Yoshiaki Norimatsu

Assessment of the localization of chondroitin sulfate in various types of endometrial carcinoma

Introduction This study aimed to assess the localization of chondroitin sulfate (CS), a primary extracellular matrix component, in the stromal region of endometrial carcinoma (EC). Methods Immunostaining was performed on 26 endometrial endometrioid carcinoma (EEC) samples of different grades and 10 endometrial serous carcinoma (ESC) samples to evaluate CS localization. This was further confirmed by Alcian Blue (AB) staining as well. Results In the G1-EEC samples, CS showed reactivity with fibrovascular stroma, supporting closely packed glandular crowding and papillary structures. As the grade increased, the original interstitial structure was re-established, and the localization of CS in the perigulandular region decreased. In the ESC samples, the thick fibrous strands supporting the papillary architecture showed reactivity with CS; however, the delicate stromal region branching into the narrow region showed poor reactivity. The AB staining results showed similar characteristics to the immunostaining ones. Conclusions The characteristic localization of CS in various EC types was elucidated. The present study provides new information on endometrial stromal assessment.

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 < 0.001) or B (44.3 ± 28.8, P < 0.05). Method B yielded significantly higher numbers of cell clusters than method A (P < 0.001). Method A (132.2 ± 107.7, p < 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.

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.

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.