Tuesday, December 15, 2020

Conditions associated with the need for additional needle passes in ultrasound‐guided thyroid fine‐needle aspiration with rapid on‐site pathology evaluation

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Abstract

Background

Rapid on‐site evaluation (ROSE) is a valuable tool for specimen adequacy assessment in thyroid ultrasound (US)‐guided fine‐needle aspiration (US‐guided FNA). To reduce the risk of nondiagnostic samples, additional needle passes may be needed at ROSE to ensure adequate sampling. Recommendations regarding the number of aspirates to ensure specimen adequacy are not well defined. Furthermore, there are limited data regarding nodule characteristics that may require increased sampling. In this study, we investigate conditions associated with requiring more than three needle passes during ROSE.

Methods

A retrospective quality review of all patients who underwent US‐guided thyroid FNA by a single board‐certified radiologist over a 1‐year period was performed. A total of 122 patients were identified: 70 with three passes performed and 52 with more than three passes to achieve adequacy.

Result

Our data demonstrate that large nodules (≥3 cm) were more likely than small nodules (≤1.1 cm) to require more than three passes to achieve adequacy. If a nodule was predominantly cystic or mixed cystic and solid, the sample was often adequate with only three passes. In cases of thyroiditis or nodules suspicious or diagnostic of neoplasia, there is a trend to require only three passes for adequacy.

Conclusion

On the basis of the data presented in this study, cytopathologists should be prepared for the potential need to obtain additional needle passes in larger (≥3 cm) nodules and provide reassurance to patients that this is an anticipated finding for these larger nodules.

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DNA methylation patterns of RAR‐β2 and RASSF1A gene promoters in FNAB samples from Greek population with benign or malignant breast lesions

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Abstract

Background

Promoter hypermethylation is common in Breast Cancer (BC) with studies mainly in histological specimens showing frequent methylation of tumor suppressor genes (TSGs) compared with normal tissues. The aim of this study was to estimate the frequency of promoter methylation of RAR‐β2 and RASSF1A genes in breast FNAB material aiming to evaluate the methylation status of these two genes as biomarker for detecting BC in Greek population.

Methods

FNAB material from 104 patients was collected for cytological evaluation and epigenetic analysis. DNA was extracted and subjected to bisulfite conversion. A methylation‐specific PCR was carried out and the final products were separated with electrophoresis in 2% agarose gels.

Results

From 104 samples, RASSF1A hypermethylation was observed in 78 (75%) and RAR‐β2 hypermethylation in 64 (61.6%). 84% and 78% of the cases diagnosed with breast malignancy (n = 50) were methylated for RASSF1A and RAR‐β2, respectively. Methylated RASSF1A and RAR‐β2 were also detected in 88.3% and 76.5% in samples diagnosed as suspicious for malignancy (n = 17) and in 57.2% of samples diagnosed with atypia (n = 14). The Odds Ratio for breast malignancy was 4.545 in patients with RASSF1A hypermethylation and 9.167 in patients with RAR‐β2 hypermethylation underlying their promoter's methylation positive correlation with breast malignancy.

Conclusion

To optimize the sensitivity and specificity of this epigenetic setting, more TSGs related to BC should be gradually imported in our evaluated methylation panel and be validated in a larger study sample with the aim that the obtained epigenetic profiles will provide clinicians with valuable tools for management of BC patients in Greece.

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Optimized liquid‐based cytology for the cellular and molecular analysis of oral keratinocytes: A promising diagnostic tool

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Abstract

Background

Liquid‐based cytology (LBC) has improved exfoliative cytology by facilitating the extraction of more precise information from epithelial cells. The aim of this study was to optimize a protocol using a conventional cytobrush to perform LBC, obtaining oral keratinocytes for their further cellular and molecular analysis.

Methods

LBC was performed in 30 healthy donors from buccal mucosa. We evaluated the use of diethyl pyrocarbonate (DEPC)‐treated Dulbecco's Modified Eagle Medium (DMEM) medium right after the collection of the cells. Cell morphology and viability were determined by Orcein staining and flow cytometry, respectively. RNA was extracted by the trizol method, and evaluated with spectrometry and electrophoresis. Finally, RNA was copied into cDNA and GAPDH and TLR2 genes were amplified by reverse transcription polymerase chain reaction (RT‐PCR) and quantitative reverse transcription polymerase chain reaction (RT‐qPCR) using specific primers.

Results

Only DEPC‐treated DMEM preserved the viability of intact intraepithelial keratinocytes. RNA quantity and quality improves in samples treated with DEPC. RNA integrity is comparable with a cell line control. GAPDH gene was successfully amplified by RT‐PCR and RT‐qPCR.

Conclusions

Therefore, LBC performed under these conditions becomes a reproducible technique for the retrieval of intraepithelial oral keratinocytes with good cell viability for cytomorphometric analysis, and extraction of good RNA quality suitable for molecular analyses such as PCR. We propose this LBC protocol as a complementary method to the cellular and molecular study of oral mucosa pathologies; however, it requires further study.

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“Single‐cell pattern” of adenocarcinoma cells in effusion cytology: Morphologic challenges of lung cancer

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Abstract

Background

Lung adenocarcinomas present as tight clusters and three‐dimensional balls in effusion specimens. Unlike carcinomas of breast and stomach where singly lying malignant cells are seen in effusion samples, lung adenocarcinomas usually show cohesive morphology. This single‐cell pattern may also be confused with reactive mesothelial cells. We studied the frequency of pulmonary adenocarcinoma with single‐cell pattern cytomorphology in pleural effusion specimens.

Materials and Methods

All cases reported as either suspicious or positive for malignancy on pleural effusion cytology (PFC) over the past 1 year were retrieved. The clinical details were obtained from requisition forms. Cases with predominant single‐cell pattern, clinically suspicious of carcinoma lung were segregated. These were de‐stained and immunocytochemistry (ICC) for TTF‐1 was performed.

Results

Of 103 cases reported as either suspicious or positive for malignancy on PFC, 29 had a predominant single‐cell pattern. Of these, 13 (44.8%) were primary lung carcinoma. The rest were metastasis from ovary (5; 17.2%), breast (2; 6.9%), stomach (2; 6.9%), lymphoma (1; 3.5%), and Ewing's sarcoma (1; 3.5%). Five (17.2%) were those with unknown primary. All cases of lung carcinoma were positive for TTF‐1 ICC.

Conclusion

Single‐cell pattern of pulmonary adenocarcinoma is commoner than popularly believed. This pattern may be difficult to differentiate from carcinoma cells of other sites as well as from reactive mesothelial cells. A high degree of suspicion is therefore needed to perform relevant ICC to clinch the correct diagnosis.

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Liquid phase human papillomavirus genotype analysis of aspirated metastatic head and neck squamous cell carcinoma: Fine needle aspiration supernatant is a rich source of tumor DNA that can increase the diagnostic yield

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Abstract

Background

Most patients with human papillomavirus (HPV)‐related head and neck squamous cell carcinoma (HNSC) present with lymph node metastasis. In these patients, fine needle aspiration (FNA) is not only a diagnostic tool, but a means for determining HPV status. HPV status, in turn, is used to determine tumor origin, prognosis, and even guide therapy. Thus, the limited sampling afforded by FNA must be optimized to meet heavy clinical demands.

Purpose

The purpose of this study was to determine whether the residual supernatant portion of the FNA could serve as a resource for reliable determination of HPV status

Design/Method

25 FNAs from 24 patients with metastatic HNSC underwent HPV genotyping of post‐centrifuged supernatant fluid from FNA needle rinses. HPV genotyping was performed using two real time PCR‐based assays, the two‐step LightCycler and the one‐step automated cobas HPV tests. HPV status of the supernatant was compared with the paired FNA cell blocks and/or surgical tissue samples.

Results

The supernatant was adequate for HPV testing in 24 (96%) of 25 cases. Of these, 14 (56%) were HPV positive and 11 (44%) negative by the LightCycler assay. HPV16 was the most commonly detected genotype (n = 12). When results of supernatant and paired cell block testing were compared, HPV status was concordant in all cases. The LightCycler method was more sensitive than the cobas assay due to its ability to detect an expanded profile of HPV variant genotypes.

Conclusion

The current standard of practice for patients with HNSC who undergo FNA is to construct a cell block and then discard the supernatant. This supernatant is a rich source of tumor DNA that can be used to detect HPV status. It should not be wasted.

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Utility of p16 and HPV testing in oropharyngeal squamous cell carcinoma

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Background

In the US, 60% to 80% of oropharyngeal squamous cell carcinomas (OPSCCs) are associated with human papillomavirus (HPV). However, until recently, no consensus existed about when and how to test for HPV in patients with head and neck cancers. We aimed to evaluate the use of p16 and HPV testing at our institution because p16 immunohistochemistry is reportedly a reliable surrogate marker for HPV detection in OPSCCs.

Methods

We identified all cases at our institution of primary or metastatic squamous cell carcinoma (SCC) of the head and neck with a concurrent p16 immunostain analysis from January 1, 2013, through August 31, 2018. Patient demographic data, tumor characteristics, p16 result, and any HPV result (in situ hybridization and E6 and E7 RNA test) were captured.

Results

We identified 104 patients. Most primary tumors (53/57 [93.0%]) and metastases (40/47 [85.1%]) were positive for p16. Thirty‐seven cases (35.6%) had reflex high‐risk HPV (HR HPV) testing performed. Of the 35 p16‐positive cases, 6 had discrepant HR HPV results (p16+/HPV). We identified 47 p16 immunostains that were performed on lymph nodes with primary tumors of unknown origin. Most were cytology cases (34/47 [72.3%]), and most were p16 positive (40/47 [85.1%]). Neither tumor differentiation nor tumor keratinization was predictive of p16 positivity. Tumors with basaloid differentiation were universally p16 positive.

Conclusion

p16 immunohistochemistry accurately identifies HPV‐positive OPSCC. Cytology specimens have an important role in characterizing SCC of unknown origin. HR HPV testing is not routinely required, and results may be discrepant with p16 findings.

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Cytomorphology and diagnostic pitfalls of sebaceous and nonsebaceous salivary gland lymphadenoma: A multi‐institutional study

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Abstract

Background

Salivary gland lymphadenoma (LAD) is a rare benign neoplasm comprising sebaceous (SLAD) and nonsebaceous (NSLAD) types. Despite established histologic criteria, limited data on cytomorphology, tumor heterogeneity, and overlap with other entities make the diagnosis of LAD by fine needle aspiration (FNA) challenging. We describe a multi‐institutional cohort of 14 LADs with cytology, clinical, radiologic, and histopathologic data.

Methods

Our cohort included nine SLAD and five NSLAD with corresponding histopathology. Mean patient age and M:F ratio were 60.4 years (range 45‐86 years) and 1:2 for SLADs and 57.4 years (range 42‐80 years) and 1:1.5 for NSLADs, respectively. One NSLAD patient had a germline predisposition for Cowden syndrome. Glass slides and whole slide images of air‐dried Diff‐Quik (DQ), alcohol‐stained Papanicolaou smears (Pap) and cellblocks were reviewed for key cytomorphologic findings.

Results

FNAs from SLAD and NSLADs demonstrated vacuolated and basaloid epithelial clusters within a lymphoid background. Vacuolated cells from SLAD showed sebaceous cells with microvesicular cytoplasm indenting a central nucleus. Vacuolated cells from NSLAD were columnar with eccentric nuclei, corresponding to abluminal glandular cells. SLADs were classified using the Milan System for Reporting Salivary Gland Cytopathology as nondiagnostic (11.1%), nonneoplastic (44.4%), atypia of uncertain significance (AUS) (22.2%), and salivary gland neoplasm of uncertain malignant potential (SUMP) (22.2%). NSLADs were classified as AUS (40%), SUMP (40%) and Benign Neoplasm (20%).

Conclusion

Although rare, knowing the cytologic features of salivary LAD is important to avoid diagnostic pitfalls. Vacuolated cells can be prominent in both SLAD and NSLAD aspirates. Diagnostic issues arise from insufficient sampling of all tumor components leading to marked variation in diagnostic classification of LAD.

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Cytologic features and immunohistochemical findings of epithelioid hemangioendothelioma (EHE) in effusion: A case series

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Abstract

Background

Epithelioid hemangioendothelioma (EHE) is a rare malignant vascular tumor characterized by WWTR1‐CAMTA1, t (1:3) (p36;q25) translocation in 90% of cases. Without prior EHE history, it can mimic other malignant effusions. Recently, CAMTA1 was published as an excellent immunohistochemical surrogate marker for molecular testing for WWTR1‐CAMTA1 fusion in surgical specimens.

Methods

A 6‐year retrospective search using our computer system was performed for cases diagnosed as EHE on effusion cytology and surgical specimens. The clinical presentation, cytologic findings and immunohistochemical stain results, including CAMTA1 were reviewed.

Results

Four pleural and one peritoneal effusions were identified. The median age was 52 years with a female to male ratio of 3:2. Most patients presented with pulmonary symptoms. The cytologic features were non‐specific easily mimicking other malignancies; especially in the absence of known prior malignancy. This was exemplified by one of our cases which was initially misdiagnosed as adenocarcinoma. Intracytoplasmic erythrocytes were present only on the cell blocks but not on cytology. The cytology cell blocks from patients with prior EHE confirmed on surgical biopsies stained positive for vascular markers (CD31, ERG) and CAMTA1.

Conclusion

The features of EHE in effusion are non‐specific and a diagnostic pitfall in cytology. In the absence of prior EHE diagnosis, inclusion of this entity in the differential diagnoses and application of immunohistochemical stain panels will be prudent in avoiding a misdiagnosis. However, in cases with prior EHE diagnosis, CAMTA1 could serve as diagnostic marker; especially on limited cytology material. Additional studies will be helpful in supporting our results.

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Concordance of HPV, conventional smear, colposcopy, and conization results in cervical dysplasia

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Abstract

Background

Cervical cancer screening algorithms are increasingly focused on Human Papillomavirus (HPV)‐based screening while the accuracy of using abnormal cytological findings to detect dysplastic lesions still remains important. This retrospective study correlated the results of conventional cervical cytology, colposcopy guided biopsy, and cold knife conization (CKC) procedures performed in a tertiary center.

Materials and Methods

Data from 9399 patients who underwent screening with conventional cervical cytology between 2010 and 2019 was obtained from the hospital registry. Abnormal cervical cytology and high‐risk HPV DNA genotypes were recorded and their colposcopic and CKC pathology was determined.

Results

Two hundred and ninety two patients underwent colposcopy for abnormal cervical cytology and/or high‐risk HPV positivity. One hundred and twenty three patients were positive for High‐risk HPV. Abnormal cervical cytology was detected in 216 patients. The most common cytological anomaly was atypical squamous cells of undetermined significance (ASCUS) found in 9399 patients (1.39%). It was determined that conventional cytology had a sensitivity of 70.8% and a specificity of 62.2% for the detection of low‐grade lesions, while it had a sensitivity of 72.4% and a specificity of 86.0% for the detection of high‐grade lesions. CKC was applied to 68 patients who were diagnosed with high‐grade squamous intraepithelial lesions (HSIL) as a result of the colposcopy. As a result of CKC, a high‐grade lesion was detected in 73.5% of these patients.

Conclusion

Conventional cervical cytology and colposcopy exhibited higher accuracy as the severity of lesions increased. Detection of HPV may prevent unnecessary surgical procedures, especially with ASCUS.

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Cytological diagnosis of angiomatoid fibrous histiocytom

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Abstract

Angiomatoid fibrous histiocytoma (AFH) is an unusual superficially located tumor primarily affecting children and young adults. It grows slowly and most often occurs on the extremities. There is a paucity of literature on the cytological findings of AFH owing to the rarity of the lesion and its superficial location which makes it easier to perform the biopsy. Here, we present a case of AFH in a 7‐year‐old girl who presented with a left upper arm swelling. The cytology of this tumor along with histopathologic correlation and review of literature is discussed.

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Cytological diagnosis of angiomatoid fibrous histiocytoma

alwin shared this article with you from Inoreader

Abstract

Angiomatoid fibrous histiocytoma (AFH) is an unusual superficially located tumor primarily affecting children and young adults. It grows slowly and most often occurs on the extremities. There is a paucity of literature on the cytological findings of AFH owing to the rarity of the lesion and its superficial location which makes it easier to perform the biopsy. Here, we present a case of AFH in a 7‐year‐old girl who presented with a left upper arm swelling. The cytology of this tumor along with histopathologic correlation and review of literature is discussed.

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Fine needle aspiration cytology of cervical lymph nodes: Comparison of liquid based cytology (SurePath) and conventional preparation

alwin shared this article with you from Inoreader

Abstract

Background

Fine needle aspiration cytology (FNAC) is the first diagnostic step in patient with cervical lymphadenopathy because of its simplicity, safety and early availability of the results. Liquid‐based cytology (LBC) is an alternative processing method which is used for both gynecological and nongynecological samples. Literature reviewed show few studies comparing LBC with conventional preparation (CP).

Aim

The present study was undertaken to evaluate the efficacy of LBC and comparison of LBC and CP in cervical lymphadenopathy.

Materials and Methods

In this prospective study, a total of 75 cases of FNAC with cervical lymphadenopathy were included. The first pass was used for CP followed by LBC with the use of SurePath (SP) technique. Both the smears were compared for cellularity, background containing blood, cell debris, lymphoglandular bodies, stromal fragments, cytoarchitectural pattern, etc., by semiquantitative scoring system.

Results

There was no statistical difference in the cellularity, cell architecture, and monolayer cells (P > .05). On the basis of background containing blood, cell debris, lympho‐glandular bodies, stromal fragments (P < .001), nuclear, and cytoplasmic details (P < .05), LBC was found to be superior to CP.

Conclusion

LBC is a relatively simple technique and superior to CP in respect of better nuclear and cytoplasmic details with loss of background blood and debris. It has a diagnostic accuracy equivalent to that of CP. However, use of both LBC and CP can result in better diagnostic accuracy.

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