Rosemary H. Tambouret

Whole-Slide Imaging Digital Pathology as a Platform for Teleconsultation: A Pilot Study Using Paired Subspecialist Correlations

CONTEXT -Whole-slide imaging technology offers promise for rapid, Internet-based telepathology consultations between institutions. Before implementation, technical issues, pathologist adaptability, and morphologic pitfalls must be well characterized. OBJECTIVE -To determine whether interpretation of whole-slide images differed from glass-slide interpretation in difficult surgical pathology cases. DESIGN -Diagnostically challenging pathology slides from a variety of anatomic sites from an outside laboratory were scanned into whole digital format. Digital and glass slides were independently diagnosed by 2 subspecialty pathologists. Reference, digital, and glass-slide interpretations were compared. Operator comments on technical issues were gathered. RESULTS -Fifty-three case pairs were analyzed. There was agreement among digital, glass, and reference diagnoses in 45 cases (85%) and between digital and glass diagnoses in 48 (91%) cases. There were 5 digital cases (9%) discordant with both reference and glass diagnoses. Further review of each of these cases indicated an incorrect digital whole-slide interpretation. Neoplastic cases showed better correlation (93%) than did cases of nonneoplastic disease (88%). Comments on discordant cases related to digital whole technology focused on issues such as fine resolution and navigating ability at high magnification. CONCLUSIONS -Overall concordance between digital whole-slide and standard glass-slide interpretations was good at 91%. Adjustments in technology, case selection, and technology familiarization should improve performance, making digital whole-slide review feasible for broader telepathology subspecialty consultation applications.

Ultrasound-guided fine-needle aspiration biopsy of the thyroid

We reviewed the Massachusetts General Hospital experience with ultrasound‐guided fine‐needle aspiration biopsies (FNABs) of the thyroid to determine the indications, rate of unsatisfactory smears, correlation with excisional biopsy results, and verification of efficient use of personnel time.

Prevalence of Abnormalities Influences Cytologists' Error Rates in Screening for Cervical Cancer

CONTEXT Medical screening tasks are often difficult, visual searches with low target prevalence (low rates of disease). Under laboratory conditions, when targets are rare, nonexpert searchers show decreases in false-positive results and increases in false-negative results compared with results when targets are common. This prevalence effect is not due to vigilance failures or target unfamiliarity. OBJECTIVE To determine whether prevalence effects could be a source of elevated false-negative errors in medical experts. DESIGN We studied 2 groups of cytologists involved in cervical cancer screening (Boston, Massachusetts, and South Wales, UK). Cytologists evaluated photomicrographs of cells at low (2% or 5%) or higher (50%) rates of abnormality prevalence. Two versions of the experiment were performed. The Boston, Massachusetts, group made decisions of normal or abnormal findings using a 4-point rating scale. Additionally, the group from South Wales localized apparent abnormalities. RESULTS In both groups, there is evidence for prevalence effects. False-negative errors were 17% (higher prevalence), rising to 30% (low prevalence) in the Boston, Massachusetts, group. The error rate was 27% (higher prevalence), rising to 42% (low prevalence) in the South Wales group. (Comparisons between the 2 groups are not meaningful because the stimulus sets were different.) CONCLUSIONS These results provide the first evidence, to our knowledge, that experts are not immune to the effects of prevalence even with stimuli from their domain of expertise. Prevalence is a factor to consider in screening for disease by human observers and has significant implications for cytology-based cervical cancer screening in the post-human papillomavirus vaccine era, when prevalence rates of high-grade lesions in the population are expected to decline.

Abdominal radical trachelectomy: Success and pitfalls in a general gynecologic oncology practice

OBJECTIVE To report our successes and complications with a series of abdominal radical trachelectomies performed to preserve fertility in young women at the Massachusetts General Hospital (MGH). METHODS Institutional review board (IRB) permission was obtained for retrospective record review. Data were collected regarding patient age and parity, tumor stage and histology, surgical time and complications, post-operative complications, follow-up, and pregnancy. RESULTS Ten patients underwent radical abdominal trachelectomy, 9 by the same surgeon (LD). Surgery was essentially identical to that of radical hysterectomy with the exception of re-anastomosis of the uterine fundus to the vagina and placement of cerclage. Pre-operative evaluation and post operative follow-up was for the most part identical for all patients. Two patients achieved pregnancy, with 1 twin delivery and 1 patient had 2 pregnancies. Two patients experienced cervical stenosis with regular menses and the same 2 patients passed their abdominal cerclage vaginally. To date there have been no cancer recurrences. Pap smear follow-up has been complicated by difficulty in reading smears from the lower uterine segment (LUS). CONCLUSION Radical abdominal trachelectomy can be successfully performed by any gynecologic oncologist who is trained in radical pelvic surgery. Pre-operative counseling is crucial in obtaining informed consent. Patients must be aware of potential post-operative complications, including pre-term delivery. Cytology department needs to be aware of potential pitfalls in reading Pap smears from the LUS.

Tissue-specific signatures of activating PIK3CA and RAS mutations in carcinosarcomas of gynecologic origin

OBJECTIVES Gynecologic carcinosarcoma is an aggressive malignancy that requires more effective treatment approaches. However, therapeutic implications regarding the specific gynecologic site of origin and the admixture of carcinomatous and sarcomatous elements that define this tumor remain uncertain. Therefore, broad genotyping was performed to identify tissue-specific somatic mutational profiles that may help direct targeted therapies in this complex neoplasia. METHODS Genotyping was conducted on primary gynecologic carcinosarcomas arising from various disease sites (uterus, ovary, fallopian tube, vagina) and within isolated histological subcomponents. Nucleic acids extracted from diagnostic tissue were used in a genotyping platform that simultaneously queried >120 common mutations across 14 cancer genes. Mutational status was correlated with clinical variables using logistic regression and Kaplan-Meier survival estimates. RESULTS Cancer gene mutations were identified in 46% of the 52 patient cohort and include TP53 (23%), PIK3CA (19%), KRAS (15%), CTNNB1 (4%) and NRAS (2%). Mutation in a single gene was observed in 31% of patient samples, while synchronous mutations involving 2 and 3 genes were noted in 13% and 2% of samples, respectively. Comparative evaluation of the carcinomatous and sarcomatous elements within a tumor demonstrated a similar mutation signature. Mutations in PIK3CA, KRAS and NRAS were exclusive to tumors of uterine origin and age-adjusted Cox proportional hazards modeling associated advanced age, stage and TP53 mutations with decreased survival in the uterine subset. CONCLUSION While carcinosarcomas across gynecologic disease sites are histologically similar, therapeutically relevant mutations in the mitogen-activated protein kinase and phosphatidylinositol 3-kinase pathways predominated in carcinosarcomas arising in the uterus.

Endometrial endometrioid adenocarcinoma with a deceptive pattern of spread to the uterine cervix: A manifestation of stage IIB endometrial carcinoma liable to be misinterpreted as an independent carcinoma or a benign lesion

The prognosis of endometrial endometrioid adenocarcinoma is determined in part by stage; endocervical stromal involvement (stage IIB) imparts a worsened prognosis. We describe a deceptive pattern of stage IIB disease that mimics a primary endocervical glandular proliferation and may lead to understaging of endometrial endometrioid adenocarcinoma. Fifteen cases of endometrial endometrioid adenocarcinoma with a peculiar pattern of cervical involvement were identified from our consultation files. All cases were referred in consultation because of doubt about the nature of the cervical process and its relation to the corpus tumor; in a few instances, the cervical proliferation was considered possibly benign and in one case was misinterpreted as mesonephric hyperplasia. The patients ranged from 49 to 84 years in age (mean age 64.9 years). There was usually a grossly evident endometrial tumor. The cervix was unremarkable grossly in at least 11 patients. The cervical tumors were composed of variably shaped, often tubular glands with little or no stromal response and mainly invaded as widely spaced glands that often appeared deceptively benign. In 14 cases luminal secretions, mainly eosinophilic, were identified, often leading to consideration of a mesonephric lesion. Ten of the endometrial tumors were grade 1, four grade 2, and one grade 3. One was noninvasive, nine superficially invasive, and five deeply invasive. In four cases myoinvasion had, at least in part, a diffusely infiltrative pattern. The tumors in the cervix showed no in situ component and no definite surface involvement. Continuity with the corpus tumor could be demonstrated in 12 cases. Ten of the cervical tumors invaded more deeply than the endometrial tumor, four invaded to a similar depth, and only one was more superficial than its endometrial counterpart. The cervical and corpus tumors had a similar immunoprofile in nine cases: all were vimentin positive, eight estrogen positive and one negative, four carcinoembryonic antigen negative, and five with focal apical or rare cytoplasmic staining. This immunoprofile in conjunction with routine morphologic similarity between the two tumors and the usual documented continuity between them indicate that the cervical process represents spread from the endometrial endometrioid adenocarcinoma. It is important for both therapeutic and prognostic reasons that the cervical abnormality is not misinterpreted as a benign or malignant primary endocervical glandular process.

Human papillomavirus testing using hybrid capture II with SurePath collection: initial evaluation and longitudinal data provide clinical validation for this method.

Testing for human papillomavirus (HPV) is an integral part of equivocal cervical cytology triage. Clinical validation of non‐FDA (Food and Drug Administration)–approved methods is therefore important because of the high volume of such tests and the implications for missed high‐grade lesions if test performance is not optimal.

Digital diffraction analysis enables low-cost molecular diagnostics on a smartphone

Significance Smartphones and wearable electronics have advanced tremendously over the last several years but fall short of allowing their use for molecular diagnostics. We herein report a generic approach to enable molecular diagnostics on smartphones. The method utilizes molecular-specific microbeads to generate unique diffraction patterns of “blurry beads” which can be recorded and deconvoluted by digital processing. We applied the system to resolve individual precancerous and cancerous cells as well as to detect cancer-associated DNA targets. Because the system is compact, easy to operate, and readily integrated with the standard, portable smartphone, this approach could enable medical diagnostics in geographically and/or socioeconomically limited settings with pathology bottlenecks. The widespread distribution of smartphones, with their integrated sensors and communication capabilities, makes them an ideal platform for point-of-care (POC) diagnosis, especially in resource-limited settings. Molecular diagnostics, however, have been difficult to implement in smartphones. We herein report a diffraction-based approach that enables molecular and cellular diagnostics. The D3 (digital diffraction diagnosis) system uses microbeads to generate unique diffraction patterns which can be acquired by smartphones and processed by a remote server. We applied the D3 platform to screen for precancerous or cancerous cells in cervical specimens and to detect human papillomavirus (HPV) DNA. The D3 assay generated readouts within 45 min and showed excellent agreement with gold-standard pathology or HPV testing, respectively. This approach could have favorable global health applications where medical access is limited or when pathology bottlenecks challenge prompt diagnostic readouts.

Longitudinal clinical evaluation of a novel antibody cocktail for detection of high-grade squamous intraepithelial lesions on cervical cytology specimens.

CONTEXT Although cervical cancer screening by cytology is successful, the test continues to show relatively poor operating characteristics. Cell cycle markers may enhance detection of high-grade squamous intraepithelial lesions. OBJECTIVE To determine the clinical usefulness of ProExC, an immunocytochemical assay for cell cycle components, performed on routine cervical cytology samples. DESIGN Cervical cytology samples were collected using the SurePath method. Residual cells remaining after preparation of the Papanicolaou-stained slide were used to make a second slide for ProExC staining using an indirect polymer-based immunoperoxidase method. Only adequately cellular slides were evaluated for the presence of nuclear staining within cytologically abnormal epithelial cells. Results were correlated with clinical follow-up. RESULTS Six hundred twenty-four samples were satisfactorily cellular and stained. Correlation with clinical follow-up for subsequent cervical intraepithelial neoplasia 2+ on biopsy/high-grade squamous intraepithelial lesion on cytology (CIN 2+/HSIL) showed that 434 results were true negative, 78 true positive, 18 false-negative, and 94 false-positive, resulting in a sensitivity/specificity of 81%/82%. When ProExC results were combined with any level of cytologic atypia, sensitivity for CIN 2+/HSIL was 92% and specificity was 84%. CONCLUSIONS ProExC shows promise as an aid in enhancing the sensitivity and specificity of cervical cytology for subsequent CIN 2+/HSIL and may be useful in identifying those cervical lesions most apt to progress.

The cytologic criteria of malignancy

Cytology and cell biology are two separate fields that share a focus on cancer. Cancer is still diagnosed based on morphology, and surprisingly little is known about the molecular basis of the defining structural features. Cytology uses the smallest possible biopsy for diagnosis by reducing morphologic “criteria of malignancy” to the smallest scale. To begin to develop common ground, members of the American Society of Cytopathology Cell Biology Liaison Working Group classify some of the “criteria of malignancy” and review their relation to current cell biology concepts. The criteria of malignancy are extremely varied, apparently reflecting many different pathophysiologies in specific microenvironments. Criteria in Group 1 comprise tissue‐level alterations that appear to relate to resistance to anoikis, alterations in cell adhesion molecules, and loss of apical–basal polarity. Criteria in Group 2 reflect genetic instability, including chromosomal and possibly epigenetic instability. Criteria in Groups 3 are subcellular structural changes involving cytoplasmic components, nuclear lamina, chromatin and nucleoli that cannot be accounted for by genetic instability. Some distinct criteria in Group 3 are known to be induced by cancer genes, but their precise structural basis remains obscure. The criteria of malignancy are not closely related to the histogenetic classification of cancers, and they appear to provide an alternative, biologically relevant framework for establishing common ground between cytologists and cell biologists. To understand the criteria of malignancy at a molecular level would improve diagnosis, and likely point to novel cell physiologies that are not encompassed by current cell biology concepts. J. Cell. Biochem. 110: 795–811, 2010.