Sheila Segura

Introduction: p53 immunohistochemistry serves as a surrogate for TP53 mutation status and plays a central role in the classification of gynecologic malignancies. However, its reliability in cytology specimens, which often constitute the initial diagnostic material, remains incompletely characterized in routine practice. Materials and Methods: After institutional review board (IRB) approval, we performed a retrospective study of 51 gynecologic malignancies and borderline tumors with paired cytology (body fluids and fine‐needle aspiration specimens) and surgical resections. p53 staining patterns were classified as wild‐type or aberrant (overexpression, null, cytoplasmic). Concordance between specimen types and correlation with TP53 mutational status were assessed. Cytology specimens were fixed in CytoRich Red fixative, while surgical specimens were formalin‐fixed. Results: Overall concordance between cytology and surgical specimens was 100% (51/51 cases), spanning multiple specimen types and tumor categories. Aberrant p53 expression was most commonly characterized by overexpression (62.8%), followed by a null pattern (23.5%). Aberrant p53 expression corresponded with TP53 mutations in all cases with available molecular data. Cytology established the initial diagnosis in 60.8% of cases, highlighting its role in early classification. Conclusion: p53 immunohistochemistry demonstrates highly reproducible, pattern‐based interpretation in cytology specimens, supporting its use as a reliable surrogate for TP53 status at the time of initial diagnosis. These findings reinforce the role of p53 in enabling early and accurate classification of gynecologic malignancies when surgical material is limited or unavailable.

Background: The emergence of personalized therapy and molecular sequencing has significantly transformed the management of advanced non–small cell lung carcinoma (NSCLC). Supernatants obtained from fine needle aspiration (FNA) have been shown to be a reliable source of high‐quality nucleic acid for molecular testing. One pre‐analytic factor that may influence the success of molecular analysis is the collection medium used for the FNA rinse. Methods: This prospective study evaluated 31 FNA specimens collected in three different media—CytoRich Red, 10% neutral buffered formalin, or 10% bovine serum albumin (BSA)—to assess their impact on DNA yield and real‐time polymerase chain reaction (RT‐PCR) outcomes for KRAS ( Kirsten rat sarcoma viral oncogene homolog) and/or EGFR (epidermal growth factor receptor) mutations. DNA extraction was performed using the QIAamp DNA FFPE (formalin‐fixed paraffin‐embedded) Tissue Kit (Qiagen), and RT‐PCR was conducted using the QIAGEN therascreen EGFR or KRAS RGQ PCR kits. Concurrent direct smears or formalin‐fixed, paraffin‐embedded tissue from a cell block or core biopsy served as standard controls. Results: In the CytoRich Red group, 11 specimens (9 adenocarcinomas and 2 NSCLC) were analyzed. All 11 specimens generated results concordant with standard controls. One specimen required repeat DNA extraction due to initial pre‐test quality control failure. The formalin group included 12 specimens (7 squamous cell carcinomas [SCC], 3 adenocarcinomas, 1 NSCLC, and 1 large cell neuroendocrine carcinoma). Nine specimens produced concordant results with controls. Two specimens were negative for EGFR mutations, although the corresponding control tissues were not tested. Six required repeat DNA extraction because of initial pre‐test failure, and one specimen remained invalid after re‐extraction. Eight specimens were collected in BSA (5 adenocarcinomas, 2 NSCLC, and 1 SCC). Five produced concordant results with standard controls, while three specimens failed PCR pre‐tests despite repeat DNA extraction. Conclusion: In this small cohort, CytoRich Red demonstrated reliable performance for RT‐PCR‐based mutation testing. FNA supernatant represents a practical alternative specimen for lung cancer molecular analysis and may optimize specimen utilization while preserving cell block material.

Background: Anal cancer rates are rising among women, immunocompromised individuals, and Men who have sex with Men (MSM), independent of HIV status. While high‐resolution anoscopy remains the diagnostic standard, limited access has increased interest in alternative screening. HPV testing now complements or replaces cytology in cervical cancer prevention, but evidence for anal screening, especially using extended genotyping platforms like BD Onclarity, remains limited. Methods: This single center cross‐sectional study enrolled high‐risk individuals undergoing anal cancer screening from May 2024 to May 2025. Anal cytology and high‐risk HPV (HR‐HPV) DNA testing were performed using the BD Onclarity assay on the BD COR system. Demographics, cytology results, HRHPV genotypes, and available histopathology were analyzed. Results: Among 117 high‐risk participants (median age 42; 106 males, 8 females, 3 transgender women), 74.3% were ‐HIV positive, predominantly MSM‐. Abnormal cytology occurred in 41% of cases including ASC‐US ( n = 29; 60.4%), LSIL ( n = 16; 33.3%), ASC‐H ( n = 2; 4.1%), and HSIL ( n = 1; 2.1%). Valid BD Onclarity results were available for 104 individuals, with 61.5% testing HR‐HPV positive. Non‐16/18 HR‐HPV types were most common overall, while HPV16 predominated in abnormal cytology cases. Twelve individuals underwent biopsy, revealing no dysplasia ( n = 2), AIN1 ( n = 5), AIN2 ( n = 1), and AIN3 ( n = 4). Conclusions: Extended HR‐HPV genotyping using the BD Onclarity assay demonstrated a high prevalence of HR‐HPV in this high‐risk population, with multiple genotype infections commonly observed. While non‐HPV16/18 genotypes predominated overall, HPV16 was more frequently associated with abnormal cytologic and high‐grade histologic findings. Given the limited number of biopsy‐confirmed high‐grade lesions, these associations should be interpreted as descriptive and hypothesis‐generating. Cytology remains central to anal cancer screening, with extended HPV genotyping serving as a complementary adjunct rather than a standalone stratification tool. Larger longitudinal studies with systematic histopathologic follow‐up are needed to clarify the clinical significance of extended HPV genotyping in this population.

Background: The 2014 Bethesda System for Reporting Cervical Cytology classifies squamous intraepithelial lesions (SILs) of cervix into two main categories: low‐grade SIL (LSIL) and high‐grade SIL (HSIL). In some clinical practices, the LSIL cannot rule out high‐grade lesion (LROH) interpretive category is used in cases with LSIL and findings that may raise the possibility of HSIL. Our purpose is to assess follow‐up histopathology and high‐risk human papillomavirus (hrHPV) results in patients with LROH, in comparison with LSIL, atypical squamous cells, cannot rule out HSIL (ASC‐H), and HSIL in our institution.Design: Cervical Papanicolaou tests with LROH, LSIL, ASC‐H and HSIL interpretation, surgical follow‐up, and hrHPV status were retrieved from the computer database from May 2014 to December 2016.Results: Of 109 963 total Papanicolaou tests, LROH comprised 0.3%, LSIL 3.1%, ASC‐H 0.2% and HSIL 0.4%. Only 3272 cases with surgical diagnoses were included in the study. The most common histological outcome for ASC‐H was cervical intraepithelial neoplasia (CIN)2/3 (32.6%); LSIL was CIN 1 (45.7%); LROH was CIN 1 (46.7%) and HSIL was CIN 2/3 (64.4%). For LROH and LSIL, 31.1% and 7.5% respectively, had CIN 2/3. Approximately 79% of cases were hrHPV positive. Of LROH cases with surgical follow‐up, 86.9% tested hrHPV positive, accounting for the second most common positive group after HSIL (92.6%).Conclusion: In our study cohort, LROH interpretation is associated with a higher number of CIN 2 or higher lesions on follow‐up compared to patients with LSIL (P < 0.0001), and is associated with a significant percentage of positive other hrHPV, supporting LROH as a useful diagnostic category that triggers appropriate follow‐up in affected women.