Dina R. Mody

Atypical epithelial cells and specimen adequacy: Current laboratory practices of participants in the College of American Pathologists Interlaboratory Comparison Program in Cervicovaginal Cytology

○ Context.-The Bethesda System for reporting cervical/ vaginal cytologic diagnoses introduced terminology for atypical squamous and glandular cells and categories for specimen adequacy. Objectives.-To analyze current laboratory reporting practices and compare trends to previous surveys. Design.-Questionnaire surveys were mailed to 2000 laboratories in 1996 and 1997. Participants.-Laboratories enrolled in the College of American Pathologists Interlaboratory Comparison Program in Cervicovaginal Cytology. Main Outcome Measures.-Laboratory policies, criteria, and reporting rates for Bethesda System categories. Results.-The 1996 specimen adequacy survey had 1166 respondents, and 768 laboratories returned the 1997 questionnaire focusing on atypical squamous cells of undetermined significance (ASCUS) and glandular cells of undetermined significance (AGUS). Nearly all laboratories (92%) routinely reported specimen adequacy, an increase from the 66% rate in 1991. The median rate for unsatisfactory specimens was 0.5% (mean 0.95%), and the median rate for the satisfactory but limited category was 5.8% (mean 9.3%). The Bethesda criteria for designating a specimen unsatisfactory were used by more than 90% of laboratories. Nearly all laboratories (97%) used the term ASCUS in 1997, and more than 80% of laboratories used the Bethesda criteria for this category. Median reporting rates for epithelial abnormalities were as follows: ASCUS, 4.5%; AGUS, 0.3%; low-grade squamous intraepithelial lesion (SIL), 1.6%; and high-grade SIL, 0.5%. The median ASCUS/SIL ratio was 2.0, with 80% of laboratories reporting ratios between 0.64 and 4.23. The median ASCUS rate and ASCUS/SIL ratio were higher than 1993 survey results. Nearly all laboratories attempted follow-up studies on patients with abnormal cytology results, and midsized laboratories achieved the highest rates of follow-up. Median rates of abnormalities following an ASCUS or AGUS diagnosis were 20% and 15%, respectively. Laboratory respondents commonly used written recommendations in ASCUS/AGUS reports. Conclusions.-Most laboratories that responded to the surveys had adopted Bethesda terminology and criteria for specimen adequacy and ASCUS/AGUS. Reporting rates for SIL and adequacy categories have remained stable, but median ASCUS rates and ASCUS/SIL ratios are higher than in 1993. The AGUS category is reported infrequently, but can be associated with significant pathology.

Innervation of the human vertebral body : A histologic study

The histology of 69 human vertebral bodies from 23 individuals was studied by hematoxylin and eosin staining using a technique that allowed the creation of complete, large histologic sections of individual vertebral bodies. Particular attention was directed toward the documentation of intraosseous nerves. The vertebral bodies were dissected free of soft tissue, and then sectioned using a diamond wafering saw into 3-mm sagittal segments. Sections were then decalcified and whole-mounted in paraffin blocks before tissue sectioning using a very-large-format microtome. One hundred thirty-eight tissue sections were prepared for evaluation. Neurovascular bundles and intraosseous nerves were routinely identified within human vertebral bone. Nerves were noted to enter the vertebral body via the centrally located posterior vascular foramen and were found to accompany the basivertebral vessels. Branches from these nerves coursed to both central and peripheral areas of the vertebral body. Nerves were also documented that entered the vertebral body by penetrating the anterior cortex to course into the marrow. Although previous studies have documented nerves within long bones, and others have described the histology of the intervertebral disc and associated soft tissues, previous literature that documents the innervation of the human vertebral body has been very sparse. The documentation of nerve tissue within normal human vertebrae further supports the proposed role of neuronal factors in the regulation of bone physiology. Furthermore, it is possible that such intraosseous nerves may play a role in the clinical problem of back pain.

Cytologic Malignancy Versus Benignancy: How Useful Are the “Newer” Markers in Body Fluid Cytology?

CONTEXT Differentiating reactive effusion, malignant mesothelioma, and metastatic adenocarcinoma in body cavity fluids can be challenging. Interpreting immunohistochemical markers in cell block preparations can be difficult because of nonspecific staining, focal staining, or poor staining quality. We selected a panel of conventional and newer markers to assess their utility in evaluating effusions. OBJECTIVE To evaluate the efficacy of 5 immunohistochemical markers in the differential diagnosis of reactive mesothelial proliferation, malignant mesothelioma, and metastatic adenocarcinoma in body cavity fluids. DESIGN A total of 72 formalin-fixed, paraffin-embedded cell block specimens from pleural and peritoneal effusions, including 5 mesotheliomas, 48 adenocarcinomas, and 19 benign effusions were stained with antibodies against calretinin, D2-40, XIAP, MOC-31, and WT1. RESULTS All benign effusions and mesotheliomas demonstrated diffuse membranous staining with D2-40. All mesotheliomas displayed calretinin positivity, whereas only 58% of benign effusions stained focally with calretinin. MOC-31 was positive in all cases of adenocarcinoma, whereas all benign effusions and mesotheliomas were negative. All cases of the metastatic adenocarcinoma were negative for calretinin and D2-40. However, background reactive mesothelial cells were positive for calretinin and D2-40. Overall, D2-40 highlighted more mesothelial cells than calretinin. WT1 was positive in 50% of benign effusions, 60% of mesotheliomas, and 27% of adenocarcinomas. XIAP stained most mesotheliomas (80%), some adenocarcinomas (51%), and rare benign effusions (11%). CONCLUSIONS MOC-31 and D2-40 were very sensitive and specific markers of epithelial and mesothelial cells, respectively. Compared with calretinin, D2-40 was a more sensitive marker of mesothelial cells. WT1 proved to be nonspecific. XIAP was not a sensitive marker for malignancy and had a limited value in cytology. We recommend using a panel to include MOC-31 and D2-40 to improve diagnostic accuracy in body cavity effusions.

Glandular lesions of the cervix: Validity of cytologic criteria used to differentiate reactive changes, glandular intraepithelial lesions and adenocarcinoma

OBJECTIVE To determine the validity and reproducibility of various cytologic criteria as discriminators between different glandular lesions of the cervix. STUDY DESIGN Seventy-three cervicovaginal smears with glandular lesions and a documented histologic and/ or clinical correlation were studied. The lesions included reactive changes, low grade glandular intraepithelial lesion (LGIL) (encompassing endocervical glandular dysplasia), high grade glandular intraepithelial lesion (HGIL) (encompassing adenocarcinoma in situ) and invasive adenocarcinoma (IA). Twenty-three cytologic criteria were used to evaluate the smears, and the results were scored and statistically analyzed. RESULTS Reactive lesions consistently showed well-defined cell borders, normal nuclear/cytoplasmic (N/C) ratio, minimal or absent nuclear overlapping, round to oval nuclei with fine chromatin and prominent nucleoli. HGIL (adenocarcinoma in situ) showed feathered edges, rosettes, cell strips, increased N/C ratio, elongated nuclei, marked nuclear overlapping and nuclei with hyperchromatic, coarse chromatin. IA shared features with HGIL but had a greater tendency to show a dirty background, single cells, mitotic figures, nuclear pleomorphism and large nucleoli. Logistic regression studies indicated that the presence of mitotic figures, a dirty background and single cells increases the odds ratio for predicting invasion. LGIL showed features similar to those of high grade lesions, but the changes were more subtle. These lesions were also less cellular and less likely to show cell strips, feathered edges and rosettes. Similar to HGIL and IA, LGIL showed nuclear overlapping, increased N/C ratio, oval to elongated nuclei and nuclear hyperchromasia. CONCLUSION There is overlap between the cytologic criteria for the various glandular lesions of the cervix. However, some of these criteria can consistently distinguish clinically significant lesions from reactive benign changes seen in the glandular epithelium.

Ber-EP4 for differentiating adenocarcinoma from reactive and neoplastic mesothelial cells in serous effusions. Comparison with carcinoembryonic antigen, B72.3 and Leu-M1.

OBJECTIVE To evaluate the efficacy of Ber-EP4 in distinguishing epithelial cells from mesothelial cells in routine cytologic preparations. STUDY DESIGN Paraffin-embedded cell blocks of serous effusions from 32 patients (11 metastatic adenocarcinomas, 16 reactive mesothelial proliferations and 5 malignant mesotheliomas) were immunostained with Ber-EP4. For comparison, cell block preparations of adenocarcinomas and mesotheliomas were also immunostained with the most commonly applied markers of adenocarcinoma: carcinoembryonic antigen (CEA), B72.3 and Leu-M1. In addition, cytocentrifuge preparations of 14 reactive effusions and 2 metastatic adenocarcinomas were stained prospectively with Ber-EP4. RESULTS All adenocarcinomas showed intense membrane staining, while all mesothelial proliferations, both benign and malignant, were negative. The Ber-EP4-positive immunostaining was remarkably clean, with very minimal nonspecific staining. CEA stained 11/11 adenocarcinomas, B72.3 stained 10/11, and Leu-M1 stained 8/11; mesotheliomas were negative with all three antibodies. CONCLUSION Ber-EP4 is at least as useful as CEA, B72.3 and Leu-M1 in the diagnosis of serous effusions. It has the advantage of high sensitivity and ease of interpretation because of the high percentage of tumor cells stained, characteristic membranous staining and lack of cross-reaction with background inflammatory cells.

Restricted κ/λ Light Chain Ratio by Flow Cytometry in Germinal Center B Cells in Hashimoto Thyroiditis

To determine the diagnostic significance of the kappa/lambda ratio in germinal center (GC) B cells in Hashimoto thyroiditis (HT), we used 4-color flow cytometry to immunophenotype 27 samples (21 patients) of well-characterized HT B-cell clonality was analyzed further by polymerase chain reaction (PCR) of the immunoglobulin heavy chain (IgH) and bcl-2/IgH fusion genes using DNA extracted from aspirate smears and/or paraffin-embedded tissues. By flow cytometric analysis, the CD10+ GC B cells had a higher mean +/- SD kappa/lambda ratio than the CD10- B cells (5.1 +/- 3.3 vs 2.0 +/- 0.8; P < .0001, Student t test). In 18 samples (67%), CD10+ GC B cells had a kappa/lambda ratio greater than 3.07 (the upper limit of kappa/lambda ratio reported in reactive nodes; range, 3.2-14.4 in the 18 cases). Cases tested by PCR showed no evidence of a clonal proliferation. None of 21 cases developed lymphoma during clinical follow-up of up to 3 years. The kappa/lambda ratio of CD10+ GC B cells in HT can be skewed markedly beyond that reported in reactive lymph nodes. This finding frequently is present in HT. Pathologists should be familiar with this phenomenon to prevent misdiagnosis of follicular lymphoma in patients with HT.

A histologic study of fractured human vertebral bodies.

Twenty-seven fractured human vertebral bodies and 24 unfractured human vertebrae from adjacent levels were studied postmortem using histologic and high-resolution radiographic techniques. The findings were compared with those in the vertebral bodies of individuals without spinal fracture. Forty-six human thoracolumbar spines were obtained from individuals at autopsy. Standard radiographs were made of all specimens. Twelve of the 46 individuals had a total of 27 fractured vertebral bodies by plain radiographic criteria. Attention was focused on these fractured vertebrae as well as on 24 unfractured vertebral bodies that were harvested from a level immediately adjacent to the fractured vertebral bodies. Twelve vertebral bodies from four individuals with no evidence of fracture or inflammatory spondyloarthropathy were also studied for comparison. The vertebral bodies were graded by their mineral density as measured by dual-energy x-ray absorptiometry and sectioned into 3-mm sagittal cuts. High-resolution contact radiographs were prepared for each section prior to decalcification and tissue sectioning on a large format microtome. Mid-and parasagittal tissue sections of each vertebra were prepared for standard hematoxylin and eosin stains. A total of 126 sections were studied. The histologic characteristics of the fractured vertebrae (n = 27) were compared with those of adjacent unfractured levels (n = 24) and with vertebrae from individuals without fracture (n = 12). Vertebral bodies with fractures secondary to osteoporosis were consistently characterized histologically by focal areas of endochondral new bone formation adjacent to avascular necrotic bone and unreactive marrow. Such ongoing new bone and new vessel formation adjacent to nonhealing areas were also documented in radiographically unfractured vertebral bodies from individuals with osteoporotic fractures at adjacent levels. No areas of endochondral new bone formation or areas of focal necrosis were found in vertebral bodies from individuals without radiographic evidence of osteoporosis. A vascular necrosis of the vertebral body is a common histologic finding in individuals with osteoporosis. Indeed, our histologic observations suggest subclinical fractures and microfractures of the vertebral body may be the underlying pathologic process leading to avascular necrosis in individuals with osteoporosis. Microtrabecular fractures and endplate fractures were commonly seen in osteoporotic vertebral bodies, often in vertebrae that appeared to be uninvolved on specimen radiographs.

Glandular lesions of the cervix on thin-layer Pap tests: Validity of cytologic criteria used in identifying significant lesions

OBJECTIVE To determine the cytologic features that are most helpful in characterizing significant glandular lesions of the cervix observed on the ThinPrep (TP) Pap test (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) and to compare these features with those published for conventional smears. STUDY DESIGN Thirty-nine TP preparations with cytologic evidence of glandular lesions of the cervix and histologic and/or clinical correlation were studied. These lesions included (1) 11 cases of benign/reactive conditions; (2) 10 cases of adenocarcinoma in situ (AIS), of which 1 had both AIS and carcinoma in situ; (3) 1 case of invasive adenocarcinoma; (4) 15 cases of squamous intraepithelial lesions and squamous cell carcinoma, including 4 with glandular involvement, and (5) 2 cases of adenosquamous cell carcinoma. These cases were reviewed by the first author without knowledge of the histologic diagnosis. Twenty-five previously published cytologic criteria were used to evaluate glandular cells on TP slides. Statistical analysis was performed using Fishers exact test to determine the significance of the features studied. RESULTS All glandular lesions had cytologic features on TP similar to those previously described on conventional smears. However, TP slides demonstrated enhanced nuclear features but less-preserved architectural patterns. Reactive lesions showed minimal overlapping without hyperchromasia or mitotic figures and with normal nuclear/cytoplasmic ratios. AIS and invasive adenocarcinoma cases had similar features. Increased cellularity and overcrowding were prominent, whereas feathering, rosettes and cell strips were present but subtle. CONCLUSION Glandular lesions of the cervix on TP slides shared many of the characteristic features reported for conventional smears. However, nuclear details were more pronounced in TP slides, while architectural patterns, although present, were relatively subtle.

Angiosarcoma at Unusual Sites

BACKGROUND: Angiosarcomas are uncommon soft tissue neoplasms with a predilection for skin and superficial soft tissues. CASES: Two cases of angiosarcoma occurred at unusual sites, the parotid gland and lung. The parotid lesion was characterized by malignant cells present singly, in loose groups, in tight three-dimensional aggregates and in acinar formation initially misinterpreted as an adenocarcinoma. The lung mass showed malignant cells in association with vascular endothelium, suggestive of angiosarcoma. Both cases were negative for Ulex europaeus and Factor VIII-related protein but demonstrated strong immunopositivity for CD31, a highly specific endothelial marker. CONCLUSION: In the absence of vasoformative structures, important diagnostic pitfalls are pseudovascular adenoid squamous cell carcinoma, poorly differentiated adenocarcinoma, melanoma and lymphoma. Immunocytochemical studies and clinical history are essential to the correct diagnosis.

A histologic study of soft-tissue reactions to spinal implants

Objectives The biologic reactions to orthopedic spinal implants were determined. Methods Biopsies of soft tissue immediately adjacent to spinal implants were done in 36 consecutive patients undergoing elective lumbar spinal hardware removal and was studied histologically. Results A fibrous tissue matrix was noted in all specimens. In 11 of 36 specimens, a discrete layer of cells with epithelial characteristics was noted on the surface immediately opposed to the metal implant. Results of immunohistochemical staining were negative using antibodies to markers, which suggests that these unique cell layers are probably of histiocytic origin, Black amorphous metallic debris was seen in nine of the specimens. In seven of these specimens, this material was associated with an inflammatory foreign-body reaction. Refractile non-biorefringent crystalline bodies were noted in five specimens. These crystalline deposits provoked a local foreign-body reaction in all cases. Conclusion The role of soft-tissue inflammatory reactions in the production of clinical symptoms of pain is discussed.