FNA needle rinses preserved in Cytolyt are acceptable specimen type for mutation testing of thyroid nodules.

INTRODUCTION: This study investigated the application of molecular testing to residual thyroid fine-needle aspiration material from needle rinses collected in Cytolyt. MATERIALS AND METHODS: Two thyroid needle rinses from 135 patients were collected in Cytolyt during routine diagnostic workup in our institution. Molecular testing was performed to detect 14 genetic alterations in BRAF, K-, H-, N-RAS genes as well as RET/PTC1, RET/PTC3, and PAX8/PPARγ and verified by next generation sequencing and correlated with cytologic diagnoses. RESULTS: Molecular testing revealed a total of 17 mutations across specimens with benign nodule (n = 5; HRAS, NRAS), Hürthle cell neoplasm (n = 2; BRAF, HRAS) and Papillary thyroid carcinoma (n = 10, 9 BRAF, 1 KRAS) cytology. No RNA gene rearrangements were detected. CONCLUSIONS: Mutations and translocations associated with thyroid cancer can be detected in thyroid fine-needle aspiration needle rinses preserved in Cytolyt specimens collected during routine patient management, which are typically discarded when a diagnosis is attained.

Genotype-specific prevalence and distribution of human papillomavirus genotypes in underserved Latino women with abnormal Papanicolaou tests.

INTRODUCTION: Knowledge about the prevalence and distribution of human papillomavirus (HPV) genotypes in cervical premalignant and malignant lesions is crucial to guide development of clinical management strategies and prophylactic vaccines. The aim of this study was to determine HPV genotype-specific prevalence and distribution in an underserved cohort of Latino women. MATERIALS AND METHODS: From December 2009 to April 2011, 808 SurePath cervicovaginal specimens were collected from women who were referred from charity clinics for abnormal Papanicolaou tests. The patients' average age was 36.5 years (range 19-85 years). The specimens were tested for HPV genotypes by DNA microarray and sequencing assays. RESULTS: The HPV infection rate was extremely high (93% for any HPV and 64% for high-risk [HR]-HPV), with frequent multiple-strain infection (39%). Younger age (<30 years) was associated with frequent HR-HPV infection, multiple strain infections, and cytologic abnormalities. When compared with previous reports, HPV 16 remained the most common genotype (44.6%) in women with high-grade squamous intraepithelial lesion; however, a significant increase in HPV 31 (17.9%) and 45 (10.7%) and a decrease in HPV 35, 52, 33, and 66 were observed. CONCLUSIONS: The HPV genotype-specific prevalence and distribution pattern in this cohort of underserved Latino women differed significantly from previously published data in the United States. Understanding the potentially changing trends in HPV distribution pattern will help guide the development of appropriate preventive and therapeutic strategies for both underserved and general populations.

The impact of LSIL-H terminology on patient follow-up patterns: a comparison with LSIL and ASC-H.

The Papanicolaou (Pap) test category of "low grade squamous intraepithelial lesion, cannot exclude high grade squamous intraepithelial lesion" (LSIL-H) is not recognized by The Bethesda System but is commonly used. It is essentially an amalgamation of the official LSIL and ASC-H categories. Since these two categories have similar follow-up algorithms, the clinical utility of the combined LSIL-H category is unclear. We have therefore studied follow-up patterns for these three entities in our laboratory to determine the real-world impact of each in our patient population. We searched our pathology database over an 18-month period to find Pap tests (predominantly ThinPrep) interpreted as LSIL-H (137), LSIL (2,189), and ASC-H (101). Like other studies, we found that the discovery rate of high grade dysplasia in biopsies after LSIL-H (31.9%) was similar to ASC-H (35.3%) and was higher than LSIL (7.6%; P < 0.0001). In women with no previous history of dysplasia, the frequency of biopsy follow-up after the initial Pap test was significantly higher for LSIL-H (68.3%) than for LSIL (49.6%; P = 0.0038) and similar to ASC-H (62.3%). We also found that women with an initial negative biopsy or a biopsy positive for low grade dysplasia were more likely to undergo an additional biopsy if the initial Pap test was LSIL-H (36.2%) than if it was LSIL (18.2%; P = 0.0023). ASC-H (26.9%) had an intermediate rate. In our patient population, the use of the terminology LSIL-H is associated with follow-up biopsy patterns much more similar to ASC-H than to LSIL.

Digital slide imaging in cervicovaginal cytology: a pilot study.

CONTEXT: Digital whole slide imaging is the anticipated future of anatomic pathology, where sign-out of glass slides will be replaced by scanned images. Whole slide imaging has been successfully used in surgical pathology, but its usefulness and clinical application have been limited in cytology for several reasons, including lack of availability of z-axis depth focusing and large file size. Recently, several systems have become available in the United States for whole slide imaging with z-axis technology. OBJECTIVE: To determine the accuracy and efficiency of whole slide imaging, as compared with traditional glass slides, for use in cervicovaginal diagnostic cytology. DESIGN: Eleven cervicovaginal cytology cases (ThinPrep and SurePath) scanned at ×20, ×40, and ×40 z-stack magnifications using the BioImagene iScan Coreo Au 3.0 scanner were evaluated by 4 cytotechnologists and 3 pathologists in a blinded study. Different magnification scans were recorded as separate cases and presented in a randomized sequence. Corresponding glass slides were also reviewed. For each case, the diagnoses and total time to reach each diagnosis were recorded. RESULTS: Diagnostic accuracy was higher and average time per case was lower with glass slides as compared with all digital images. Among the digital images, the ×40 or ×40 z-stack had the highest diagnostic accuracy and lowest interpretation time. CONCLUSIONS: Whole slide imaging is a viable option for the purposes of teaching and consultations, and as a means of archiving cases. However, considering the large file size and total time to reach diagnosis on digital images, whole slide imaging is not yet ready for daily cervicovaginal diagnostic cytology screening use.

Only a small fraction of high-grade cervical lesions are discovered after an interpretation of atypical squamous cells of undetermined significance when using imager-assisted, liquid-based papanicolaou tests and the Bethesda 2001 system.

CONTEXT: Previous work has reported that most high-grade cervical neoplasia is seen in patients with preceding Papanicolaou test results of atypical squamous cells of undetermined significance. This information was based on conventional test results and the Bethesda 1991 reporting system and was determined before the current treatment guidelines. OBJECTIVE: Our objective was to perform a retrospective review of all histologically confirmed, high-grade cervical neoplasia to determine the diagnosis of the preceding liquid-based Papanicolaou test. DESIGN: A total of 189 histologically confirmed, high-grade cervical intraepithelial neoplasia (CIN) cases grade 2 and greater were identified for a 1-year period. RESULTS: Of the 189 cases, 10 (5.3%) had a previous diagnosis of atypical squamous cells of undetermined significance; 55 (29.1%) had low-grade squamous intraepithelial lesions; 31 (16.4%) had low-grade squamous intraepithelial lesions, unable to rule out a high-grade squamous intraepithelial lesion; 21 (11.1%) had atypical squamous cells, unable to rule out a high-grade squamous intraepithelial lesion; 68 (36%) had high-grade squamous intraepithelial lesions; 1 (0.5%) had atypical glandular cells; 1 (0.5%) had adenocarcinoma in situ; and 2 (1%) had invasive carcinoma. Combined "low grade" Papanicolaou test results (atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesion) preceded 51 of 103 cases of CIN 2 (49.5%) and 14 of 103 cases (13.6%) of CIN 3/cancer, whereas "high grade" Papanicolaou test results (atypical squamous cells, unable to rule out a high-grade squamous intraepithelial lesion; low-grade squamous intraepithelial lesions, unable to rule out a high-grade squamous intraepithelial lesion; high-grade squamous intraepithelial lesions; atypical glandular cells; adenocarcinoma in situ; and invasive carcinoma) preceded 52 of 103 CIN 2 cases (50.5%) and 72 of 103 CIN 3/cancer cases (69.9%). CONCLUSIONS: Our data show that we can now more-reliably predict high-grade dysplasia on routine Papanicolaou tests. Only a small fraction of histologically confirmed CIN 2/3 cases are found following a Papanicolaou test diagnosis of atypical squamous cells of undetermined significance.

Should LSIL-H be a distinct cytology category?: A study on the frequency and distribution of 40 human papillomavirus genotypes in 808 women.

BACKGROUND: The 2001 Bethesda System for gynecologic cervical cytology reporting classifies squamous intraepithelial lesions into low-grade (LSIL) and high-grade (HSIL) lesions. An intermediate term, "low-grade squamous intraepithelial lesion, cannot exclude high-grade squamous intraepithelial lesion (LSIL-H)," has been used in a small percentage of LSIL cases. To the authors' knowledge, little is known regarding the human papillomavirus (HPV) status in patients with LSIL-H. METHODS: A total of 808 SurePath specimens obtained between December 2009 and April 2011 were tested for 40 HPV genotypes using DNA microarray, followed by a confirmatory DNA sequencing assay. RESULTS: The infection rate for high-risk HPV in women with LSIL-H (92%) was strikingly close to that for women with HSIL (91%), which was higher than that for those with LSIL (74%); atypical squamous cells, cannot rule out high-grade lesion (ASC-H) (78%); or LSIL and ASC-H combined (74%). HPV type 16, the most common carcinogenic HPV genotype, was detected in 36% of women with LSIL-H, which was significantly higher than that in women with LSIL and ASC-H combined (13.8%), but less than that in women with HSIL (44.6%). Patients with LSIL-H and HSIL had similar infection rates for low-risk/intermediate-risk HPV genotypes, which were lower than those in LSIL or LSIL and ASC-H combined. CONCLUSIONS: Women found to have LSIL-H on a Papanicolaou test appear to have a unique HPV distribution pattern that clearly differs from LSIL and is comparable to that for HSIL, suggesting an increased risk of high-grade lesions over that of women with LSIL. Recognizing LSIL-H as an independent diagnostic category may help in the early identification of the high-risk subgroup that may require a management algorithm comparable to that for patients with HSIL. Cancer (Cancer Cytopathol) 2012. © 2012 American Cancer Society.

p16(INK4a) and ProEx C immunostains facilitate differential diagnosis of hyperchromatic crowded groups in liquid-based Papanicolaou tests with menstrual contamination.

OBJECTIVE: Evaluation of hyperchromatic crowded groups in Papanicolaou (Pap) tests from women during menstruation can be a diagnostic pitfall due to similar morphological appearances with significant cervical lesions. We studied the results of p16(INK4a) and ProEx C on cell blocks from Pap tests during menstruation in an attempt to facilitate the differentiation. STUDY DESIGN: Immunohistochemical stains for p16(INK4a) and ProEx C were performed on 25 cell blocks prepared from residual liquid-based cervical material with menstrual contamination. RESULTS: Strong, diffuse, and full thickness staining pattern for p16(INK4a) and ProEx C was observed in cases of high-grade squamous intraepithelial lesion (HSIL) and small cell carcinoma of the cervix. The low-grade squamous intraepithelial lesion cases and cases negative for intraepithelial lesion or malignancy were negative for ProEx C, with focal staining for p16(INK4a). The benign endometrial cells had either negative or focal patchy staining, which is often associated with tubal metaplasia. CONCLUSION: p16(INK4a) and ProEx C are sensitive markers for identifying significant lesions in Pap test specimens with menstrual contamination. Patchy/mosaic staining may be seen in benign endometrial tissue with tubal metaplasia, but strong, diffuse staining likely indicates HSIL or carcinoma. These findings can be helpful in interpreting hyperchromatic crowded groups in menstrual Pap specimens. Further study may be prudent, being aware of the small study group.

Women >or=30 years of age with low grade squamous intraepithelial lesion (LSIL) have low positivity rates when cotested for high-risk human papillomavirus: should we reconsider HPV triage for LSIL in older women?

High-risk human papillomavirus (HR-HPV) testing for colposcopy triage of low grade squamous intraepithelial lesion (LSIL) is not recommended because of high positive rates in young women. It remains unclear whether HR-HPV testing may be useful for triage of older women. We compiled HR-HPV data for women aged >or=30 years with LSIL for the period March 1, 2006 to February 28, 2008. Follow-up cervical biopsy information was collected for the period March 1, 2006 to August 15, 2008. We used the Hybrid Capture II test performed on residual material from liquid-based Pap tests. Of 735 women, 254 had HR-HPV testing, and of these 144 had positive HR-HPV results. Among women with positive HR-HPV results 79 underwent biopsy (54.9%) and 11 had cervical intraepithelial neoplasia (CIN) 2 or 3 (13.9% of women with biopsy follow-up). A total of 481 women did not undergo HR-HPV testing, of whom 192 underwent biopsy (39.9%) and 11 had CIN 2 or 3 (5.7% of biopsied women [P = 0.04]). Among women who tested negative for HR-HPV and had follow-up biopsies, only one had a high grade lesion found (CIN 2). The overall HR-HPV positive rate in tested women >or=30 years old with LSIL was 56.7% if women who had reflex HR-HPV testing for ASC-US are included. The HR-HPV positive rate in residual material from Pap tests interpreted as LSIL was 63.8%. Among women >or=30 years of age with LSIL, CIN 2-3 is significantly more likely in HR-HPV positive women. Relatively few older women with LSIL test positive for HR-HPV. Colposcopy triage using HR-HPV may be justified in this population.

Image-guided ThinPrep Papanicolaou tests and cotesting with high-risk human papillomavirus in women aged 30 years and older in a low-risk private practice population.

BACKGROUND: Screening for cervical cancer precursors has evolved considerably with the introduction of new technologies to improve the early detection of disease. The objective of this study was to analyze the accuracy and effectiveness of combined screening with cytology and high-risk human papillomavirus (HR-HPV) testing in a low-risk population of women aged >or=30 years. METHODS: Consecutive unselected samples from a group of 1871 women aged >or=30 years were screened with image-guided ThinPrep tests and HR-HPV tests during a 6-month period. Histologic follow-up was reviewed among women with positive HR-HPV tests. RESULTS: A total of 85 (4.5%) women had positive HR-HPV tests. In 48 HR-HPV-positive women with follow-up biopsies, 41 (85%) were found to have histologic abnormalities. Thirty-three (1.9%) women with cytologically normal Papanicolaou (Pap) tests harbored HR-HPV, and a cervical intraepithelial neoplasia (CIN) 2+ lesion was detected in 1 (16%) of 6 women with histologic follow-up. Conversely, 2 (28%) of 7 women with high-grade intraepithelial lesion on cytology tested negative for HR-HPV during the same period. A case of serous carcinoma with atypical glandular cells on cytology was also negative for HR-HPV, as expected. CONCLUSIONS: In this low-risk population of women aged >or=30 years, histology-confirmed CIN2+ lesions were identified in women with negative cytology and positive HR-HPV tests, as well as in those with positive cytology and negative HR-HPV tests. Because both cytology and HPV testing alone missed significant lesions, cotesting with Pap and HR-HPV in women aged >or=30 years appears to be a reasonable option in a low-risk population.

Thyroid fine-needle aspiration biopsy in children and adolescents: experience with 218 aspirates.

To evaluate the role of fine-needle aspiration (FNA) biopsy of thyroid nodules in pediatric and adolescent patients, the cytology reports of 218 thyroid FNA biopsies performed on children and adolescents ranging from 10 to 21 yr of age were reviewed. The cytology diagnoses were categorized into four groups: unsatisfactory, benign, suspicious, and malignant. One hundred nineteen (54%) of the aspirates were diagnosed as "benign," 20 (9%) were diagnosed as suspicious for malignancy; and 17 (8%) were diagnosed as malignant. Sixty-two (28%) of the aspirates were read as unsatisfactory for interpretation. Sensitivity of thyroid FNA in diagnosing thyroid malignancy relative to final histological diagnoses was 100%, and specificity was 65%. FNA of thyroid nodules in the pediatric and adolescent population is comparably as sensitive and specific as in the adult population. The acceptance of this procedure in the routine evaluation of young patients' thyroid nodules should reduce the number of unnecessary surgeries for benign thyroid disease.

The utility of p16INK4a and Ki-67 staining on cell blocks prepared from residual thin-layer cervicovaginal material.

BACKGROUND: Cell blocks can be prepared from residual thin-layer cervicovaginal (ThinPrep) material and can be used in immunohistochemical staining assays for p16INK4a and Ki-67, which are surrogate markers related to human papillomavirus infection and cell proliferation, respectively. The objectives of the current study were 1) to investigate the feasibility and the role of cell block preparations in identifying significant neoplastic and preneoplastic lesions of the uterine cervix and 2) to assess the feasibility of using p16INK4a and Ki-67 immunohistochemical staining patterns on cell blocks to identify significant preneoplastic cervical lesions. METHODS: Cervicovaginal cytology specimens from 85 patients were analyzed. Cytologic diagnoses based on ThinPrep Papanicolaou test results were as follows: squamous cell carcinoma was diagnosed in 3 specimens, high-grade squamous intraepithelial lesions (HSIL) were diagnosed in 27 specimens, low-grade squamous intraepithelial lesions (LSIL) were diagnosed in 20 specimens, and atypical squamous cells of uncertain significance (ASCUS) were diagnosed in 11 specimens. Diagnoses of negativity for intraepithelial lesions or malignancy (NILM) were made in 24 specimens. Cell block sections were stained with hematoxylin and eosin and were immunostained with antibodies against p16INK4a protein and Ki-67 antigen. RESULTS: The cytomorphologic diagnoses made using cell block preparations were as follows: SCC in 2 specimens, HSIL in 20 specimens, LSIL in 30 specimens, NILM in 32 specimens, and no diagnosis in 1 specimen. In 62 cases (73%), the diagnoses made using cell block preparations were in agreement with the ThinPrep diagnoses. Immunostaining of cell blocks for p16INK4a and Ki-67 exhibited a statistically significant association (P < 0.05) with the presence of significant lesions on either cell block or ThinPrep analysis. CONCLUSIONS: To the authors' knowledge, p16INK4a has not been analyzed previously in ThinPrep cell blocks, and the correlation between Ki-67 expression and cell block diagnoses also has not been reported previously. The current results indicate that cell blocks prepared from residual ThinPrep material represent an additional reliable diagnostic tool in the evaluation of cervical samples. Furthermore, immunohistochemical studies may be helpful in differentiating significant preneoplastic changes from other cervical lesions, such as atrophy.

Endomiocardiofibrose com calcificaçäo maciça do ventrículo esquerdo / Endomyocardial fibrosis associated with massive calcification of the left ventricle

This is the report of a rare case of endomyocardial fibrosis associated with massive calcification of the left ventricle in a male patient with dyspnea on great exertion, which began 5 years earlier and rapidly evolved. Due to lack of information and the absence of clinical signs that could characterize impairment of other organs, the case was initially managed as a disease with a pulmonary origin. With the evolution of the disease and in the presence of radiological images of heterogeneous opacification in the projection of the left ventricle, the diagnostic hypothesis of endomyocardial disease was established. This hypothesis was later confirmed on chest computed tomography. The patient died on the 16th day of the hospital stay, probably because of lack of myocardial reserve, with clinical findings of refractory heart failure, possibly aggravated by pulmonary infection. This shows that a rare disease such as endomyocardial fibrosis associated with massive calcification of the left ventricle may be suspected on a simple chest X-ray and confirmed by computed tomography.