Stratified Mucin-Producing Intraepithelial Lesion (SMILE) of the Uterine Cervix: High-Risk HPV Genotype Predominance and p40 Immunophenotype.

Stratified mucin-producing intraepithelial lesion (SMILE) is a rare high-grade cervical precancerous lesion designated a variant of adenocarcinoma in situ (AIS) in the WHO classification. We aimed to determine HPV genotypes, immunohistochemical phenotype and mucin presence in SMILE. Between 2010 and 2018, SMILE was diagnosed in 34 out of 6958 (0.5%) cervical biopsies, in 23 patients. Twenty-six tissue samples from twenty-one patients were available for further analysis, including 13 with SMILE alone, 12 with SIL and/or AIS and one with HSIL, AIS and endocervical adenocarcinoma. HPV genotyping was performed using the Seegene Anyplex II HPV 28 assay. Of the 26 samples, a single HPV genotype was identified in the majority of cases (n = 22), including 12/13 SMILEs associated with SIL/AIS. All but one were high-risk HPV genotypes (23/24; 96.8%). We identified seven different HPV genotypes, the most common being HPV16 (n = 10; 43.5%), HPV18 (n = 8, 34.8%) and HPV 31 (n = 5, 21.7%). All SMILEs showed a strong positive reaction to p16, CK7, CK19 and high Ki67 expression comparable to adjacent HSIL and/or AIS if present. SMILE showed variable mucin presence and p40-positive squamous differentiation suggesting phenotypic diversity in cervical precancerous lesions infected by single HPV.

A novel combined animal tissue model for freehand and ultrasound-guided fine needle aspiration biopsy and smear preparation techniques training.

OBJECTIVE: A variety of models are used for fine needle aspiration biopsy (FNAB) and smear preparation techniques training: human, animal and silicon models or combined models. We present fresh animal tissues as models for freehand and ultrasound (US)-guided FNAB technique training, enabling an integrated approach from tumour detection to smear evaluation. METHODS: We introduced a novel combined animal tissue model using dietary animal meat with covering skin as a substrate. Animal liver tissue of various sizes, representing tumour, was inserted into the various layers of the substrate (subcutaneous fat, muscle tissue, proximity of bone). Freehand and US-guided FNAB smear preparation, including fixation, was then performed and assessed. RESULTS: The use of a combined animal tissue model for 6 freehand and 3 US-guided FNAB sessions showed a statistically significant improvement in the US-guided FNAB retrieval of liver tissue (Fisher's exact test, p =  .0216), in smear preparation technique reflected in a decrease in the number of too thick smears after freehand FNAB (Fisher's exact test, p  =  .0070), in the overall number of smears satisfactory for evaluation by US-guided FNAB (Fisher's exact test, p =  .0206) and in the number of flawless smears obtained in the freehand FNAB training sessions (Fisher's exact test, p =  .0020). CONCLUSIONS: A unique advantage of the presented model encompassing various layers of animal tissues with covering skin, offers an integrated approach for FNAB training from "tumour" detection, puncture precision, to smear preparation and cytological evaluation for a wider audience and does not compromise patient safety.

Histomorphologic assessment and distribution of high-risk human papillomavirus (HPV) types in cervical high-grade squamous intraepithelial lesions with unusual histomorphologic features.

In rare cases, equivocal histomorphology ('deceiving dysplasia') does not allow immediate diagnosis of cervical high-grade squamous intraepithelial lesion (HSIL). We studied whether these cases are correlated with specific high-risk human papillomavirus (hr HPV) types. During 2011-2017, 39 cases of p16-positive cervical tissue biopsies with unusual ('deceiving') dysplastic histomorphology were identified and matched with the same number of controls (typical HSIL samples). Histomorphological characteristics were reviewed blindly and HPV testing was performed using the clinically validated RealTime test (Abbott) and Anyplex HPV 28 (Seegene). HPV 16 and HPV 31 were the two most frequent HPV types in both groups, although minimum, proportional, hierarchical and any etiological attribution estimates for HPV 16 were significantly lower in the deceiving group (13.2%, 21.3%, 23.7% and 23.7%) than in the control group (32.4%, 48.1%, 48.6% and 48.6%). In addition, the distribution of other hr HPV types differed between the two study groups, with five HPV types (HPV 56, 58, 59, 73 and 82) detected only in the deceiving group. Histomorphologic review of both groups (regardless of HPV type) confirmed significant differences in nuclear atypia, maximum lesion thickness and cellularity, although these were diminished when cross-comparisons between HPV16/18 and non-HPV16/18 cases pooled from both study groups were evaluated. Different attribution estimates for HPV 16, HPV 16/18 and non-16/18 hr HPV types in deceiving and control groups were observed, in particular for HPV 16. However, an unusual (deceiving) histomorphology may also depend on unknown HPV-related molecular changes.

Use of vimentin immunocytochemical staining for evaluation of atypical cells in voided urine samples.

BACKGROUND: Cytomorphology of exfoliated atypical reactive/repair renal tubular cells (RRTC) can resemble atypical urothelial cells thus suggesting a differential diagnostic question of urothelial neoplasia in urinary cytology. Vimentin expression has been shown in RRTC and used for differentiation from atypical urothelial cells. METHODS: The institutional computer database was searched for urinary cytology cases with vimentin immunocytochemical staining (2008-2012). Original cytopathological diagnoses based on cytomorphology and the results of vimentin immunostaining were compared to follow-up data, including histopathological diagnosis, subsequent urinary cytopathology reports, and clinical findings. RESULTS: Of the 42 cases with vimentin immunocytochemical staining, 33 were positive and 9 negative. Consequently, significant renal disease was found in 9/33 (27%) of vimentin positive cases and nehrolithiasis in 4/33 (12%) of vimentin positive and 1/9 (11%) of vimentin negative cases. Erythrocyturia of undetermined origin was diagnosed in nine cases (seven vimentin positive and two negative). Urinary cytology follow-up was negative in three vimentin positive cases. Urothelial carcinoma was found in 3/9 (30%) of vimentin negative cases. Thirteen patients were lost to follow-up. CONCLUSIONS: Vimentin immunocytochemical staining could be used as an ancillary method for evaluation of atypical cells in urinary specimens in selected cases with RRTC exhibiting cytological atypia. Unnecessary diagnostic procedures for evaluation of urothelial carcinoma could be avoided in vimentin positive cases and further diagnostic work-up for evaluation of a significant renal disease could be suggested in vimentin positive cases. Diagn. Cytopathol. 2017;45:85-90. © 2016 Wiley Periodicals, Inc.

Microinvasive cervical squamous cell carcinoma in Slovenia during the period 2001-2007.

BACKGROUND: Microinvasive squamous cell carcinoma (MISCC) comprises a significant portion of all cervical cancers in Slovenia. Criteria of carcinomatous invasion are well described in the literature, however histopathological assessment of MISCC is difficult, because morphological characteristics can overlap with cervical intraepithelial neoplasia grade 3 (CIN 3) and other pathological changes. The aim of our study was to evaluate the reliability of the histopathological diagnosis of MISCC in Slovenia during the period from 2001 to 2007. MATERIALS AND METHODS: Data on patients with a histopathological diagnosis of cervical MISCC (FIGO stage IA) in the period of 2001 to 2007 were obtained from the Cancer Registry of Slovenia. Histological slides were obtained from the majority of pathology laboratories in Slovenia. We received 250 cases (69% of all MISCC) for the review; 30 control cases with CIN 3 and invasive squamous cell carcinoma FIGO stage IB were intermixed. The slides were coded and reviewed. RESULTS: Among 250 cases originally diagnosed as MISCC, there was an agreement with MISCC diagnosis in 184 (73.6%) cases (of these 179/184 (97.3%) cases were FIGO stage IA1 and 5/184 (2.7%) cases were FIGO stage IA2). Among 179 FIGO stage IA1 cases 117 (65.4%) showed only early stromal invasion. CONCLUSIONS: The retrospective review of cases diagnosed as MISCC during the period 2001-2007 in Slovenia showed a considerable number of overdiagnosed cases. Amongst cases with MISCC confirmed on review, there was a significant proportion with early stromal invasion (depth of invasion less than 1 mm).

Typing of renal tumors by morphological and immunocytochemical evaluation of fine needle aspirates.

Image-guided fine needle aspiration biopsy (FNAB) of renal masses can accurately evaluate malignancy. Adjunct methods are needed for accurate typing of renal cell carcinomas (RCC) and benign neoplasms. Cytopathological diagnoses of 79 routine ultrasound-guided FNAB of renal lesions were compared to consequent histological diagnosis and size of tumors. Cytology samples were sufficient for immunocytochemical subtyping in 43 cases (54.4%). The median tumor size was 2.8 cm, with 57 cases (76%) smaller than or equal to 4 cm. When a panel of immunocytochemical stainings (vimentin, CK7, CD117, P504S) was applied, accurate diagnoses were obtained in 11/12 (91.7%) of clear cell RCC (CRCC), 14/17 (82.3%) of papillary RCC (PRCC) and 5/7 (71.4%) of chromophobe RCC (ChRCC), respectively. Substantial cell pleomorphism with unusual immunostainings led to erroneous diagnosis of pheochromocytoma in CRCC with eosinophilic cytoplasm. Only 30% of CRCC were correctly diagnosed in the group without immunostaining, seven were suspicious for CRCC, and the remainder had unrepresentative material for CRCC. Cytopathological diagnoses were less accurate in oncocytomas (n=11), regardless of immunocytochemical staining. Cystic nephromas (n=2) and MEST (n=1) were overdiagnosed as suspicious and positive for PRCC, respectively, with immunocytochemical staining not assisting in correct diagnosis. RCC can be accurately typed as CRCC, PRCC or ChRCC in fine needle aspirates in a routine clinical setting if the cellular material is sufficient and a panel of antibodies is used (vimentin, CK7, P504S, CD117). The classification of oncocytomas and cystic nephromas is not reliable since atypical morphology and immunocytochemical reactions overlap with RCC.

Impact of different sectioning of excision biopsies of the cervical transformation zone for detection of invasive cervical cancer in 2 European centers related to national cervical screening programs.

BACKGROUND: The impact of the number of sections used in cervical excision biopsies of transformation zone on sensitivity of histological detection of cervical cancer is poorly documented. AIM: To assess whether different techniques of sectioning cervical excision biopsies in 2 European laboratories influence the finding of cervical cancer by histopathological examination. MATERIALS AND METHODS: The routine assessment at the Department of Histopathology and Cytology, Sheffield Teaching Hospitals NHS Foundation Trust, Sheffield, UK (Sheffield), encompassed sectioning a cone in 3-mm tissue blocks and cutting 3 levels per block. At the Institute of Pathology, Faculty of Medicine, University of Ljubljana, Slovenia (Ljubljana), cones were cut in a 3- to 4-mm-thick tissue block, and 10 levels were cut per block, spaced every 100 microm. The number of blocks and levels per block were assessed for each cone. Histopathological detection of cervical carcinoma between the laboratories was compared. RESULTS: Among 820 cones in the Sheffield laboratory, we detected 35 invasive carcinomas, whereas 6 invasive carcinomas were detected among 94 cones in the Ljubljana laboratory. Although the Slovenian laboratory examined a significantly larger number of levels per cone biopsy (64 vs 24 in Sheffield), this was not associated with a larger proportion of invasive cervical carcinoma, especially foci of stage IA1 or high-grade cervical intraepithelial neoplasia diagnosed on excision biopsies: 6.4% of invasive cervical carcinomas (5 [83.3%] of stage IA1) in Ljubljana and 4.3% (30 [85.7%] of stage IA1) in Sheffield laboratory (P > 0.05); 71.2% of high-grade cervical intraepithelial neoplasia in Ljubljana and 75.5% in Sheffield (P > 0.05). CONCLUSIONS: Our assessment showed that a method with a large number of levels per cervical cone or large loop excision of the transformation zone biopsy did not increase sensitivity for the detection of cervical cancer. However, extensive sectioning substantially affects the pathologist's workload, and this may need to be reconsidered.