Utilization of Dynamic Telecytopathology for Rapid Onsite Evaluation of Touch Imprint Cytology of Needle Core Biopsy: Diagnostic Accuracy and Pitfalls.

Background: In the era of personalized medicine, there has been an increase in demand on cytopathology service to perform rapid onsite evaluation of touch imprints (ROSETIs) of needle core biopsies (NCBs) to ensure sample adequacy and provide preliminary diagnosis. Limited publications have addressed use of telecytopathology for ROSETIs. We present our experience with telecytopathology-guided ROSETIs of NCBs. Materials and Methods: Cytotechnologist onsite transmitted real-time images of Diff-QUIK stained touch imprints of NCBs using an Olympus digital camera through Ethernet to a pathologist. The preliminary diagnosis from telecytopathology-guided ROSETIs was compared with that of the final NCB for diagnostic accuracy. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated. Results: Of 186 cases, 163 (87%) were concordant and 23 (13%) were discordant. Of the 23 discordant cases, 14 (58%) were diagnosed as benign with final NCB diagnoses as malignant and 9 (41%) were suspicious on ROSETIs with final NCB diagnoses as benign. The causes of discordance among cases categorized as benign on preliminary and malignant on final included interpretative error (9) and sampling (6). Six cases categorized as suspicious on preliminary and negative on final biopsy diagnosis correlated with concurrent fine-needle aspiration, raising the possibility of loss of diagnostic areas in processed tissue cores. Remaining three cases in this category represented misinterpretation of reactive cells. Sensitivity, specificity, PPV, and NPV were 87.9%, 95.7%, 97.1%, and 82.7%, respectively. Conclusions: Telecytopathology-guided ROSETI yields high accuracy for preliminary interpretation of NCBs and may be utilized as an effective substitute for conventional microscopy.

Dynamic Telecytopathology-Guided Rapid On-Site Assessment of Percutaneous Image-Guided Fine-Needle Aspiration of Hepatic Lesions: An Institutional Review of 178 Cases.

Background: Rapid on-site evaluation (ROSE) of image-guided fine-needle aspiration (IGFNA) of hepatic lesions is necessary to ensure adequate sample collection and render preliminary diagnosis. We review our experience with utilization of telecytopathology for ROSE of IGFNA of liver lesions. Materials and Methods: Cytotechnologists at a remote site transmitted real-time images of Diff-Quik-stained cytology smears of hepatic lesions obtained using an Olympus DP70 digital camera via Ethernet to a pathologist. All live communications were conducted using the Vocera voice communication system. Preliminary telecytopathology ROSE diagnosis was compared with the final diagnosis for accuracy. Results: ROSE using telecytopathology of IGFNAs of hepatic lesion was performed on a total of 178 cases. Preliminary diagnosis of unsatisfactory, benign, suspicious for malignancy, and positive for malignancy was 2%, 28%, 15%, and 55%, respectively, for telecytopathology. The overall concordance between the preliminary and final cytology diagnoses was 94.4%. Nine of the cases were upgraded to suspicious/malignant final diagnosis contributing to an upgrade rate of 5%. One case with preliminary evaluation of positive for malignancy was downgraded to benign cytology on final diagnosis resulting in a downgrade rate of 0.1%. Conclusion: Utilization of telecytopathology is highly effective and accurate for ROSE of IGFNAs of hepatic lesions and may serve as a substitute for conventional microscopy.

Telecytopathology for on-site adequacy evaluation decreases the nondiagnostic rate in endoscopic ultrasound-guided fine-needle aspiration of pancreatic lesions.

OBJECTIVE: Advances in digital imaging methods have resulted in use of telecytology in the immediate assessment of fine-needle aspiration (FNA) specimens. We retrospectively compared the nondiagnostic rate for endoscopic ultrasound-guided (EUS) FNA of pancreatic lesions in two groups: one with on-site evaluation for adequacy via telecytopathology and the other without on-site adequacy evaluation. SUBJECTS AND METHODS: All patients undergoing EUS-FNA of pancreatic lesions over a 2-year period were included. Direct smears were immediately wet-fixed or air-dried, and any residual material was rinsed in saline for cell block or cytospin preparation. Patients were divided into two groups: Group 1 had on-site telecytopathology evaluation for adequacy by a cytopathologist, and Group 2 had no on-site adequacy evaluation. The cytologic diagnoses were reviewed, and the nondiagnostic rates for each group were calculated. The age, sex, and characteristics of pancreatic lesions (solid versus cystic) between the two groups were compared. RESULTS: In total, 217 patients were included. Telecytopathology on-site evaluation was provided for 95 (43.8%) cases. There was no difference between the groups in terms of age and sex. Pancreatic lesions were predominantly solid in the group that underwent telecytopathology on-site evaluation (p<0.005). The nondiagnostic rates for solid lesions in Group 1 and Group 2 were 3.7% and 25.6%, respectively (p<0.0001). Although the nondiagnostic rate for cystic lesion was higher in Group 2, it did not reach a level of statistical significance (16.5% versus 7.1%; p=0.249). After adjusting for the effects of sex and lesion characteristics (solid versus cystic lesion) with multivariate logistic regression, the odds of having a nondiagnostic specimen in Group 2 was 6.9 times greater than in Group 1, and the result was statistically significant (p=0.0013). CONCLUSIONS: Telecytopathology on-site evaluation of EUS-FNA of pancreatic lesions reduces the nondiagnostic rate, especially in lesions with solid characteristics, and may serve as an effective substitute for on-site evaluation by a cytopathologist.

Intraductal papillary mucinous neoplasms of the pancreas: Cytologic-histologic correlation study and evaluation of the cytologic accuracy in identifying high-grade dysplasia/invasive adenocarcinoma.

Objective: Intraductal papillary mucinous neoplasms (IPMNs) may be associated with invasive adenocarcinoma, low-grade dysplasia (LGD), or high-grade dysplasia (HGD). We aimed to review the cytologic-histologic correlation of cases with a histologic diagnosis of IPMN. Material and Methods: A database search (January 2010-January 2021) was performed for resected IPMNs with preceding endoscopic ultrasound-guided fine-needle aspiration (FNA). Cytology slides were reviewed for the presence of benign, atypical, or malignant cells, and necrosis. Histologically, IPMNs were classified as benign (LGD) or malignant (HGD or adenocarcinoma). Results: There were 41 patients with IPMN; 24 malignant and 17 benign. Sixteen of the 24 malignant IPMNs were accurately classified as malignant on cytology. There were eight false negatives and one false positive. Cytology yielded a sensitivity of 67% and a specificity of 94%. Among the 16 true positives with FNA diagnosis of adenocarcinoma, seven were IPMNs with HGD, and nine had invasive adenocarcinomas on histology. Cellular morphology and absence or presence of necrosis did not help distinguish HGD from adenocarcinoma on cytology (P > 0.5). Sampling errors and interpretative errors resulted in false-negative cases. Cytology yielded diagnoses related to IPMN in 73% of cases (30/41) and lack of identification of mucinous cells/mucinous background resulted in interpretative errors (9). Conclusion: This study shows that there is a good correlation between cytopathology and surgical pathology diagnoses of IPMNs and that cytology is mostly able to recognize IPMN with HGD/adenocarcinoma. However, heterogeneity in areas of IPMN with HGD/adenocarcinoma may result in sampling errors yielding false-negative cases. Mucinous cells/background should raise the suspicion of IPMN on cytology, even when no neoplastic epithelium is present for the evaluation of dysplasia.

Feasibility of dynamic telecytopathology for rapid on-site evaluation of endobronchial ultrasound-guided transbronchial fine needle aspiration.

OBJECTIVE: Rapid on-site evaluation (ROSE) at the time of endobronchial ultrasound-guided transbronchial fine needle aspiration (EBUS-TBFNA) is useful in obtaining adequate samples and providing preliminary diagnosis. We present our experience with ROSE of EBUS-TBFNA using telecytopathology. MATERIALS AND METHODS: Real-time images of Diff-Quik (Mercedes Medical, Sarasota, FL)-stained cytology smears were obtained with an Olympus (Olympus America, Center Valley, PA) digital camera attached to an Olympus CX41 microscope and transmitted via ethernet by a cytotechnologist to a cytopathologist in a cytopathology laboratory who rendered a preliminary diagnosis while communicating with an on-site cytotechnologist via the Vocera (San Jose, CA) voice communication system. The endoscopy suite was located a block away from the cytopathology laboratory. Accuracy of ROSE via telecytopathology was compared with an equal number of cases that received ROSE, prior to introduction of telecytopathology, via conventional microscopy. RESULTS: ROSE was performed on a total of 200 EBUS-TBFNAs. The telecytopathology system and conventional microscopy were used to evaluate equal numbers of cases (100 each). Preliminary diagnoses of negative/benign, atypical/suspicious, and positive for malignancy were 58%, 14%, and 24% for telecytopathology and 57%, 10%, and 31% for conventional microscopy. Four percent of telecytopathology cases and 2% of conventional microscopy cases were deemed unsatisfactory at the time of ROSE. The overall concordance between the preliminary and final diagnoses was 96% for telecytopathology and 93% for conventional microscopy. The causes of discordant preliminary and final diagnoses could be mainly attributed to difficulty in distinguishing small cell carcinoma versus reactive lymph node due to crush artifact, atypia related to reactive bronchial epithelial cells, and availability of cell block material and Papanicolaou-stained slides for review at the time of final cytologic sign out. CONCLUSIONS: Telecytopathology is comparable with conventional microscopy in ROSE of EBUS-TBFNA. It can serve as a valid substitute for conventional microscopy for on-site assessment of EBUS-TBFNA.

Financial and educational impact of the COVID-19 pandemic in an academic hospital-based tertiary cytopathology practice.

BACKGROUND: The ongoing COVID-19 pandemic has led to a dramatic shift in volumes and practice patterns for hospitals around the globe. We analyzed its effect on the cytopathology subspecialty practice and resident education at our institution. DESIGN: Specimen volumes were analyzed for the cytology practice for 2019 and 2020. Patient registration and elective and scheduled surgery volumes were also included in the analysis for 2020. The impact of innovative concepts, such as virtual teaching, on resident teaching was evaluated using a survey consisting of 5 multiple choice questions with 4 possible responses each. RESULTS: The total number of specimens decreased by 28% in March 2020 (P < 0.00001), with a continuing decline in April (66% decrease year-over-year, P < 0.00001), followed by recovery in May and return to baseline within June 2020. Specimen volumes continued to show an upward trend thereafter. Improved specimen volumes correlated with patient registration and surgical volumes. The majority of residents considered virtual teaching conferences (75%) and self-study sets (58%) as beneficial and did not view absence of one-on-one microscope learning (58%) as significantly affecting their education. CONCLUSION: The recovery curve for our cytopathology service was V-shaped, essentially the most ideal response to an economic downturn. The majority of residents viewed virtual teaching conferences and self-study sets favorably and did not regard absence of one-on-one microscope learning as adversely affecting their education.

Telecytopathology for rapid preliminary diagnosis of ultrasound-guided fine-needle aspiration of thyroid nodules.

BACKGROUND: Onsite evaluation of ultrasound-guided fine-needle aspiration (USGFNA) of thyroid nodules is essential to procure adequate samples and provide initial assessment. We present our experience with onsite evaluation of USGFNA of thyroid nodules using telecytopathology. METHODS: Real-time images of Diff-Quik-stained cytology smears were obtained with an Olympus Digital camera attached to an Olympus CX41 microscope and transmitted via ethernet by a cytotechnologist to a pathologist who rendered a preliminary diagnosis while communicating with an onsite cytotechnologist over the phone. Accuracy of preliminary diagnosis was compared with final diagnosis, retrospectively. RESULTS: A total of 79 patients (mean age 48.9 year) underwent USGFNA of 100 thyroid nodules. Preliminary diagnoses of benign, suspicious/malignant, and unsatisfactory were 72%, 7%, and 21%, respectively. Of the 72 cases initially reported as benign all remained benign on the final cytology. Of the seven suspicious/malignant cases on initial cytology, five were suspicious/malignant and two were benign on final cytology. Of the 21 cases that were initially interpreted as unsatisfactory only four were reclassified as benign on final diagnosis. The accuracy rate between the final cytology and preliminary telecytopathology diagnosis was 94%. Presence of additional material on Papanicolaou-stained slides and cellblock sections was the main reason for discrepancy that accounted for six discrepant cases. CONCLUSIONS: On-site telecytopathology for thyroid USGFNAs is highly accurate compared with final cytologic evaluation. It allows pathologists to use their time more efficiently and makes onsite evaluation at a remote site possible.

To whom the specimen goes: a look at how touch preparations and core needle biopsies are handled in different practices and the effect on fellowship education.

INTRODUCTION: Core needle biopsies (CNBs) have proven to be an excellent source of tissue for diagnosis and ancillary testing in the era of personalized medicine, commonly yielding sufficient material for testing via a relatively minimally invasive technique. Thus, there has been an increase in touch preparations (TPs) evaluated with rapid onsite evaluation (ROSE) of these small biopsies either in isolation or with concurrent fine needle aspiration (FNA). This in turn has forced cytopathology practices to make decisions with regard to processing and workflow of CNBs, which affects cytopathology fellowship education substantially. STUDY DESIGN: The present review is based on a review of recent literature and an evaluation of the authors' personal experiences. RESULTS AND CONCLUSIONS: Deciding whether CNBs with associated TPs should be assigned to the cytology service, the subspecialty or general surgical pathology service, or a split between cytopathology and surgical pathology, is complicated. The workflow is variable at different institutions depending on multiple factors. Each of these routes has benefits and disadvantages that can affect patient care and laboratory workflow, in addition to having downstream effects on the quality and type of education our pathology trainees receive. Herein, the advantages and disadvantages of the different approaches for CNB triage are discussed, with an emphasis on the impact upon cytopathology fellowship education.

PAX-8 Expression in Salivary Duct Carcinoma.

Salivary duct carcinoma (SDC) is a high-grade adenocarcinoma resembling breast ductal carcinoma. It accounts for ~10% of malignant tumors of the salivary glands. Most cases show expression of CK7 and androgen receptor. PAX-8 is a transcription factor, with expression reported in renal, Müllerian, and thyroid carcinomas. Previous studies have described an absence of PAX-8 immunostaining in most primary salivary gland neoplasms, including SDCs. However, PAX-8 expression is frequently found in neoplasms that can metastasize to salivary glands, suggesting the possibility that this protein can be used to differentiate SDC from secondary neoplastic involvement of the salivary gland. We evaluated the expression of PAX-8 in 14 cases of SDC from our institution. One case showed diffuse moderate to strong PAX-8 positivity, while 2 tumors showed focal weak staining. Therefore, we conclude that although the majority of SDC are negative for PAX-8, rare diffuse positivity can be seen in these primary salivary gland tumors. This could potentially pose difficulty in ruling out metastatic disease from another PAX-8-positive primary neoplasm.

Reparative change with extensive squamous metaplasia: a potential diagnostic pitfall on thyroid aspiration.

Squamous metaplasia and cystic change can appear in the thyroid and mimic a variety of lesions. In this report, a patient with two dominant left thyroid nodules underwent fine needle aspiration (FNA) biopsy for each nodule. One of the nodules was diagnosed as consistent with papillary carcinoma and the other as suspicious for papillary carcinoma. Subsequent near total thyroidectomy revealed both lesions to be part of a goiter with extensive squamous metaplasia. This article discusses a potential pitfall in the interpretation of fine needle aspirates of squamous metaplastic cells in the thyroid.

Incidence of Non-Salivary Gland Neoplasms in Patients with Warthin Tumor: A Study of 73 Cases.

Warthin tumor is the second most common benign parotid neoplasm. Its association with non-salivary gland neoplasms has been sporadically reported. We reviewed clinical records of Warthin tumor diagnosed on aspiration cytology and surgical pathology to determine if there is any association with other extra-salivary gland malignant neoplasms. Computer search was made for all cases of Warthin tumor diagnosed in the parotid gland by aspiration cytology and surgical pathology at our institution between January 2007 and August 2016. Clinical records of all cases were reviewed for any associated malignant neoplasms and any surgical follow up. All available cytology and histologic material was reviewed. Seventy-three patients (mean 66.9, M:F 1.1:1, age range 43 to 87 years) with Warthin tumor were identified. 45 (62%) were diagnosed on aspiration cytology only, 19 (26%) had cytologic diagnosis as well as concordant surgical follow up, and 9 (12%) were diagnosed based on surgical pathology only. Average age for patients with and without secondary malignancy was 70.5-years, and 63.4-years, respectively (p < 0.05). Average pack years for patients with and without secondary malignancy was 45.4, and 39.8, respectively (p > 0.05). Twenty-seven (37.0%) patients harbored a malignant neoplasm. Association of extra salivary gland malignant neoplasms in 37.0% of our cases suggest that the prevalence of secondary non-salivary neoplasms in patients harboring Warthin tumor might have been underestimated. Squamous cell carcinoma was the most commonly associated non-salivary malignant neoplasm. The association of Warthin tumor with smoking plays an important role in this increased rate of malignancy, and this is supported by the fact that smoking is highly associated with head and neck and lung cancers.

Cytomorphologic findings of cervical Pap smears from female-to-male transgender patients on testosterone therapy.

BACKGROUND: The cervical cancer screening recommendation for transgender female-to-male (FTM) patients is the same as that for cisgender females. A lack of literature on testosterone-induced changes in cervical cytology in these patients may result in interpretation errors, especially without a proper clinical history. The aim of this study was to delineate the Papanicolaou (Pap) test findings in this patient population. METHODS: A pathology laboratory information system was used to obtain a cohort of FTM transgender patients on testosterone therapy (2009-2019). A cohort of age-matched, atrophic, control cisgender female patients (postpartum or menopausal) was selected. A retrospective review of the cytomorphologic findings on cervical Pap smears, pertinent follow-up, and human papillomavirus (HPV) test results was performed. RESULTS: Fourteen transgender patients (age range, 21-64 years; mean age, 42.5 years) receiving testosterone therapy with 17 Pap smears were identified. One of the 5 available HPV tests was positive for HPV, and 4 were negative. A Pap smear review revealed the following: negative for intraepithelial lesion (NILM; 82.4%), unsatisfactory (5.9%), atypical squamous cells of undetermined significance (ASCUS; 5.9%), and low-grade squamous intraepithelial lesion (5.9%). The Pap smears of the atrophic cisgender cohort (102 patients) revealed the following: NILM (92.5%), unsatisfactory (0.9%), ASCUS (5.6%), and high-grade squamous intraepithelial lesion (0.9%). The difference between the rates of epithelial cell abnormality in the 2 cohorts was not statistically significant. Although atrophy was noted in both groups, cytomorphologic findings of transitional cell metaplasia (TCM; 88.2%) and "small cells" (82.4%) were characteristic of the testosterone-treated transgender cohort. Histologic correlates of TCM and small cells were noted in hysterectomy specimens from 6 patients. CONCLUSIONS: Small cells and TCM are common cytomorphologic findings in Pap smears of testosterone-treated transgender (FTM) patients. On the basis of histologic follow-up, small cells most likely represent atrophic parabasal cells of cervical-vaginal epithelium.

Anti-NMDAR-Positive Small-Cell Lung Cancer Paraneoplastic Limbic Encephalitis: A Case Report and Literature Review.

INTRODUCTION: Paraneoplastic limbic encephalitis (PLE) is a rare disease that presents as rapid onset dementia characterized by short-term memory loss (STM), anxiety, and behavioral changes. Anti-NMDAR antibodies are unfrequently reported in PLE associated with small-cell lung cancer (SCLC). Given that PLE can precede the diagnosis of cancer, it is very important that once infectious, metabolic, nutritional, or structural disorders associated with short-term memory loss are ruled out that vigorous effort must be made to rule out underlying malignancy. CASE: We report a rare case of PLE as the presenting symptom of SCLC. A 72-year-old male with history of COPD was brought to the ED by his wife after he was found to have short-term memory loss, including forgetfulness of his wedding anniversary the day before, and anxiety. Neurological exam showed impaired short-term recall on MOCA. CT head showed no evidence of infarct. Lumbar puncture was performed which showed lymphocytic pleocytosis, a nonspecific inflammatory change. CSF panel was negative for HSV, Neisseria, Hemophilus, E. coli, and HIV. Initial EEG was unremarkable, though a repeat EEG showed mild slowing of the posterior dominant rhythm consistent with mild encephalopathy. MRI showed equivocal increased FLAIR on T2-weighted images in the bilateral temporal lobes, left greater than right. CTA thorax showed bulky mediastinal and right hilar LAD. FNA of the R4 lymph node revealed SCLC. The NM bone scan showed no osteoblastic lesions. While the serum autoantibody panel was positive for anti-NMDAR, the CSF autoantibody panel returned entirely negative. Chemotherapy with etoposide and cisplatin was started on Day 4 of admission. The patient's neurological symptoms showed improvement following chemotherapy. CONCLUSION: This case highlights the importance of recognizing short-term memory loss as a feature of PLE.

FNA of misclassified primary malignant neoplasms of the thyroid: Impact on clinical management.

BACKGROUND: Fine needle aspiration (FNA) cytology is a popular, reliable and cost effective technique for the diagnosis of thyroid lesions. The aim of our study was to review cases of misclassified primary malignant neoplasms of the thyroid by FNA, and assess the causes of cytologic misdiagnosis and their impact on clinical management. METHODS: Clinical data, FNA smears and follow-up surgical specimens of cases diagnosed with primary thyroid carcinoma were reviewed. RESULTS: Of the 365 cases with a malignant diagnosis by FNA over a period of 11 years, nine (2.4 %) were identified with discrepant histologic diagnosis with regard to the type of primary thyroid malignancy. In addition, four cases were added from the consultation files of the Massachusetts General Hospital. Areas of difficulty contributing to misclassification included overlapping cytologic features (n = 6), rarity of tumors (n = 3), and sampling limitations (n = 4). Of the 13 cases, 12 underwent total or near total thyroidectomy and one patient had concurrent surgical biopsy. Measurement of serum calcitonin levels in one case, with an initial cytologic diagnosis of medullary carcinoma, prevented unnecessary lymph node dissection. Misclassification of medullary carcinoma as papillary carcinoma precluded lymph node dissection in one case. Further management decisions were based on the final histologic diagnosis and did not require additional surgery. Two cases of undifferentiated (anaplastic) thyroid carcinoma were misdiagnosed as papillary thyroid carcinoma. Both patients received total thyroidectomies, which may not otherwise have been performed. CONCLUSIONS: A small subset of primary malignant neoplasms of the thyroid may be misclassified with regard to the type of malignancy on FNA. The majority of primary malignant neoplasms diagnosed on FNA require thyroidectomy. However, initial cytologic misclassification of medullary carcinoma or undifferentiated carcinoma as other malignant neoplasms or vice versa may have an impact on clinical management.

Calcium pyrophosphate dihydrate deposition disease (CPPD)/Pseudogout of the temporomandibular joint - FNA findings and microanalysis.

We report a case of a Calcium pyrophosphate dihydrate deposition disease (CPPD) presenting as a mass in the parotid and temporomandibular joint (TMJ) that simulated a parotid tumor. A 35 year-old man presented with pain in the left ear area. A CT Scan of the area showed a large, calcified mass surrounding the left condylar head, and extending into the infratemporal fossa. FNA of the mass showed birefringent crystals, most of which were rhomboid with occasional ones being needle shaped, embedded in an amorphous pink substance. Scanning electron microscopy (SEM) with energy dispersive x-ray spectroscopy (EDS) of these crystals showed peaks corresponding to calcium and phosphorus. SEM/EDS is a rapid method of diagnosing calcium pyrophosphate dihydrate deposition disease (CPPD) and an alternative to more commonly used method of special staining of cell block sections coupled with polarizing microscopy.

HPV16-Immortalized Cells from Human Transformation Zone and Endocervix are More Dysplastic than Ectocervical Cells in Organotypic Culture.

A major risk factor for cervical cancer is persistent infection with high-risk human papillomaviruses (HPV) which can cause cervical intraepithelial neoplasia. Greater than 90% of cervical cancers develop in the transformation zone (TZ), a small region of metaplastic squamous epithelium at the squamocolumnar junction between endocervix and ectocervix. However, it is unclear why this region is highly susceptible to malignant progression. We hypothesized that cells from TZ were more susceptible to dysplastic differentiation, a precursor to cervical cancer. We used three-dimensional organotypic culture to compare differentiation of HPV16-immortalized epithelial cell lines derived from ectocervix, TZ, and endocervix. We show that immortal cells from TZ or endocervix form epithelia that are more dysplastic than immortal cells from ectocervix. A higher percentage of immortal cells from TZ and endocervix express the proliferation marker Ki-67 and are positive for phospho-Akt. Immortal cells from TZ and endocervix invade collagen rafts and express increased levels of matrix metalloproteinase-1. Inhibition of MMP-1 or Akt activity blocks invasion. We conclude that HPV16-immortalized cells cultured from TZ or endocervix are more susceptible to dysplastic differentiation, and this might enhance their susceptibility to cervical cancer.

Histopathologic and Cytologic Follow-Up in High Risk Male Patients with Unsatisfactory Anal Cytology.

OBJECTIVE: Anal cytology is being increasingly used as part of anal cancer screening in patients at high risk for anal neoplasia. Most studies in anal cytology have focused on correlating the abnormal anal Pap smear with histopathologic outcomes. The aim of this study was to document histopathologic or repeat anal cytology outcomes in patients with unsatisfactory cytology. MATERIALS AND METHODS: Unsatisfactory anal Pap tests in high risk male patients were correlated with follow-up histopathologic diagnoses or cytology. RESULTS: 1205 anal tests were performed during the study period and 214 (17.8%) were unsatisfactory. Adequate follow-up cytology was available in 75 cases and revealed epithelial cell abnormality (ECA) in 40% [30/75] (atypical squamous cells of undetermined significance (ASCUS) [20%] and low-grade squamous intraepithelial lesions (LGSIL) [20%]) and was negative for intraepithelial lesion or malignancy (NILM) in 60% [45/75] of cases. 28.7% of unsatisfactory Pap smears had unsatisfactory repeat cytology. Histopathological follow-up on these unsatisfactory Pap smears revealed anal intraepithelial neoplasia (AIN) 1 and AIN 2/3 or 2/3+ in 39% and 18% of the total number of biopsy cases, respectively. CONCLUSIONS: High risk male patients with unsatisfactory Pap smears are at significant risk of epithelial cell abnormality and histopathologically verifiable anal intraepithelial lesions.

Utility of Immunohistochemistry and ETV6 (12p13) Gene Rearrangement in Identifying Secretory Carcinoma of Salivary Gland among Previously Diagnosed Cases of Acinic Cell Carcinoma.

Objective. Secretory carcinoma is a recently described entity with characteristic immunoprofile and ETV6 (12p13) rearrangement. Before its initial description, it was generally diagnosed as acinic cell carcinoma (ACCi). We evaluated immunoprofile and ETV6 rearrangement in cytological and surgical cases of previously diagnosed ACCi, in an attempt to identify any misclassified SC. Methods. Fifteen cytology and surgical cases of ACCi diagnosed over a 13-year period were retrieved and subjected to immunohistochemistry for S-100, mammaglobin, GATA-3 and DOG-1 as well as FISH for ETV6 (12p13). Results. Of the 8 cytology cases, only 1 was positive for S100, GATA-3, and mammaglobin, and negative for DOG-1. It also demonstrated ETV6 rearrangement and was reclassified as SC. The same immunoprofile was present in 2 of the 13 surgical cases. ETV6 rearrangement characterized by 3' interstitial deletion was detected in one of these cases and was reclassified as SC. Immunohistochemistry and ETV6 rearrangement were useful in identifying 2 (13.3%) cases misclassified as ACCi. Conclusions. Characteristic immunoprofile and ETV6 gene rearrangement may prove useful in identifying cases of SC. The presence of ETV6 3' interstitial deletion in one of our cases suggests that there may be additional ETV6 related genetic alterations contributing to the pathogenesis of SC.

Destaining of Diff-Quik stained cytologic smears is not necessary for the detection of anaplastic lymphoma kinase gene rearrangement in lung adenocarcinoma by fluorescence in situ hybridization.

BACKGROUND: Anaplastic lymphoma kinase (ALK) gene rearrangement analysis by fluorescence in situ hybridization (FISH) is one of the standard molecular tests for targeted therapy of lung adenocarcinoma. However, insufficient cell block cellularity may impede molecular testing. A recent study showed that Diff-Quik (DQ) stained cytology smear is suitable for ALK by FISH. AIMS: The aim of our study was to observe the impact of destaining intervals on the quality of FISH signals and determine if DQ smears without destaining would allow FISH analysis. MATERIALS AND METHODS: Thirty-five DQ smears from 27 cases of lung adenocarcinoma were analyzed for ALK gene rearrangement by FISH. Twenty three DQ smears were destained for different intervals, including 30 s (13 cases), 1 min (6 cases), or 2 min (4 cases). Twelve DQ smears were not subjected to destaining. For further validation, FISH signals in 8 smears and 6 cell blocks were compared with the paired destained DQ smears. The signal quality was semi-quantified and analyzed with Chi-squared test. RESULTS: Of the total 27 selected cases, three (11%) were positive for ALK gene rearrangement, whereas 24 (89%) were negative. FISH signal was satisfactory in all DQ smears. There was no significant difference in the quality of signal among smears with different destaining intervals (P = 0.55) or between smears with and without destaining (P = 0.41). DQ smears without destaining showed identical FISH results and similar or better signals as compared with paired destained smears and cell blocks in all cases. CONCLUSIONS: Duration of destaining intervals does not impact the quality of FISH signal on DQ smears. Destaining of DQ smears is not necessary for ALK by FISH.

Rapid on-site evaluation with dynamic telecytopathology for ultrasound-guided fine-needle aspiration of head and neck nonthyroid lesions.

BACKGROUND: Rapid on-site evaluation (ROSE) at the time of ultrasound-guided fine-needle aspiration (USGFNA) of head and neck lesion is essential for obtaining adequate samples and providing the preliminary diagnosis. We summarize our experience with ROSE of USGFNA on head and neck nonthyroid lesions using telecytopathology. MATERIALS AND METHODS: Real-time images of Diff-Quik stained cytology smears were obtained at ultrasound suite with an Olympus DP-70 digital camera attached to an Olympus CX41 microscope, and transmitted via ethernet by a cytotechnologist to a cytopathologist in cytopathology laboratory who rendered a preliminary diagnosis. Live communication was conducted with Vocera voice communication system. The ultrasound suite was located on different floor from the cytopathology laboratory. Accuracy of ROSE via telecytopathology was compared with an equal number of cases that received ROSE, prior to introduction of telecytopathology, via conventional microscopy. RESULTS: Rapid on-site evaluation was performed on a total of 116 USGFNA of head and neck nonthyroid lesions. The telecytopathology system and conventional microscopy was used to evaluate equal number of cases (58 each). Preliminary diagnoses of benign, atypical/suspicious for malignancy, and positive for malignancy were 72.4%, 17.2% and 10.3% for telecytopathology, and 69.0%, 10.3% and 20.7% for conventional microscopy. None of the cases were deemed unsatisfactory. The overall concordance between the preliminary and final diagnoses was 94.8% for telecytopathology and 98.3% for conventional microscopy and was not statistically significant (P = 0.309). The causes of discordant preliminary and final diagnoses were mainly attributed to availability of cell block and Papanicolaou-stained slides for review or flow cytometry results for lymphoma cases at the time of final sign out. CONCLUSIONS: Telecytopathology is comparable with conventional microscopy in ROSE of USGFNA of head and neck nonthyroid lesions.