The Founding Pioneer Cytotechnologists: The Women Who Assisted George N. Papanicolaou, M.D., Ph.D. develop the Pap Test for Cervical Cancer Prevention.

BACKGROUND: The earliest cytotechnologists are largely unknown. SUMMARY: In 1943, the book "Diagnosis of Uterine Cancer by the Vaginal Smear" by Papanicolaou and Traut recognized several women who have largely faded from memory. While Mary Papanicolaou and Charlotte Street are familiar names, others like Alberta Kuder and Huldah Boerker, who inadvertently laid the groundwork for the field of Cytotechnology, remain obscure. There were also women like Christine Rassias and Adele Reboul who did not receive recognition. Notably, Mrs. Lady Mary G. Papanicolaou, despite her significant contributions both in the lab and at home since 1914, was not acknowledged in her husband's work until the publication of his Atlas in 1954. KEY MESSAGE: These women set the benchmark for future Cytotechnologists, unknowingly shaping the profession as we know it today.

The utility of 'low-grade intraepithelial lesion, cannot exclude high grade' diagnosis: a single institution's experience.

OBJECTIVE: Squamous intraepithelial lesions (SILs) are classified as low-grade SIL (LGSIL) and high-grade SIL (HGSIL). 'LGSIL cannot exclude high grade' (LGSIL-H) interpretive category has been used in cases where findings exceed criteria for LGSIL, but do not fulfill the criteria for HGSIL. This study analyzed follow-up histology of LGSIL-H cases and compared the follow-up results of LGSIL-H with LGSIL to determine the utility of LGSIL-H category using a single institution's experience. STUDY DESIGN: Pap smears with LGSIL-H interpretation from 2005 to 2008 were retrieved. Histological follow-up results for LGSIL-H cases were analyzed and compared to the follow-up results of LGSIL cases. RESULTS: Cases with LGSIL-H interpretation (311) comprised 0.18% of all cases (170,307). Follow-up was available for 144 patients and 13.2% had benign findings, 51.4% had cervical intraepithelial neoplasia (CIN) 1, and 35.4% had CIN 2 or higher. In comparison, of 425 patients with LGSIL, 22.6% had benign findings, 71% had CIN 1 and 6.4% had CIN 2 or higher. CONCLUSION: A significantly greater number of patients with LGSIL-H interpretation had a CIN 2 or higher lesion on follow-up compared to patients with LGSIL. This suggests LGSIL-H may be a useful diagnostic category.

Reporting of Benign Endometrial Cells in Papanicolaou Tests.

OBJECTIVES: The 2014 Bethesda System (TBS 2014) guidelines for reporting cervical cytology revised the age for reporting benign endometrial cells (BECs) from 40 years or older to age 45 years or older. We evaluated this change and further investigated if extending the reporting age to 50 years or older may be acceptable. METHODS: We reviewed cases with BECs reported on Papanicolaou tests in women age 40 years or older and 45 years or older before and after implementation of TBS 2014. Follow-up endometrial biopsy/curettage results were categorized as benign, endometrial hyperplasia with or without atypia, or malignant. Hyperplasia and malignant follow-up were considered clinically significant. Clinical data were documented. Results were compared for women age 40 to 44, 45 to 49, and 50 years or older. RESULTS: Follow-up in 15 (100%) women age 40 to 44 years was benign. In women age 45 to 49 years, 61 (96.8%) had benign follow-up, one (1.6%) had atypical hyperplasia, and one (1.6%) had malignant follow-up. In women age 50 years or older, 57 (86.5%) had benign follow-up, four (6%) had malignant follow-up, and seven (7.5%) had atypical or nonatypical hyperplasia. There was a significant difference in follow-up between the age groups of 40 to 49 and 50 or older (P = .023). CONCLUSIONS: We conclude that the TBS 2014 revision was justified. Our data suggest that age 50 years or older rather than age 45 years or older may be an acceptable cutoff for reporting BECs.

Diagnosis of Schistosoma haematobium on voided urine cytology: a case report with radiologic correlation.

BACKGROUND: Infection with schistosomal species is becoming a more frequent finding in hospitals throughout the United States. Some causes that can be attributed to the rise include increased immigration from and travel to endemic areas. CASE: We report a case of urinary schistosomiasis diagnosed on urine cytology in a 7-year-old Nigerian boy. Infection was suspected after review of the clinical history and correlation with radiologic images. CONCLUSION: The rise in incidence has made it necessary for cytopathologists to be increasingly aware of these infections, in particular, Schistosoma haematobium, because it is the most frequent agent to be encountered on a cytology specimen, particularly urine. Similar cases have been published, one with specimen concentration and one without. However, no cases of cytology diagnosis with radiologic correlation are seen in the English literature.

Origin, frequency and clinical significance of glandular cells in liquid-based pap tests from patients posthysterectomy.

OBJECTIVE: To quantify the frequency and significance of glandular cells in posthysterectomy liquid-based (SurePath, TriPath Imaging, Burlington, North Carolina, U.S.A.) vaginal Pap tests. STUDY DESIGN: The presence of benign glandular cells in vaginal Pap tests from posthysterectomy patients represents a diagnostic challenge and may pose management issues. We investigated the presence, frequency and significance of glandular cells in 52 liquid-based (SurePath) vaginal Pap tests from posthysterectomy patients by combining cytomorphologic findings with adjunctive immunohistochemistry and mucin stains performed on cell block preparations and correlated the findings with clinical data. RESULTS: After performing these special studies, the frequency of reporting glandular cells in posthysterectomy Pap tests decreased from 3.5% to 1.2% of all vaginal Pap tests performed in a 6-month period. CONCLUSION: A strong association of the presence of benign appearing glandular cells and a previous history of gynecological malignancy (71%) and chemotherapy/radiation (59%) were noted, likely representing a regenerative process in response to injury or therapy.

Cytomegalovirus (CMV) in cervical cancer screening tests: A series of 8 cases and review of the literature.

OBJECTIVE: Cytomegalovirus (CMV) is a ubiquitous infection typically affecting over 50% of the US population by age 40. We report 8 cases of CMV infections detected in cervical cancer screening tests, the largest series of cases thus far reported in gynecologic cytology specimens. METHODS: A retrospective review of our pathology archival computer database was performed from January 1, 1994 through December 31, 2016 for CMV infections reported in cervical cytology specimens. The slides were retrieved for review if available. RESULTS: The eight patients ranged in age from 21-46 years, with a median age of 27 years and average age of 29.5 years. Two patients were significantly immunocompromised with one patient having AIDS and one patient diagnosed with autoimmune disease. The remaining six patients are considered immunocompetent. Cases were identified most often in the fall and winter months (6 of 8 cases). Seven cases were found using Surepath Pap (SP) liquid-based processing and one case was reported in a classic Papanicolaou smear (CPS). CONCLUSION: There was no correlation of cytologic presentation of CMV with a distinct cytohormonal pattern or inflammatory constituents. Rare diagnostic cells with changes of CMV infection were seen in 75% of the cases. The presence of CMV did not usually result in adverse patient outcome, except where CMV was one of the multiple opportunistic infections detected in the patient with AIDS.

Self-clotting method improves cell block preparation.

BACKGROUND: The success of cell block preparation is crucial for ancillary diagnostic tests in cytology. However, achieving an optimal cell block can be challenging. The current study describes a self-clotting-based technique for fine-needle aspiration (FNA) cell block preparations and evaluates its usefulness in comparison with the conventional needle wash technique. METHODS: The clinical data, FNA procedure, and cellularity of cell blocks of the self-clotting group (37 cases) and the conventional needle wash group (33 cases) were compared. The cellularity was evaluated using a scoring system (0 indicated acellular, 1 indicated 1-50 cells, and 2 indicated >50 cells). RESULTS: Approximately 76% of cases in the self-clotting group received a score of 2 versus 36% in the conventional needle wash group. Approximately 14% received a score of 1 in the self-clotting group compared with 9% in the conventional needle wash group, whereas 11% in the self-clotting group received a score of 0 versus 55% in the conventional needle wash group. The differences between the 2 methods were statistically significant. CONCLUSIONS: The results of the current study demonstrate that the self-clotting method is superior to the conventional needle wash method for FNA samples. Cancer Cytopathol 2018;126:190-9. © 2017 American Cancer Society.

"Low-grade squamous intraepithelial lesion, cannot exclude high-grade:" TBS says "Don't Use It!" should I really stop it?

BACKGROUND: The Bethesda System uses a two-tiered approach in the diagnosis of cervical squamous intraepithelial lesions (SILs). Occasionally, Papanicolaou (Pap) tests with evident low-grade SIL (LSIL) also have some features suggestive but not diagnostic of high-grade SIL (HSIL). This study reviews our experience with "Low-grade Squamous Intraepithelial Lesion, Cannot Exclude High-grade" (LSIL-H) and discusses the best approach to report such Paps if the LSIL-H interpretation is abandoned. METHODS: Abnormal Paps were identified between January and December 2014 that had surgical follow-up within 6 months. Their biopsy outcomes were compared. Statistical analysis was performed using Pearson's Chi-square and McNemar tests in SPSS software version 23. Statistical significance was defined as P ≤ 0.05. RESULTS: There were a total of 1049 abnormal Paps with follow-up. High-grade dysplasia/carcinoma (HGD+) was found in 8% of LSIL, 30% of LSIL-H, 52% of atypical squamous cells (ASCs), cannot rule out HSIL (ASC-H), and 77% of HSIL Paps. The detection rate of HGD+ for LSIL-H was between that of LSIL (Pearson's Chi-square test, P = 0.000) and ASC-H (P = 0.04). If LSIL-H cases are reported as ASC-H, the rate of HGD+ for the ASC-H category would decrease from 51.5% to 37.4% (McNemar test, P = 0.000). Alternatively, if LSIL-H cases are downgraded to LSIL, the rate of HGD+ for the LSIL category would rise from 7.7% to 10.4% (McNemar test, P = 0.000). Nearly 86.7% of LSIL-H cases were positive for high-risk HPV (HR-HPV) in comparison to 77.5% of LSILs, 100% of ASC-Hs, and 75% of HSILs. The sample size for HR-HPV and LSIL-H was too small for meaningful statistical analysis. CONCLUSIONS: "LSIL-H" category detects more HGD+ than LSIL, and fewer than ASC-H and HSIL. If LSIL-H is eliminated, Paps with this finding are best reported as ASC-H to ensure that women with potential HGD+ undergo colposcopy in a timely manner. Reporting LSIL-H as LSIL may delay colposcopy since management of LSIL Paps depends on multiple factors (age, HPV status, etc.).

Effect of Bethesda 2001 on reporting of atypical squamous cells (ASC) with special emphasis on atypical squamous cells-cannot rule out high grade (ASC-H).

We compared the overall ASC rate and the outcomes for women with different categories of ASC before and after TBS 2001 to evaluate the impact of TBS 2001. Our laboratory reported ASC in four subcategories before TBS 2001; ASC, favor reactive (ASC-R), favor low-grade squamous intraepithelial lesion (SIL) (ASC-L), undetermined significance (ASC-US), and ASC-H. Since the implementation of TBS 2001, we have been reporting ASC as ASC-US and ASC-H. After TBS 2001, our ASC rate decreased from 6.2% to 4% (P < 0.0001). Before TBS 2001, ASC-R carried the same risk as ASC-US for underlying condyloma/CIN 1. Moreover, ASC-R was rarely associated with CIN 2 or 3. Before TBS 2001, ASC-H captured more low-grade (condyloma/CIN 1) and less high-grade (CIN 2, 3, and SCC) lesions compared to after TBS 2001. More women with ASC-H after TBS 2001 underwent colposcopy (80% vs. 71%, P < 0.05). Twenty-two percent of women with ASC-H required 2-4 colposcopies to identify the high-grade lesion. Our results confirm that consistent application of TBS 2001 terminology for ASC reduces the ASC rate, and better identifies women at high risk for CIN 2, 3, and SCC. In addition, ASC-H requires very close clinical follow-up as a significant percentage of women require more than one colposcopy to identify the high-grade lesion.

Reporting endometrial cells in women 40 years and older: assessing the clinical usefulness of Bethesda 2001.

We assessed the usefulness of revised Bethesda System reporting of exfoliated benign endometrial cells (EMs) in postmenopausal women. Cervicovaginal cytology specimens with benign EMs in postmenopausal women and "out-of-phase EMs" in premenopausal women 40 years and older were identified. Cases with histologic follow-up within 12 months were selected. There was tissue follow-up for 130 postmenopausal women: 10 (7.7%) had significant findings (endometrial adenocarcinoma, 6 [2 (33%) in asymptomatic women]; complex atypical endometrial hyperplasia [CAH], 3; leiomyosarcoma, 1); 20 were receiving hormone replacement therapy (HRT; n = 15) or tamoxifen (n = 5); 2 (10%) had significant pathology (endometrial adenocarcinoma, 1; CAH, 1). Eight not taking hormones (7.3%) had significant pathology (adenocarcinoma, 5; CAH, 2; leiomyosarcoma, 1). There were follow-up data for 96 premenopausal women; only 1 (who had vaginal bleeding) had significant pathology (CAH). The difference in incidence of preneoplastic and neoplastic conditions after a cytologic interpretation of "benign EM" between postmenopausal and premenopausal women was significant (P pound .025); There was no difference between postmenopausal women receiving or not receiving HRT (P > .05). Reporting benign EMs for premenopausal women 40 years and older has no clinical significance but does for postmenopausal women, regardless of HRT and symptoms.

Atypical repair on Pap smears: clinicopathologic correlates in 647 cases.

The Bethesda system separates atypical reparative changes (ARC) from "typical" repair and places it into the atypical squamous cells of undetermined significance (ASC-US) category. The cytologic diagnosis of ARC represents both diagnostic and management challenges because its clinical significance is controversial and has not been fully investigated. On the basis of scant literature on follow-up of women with ARC on Papanicolaou (Pap) test, we reviewed data from our patient population, which consists of a mixture of low- and high-risk women. Six hundred forty-seven patients with ARC on their Pap tests were identified in a 7-yr period. Of this, 189 (29%) women were lost to follow-up. Of the 458 women with follow-up, 31% had cervical biopsies and 69% were followed by repeated Pap tests. The age ranged from 16 to 86 (mean 47 yr). The incidence of squamous intraepithelial lesion was 5.2% (5% low-grade and 0.2% high-grade). Most women (62%) with ARC on Pap test had a benign condition on follow-up. For some women, the immediate cause was not known and the Pap abnormality resolved spontaneously. Our study questions the validity of reporting ARC within the ASC-US category.

Metastases to the kidney: eleven cases diagnosed by aspiration biopsy with histological correlation.

Metastases to the kidney from extrarenal primary tumors are uncommon and may mimic renal-cell carcinoma clinically when presenting as a single mass with hematuria. Fine-needle aspiration biopsy (FNAB) is a useful diagnostic method for the evaluation of primary renal tumors. Only a few studies have investigated the value of cytological evaluation of secondary renal tumors. We report our experience with these tumors. Eleven cases of extrarenal primary tumors metastatic to the kidney, diagnosed by aspiration biopsy with histological correlation, are discussed. The diagnosis of metastatic disease to the kidney was accurately made by aspiration biopsy. Knowledge of the patients' history, histological correlation with the primary tumor, and the radiological characteristics of the renal masses were helpful in achieving a correct diagnosis. FNA cytology (FNAC) is an accurate method for the diagnosis of tumors metastatic to the kidney. Distinction between primary and secondary tumors of the kidney is crucial to guide management and prevent unnecessary surgery.

Sensitivity of high-risk HPV Hybrid Capture II (hrHPV HC2) test using SurePath(TM) specimens in the prediction of cervical high-grade squamous lesions.

INTRODUCTION: High-risk HPV (hrHPV) testing is now considered standard of care in the detection and management of cervical high-grade squamous intraepithelial lesions (HSIL/CIN 2-3) and their precursors. Recently, there has been concern in the scientific literature and lay media about the lack of data regarding the false-negative rate (FNR) of HPV testing on SurePath(TM) cytology specimens. This is a critical issue, since guidelines on the management of Pap test abnormalities rely heavily on HPV status. We undertook this study to determine whether HPV testing on SurePath(TM) specimens is less sensitive compared to reports in the literature for ThinPrep®. METHODS: We identified women with new diagnoses of CIN 2, CIN 3, and squamous cell carcinoma (SCC) on biopsy or excision in 2009-2013. For each patient, we recorded all SurePath(TM) cytology and hrHPV HC2 (high-risk HPV Hybrid Capture 2) test results from within 5 years prior to histologic diagnosis. Using the histologic diagnosis as the gold standard, we calculated the sensitivities of cytology and hrHPV HC2 tests for the detection of CIN 2, 3, and SCC. Our findings are based only on women who underwent biopsy or excision after having an abnormal cytology and/or positive HPV result. RESULTS: In our cohort, the sensitivity of testing in the 5 years prior to histologic diagnosis of CIN 2, 3, and SCC (combined as a single group) is 98.4% for SurePath(TM) cytology, 95.3% for hrHPV HC2, and 100% if both tests are used together. No conclusion can be drawn regarding testing for SCC alone, because there was only one case of SCC. CONCLUSION: Our results show that the false-negative rate of hrHPV HC2 testing on SurePath(TM) specimens for the detection of CIN 2 and CIN 3 is low and comparable to that of ThinPrep® specimens.

Interobserver variability in assessing adequacy of the squamous component in conventional cervicovaginal smears.

We compared the interobserver reproducibility of estimating the adequacy of the squamous component of conventional Papanicolaou (Pap) smears using traditional and newly proposed criteria. Forty conventional Pap smears with varying degrees of squamous cellularity were reviewed by 13 observers who evaluated adequacy (satisfactory vs unsatisfactory) based on the traditional criterion of estimating 10% slide coverage. After being introduced to the new criterion and the reference images, the observers reevaluated adequacy on the same set of smears, using the new criterion and the reference images. With the original criterion of 10% slide coverage, 15 smears had a unanimous designation; the overall kappa value was 0.49 (P < .001). With the newly proposed adequacy criterion and reference images, 17 smears had a unanimous designation; the overall kappa value was 0.60 (P < .001). The difference in the kappa correlation coefficients was statistically significant (P = .007). While traditional and newly proposed criteria resulted in fair interobserver agreement, it seemed that the newly proposed criterion, along with the use of reference images, for evaluating adequacy of the squamous component of conventional Pap smears results in better interobserver reproducibility.

ASCCP patient management guidelines: Pap test specimen adequacy and quality indicators.

Our objective was to provide management guidelines according to Papanicolaou (Pap) test specimen adequacy based on literature review and expert opinion. A task force named by the American Society for Colposcopy and Cervical Pathology (ASCCP) conducted a literature review and discussed appropriate management. The Steering Committee of the ASCCP and other experts reviewed the guidelines. The guidelines recommend a repeated Pap test in 12 months for most women undergoing routine annual/biennial screening if the current Pap test is negative but either lacks an endocervical/ transformation zone component or is partially obscured. Indications for considering an earlier repeat are also provided. The preferred managementfor unsatisfactory Pap tests is a repeated Pap test within a short interval of 2 to 4 months. The management guidelines will help promote optimal and uniform follow-up of women according to Pap test specimen adequacy.

False-positive squamous cell carcinoma in cervical smears: cytologic-histologic correlation in 19 cases.

Cytologic features of squamous intraepithelial lesions (SIL) can mimic those of invasive squamous-cell carcinoma. We compare and correlate the cytological findings of 19 false-positive squamous-cell carcinomas with follow-up cone biopsies or hysterectomy specimens to define which type of dysplasia is more prone to diagnostic errors on cervical Papanicolaou (Pap) smears. Out of 128 patients diagnosed with invasive squamous-cell carcinoma from 1994-2000, 19 (14.8%) with follow-up cone biopsies or hysterectomy specimens were false-positive cases, showing only cervical intraepithelial neoplasia (CIN). We reviewed tissue sections from these 19 cases of CIN for cytologic features of squamous-cell carcinoma, such as markedly pleomorphic and/or dysplastic squamous cells, necrosis, and nucleoli. Twelve of 19 patients (63%) were menopausal. The mean age was 50.5 yr. On review of cervical smears, 18 cases qualified for the cytologic diagnosis of squamous-cell carcinoma, keratinizing type, and one case qualified for squamous-cell carcinoma, nonkeratinizing type. Pleomorphic and/or keratinizing dysplasia was found in 15 out of 19 patients (79%), necrosis within superficial endocervical glands in 9 out of 19 patients (47%), and conspicuous nucleoli in 12 out of 19 patients (63%). One or more of these changes were seen in all but 2 patients (89%). Endocervical gland involvement was present and extensive in 18 of the 19 cases (94%). The mean age was older than expected for SIL (50.5 vs. a reported 40), and matched the mean age found in patients with invasive squamous-cell carcinoma. Pleomorphic and/or keratinizing dysplasia involving endocervical glands may exhibit the cytologic features of squamous-cell carcinoma on cervical Pap smears.

Cytology of primary central nervous system neoplasms in cerebrospinal fluid specimens.

Although two-thirds of tumors occurring in the central nervous system (CNS) are primary neoplasms, only 10% of positive cerebrospinal fluid (CSF) specimens are from primary CNS tumors. In this study, we reviewed the cytologic findings of 21 positive CSF specimens from primary CNS tumors. A computer search identified 21 cases of positive CSF specimens from patients with primary CNS tumors from the archives. Follow-up included review of medical charts and histologic correlation. The specimens were from 20 patients (9 females and 11 males). Their ages ranged from 6-83 yr, old with a mean of 30 yr. The cases included 9 medulloblastomas, 7 gliomas (3 glioblastoma multiformes, 2 anaplastic astrocytomas, and 2 ependymomas), 2 germinomas, 2 non-Hodgkin's large B-cell lymphomas, and 1 ganglioneurocytoma. Two cases were classified as suspicious and the remaining as positive for malignancy. Immunocytochemistry was employed in 3 cases to support the cytologic diagnosis. These cases included one large-cell lymphoma (leukocyte-common antigen-positive), one germinoma (placental alkaline phosphatase-positive), and the ganglioneurocytoma (neuron-specific enolase- and synaptophysin-positive). There were no false-positive cases. Our results suggest that positive CSF cytology in patients with a primary CNS tumor is a reliable indicator of malignancy and reflects leptomeningeal involvement. The use of immunocytochemistry is helpful in confirming the cytologic impression in some cases.