cytology or diagnosed with cervical AIS after biopsy or excision, from January 2014 until December 2018 in a London district hospital.ResultsOf 55 women referred with possible glandular neoplasia of endocervical type, 47 (85.4%) had a significant pathology on histopathological analysis: AIS (n = 22); invasive cancer (n = 7); high‐grade cervical intraepithelial neoplasia (n = 18). Women with a history of borderline abnormality on cervical cytology within the last 5 years were significantly more likely to be diagnosed with AIS or invasive cancer (P < .05). For the same period 49 women had histologically proven AIS. Among these 22 (44.8%) were referred as possible cervical glandular intraepithelial neoplasia. Other reasons for referral were the following indications: borderline dyskaryosis (n = 13); high‐grade dyskaryosis (n = 8); low‐grade dyskaryosis (n = 4); postcoital bleeding (n = 2).ConclusionDue to the raised risk of significant gynaecological pathology in women with possible glandular neoplasia of endocervical type on cervical cytology, excisional biopsy is essential. Colposcopic impression varies significantly and complete excision of the abnormal lesions should be achieved. AIS is a histological diagnosis and should always be considered during colposcopical and cytopathological assessment.

Risk adaptive triage in cervical screening: challenges and opportunities

This is a perspective on some of the recent developments in molecular triage strategies for the risk stratification of High Risk HPV infection. The article was created to expand on some of the triage approaches described in a EUROGIN seminar by Prof Jack Cuzick in February 2021.

Atypical glandular cells in Papanicolaou test: Which is more important in the detection of malignancy, architectural or nuclear features?

AbstractObjectiveAtypical glandular cells (AGCs) in Pap (Papanicolaou) smears are uncommon but may represent various benign and malignant lesions. The aim of this study was to evaluate the AGC incidence in Pap smears, analyse the relationship between AGC and malignancy, and reveal the importance of architectural and nuclear features observed cytologically in malignancies.MethodsPatients diagnosed with AGC on the basis of cervicovaginal cytology between May 2011 and July 2018 were included in this study. All slides were retrospectively reviewed and subclassified according to the Bethesda 2001 classification system. The cytomorphological features observed in the smears were recorded. Cytohistological correlations were evaluated, and the significant clinicopathological findings for malignancy were determined.ResultsOf 87 536 Pap smears, 195 (0.22%) had AGC results and 156 had tissue follow‐up. Among the 156 smears with AGC, 80 (51.3%) were diagnosed as AGC‐NOS (atypical glandular cells, not otherwise specified) and 76 (48.7%) as AGC‐FN (atypical glandular cells, favour neoplastic). Follow‐up biopsies revealed benign pathologies in 49 cases (31.4%) and malignant pathologies in 107 (68.6%). The rate of malignancy observed in AGC‐FN cases (89.5%) was higher than the rate of malignancy in AGC‐NOS cases (48.8%). Among the cytomorphological features, nuclear irregularity, presence of macronucleoli, feathering, loss of polarity, papillary pattern, and three‐dimensional formation were found to be significant indicators of malignancy.ConclusionAs AGC in Pap smear was associated with a clinically significant diagnosis in 68.6% of the cases in our study, we suggest that all patients with AGC should undergo further clinical assessment.

Cellular debris on negative liquid‐based cytology cervicovaginal smears

AbstractObjectivesTo determine the prevalence of cellular debris (CD) on benign cervicovaginal liquid‐based cytology (LBC) smears and which factors predict the presence and larger amount of CD.MethodsCervicovaginal smears evaluated as negative for intraepithelial lesion or malignancy (NILM) between 1 January and 31 March 2020 were retrospectively reviewed to record the presence and amount of CD. All smears were prepared with the SurePath platform. Patient ages and past medical and surgical histories were also retrieved. Multivariate regression analyses were performed to find positive predictors of a larger amount of CD.ResultsThree hundred forty‐nine NILM smears were included in this study. The cohort consisted of 222 cervical smears (CS), and 127 vaginal smears (VS) taken from patients who had undergone hysterectomy. Overall, CD was observed in 111 (31.8%) cases. The positive predictors of CD were increasing age, specimen type (VS compared to CS), history of chemotherapy or radiation therapy (CRT), and more than mild background inflammation. Among the VS group, CD was present in 64 cases (50.4%) regardless of the time between the hysterectomy and specimen collection. Positive predictors in the VS group were age and more than mild inflammation. By contrast, the prevalence of CD in the CS group was 21.2%, and age was the only positive predictor. Histories of CRT, conisation, and inflammation were not statistically significant positive predictors for CD among CS.ConclusionsCellular debris could be seen in as much as 50% of NILM smears taken after hysterectomy, regardless of the time since the procedure. Increasing age was a positive predictor of the presence and a larger quantity of CD. These findings are helpful when evaluating smears with moderate to abundant debris in the background with questionable cellular atypia.

DNA image cytometry parameters to identify high‐grade cervical lesions

AbstractObjectiveEvaluate the performance of different DNA image cytometry (DNA‐ICM) ploidy parameters in the categorisation of DNA‐ICM results and identification of high‐grade cervical intraepithelial neoplasia or worse (≥ CIN2).MethodsCervical samples from 232 women were collected for DNA‐ICM analysis and biopsy confirmation. Five DNA parameters were used to define DNA aneuploidy: number of cells with exceeding events (EE) over 2.5cEE, 4cEE, 5cEE and 9cEE, and aneuploid stemlines. DNA‐ICM results were categorised as normal, suspicious, and abnormal.ResultsFor individual DNA ploidy parameters, sensitivity values for 50 cells with 2.5cEE, 45 cells with 4cEE, 1 cell with 9cEE and aneuploid stemline were 72.95%. 54.1%, 69.67% and 54.1%, while specificity values were 80.0%, 90.0%, 89.09% and 95.45%, respectively. For the 5cEE parameter, the sensitivity values for 1, 2, 3, 4 and 5 cells were 93.44%, 85.25%, 81.97%, 77.87% and 75.41%, while specificity values were 46.36%, 63.64%, 74.55%, 76.36% and 80.91%, respectively. For categorised DNA‐ICM results, a suspicious result showed superior sensitivity than an abnormal result (87.70% vs 82.79%, P = 0.031), but lower specificity (54.55% vs 75.45%, P < 0.001). Both types of DNA‐ICM result were statistically significantly different from a normal result (P < 0.05).ConclusionFor prognostic purposes, 1 cell with 9cEE, 45 cells with 4cEE and aneuploid stemline are the best parameters with which to categorise an abnormal DNA‐ICM result, followed by 50 cells with 2.5cEE and 4 cells with 5cEE. For screening purposes, 10 cells with 2.5cEE, 10 cells with 4cEE, and 2 cells with 5cEE are suitable parameters with which to categorise a suspicious DNA‐ICM result.

A review of the directly sampled endometrial cytology on LBC samples: Classification, microscopic criteria and beyond

AbstractThe Yokohama System for Reporting Endometrial Cytology (TYS) has been proposed by an expert meeting under the auspices of the International Academy of Cytology (IAC) in May 2016 at the IAC in Yokohama. Since its introduction, the TYS has been receiving worldwide acceptance, and this review aims to assess its global impact. The adoption of endometrial cytology as a diagnostic procedure has been hampered in the past by difficulties arising in interpreting the cellular findings due to a number of factors (such as excess blood, cellular overlapping and the complex physiology of endometrium). Recently, the use of liquid‐based cytology (LBC), with its ability to remove blood and mucus and to distribute cells uniformly in a thin layer on the slide, has provided an opportunity to re‐evaluate the role of endometrial cytology. LBC is a useful tool in the cytologic diagnosis and follow‐up of endometrial abnormalities, which remains complementary to the emerging molecular diagnostic cytopathology. The study of LBC from endometrial cytology could be challenging since it is affected by numerous look‐alikes and diagnostic pitfalls. This review discusses these various entities and takes into consideration the ancillary techniques that may be useful in the diagnostic procedure. In conclusion, our review of the published data suggests that the TYS is a valid classification scheme that has been widely accepted by cytopathologists globally, is highly reproducible and makes a valuable contribution to clinical therapeutic management. At present, molecular cytopathology is a rapidly evolving field of modern cytopathology, which underlines the effective interplay between genomics and cytology. This review aims to provide a comprehensive review of the drawbacks of endometrial cytopathology, particularly in terms of endometrial cancer diagnosis and molecular testing.

Cytological and Immunocytochemistry Findings in Fallopian Tube Brush Specimens and Their Correlation With Histology

ABSTRACTContextHigh‐grade serous carcinoma (HGSC), the most prevalent and lethal form of ovarian cancer, is increasingly recognised to originate in the fimbrial end of the fallopian tube (FT). Timely detection remains a critical unmet clinical need due to ineffective screening methods. This prospective observational study assesses the diagnostic potential of FT brush cytology by correlating cytomorphological and immunocytochemical findings with histologically confirmed HGSC.Methods and MaterialA total of 134 FT from 89 patients undergoing salpingo‐oophorectomy (with or without hysterectomy) were analysed. Liquid‐based cytology samples were evaluated for morphological abnormalities and subjected to immunocytochemistry using p53 and Ki‐67 markers. Cytological results were classified as benign, suspicious or malignant. Statistical analyses included sensitivity, specificity and odds ratio calculations via logistic regression (α = 0.05), performed using the R software.ResultsHistopathology confirmed HGSC in 15 patients. Of these, brush cytology identified 12 as suspicious or malignant, demonstrating high diagnostic concordance. Aberrant p53 expression was found in 11 cases, and a high Ki‐67 proliferation index was observed in 10. These findings underscore the strong correlation between cytological, immunocytochemical and histological features of tubal HGSC.ConclusionsIn conclusion, FT brush cytology combined with p53 and Ki‐67 immunocytochemistry shows promise as a minimally invasive approach for early HGSC detection. Future research should focus on larger prospective cohorts, ideally incorporating in vivo hysteroscopic sampling.

The Diagnostic Role of TFF1, TFF3, FOXA1, CA XII and TRPS1 in Serous Effusions

ABSTRACTObjectiveTo analyse the diagnostic role of trefoil factor‐1 and ‐3 (TFF1, TFF3), forkhead box protein A1 (FOXA1), carbonic anhydrase XII (CA XII) and trichorhinophalangeal syndrome type 1 (TRPS1) in serous effusions. The prognostic role of these markers in breast carcinoma was additionally studied.MethodsProtein expression by immunohistochemistry was analysed in 247 effusions, consisting of 60 breast carcinomas, 54 tubo‐ovarian carcinomas, 47 mesotheliomas, 44 lung carcinomas, 20 uterine corpus and cervical carcinomas, 17 gastrointestinal carcinomas and 5 cancers of other origin.ResultsTFF1, TFF3, FOXA1, CA XII and TRPS1 expression was found in 67%, 70%, 88%, 82% and 83% of breast carcinomas, respectively. Expression of all markers was seen in some carcinomas of other origin, most commonly in GI metastases, but was least frequent for TRPS1. CA XII expression was additionally seen in mesotheliomas and reactive mesothelial cells. All 5 markers were significantly overexpressed in breast compared to tubo‐ovarian carcinoma (all p < 0.001) and lung carcinoma (all p < 0.001 except for FOXA1, p = 0.023). TFF1 (p = 0.003), TFF3 (p = 0.008) and FOXA1 (p = 0.017) expression was significantly higher in receptor‐positive compared to receptor‐negative primary breast carcinomas. In survival analysis for 44 breast carcinoma patients with clinical data, TFF1 expression was associated with a trend for longer overall (p = 0.096) and disease‐free (p = 0.06) survival.ConclusionTFF1, TFF3, FOXA1, CA XII and TRPS1 are sensitive breast carcinoma markers, with FOXA1 performing best in terms of sensitivity and TRPS1 being the most specific. Whether the expression of these markers in breast carcinoma effusions is informative of survival merits further research.

Cytopathological features associated with POLE mutation in endometrial cancer

AbstractObjectiveFor patients with endometrial cancer, the POLE (polymerase epsilon) mutation (POLEmut)‐subtype, one of four molecular‐analysis‐based categories in the Cancer Genome Atlas (TCGA), has the best prognosis. The following histological characteristics are typically observed in endometroid carcinoma cases with the POLEmut‐subtype: (1) the presence of tumour giant cells, (2) numerous tumour‐infiltrating lymphocytes (TILs) and/or peri‐tumoral lymphocytes, and (3) a high grade. However, in the context of cytology, the morphological characteristics of this subtype remain unknown.MethodsDNA extracted from formalin‐fixed paraffin‐embedded (FFPE) tissues was subjected to next‐generation sequencing analysis and categorised according to the TCGA classifications. Genomic mutation, tumour mutation burden (TMB), and microsatellite instability were also assessed. Cytological specimens of resected uteri obtained using the Papanicolaou method were histologically separated into three types.ResultsSeven out of 112 patients (6%) with endometrial cancer were diagnosed with the POLEmut‐subtype between January 2019 and August 2021. Tumour giant cells were observed in three cases (43%) on histology and cytology. TIL and/or peritumoral lymphocytes with inflammatory cells were detected in five cases (71%) on histology and three cases (43%) on cytology. Cases in which these three characteristics were observed on both cytology and histology may have belonged to the POLEmut‐subtype. There were no cases in which these characteristics were absent on histology but present on cytology. TMB tended to be higher in cases when the three characteristics were observed in both cytological and histological findings.ConclusionsPreoperative endometrial cytology highlighted the characteristics of the POLEmut‐subtype in the histological analysis of resected uterine specimens and has the potential to play an important role in treatment decisions.

A sub‐hepatic nodule in a young female: Chase the case

Sertoli-Leydig cell tumours (SLCTs) are rare, mixed sex-cord stromal tumours composed of varying proportions of both Sertoli and Leydig cells, which account for <0.5% of all ovarian tumours. The cytomorphologic features of SLCTs are not well described in literature. Herein, we describe the cytomorphologic features of an SLCT at an uncommon metastatic site in a young female. Sertoli-Leydig cell tumours (SLCTs) are rare, mixed sex-cord stromal tumours composed of varying proportions of both Sertoli and Leydig cells, which account for <0.5% of all ovarian tumours. The cytomorphologic features of SLCTs are not well described in literature. Herein, we describe the cytomorphologic features of an SLCT at an uncommon metastatic site in a young female.

Diagnostic Value of Liquid‐Based Cervical Cytology Test in Endometrial Carcinoma

ABSTRACTBackgroundAbnormal endometrial cells could be detected by liquid‐based cervical cytology test due to the anatomical continuity of the uterine cavity with the cervix. Our aim was to evaluate the diagnostic value of Papanicolaou (Pap) test in endometrial cancer (ECa).MethodsECa cases with liquid‐based Pap smears within 6 months before the histological diagnosis were retrospectively searched from an academic women's and children's hospital pathology archive. Corresponding smears were reviewed, and clinicopathological parameters and Pap test results were recorded.ResultsA total of 171 cases were identified. Among these patients, 43 (25.1%) cases were interpreted as abnormal, 32 (74.4%) of which suggested glandular abnormality in the routine screening (initial Pap smear reports). After re‐examining all slides, 123 (72.5%) cases showed the presence of endometrial abnormality. Among these, 87 cases (70.7%) were diagnosed as atypical endometrial cells (AEC), 33 cases (26.8%) as adenocarcinoma (ADC), and 3 cases (2.4%) as benign endometrial cells. The cases with non‐endometrioid histology were more likely to be interpreted cytologically as ADC, while endometrioid subtypes were often diagnosed as AEC (p &lt; 0.05). Except for the tumour type (p &lt; 0.05), the other clinicopathological factors were not significantly related to a higher percentage of positive Pap test results, including age, histological grade, myometrial invasion, cervical involvement, lymphovascular invasion, and FIGO stage(p &gt; 0.05).ConclusionPap test plays a role in the detection of ECa. Detection could be improved in routine screening by paying more attention to endometrial lesions in high‐risk populations.

A study on preserving endometrial glandular architecture during preparation using BD SurePath™ liquid‐based cytology reagents: Cellular fixation with preservative fluid requires at least 18 h

AbstractIntroductionThis study aimed to determine the causes of disruption of the three‐dimensional architecture of endometrial glands prepared using BD SurePath™ liquid‐based cytology (SP‐LBC) reagents. One sample preparation method for endometrial cytology is presented in which this three‐dimensional architecture can be retained.MethodsSP‐LBC specimens were prepared by the following three methods: (1) using the BD PrepMateTM (PrepMate) System after cellular fixation for 1‐6 h (method A); (2) without using the PrepMate System after cellular fixation for 1‐6 h (method B); and (3) using the PrepMate System after cellular fixation for at least 18 h (method C). Size and numbers of endometrial cell clusters and numbers of solitary scattered cells were then evaluated.ResultsSignificantly higher numbers of cell clusters with a major axis of 200 μm or more were yielded by method C (71.3 ± 57.2) than methods A (9.3 ± 5.9, P &lt; 0.001) or B (44.3 ± 28.8, P &lt; 0.05). Method B yielded significantly higher numbers of cell clusters than method A (P &lt; 0.001). Method A (132.2 ± 107.7, p &lt; 0.001) yielded significantly higher numbers of solitary scattered cells than methods B (29.1 ± 14.8) and C (35.7 ± 23.3). No significant difference in solitary cell numbers was found between methods B and C.ConclusionsRetention of endometrial glandular architecture is rendered possible by allowing sample fixation times of 18 h or more when preparing specimens using the PrepMate System.

The expression pattern of CD10 and CD31 identifies fine fibrovascular stroma of grade 1‐endometrial endometrioid carcinomas in cytology

AbstractIntroductionThe objective of this study was to assess the diagnostic utility of CD10 in the differential diagnosis of grade 1‐endometrial endometrioid carcinoma (G1‐EEC) and the metaplastic changes associated with the endometrial glandular and stromal breakdown (EGBD) on liquid‐based cytological (LBC) samples.Methods(1) The type and distribution of CD10‐positive cells in EGBD and G1‐EEC patients were evaluated. (2) Based on the results from (1), histological and cytological specimens were double‐immunostained with CD31 and CD10 to confirm whether CD10‐positive tubular‐canalicular material found in (1) was represented by fine threads of endometrial‐type fibrovascular stroma. (3) Based on the results from (2), additional immunostaining of histological specimens was performed for CD146 and αSMA as markers of perivascular cells.Results(1) CD10 positive cells showed two main patterns of expression: cytoplasmic immunoreactivity in the form of dense brown granules in EGBD and tubular‐canalicular branching patterns in G1‐EEC. (2) The tubular‐canalicular material observed in cytological specimens of G1‐EEC samples co‐expressed CD10 and CD31, and was interpreted as representing fine threads of endometrial fibrovascular stroma in the corresponding histological samples. Conversely, metaplastic changes in EGBD cases, only a few CD31‐positive signals were found inside the condensed stromal clusters with CD10‐positive. (3) Cells surrounding the CD31‐positive vascular endothelial cells expressed CD146 and αSMA; moreover, some of the thin CD10‐positive fibrous stromal strands also co‐expressed αSMA.ConclusionsCD10 is a very useful immunomarker for distinguishing between G1‐EEC and the metaplastic changes of EGBD in LBC samples.

Cervical cytology in the detection of uterine clear cell carcinoma: Diagnostic predictors from a case‐control study

AbstractIntroductionUterine clear cell adenocarcinoma (CCC) is a rare, aggressive malignancy with poor prognosis. The present study aimed to identify and describe its characteristic morphological features in cervical cytology.MethodsThis was a 3‐year retrospective case‐control study. Cases included cervical samples of histopathologically proven endometrial and cervical CCC. Controls included cervical samples of histopathologically proven endometrial serous carcinoma (n = 15), endometrioid adenocarcinoma (n = 20), and endocervical adenocarcinoma (n = 15). Twenty‐eight cytomorphological features were evaluated; the strength of association was determined by odds ratio (OR) and Cramer's V, and the diagnostic accuracy of statistically significant features was assessed.ResultsCases consisted of histopathologically proven 25 (34.7%) endometrial and 13 (18.0%) cervical CCC. Corresponding cervical samples were available for a total of 14 (36.8%) patients, of which 13 (92.8%) were positive for epithelial cell abnormality. On univariate analysis, three cytomorphological variables were significant predictors of uterine CCC: presence of dense cytoplasm (OR = 88; V = 0.72), deep nuclear membrane irregularities (OR = 17.5; V = 0.55), and coarse chromatin (OR = 21.3; V = 0.46). Dense cytoplasm had the highest positive predictive value (92%) and high specificity (97.8%), whereas coarse chromatin had the highest sensitivity (92.3%) and negative predictive value (96.7%).ConclusionsThe presence of dense cytoplasm and deep nuclear membrane irregularities in the tumour cells were strong predictors, and coarse chromatin a moderate predictor, of uterine CCC compared to its close cytological mimics.

Primary squamous cell carcinoma of the endometrium—Case report with cytological characteristics in direct and indirect endometrial samples

This case presents Primary Squamous Cell Carcinoma of The Endometrium with cytological and immunocytological characteristics in specimens from endometrial aspirate, Pap smear and peritoneal washing. Initial TVUS and clinical examination showed no abnormalities, and the first curettage sample was interpreted as benign while the second was histopathologically inconclusive. The final histopathology diagnosis in the hysterectomy specimen confirmed the diagnosis of this rare tumour.

Lesion‐targeted cytology to improve cytological sampling for atypical polypoid adenomyomas of the uterus: A case series and review of the literature

AbstractObjectiveAtypical polypoid adenomyomas (APAs) are uncommon tumours consisting of atypical endometrioid glands and fibromyomatous stroma. Identifying the biphasic nature of atypical glandular components and spindle mesenchymal components without atypia is crucial for the cytological diagnosis of APA. We investigated the utility of lesion‐targeted cytology (LTC) to directly collect firm spindle components.MethodsWe recruited seven consecutive surgical patients who underwent cytological examinations before surgery and were diagnosed with APA on postoperative histological examinations. Cytological smears were obtained by routine sampling in five cases and by targeted sampling using transvaginal ultrasonography, that is, LTC, in two cases. We retrospectively analysed the cytological findings from our cases and compared them to those of APA cases previously reported in the English literature.ResultsAmong 5/7 cases that involved routine cytological sampling, normal cytological findings were found in 2 and atypical glandular cells were found in 3, but spindle cells from mesenchymal components were not detected. In contrast, among 2/7 cases in which sampling involved LTC, spindle cells without atypia, in addition to atypical glandular cells were found.ConclusionsLesion‐targeted cytology is useful to assess mesenchymal components of APAs and may improve the cytological diagnosis of APA.

Nuclear morphometry as an adjunct to cytopathologic examination of endometrial brushings on LBC samples: A prospective approach to combined evaluation in endometrial neoplasms and look alikes

AbstractObjectiveIn this study, we aimed to retrospectively investigate and confirm whether atypical nuclear findings in endometrial cytology are useful when assessed by image morphometry in liquid‐based cytology (LBC) and compared with microscopic evaluation.MethodsIn total, 53 cases were selected for this study, including 11 presenting proliferative endometrium, 12 with surface papillary syncytial change with endometrial glandular and stromal breakdown (EGBD‐SPSC), 10 endometrioid carcinoma grade 1 (G1‐EEC), 10 EEC grade 3 (G3‐EEC), and 10 endometrial serous carcinomas (ESC). Nuclear image morphometry for nuclear geometric features (area, grey value, aspect ratio, internuclear distance, nucleolar diameter) was performed using ImageJ computer software. For assessing nucleoli, 3861 nuclei were measured, and for nuclear findings, except for nucleoli, 4036 nuclei were measured in total.Results(a) Compared with G1‐EEC, G3‐EEC and ESC presented a marked increase in all six parameters (nuclear enlargement, anisonucleosis, nuclear shade, nuclear shape, irregularity of nuclear arrangement, and nucleolar size). (b) EGBD‐SPSC presented a marked increase in two parameters (nuclear shade, nuclear shape) when compared with G1/G3‐EEC and ESC. (c) Compared with EGBD‐SPSC, EEC and ESC demonstrated a marked increase in nucleolar size (≥2.0 μm). (d) ESC presented a marked increase in nucleolar size (≥3.0 μm) when compared with G3‐EEC.ConclusionsHere we confirmed that atypical nuclear findings evaluated by image morphometry are as useful as microscopic evaluations in endometrial cytology. We believe that the objective evaluation of nucleolar size could contribute to an accurate diagnosis of endometrial‐LBC samples.

Role of fractal dimension in distinguishing benign from malignant endometrial clusters in liquid‐based cervical samples

AbstractIntroductionExfoliated endometrial cells are often seen as hyperchromatic crowded groups (HCGs) in cervical samples. It is challenging for the cytopathologists to discriminate between HCGs of benign and malignant endometrial cells. Fractal dimension (FD) analysis has proved to be a useful tool in discriminating between different types of cell groups in previous studies.AimsThis study was conducted to evaluate the utility of FD for differentiating between benign and malignant endometrial HCGs, in liquid‐based cervical samples.MethodsTwo groups of cervical samples, with subsequent histopathology, were selected: Group A: 30 cases with benign endometrial HCGs; and Group B: 39 cases with malignant endometrial HCGs. Image J, NIH and FracLac software were used for selecting and measuring the FD of the HCGs. Student t‐test was used for statistical analysis.ResultsThe mean FD for benign endometrial HCGs (1.066943 ± 0.0699) was significantly lower than that of the malignant endometrial HCGs (1.086271 ± 0.05121; P = .001). Using receiver operator characteristic curve analysis, we determined that an FD cut‐off value of 1.01 would yield sensitivity of 90.3%, specificity of 26.1%, positive predictive value of 47.3% and negative predictive value of 78.6%.ConclusionThe measurement of FD of HCGs in cervical samples can serve as a useful screening adjunct to differentiate malignant from benign HCGs, owing to its high sensitivity. However, in view of its low specificity and positive predictive value, we recommend that cases labelled as malignant by the FD value be confirmed for malignancy by other methods.

Large Cell Neuroendocrine Carcinoma of the Uterine Cervix With a Biphasic Synaptophysin Staining Pattern and Followed Vaginal Intraepithelial Recurrence: A Case Report

This case highlights the importance of early, aggressive surgery for large cell neuroendocrine carcinoma and continuous cytological monitoring in detecting recurrence at the early intraepithelial stage. The pagetoid spread pattern with a biphasic synaptophysin staining differentiated LCNEC from CIN3 and AIS.

Commercially Available Artificial Intelligence Solutions for Gynaecologic Cytology Screening and Their Integration Into Clinical Workflow

ABSTRACT Historically, gynaecologic cytology, particularly cervical screening through Pap smear tests, has been instrumental in early cancer detection, but not without its challenges. These include variability in interpretation and the labour‐intensive nature of manual screening processes. The advent of artificial intelligence (AI) technologies, especially machine learning and deep learning, heralds a new era in cytology, offering enhanced accuracy, consistency, and efficiency. These advancements promise to mitigate traditional limitations by automating routine analyses, aiding early cancer detection, and reducing the workload of laboratory personnel. This review thoroughly examines the current status of commercial AI software in gynaecologic cytology screening. We critically assess the capabilities, performance, and impact of these AI tools in a clinical context. Additionally, the review addresses the integration challenges and potential of AI in clinical practice, including workflow integration, regulatory compliance, and ethical considerations. Through this comprehensive analysis, we aim to provide insights into how AI is reshaping gynaecologic cytology, paving the way for more effective disease management and enhanced patient care in women's health.

A Sneak Peek Into a Rare Case of Metastatic Cervix Adenocarcinoma in Pleural Fluid With an Approach to Diagnosis

ABSTRACTEffusion samples are the minimally invasive cytology specimens, harbouring metastases from variable primary sites. Pleural metastasis from cervical carcinoma is quite rare and comprises 2.1%–6.1% of all the tumours. Endocervical adenocarcinoma with pleural metastasis is even rarer. Through this, we try to emphasise the cytomorphological and immunohistochemical approach along with histopathological correlation, all of which were indispensable in such a rare case.

Metastatic pleomorphic undifferentiated uterine sarcoma detected in pleural effusion

Undifferentiated uterine sarcoma is an extremely rare cytologically challenging finding in pleural effusions undistinguished from the more common causes of malignant effusions. Cellblock is a valuable diagnostic tool when combined with certain diagnostic clues, relevant immunoprofile and knowledge of the patient's history.

Villoglandular Carcinoma of the Uterine Cervix: Clinic‐Cytopathological‐Histological Features of a Rare Case With Brief Review of the Literature

ABSTRACTVilloglandular adenocarcinoma (VGA) of the cervix is a rare tumour with very few reports of cytological diagnosis on a cervical smear. Since the usual prognosis of VGA is more favourable than that of a conventional cervical adenocarcinoma, a pre‐operative diagnosis is essential for appropriate therapeutic decisions. We report the clinical, cytological, and histopathological features of a case of VGA in an elderly woman. A 60‐year‐old female presented with postmenopausal bleeding and was found to have a hypertrophied cervix. Conventional smears from the cervix demonstrated features of atypical glandular cells with a villoglandular configuration. Endocervical cell nuclei were enlarged with overlapping, hyperchromasia and mild to moderate anisonucleosis. The cytological impression of villoglandular carcinoma was confirmed on subsequent hysterectomy and histopathology. Hence, cytopathologists need to be aware of the entity of villoglandular carcinoma of the cervix, its relatively bland nuclear features, and the diagnostic clues to allow for an accurate diagnosis on cervical smear. An accurate pre‐operative diagnosis can assist the gynaecologists in arriving at appropriate therapeutic and prognostic decisions.

Digital Cytology Combined With Artificial Intelligence Compared to Conventional Microscopy for Anal Cytology: A Preliminary Study

ABSTRACTIntroductionRecent studies have shown that digital cytology (DC) coupled with artificial intelligence (AI) algorithms is a valid approach to the diagnosis of cervico‐vaginal lesions using liquid‐based cytology (LBC). We evaluated the use of these methods for anal LBC specimens.MethodsA series of 124 anal LBC slides previously diagnosed by conventional microscopy (CC) were reviewed with a DC/AI system that generated a gallery of images. Diagnoses based on the selected images, according to the 2014 Bethesda System for Reporting Cervical Cytology, were compared to CC.ResultsOverall, CC and DC/AI approaches detected a similar number of abnormal (ASC‐US+) cases (63 and 62 cases, respectively). We observed an exact concordance between CC and DC in 70 (57.9%) cases, corresponding to a moderate agreement between the two approaches (κ = 0.41, p &lt; 0.001). A moderate agreement (κ = 0.48, p &lt; 0.001) was also found when positive cases were stratified into ‘low‐grade’ (ASC‐US, LSIL) and ‘high‐grade’ lesions (ASC‐H, HSIL). The DC/AI system detected more cases of higher severity (ASC‐H, HSIL: 9 and 2 cases, respectively) than CC (3 cases classified as HSIL).ConclusionsThe number of ASC‐US+ cases detected by both systems was similar. The DC/AI system detected more cases of higher severity compared to the CC.

Integration of AI‐Assisted in Digital Cervical Cytology Training: A Comparative Study

ABSTRACTObjectiveThis study aimed to investigate the supporting role of artificial intelligence (AI) in digital cervical cytology training.MethodsA total of 104 trainees completed both manual reading and AI‐assisted reading tests following the AI‐assisted digital training regimen. The interpretation scores and the testing time in different groups were compared. Also, the consistency, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of diagnoses were further analysed through the confusion matrix and inconsistency evaluation.ResultsThe mean interpretation scores were significantly higher in the AI‐assisted group compared with the manual reading group (81.97 ± 16.670 vs. 67.98 ± 21.469, p &lt; 0.001), accompanied by a reduction in mean interpretation time (32.13 ± 11.740 min vs. 11.36 ± 4.782 min, p &lt; 0.001). The proportion of trainees' results with complete consistence (Category O) were improved from 0.645 to 0.803 and the averaged pairwise κ scores were improved from 0.535 (moderate) to 0.731 (good) with AI assistance. The number of correct answers, accuracies, sensitivities, specificities, PPV, NPV and κ scores of most class‐specific diagnoses (NILM, Fungi, HSV, LSIL, HSIL, AIS, AC) also improved with AI assistance. Moreover, 97.8% (89/91) of trainees reported substantial improvement in cervical cytology interpretation ability, and all participants (100%, 91/91) expressed a strong willingness to integrate AI‐assisted diagnosis into their future practice.ConclusionsThe utilisation of an AI‐assisted digital cervical cytology training platform positively impacted trainee performance and received high satisfaction and acceptance among clinicians, suggesting its potential as a valuable adjunct to medical education.

Tall‐columnar glandular cells in SurePath™ liquid‐based cytology Pap sample: Learning from mimics/pitfalls

We offer a comprehensive depiction of the cytomorphological characteristics of lobular endocervical glandular hyperplasia (LEGH) as observed in SurePath™ liquid-based cytology (LBC), subsequently confirmed on cone biopsy. Lobular endocervical glandular hyperplasia (LEGH), a precursor to gastric-type adenocarcinoma (GAE) of the endocervix, is rare and reports of it in cervical cytology are scarce. We provide a thorough description of the cytomorphological features of LEGH observed in SurePath™ liquid-based cytology (LBC), later confirmed by cone biopsy. To the best of our knowledge, this is the first report documenting cytology of LEGH in LBC of a Pap sample.

Atypical glandular cells and predictive features of malignancy in Pap smears: A retrospective monocentric study

Abstract Objective The introduction of cytological screening with the Papanicolau smear significantly reduced cervical cancer mortality. However, Pap smear examination can be challenging, being based on the observer ability to decode different cytological and architectural features. This study aims to evaluate the malignancy rate of AGC (atypical glandular cells) category, investigating the relationships between cytological and histological diagnosis. Methods Eighty‐nine patients, diagnosed as AGC at cytological evaluation and followed up with biopsy or surgical procedure at Policlinico Gemelli Hospital, Rome, Italy, were included in the study. The cytopathological architectural (feathering, rosette formation, overlapping, loss of polarity, papillary formation, three‐dimensional formation) and nuclear (N/C ratio, nuclear enlargement and hyperchromasia, mitoses, nuclei irregularity, evident nucleoli) features of AGC were evaluated. Statistical analyses were performed to assess cyto‐histological correlation and determine the relevance of architectural and nuclear features in the diagnosis of malignancy. Results Of the 89 AGC patients, 48 cases (53.93%) were diagnosed as AGC‐NOS and 41 (46.07%) were diagnosed as AGC‐FN, according to the Bethesda classification system. The follow‐up biopsies or surgical resections revealed malignancy in 46 patients (51.69%). The rates of malignancy for AGC‐NOS and AGC‐FN were 35.41% and 70.73% respectively. Furthermore, analysing cytopathological features, we found that both architectural and nuclear criteria were statistically significant ( p  &lt; 0.05). Only overlapping, nuclear irregularity and increased N/C ratio were not found to be statistically significant for detecting malignancy. Conclusions Cytological diagnosis of glandular lesions remains a valid tool, when appropriate clinical correlation and expert evaluation are available.

The Methylation Test Is Highly Sensitive for HPV‐Associated Endocervical Adenocarcinoma and Could Be Helpful as a Cytological Ancillary Test in Women With a PAP‐Smear Diagnosis of Severe Glandular Lesion (AGC–NEO+)

ABSTRACTObjectiveThe limitations of PAP smears in diagnosing severe cervical glandular lesions motivate the development of ancillary methods to facilitate their detection. This prospective cytology–histology and molecular study aims to investigate methylation test performance by establishing the test sensitivity and its relevance to patient management.MethodLBC samples were prospectively acquired after 3 months following the primary conventional PAP‐smear screening diagnosis of AGC–NEO (atypical glandular cells, favour neoplastic) and AIS. An HPV test and methylation test were performed. Corresponding subsequent biopsy reports were collected.ResultsSeven hundred and seventy Pap tests were signed out as AGC–NEO+. Complete study data were available for 85 cases (AIS in 46 cases, EA in 39 cases) that were further analysed. The methylation test was positive in 95.3% (81/85) cases, negative in 3.5% (3/85, AIS in two cases, EA in one case) and unanalysable in one sample (1.2%). HPV genotyping revealed a multi‐infection rate of 29%. The presence of HPV types 16 and 18 was detected in 84% (72/85) of lesions, and HPV type 45 in 12% (10/85).ConclusionsThe methylation test shows a high sensitivity of 95.3% and reliably identifies histologically confirmed AIS+ lesions. This argues for further investigation into its performance characteristics and consideration of its use, especially as a pre‐surgical triaging test in sensitive cases if the AIS+ lesion is suspected in pregnant women or nullipara.

From the Daily Peer Review of Abnormal Pap Test Slides to the Monitoring of Individual and Laboratory Performances: 5 Years of Data Collection and New Potential (Key) Performance Indicators

ABSTRACTObjectivesThe Peer Review (PR) consists of the daily examination, by all cytologists, of Pap slides that resulted abnormal/difficult, in order to reach a consensus on the final diagnosis (FD). We explore data from 5 years (2017–2021) of PR to: (i) evaluate the agreement (both inter‐observer and versus FD) over time; (ii) identify new quality indicators.Methods5673 slides were submitted to PR and examined by an average of 8 cytologists (range: 4–13). The agreement between cytologists and between the individual diagnosis with FD were evaluated by Kappa (k) and weighted Kappa (wK) and compared between ‘experts’ and ‘less experienced’ readers.ResultsThe inter‐observer agreement showed a moderate agreement among readers (whole team k = 0.44; experts k = 0.48). The highest and the lowest agreement was reported in HSIL and ASC‐H, respectively. In 2018 and 2021, a significant reduction of kappa was observed, likely attributable to team turnover. The laboratory agreement versus FD resulted in significantly higher scores in experts (wk = 0.73, 95% CI 0.73–0.74) compared to less experienced individuals (wk = 0.65, 95% CI 0.64–0.66), with a general reduction of wk recorded in 2021. The individual agreement versus FD (calculated for 16 cytologists) achieved a moderate/substantial level of agreement (wK range: 0.57–0.80), with a shift toward higher wk in experts.ConclusionThe levels of agreement are influenced by cytologist experience and team turnover. We propose new potential (key) performance indicators to strictly monitor the occurrence of systematic differences in interpretation criteria among cytologists. The proposed reference values are based on preliminary data and should be validated prospectively over a longer monitoring period.

Role of DNA ploidy in diagnosis and prognosis of high‐grade cervical intraepithelial neoplasia: A prospective cohort study

AbstractObjectiveTo compare the sensitivity and specificity of DNA ploidy with cytology, human papillomavirus (HPV) testing and colposcopy in diagnosis of high‐grade cervical intraepithelial neoplasia (CIN) and to assess the role of aneuploidy in cervical lesions with the worst prognosis. A prospective observational cohort study was conducted on 254 women with altered colpocytology.MethodsColposcopy, biopsy, DNA‐ICM and HPV examinations were applied to cervical cytological and histological samples. Participants were evaluated every 6 months and divided into two groups: ‘Harm’ and ‘No‐harm’. Logistic regression and multivariate COX model were used to identify independent risk factors for diagnosis and prognosis of high‐grade CIN, and ROC curve to assess the sensitivity and specificity of methods.ResultsVariables ‘age greater than or equal to 30 years’, ‘lesion size greater than 20%’, ‘aneuploidy’ and ‘HPV 16’ were associated with diagnosis of high‐grade CIN and ‘aneuploidy’ and ‘women living with HIV’, with a worse prognosis. Agreement for colposcopy was good, with a sensitivity of 79.3% and specificity of 94.4%; DNA‐ICM and cytology were moderate, with sensitivity of 74.6% and 72.3% and specificity of 85.3% and 76.1%, respectively. High‐risk HPV and HPV 16 tests were weak, with sensitivity of 75.0% and 43.75% and specificity of 50.0% and 88.64%, respectively.ConclusionsIn relation to high‐grade CIN diagnosis, DNA‐ICM presented similar sensitivity and specificity to cytology and high‐risk HPV test when associated with HPV 16. Regarding prognosis, this research certifies that aneuploidy is considered a predictor of more severe cervical injury.

Challenging lesions in cervical cytology: The elusive HSIL

AbstractCervical cytology has been an integral part of cervical cancer screening since the mid‐20th century with the implementation of screening protocols utilising Pap testing. During that time, cervical cancer has gone from the leading cause of cancer deaths in women to not even appearing in the top 10 causes of US cancer deaths. However, despite its long and widespread use, cervical cytology remains a diagnostically challenging area in the practice of cytopathology. Of particular importance for diagnosticians is the accurate diagnosis of high‐grade squamous intraepithelial lesions (HSILs), given the significant risk of progression to invasive cervical cancer and the importance to patient management. Therefore, this review is presented in order to highlight the diagnostic features of HSIL, its various appearances, and important benign and neoplastic differential considerations with an emphasis on morphological clues that can aid in distinguishing between these different processes.

Urothelial bladder carcinoma recurrence in the uterine cervix: Cytological findings on a cervical cytology test

The authors discuss a rare case of cervical asymptomatic metastasis of a primary high‐grade poorly differentiated urothelial bladder carcinoma, diagnosed by cervical cytology test.

Molecular characteristics of low‐grade serous carcinoma in effusions

AbstractObjectiveThe molecular characteristics of low‐grade serous carcinoma (LGSC) in serous effusions have not been studied previously. The present study analysed the molecular profile of LGSC at this anatomical site.MethodsSpecimens consisted of a series of 17 serous effusions (15 peritoneal, 2 pleural) from 16 patients, of which 15 were LGSC and 2 serous borderline tumour (SBT) who later progressed to LGSC. For comparative purposes, 9 surgical specimens from 6 patients with LGSC were analysed. Fresh‐frozen cell pellets and surgical specimens underwent targeted next‐generation sequencing covering 50 unique genes.ResultsMutations were found in tumours from 14 of the 22 patients, of whom 4 had 2 different mutations and 10 had a single mutation. Overall, the most common mutations were in KRAS (n = 3) and BRAF (n = 3), followed by NRAS (n = 2), CDK2NA (n = 2), TP53 (n = 2), ATM (n = 2). Mutations in MET, STK11, ERBB2 and FLT3 were found in one case each. Patient‐matched specimens had the same molecular profile. Both effusions with TP53 mutation had concomitant ATM mutation, and both stained immunohistochemically with a wild‐type pattern. The absence of mutations was associated with a trend for shorter overall survival in univariate analysis (p = 0.072).ConclusionsThe molecular alterations in LGSCs in serous effusions are consistent with those found in solid tumours, with frequent alterations in the mitogen‐activated protein kinase pathway. Mutations in LGSC may be a marker of better outcomes.

An audit of liquid‐based cytology samples reported as high‐risk human papillomavirus and borderline nuclear change in endocervical cells

AbstractIntroductionPrimary human papillomavirus (HPV) screening, testing for the virus responsible for 99% of cervical cancers, was introduced in 2018‐2020 in the UK. This was preceded by HPV triage of low‐grade cytology from 2012. Much of the evidence incorporated into current National Health Service (NHS) colposcopy guidance assessed outcomes prior to this change in screening. The aim of this paper is to assess adherence to NHS cervical screening programme standards, determine the incidence of cases reported as high‐risk HPV plus borderline nuclear change in endocervical cells, to calculate colposcopic accuracy and assess histological outcomes in this cohort.MethodA retrospective audit of women referred to a colposcopy clinic in one NHS trust from 2016 to 2018. Data relating to histological outcomes, cytological follow‐up and demographics were collected.ResultsOf 2001 referrals, 22 data sets identifying HPV‐positive borderline endocervical change were eligible for analysis (1.2% incidence). Median age was 29.5. Two‐thirds (68.2%, n = 15) had high‐grade dysplasia at diagnostic biopsy. Those women with reassuring histology had normal cytological follow‐up. Colposcopic accuracy was moderate (positive predictive value 43.8%, negative predictive value 100%).ConclusionsBorderline nuclear change in endocervical cells is an uncommon condition but should be treated as a high‐grade referral. All women should be offered a diagnostic biopsy at the initial colposcopy; if no histopathological abnormality is identified, alternative sources of pathology should be considered. Excisional treatment should be recommended to unreliable attenders, those with a complete family and inadequate colposcopy (TZ3) and considered in younger women with a TZ3.

Effusion cytology of a mucinous borderline ovarian tumour: Pitfall or controversy? A case report with insight into the newly proposed International System for Reporting Serous Fluid Cytology

A case report of effusion cytology of a mucinous neoplasm is illustrated. The authors share their insight into adapting the newly proposed international system for reporting serous fluid cytology.

The phosphatase PTPN1/PTP1B is a candidate marker of better chemotherapy response in metastatic high‐grade serous carcinoma

AbstractObjectiveTo analyse the expression and clinical role of the phosphatase PTPN1 (PTP1B) in serous effusions.MethodsPTPN1 mRNA expression by quantitative RT‐PCR was analysed in 83 high‐grade serous carcinoma (HGSC) and 15 malignant mesothelioma (MM) effusions. PTP1B and phospho‐PTP1B (pPTP1B) protein expression by immunohistochemistry was analysed in 62 HGSC and 44 MM effusions.ResultsPTPN1 mRNA (P = .048), PTP1B protein (P = .047) and pPTP1B protein (P &lt; .001) were overexpressed in HGSC compared to MM effusions. PTPN1 mRNA was additionally overexpressed in post‐chemotherapy HGSC effusions compared to chemo‐naïve effusions (P = .005). However, pPTP1B protein expression was higher in effusions from patients with FIGO stage III compared to stage IV (P = .006), and higher expressions of both PTPN1 mRNA (P = .041) and PTP1B protein (P = .035) in HGSC effusions were associated with better (complete) chemotherapy response at diagnosis. PTPN1 RNA and protein expression was unrelated to survival in HGSC, whereas a trend for shorter overall survival (P = .06) was found for MM patients whose tumours expressed pPTP1B protein.ConclusionPTPN1 is overexpressed in HGSC compared to MM effusions, and may be a marker of better chemotherapy response in the former. Whether PTPN1 activation is informative of adverse outcome in MM merits further investigation.

Ovarian pulmonary‐type small cell carcinoma diagnosed by means of ascitic fluid cytology: A case report with immunocytochemical analysis

Primary ovarian small cell carcinomas of pulmonary type are very rare neoplasms. This report describes an extraordinary case of small cell carcinoma of pulmonary type accurately diagnosed on cytology.

Clinicopathological prognostic parameters in patients with tubo‐ovarian carcinoma effusions

AbstractObjectiveTo analyse the predictive and prognostic role of clinicopathological parameters in patients with tubo‐ovarian carcinoma and malignant effusion.MethodsA retrospective series of 700 malignant peritoneal (n = 610) and pleural (n = 90) effusions from 558 patients was revised for histotype based on the 2014 World Health Organization criteria. The role of clinicopathological parameters in determining outcome was assessed.ResultsThe majority of specimens (597 effusions from 473 patients) were high‐grade serous carcinomas (HGSC), followed by low‐grade serous carcinoma (LGSC; 48 effusions, 37 patients), clear cell carcinoma (CCC; 23 effusions, 19 patients) and carcinosarcoma (CS; 16 effusions, 16 patients). Patients with CCC and CS had the shortest, those with HGSC intermediate, and those with LGSC longest overall and progression‐free survival (both P &lt; 0.001). For patients with HGSC, older age (P = 0.002), more advanced FIGO stage (IV vs III; P &lt; 0.001), delayed/no surgery (P &lt; 0.001), larger residual disease volume (RD; P &lt; 0.001), non‐complete response to chemotherapy at diagnosis (P &lt; 0.001), and primary platinum resistance (P &lt; 0.001) were associated with shorter overall survival. In Cox multivariate analysis, FIGO stage (P = 0.002) and primary platinum resistance (P &lt; 0.001) were independent prognosticators. Significant association was additionally found for parameters analysed for progression‐free survival in HGSC (previous chemotherapy: P = 0.029; age: P = 0.046; FIGO stage, upfront therapy, RD: P &lt; 0.001), of which previous chemotherapy, upfront therapy, and RD were independent prognosticators (all P &lt; 0.001).ConclusionsThe vast majority of malignant effusions in patients with tubo‐ovarian carcinoma are derived from serous carcinoma or related tumours, such as CS. Histology is a powerful prognostic factor in this patient group, as are established clinical parameters.

The early detection of cervical cancer. The current and changing landscape of cervical disease detection

AbstractCervical cancer prevention has undergone dramatic changes over the past decade. With the introduction of human papillomavirus (HPV) vaccination, some countries have seen a dramatic decline in HPV‐mediated cervical disease. However, widespread implementation has been limited by economic considerations and the varying healthcare priorities of different countries, as well as by vaccine availability and, in some instances, vaccine hesitancy amongst the population/government. In this environment, it is clear that cervical screening will retain a critical role in the prevention of cervical cancer and will in due course need to adapt to the changing incidence of HPV‐associated neoplasia. Cervical screening has, for many years, been performed using Papanicolaou staining of cytology samples. As our understanding of the role of HPV in cervical cancer progression has advanced, and with the availability of sensitive detection systems, cervical screening now incorporates HPV testing. Although such tests improve disease detection, they are not specific, and cannot discriminate high‐grade from low‐grade disease. This has necessitated the development of effective triage approaches to stratify HPV‐positive women according to their risk of cancer progression. Although cytology triage remains the mainstay of screening, novel strategies under evaluation include DNA methylation, biomarker detection and the incorporation of artificial intelligence systems to detect cervical abnormalities. These tests, which can be partially anchored in a molecular understanding of HPV pathogenesis, will enhance the sensitivity of disease detection and improve patient outcomes. This review will provide insight on these innovative methodologies while explaining their scientific basis drawing from our understanding of HPV tumour biology.

Detection of HIV mRNA in routine liquid‐based cytology specimens of HIV‐infected women

AbstractObjectiveHuman immunodeficiency virus‐infected women have a high incidence of HPV infection, and HIV and HPV coinfection is associated with high incidence of cervical intraepithelial lesions and cervical cancer. This study investigated the ability to detect HIV mRNA in routine screening cervical liquid‐based cytology (LBC) samples and its correlation with HPV coinfection and cervical intraepithelial lesions.MethodsLiquid‐based cytology samples from 80 HIV‐infected women under combined antiretroviral therapy (cART) were studied for detection of HIV and HPV mRNA using Aptima® tests and for cytology diagnosis according to the 2014 Bethesda System for Reporting Cervical Cytology. Peripheral blood (PB) HIV mRNAs were assessed by real‐time polymerase chain reaction (RT‐PCR). Statistical analysis used Fisher's exact or Chi‐square test to compare frequencies among groups and the Mann‐Whitney U test to compare continuous variables.ResultsHuman immunodeficiency virus mRNA was present in 21.3% of routine LBC samples in HIV‐infected women, 12.5% of which had no detectable PB viral load. Among 10 patients diagnosed with high‐grade squamous intraepithelial lesion (HSIL), 50% had detectable HIV viral load. The occurrence of HSIL vs low‐grade intraepithelial lesion/negative intraepithelial lesion or malignancy in LBC samples was significantly higher in women with detectable HIV viral load (P = .029).ConclusionsHuman immunodeficiency virus mRNA was present in routine LBC samples in HIV‐positive women under cART. Detection of HIV viral load in LBC is significantly associated with cervical HSIL. This suggests the relevance of HIV mRNA viral load assessment in routine LBC, to evaluate patients' infectious potential and monitor efficacy of the cART scheme.