The spectrum of cytologic features in nonproliferative breast lesions.

BACKGROUND: The reliability of cytologic criteria to classify nonproliferative breast lesions (NPBL) is still debated. Sampling error and heterogeneity of breast lesions complicates the histologic correlation of fine-needle aspiration results further. METHODS: To provide optimal cytohistologic correlation, two smears (one that was stained with hematoxylin and eosin and one that was stained with Diff-Quik [American Scientific Products, McGraw Park, IL]) were prepared from specific tissue sections from breast biopsies without mass lesions. The 42 cases classified as NPBL histologically were included in the current study. The cytologic features of the smears were evaluated. RESULTS: Cellularity ranged from low (40% of cases) to moderate (50% of cases) to high (7% of cases). The cells were arranged in small clusters in 79% of cases, were mixed with large sheets in 17% of cases, and were in large sheets in 2% of cases. Intact lobules were noted in 31%. The configuration of the epithelial groups was complex in 62% of cases. Myoepithelial cells in the background and within the epithelial groups were noted in all the specimens. The percentage of single epithelial cells was < 10 in 38% of cases, 10-20 in 41%, and 20-30 in 19%. Mild nuclear enlargement and overlap, micronucleoli, and mild chromatin clumping were noted in a significant number of cases. CONCLUSIONS: NPBL have been found to have a wide spectrum of cytologic appearances. At one end of the spectrum, smears are cellular with up to 30% single cells and large sheets in a complex configuration and exhibit nuclear enlargement and overlap and prominent nucleoli, features that overlap with those described in proliferative breast lesions.

Review of the Bethesda System atlas does not improve reproducibility or accuracy in the classification of atypical squamous cells of undetermined significance smears.

BACKGROUND: The Bethesda System (TBS) and its accompanying atlas were developed to promote uniform diagnosis and reporting of cervical and vaginal cytology, especially with respect to borderline abnormal smears. The authors assessed whether group study of TBS atlas improves the reproducibility and accuracy of the cytopathologic diagnosis of equivocal Papanicolaou smears. METHODS: One hundred "atypical" smears were divided into pretest and posttest sets containing equal numbers of negative, atypical squamous cells of undetermined significance (ASCUS), and squamous intraepithelial lesion (SIL) diagnoses based on a five-member panel review. Two comparable teams of four pathologists from George Washington University Medical Center (Washington, DC) and Kaiser Permanente (Portland, OR), each comprised of two more experienced cytopathologists and two less experienced pathologists, independently reviewed the 50 pretest slides and classified the slides according to TBS as negative, ASCUS, or SIL. The teams then conducted group study sessions using TBS atlas. After the review, the pathologists independently classified the 50 posttest slides in a similar manner. RESULTS: Pretest, pair-wise interobserver agreement ranged from 30% to 66% compared with 34-62% for posttest agreement. Absolute percent agreement of reviewers' diagnoses with a previously developed consensus diagnosis based on opinions of a five-expert panel (cytopathologic certainty scale) ranged from 44% to 62% for the pretest set and from 40% to 60% for the posttest set. Comparison of the detection of oncogenic human papilloma virus (HPV) DNA by hybrid capture tube test with smears classified as negative, ASCUS, or SIL revealed that seven of eight reviewers did not demonstrate a stronger association between HPV detection and cytologic diagnosis in the posttest set. CONCLUSIONS: Review of TBS atlas by itself does not appear to improve the reproducibility or accuracy of cytologic diagnoses. The lack of improvement was similar among the pathologists involved regardless of experience level or whether they had a close working relation. Cancer (Cancer Cytopathol)

Overexpression of the glucose-regulated stress gene GRP78 in malignant but not benign human breast lesions.

The 78 kDa glucose-regulated stress protein GRP78 is induced by physiological stress conditions such as hypoxia, low pH, and glucose deprivation which often exist in the microenvironments of solid tumors. Activation of this stress pathway occurs in response to several pro-apoptotic stimuli. In vitro studies have demonstrated a correlation between induced expression of GRP78 and resistance to apoptotic death induced by topoisomerase II-directed drugs. We were interested in characterizing this protein in human breast lesions for potential implications in chemotherapeutic intervention. Surgical specimens of human breast lesions and paired normal tissues from the same patients were flash frozen for these studies. Total RNA and/or protein were extracted from these tissues and used in northern and/or western blot analyses, respectively, to quantify the relative expression of GRP78. Northern blot analysis indicated that 0/5 benign breast lesions, 3/5 estrogen receptor positive (ER+) breast tumors, and 6/9 estrogen receptor negative (ER-) breast tumors exhibited overexpression of GRP78 mRNA compared to paired normal tissues, with fold overexpressions ranging from 1.8 to 20. Western blot analyses correlated with these findings since 0/5 benign breast lesions, 4/6 ER+ breast tumors, and 3/3 ER- breast tumors overexpressed GRP78 protein with fold overexpressions ranging from 1.8 to 19. Immunohistochemical analysis of these tissues demonstrated that the expression of GRP78 was heterogeneous among the cells comprising different normal and malignant glands, but confirmed the overexpression of GRP78 in most of the more aggressive ER- tumors. These results suggest that some breast tumors exhibit adverse microenvironment conditions that induce the overexpression of specific stress genes that may play a role in resistance to apoptosis and decreased chemotherapeutic efficacy.

Cytologic features of proliferative breast disease: a study designed to minimize sampling error.

BACKGROUND: Assessment of cytologic features that allow accurate classification of proliferative breast disease has been hampered by sampling errors when fine-needle aspirations have been compared with their corresponding histologic sections. METHODS: To allow for optimal cytohistologic correlation, 2 smears (1 hematoxylin and eosin-stained and 1 Diff-Quik-stained) were prepared from each of 98 breast biopsies without mass lesions and compared with the corresponding histologic sections of the scraped area. Each smear was reviewed in a blinded fashion and assessed for cellularity, background elements, cytoarchitectural features of cell groups, and nuclear features by 2 reviewers. Smears were then classified as nonproliferative breast disease (NPBD), proliferative breast disease without atypia (PBD) or with atypia (PBDA), or DCIS, based on review of the corresponding histologic sections. RESULTS: When comparing NPBD/PBD (n = 86) with PBDA/DCIS (n = 12), smears from PBDA/DCIS were significantly (by the Fisher exact test or Wilcoxon rank sum P values with adjustment for multiple comparisons) more likely to be cellular; contain single cells and necrosis; exhibit nuclear overlap and cytoplasmic vacuoles; have large nuclei, macronucleoli, pleomorphism, clumped chromatin, and hyperchromasia; and were less likely to have complex cell groups, monolayers, swirling, cohesion, and myoepithelial cells in epithelial sheets and the smear background. When NPBD (n = 53) and PBD (n = 33) were similarly compared, smears from PBD were more likely to exhibit larger and more complex cell groups, but they were otherwise similar to smears from NPBD. CONCLUSIONS: There are many cytologic features that will allow a distinction of NPBD/PBD from PBDA/DCIS, but relatively few that can aid in separating NPBD from PBD.

Changing trends in breast fine-needle aspiration: results of the Papanicolaou Society of Cytopathology Survey.

Following the NCI-sponsored consensus conference on fine-needle aspiration of the breast, the Criteria and Nomenclature Task Force of the Papanicolaou Society of Cytopathology undertook a survey to assess the status of these issues and recommendations among practicing cytopathologists. The survey was designed to assess the impact of the changing trends in the diagnosis of breast lesions on cytopathology laboratories. It also intended to assess the impact of the recommendations of the consensus conference concerning the inclusion of a statement in breast FNA reports recommending the use of the triple test, the use of the proposed diagnostic terminology, and to evaluate criteria for specimen adequacy in breast FNAs used in different institutions. The results of this survey indicate the impact of an increasing use of core biopsies on the number of breast FNAs performed over the last several years. The recently recommended diagnostic terminology for breast FNA has quickly gained wide acceptance, as has the fundamental concept of the triple test. The issue of specimen adequacy, however, remains controversial, with some laboratories utilizing quantitative criteria, while the majority do not. Diagn. Cytopathol. 2000;22:126-130.

"ASCUS, rule out HSIL": cytologic features, histologic correlates, and human papillomavirus detection.

Risk factors, cytologic and histopathologic features, and human papillomavirus (HPV) detection associated with 75 cervical smears classified as atypical squamous cells of undetermined significance, rule out high-grade squamous intraepithelial lesion (ASCUS, rule out HSIL) were reviewed. Cases were identified in a pathology panel review of material collected from 1953 women participating in a 5-year prospective study of HPV infection and squamous intraepithelial lesions at Kaiser Permanente, Portland, Oregon, sponsored by the National Cancer Institute. Initial abnormal smears diagnosed as ASCUS, rule out HSIL by one panelist or diagnosed as ASCUS by one pathologist and as HSIL by another were included. The 75 ASCUS, rule out HSIL smears identified were examined again by two pathologists after the study. These cases were compared with cases of ASCUS, not otherwise specified (ASCUS, NOS) and HSIL identified in the same group of 1953 women. Findings in ASCUS, rule out HSIL included tissue fragments (21%); atypical immature metaplasia (17%); atypical mature metaplasia (15%); small atypical cells (9%); and atypical repair (4%). A final patient classification of HSIL, reflecting all available data, was assigned to 11 (24%) of 46 women with ASCUS, rule out HSIL and to 1(1%) of 80 women with ASCUS, NOS in the original review (P < .001). Detection of oncogenic HPV types at diagnosis in ASCUS, rule out HSIL; ASCUS, NOS; and HSIL was similar, but data were unavailable for many subjects. Among women not tested at diagnosis, enrollment testing (1 to 4 years earlier) revealed that HPV detection in women with ASCUS, rule out HSIL was intermediate in frequency between ASCUS, NOS and HSIL. These data suggest that ASCUS, rule out HSIL is a distinct diagnosis from ASCUS, NOS because it is more often associated with an underlying HSIL. Consequently, women with ASCUS, rule out HSIL should be referred for colposcopic examination.

Utility of thin-layer preparations in thyroid fine-needle aspiration: diagnostic accuracy, cytomorphology, and optimal sample preparation.

BACKGROUND: The efficacy of preparing thyroid fine-needle aspirations (FNAs) as thin-layer slides has not been evaluated extensively. METHODS: To evaluate the efficacy of thin-layer cytology, the authors examined thyroid FNAs from 135 unselected palpable lesions that were aspirated using uniform procedures and prepared as air-dried, Diff-Quik-stained direct smears (DS) and Papanicolaou-stained thin-layer slides (TL). Diagnoses rendered independently on masked slides from each thyroid nodule were compared with the reported final cytologic or histologic diagnoses based on all available pathologic material. Slides were analyzed for diagnostically important cytologic features and the optimal number of TL slides per sample was assessed. RESULTS: TL diagnoses agreed with final diagnoses in 85% of cases compared with 96% for DS. All 11 neoplasms were recognized in both preparations. A final diagnosis of chronic lymphocytic thyroiditis (CLT) was missed in 10 of 26 TL (39%) compared with 2 of 26 DS (8%) (P = 0.013, McNemar's test). Cytologic features appreciated less often in TL than DS (McNemar's test) included diffuse colloid, spherules, tissue fragments, and lymphocytes. Multinucleated giant cells were identified more often in TL than DS. Two TL slides were sufficient for diagnosis in 83% of cases. CONCLUSIONS: The diagnostic accuracy of TL was 85% compared with 96% for DS. CLT was diagnosed accurately in 62% of TL compared with 92% of DS. Cytologic features in TL and DS may differ. Preparation of only two TL slides is adequate for definitive diagnosis in most cases.

Malignant osteoclast-like giant cell tumor of the kidney.

Osteoclast-like giant cell tumors are rare mesenchymal tumors that typically present in osseous tissue and are remarkable because of their slow growth. We report the sixth documented case of a malignant osteoclast-like giant-cell tumor primary to the kidney of an 81-year-old man with a 5-month history of hematuria. Postnephrectomy analysis of the tumor, including immunohistochemical stains and electron microscopy, confirmed the diagnosis. Although the patient died of unrelated complications, metastatic disease was suspected.

The stability of estrogen and progesterone receptor expression on breast carcinoma cells stored as PreservCyt suspensions and as ThinPrep slides.

BACKGROUND: Analysis of estrogen receptor (ER) and progesterone receptor (PR) status is an important ancillary test in the evaluation of positive breast fine-needle aspirates. This study compares the detection of ER and PR in breast carcinoma cells suspended in PreservCyt with that achieved with stored ThinPrep slides (TP). METHODS: ER and PR positive mammary tumor cells (cell line ZR-75-1 spiked in PreservCyt by the American Type Culture Collection) were used to evaluate the stability of immunodetection of ER and PR under two conditions: 1) TP slides prepared immediately from PreservCyt and stored air-dried (stored TP) for up to 56 days, and 2) TP prepared from cells suspended in PreservCyt (newly prepared TP) on Days 1, 2, 5, 14, 21, 42, and 56. At each of the time periods, stored TP and newly prepared TP were analyzed for ER and PR using the same immunocytochemical staining protocol. The percentage of positive cells was calculated by counting 1000 cells/TP. RESULTS: Positivity for ER and PR was demonstrated in both stored TP and newly prepared TP on Days 1, 2, 5, 14, 21, 42, and 56. Over the 56-day period, the number of ER positive cells ranged from 41% to 57% in stored TP and from 38% to 58% in newly prepared TP. The number of PR positive cells ranged from 31% to 41% in stored TP and from 26% to 37% in newly prepared TP. Mild, nonspecific cytoplasmic and nuclear staining occurred in all newly prepared TP (PR > ER). CONCLUSIONS: ER and PR antigenicity was preserved in both stored TP and newly prepared TP of mammary tumor cells over a 56-day storage period. This demonstrates that ER and PR status can be evaluated in cytologic material from breast carcinoma using the ThinPrep technique.

Cytomorphology of cystic parathyroid lesions: report of four cases evaluated preoperatively by fine-needle aspiration.

Cystic parathyroid lesions (CPL) account for 1-5% of neck cysts. They are seldom palpable; however, they may present as neck swellings that are mistaken both clinically and cytologically for thyroid nodules and subsequently referred for evaluation by fine-needle aspiration. We present 4 cases of histologically confirmed CPL (one simple cyst, one hyperplasia, and two adenomas), 2 of which were misdiagnosed as adenomatoid thyroid nodules by fine-needle aspiration. Aspirated fluid from one patient was clear and colorless, a classic finding for parathyroid cysts, and contained high levels of C-terminal/midmolecule parathyroid hormone (CMPH). Fluids obtained from the remaining 3 patients were bloody to brown, resembling thyroid cyst fluid. In only 1 of the 3 patients, the fluid was analyzed and contained high levels of CMPH. Though cytologic features of parathyroid epithelium overlap with those of thyroid epithelium, distinguishing features such as unusual cytoplasmic vacuolization and granularity, and the absence of colloid, should raise suspicions of a parathyroid lesion. Fluid from CPL may not possess the classic appearance; however, identification of the subtler cytologic features with knowledge of relevant clinical data should prompt analysis of cyst fluid for CMPH, thus confirming the diagnosis and avoiding inappropriate therapy.

Multinucleated giant cells in thyroid neoplasms. A cytologic, histologic and immunohistochemical study.

OBJECTIVE: To evaluate the presence of multinucleated foreign body-type giant cells (MGCs) in papillary carcinoma (PC), follicular variant of papillary carcinoma (FV) and follicular adenomas (FA) and to investigate their origin by immunoperoxidase studies. STUDY DESIGN: The cytologic and histologic material of 10 PCs, 8 FVs, and 11 FAs were evaluated for the presence of MGCs. The immunohistochemical staining pattern of MGCs in paraffin-embedded tissue was studied using cytokeratin, epithelial membrane antigen, thyroglobulin, vimentin, A-1 antichymotrypsin, lysozyme and CD68. RESULTS: MGCs were demonstrated in 10/10 PCs, 5/8 FVs and 0/11 FAs. In the immunostained sections, MGCs were negative for epithelial markers and positive for histiocytic markers. CONCLUSION: These results indicate that MGCs are histiocytic and frequently present in PCs and FVs and usually absent from FAs.

Evaluation of the 5-year review of negative cervical smears in patients with high grade squamous intraepithelial lesions.

Federal regulations require laboratories to re-examine negative cervical smears from the 5 yr preceding a diagnosis of high-grade squamous intraepithelial lesion (HSIL) or carcinoma. To assess the value of this regulation as a quality assurance measure, we evaluated the previous cervical smears from 47 patients with a diagnosis of HSIL rendered during a 9-mo period. Fifteen patients had a total of 24 previous smears initially reported as squamous intraepithelial lesions (SIL). The remaining 32 patients had a total of 58 smears that carried a diagnosis lesser than SIL. Upon re-screening 22/58 (38%) smears, from 16 patients, were upgraded to SIL (8 low grade, 14 high grade). Underdiagnosed SIL was detected in seven (44%) patients with the 1-yr review, 12 (75%) with the 2-yr review, 15 (94%) with the 3-yr review, and 16 (100%) with the 5-yr review. The authors conclude that the negative smear review is a beneficial quality assurance and teaching method that may lead to quality improvement. A 3-yr retrospective review is effective and detects 94% of the undercalls.

Evaluation of the 10% rescreen of negative gynecologic smears as a quality assurance measure.

The CLIA '88 regulations require the laboratory to review at least 10% of the gynecologic cases interpreted as negative that are examined by each cytotechnologist. The review must include negative cases selected at random and from patients identified as a high risk group. To evaluate the validity of this regulation, we reviewed the results of the 10% rescreen over a 24-mo period. The rescreening by pathologists detected epithelial abnormalities in 4/2,124 negative cases (0.18%). Subsequently, biopsy-proven squamous intraepithelial lesions were found in all four cases. Three different cytotechnologists were responsible for the four false-negative interpretations. Our results show that 10% rescreen of negative smears may detect a few false-negative cases. However, it is a time-consuming and inefficient mean to evaluate the performance of cytotechnologists. Quality control in gynecologic cytopathology is essential, but it is equally important that laboratories concentrate their resources and energy on methods proven to be effective.

Increased numbers of mast cells in pterygia.

PURPOSE/METHODS: We examined the mast cells in 12 pterygium specimens of patients who underwent primary excisions and the conjunctival specimens of ten normal age-matched control subjects. RESULTS/CONCLUSION: The mean mast cell count per cubic millimeter was twice as high in the pterygium specimens as in the control specimens. Mast cell proliferation and activation may contribute to the pathogenesis of pterygium formation.

ACTH-producing pituitary carcinoma presenting as the cauda equina syndrome.

A 43-year-old woman presented with incontinence, weakness, and paresthesia, consistent with the cauda equina syndrome, 10 years after having a pituitary tumor surgically removed and 4 years after excision of two "meningiomas" of the cervical cord. The patient was also hypertensive and had a cushingoid habitus. Emergent surgical decompression of the spinal cord revealed intradural metastatic adrenocorticotropic hormone-producing pituitary carcinoma. Pituitary carcinomas are rare. The majority of reported cases of adrenocorticotropic hormone-producing carcinoma have exhibited metastases outside the central nervous system. To our knowledge, this represents the first case of an adrenocorticotropic hormone-producing pituitary carcinoma presenting with the cauda equina syndrome. A review of all reported cases of pituitary carcinoma indicated that central nervous system metastases were more common than metastases to distant sites, and patients with distant metastases experienced a shorter duration of disease than did those with central nervous system metastases.

The adequacy of the one-slide cervical smear in the detection of squamous intraepithelial lesions.

This prospective study evaluated the impact of replacing the two-slide cervical smear by a one-slide smear to detect squamous intraepithelial lesions (SIL). The study population consisted of 87 patients with biopsy-proven SILs, a "referral" (precolposcopy) two-slide cervical smear, and a "one-slide repeat" smear done at the time of biopsy. The detection rate of SIL was compared with that of a control group of 85 patients who fulfilled similar criteria; they had a two-slide referral smear, but a "two-slide repeat" smear. Smears nondiagnostic of SIL occurred in 5.7%, 11.8%, 27.6%, and 15.3% in the referral group of the study population, the referral group of the control population, the one-slide repeat smears, and the two-slide repeat smears, respectively. Statistical analysis showed a higher rate of nondiagnostic smears in the one-slide repeat smear (P = .05). Although this may be attributed to the less-than-optimal quality of the preparation, the above findings do not support the use of a one-slide technique for sampling the ectocervix and endocervix.

Reactive change and atypical squamous cells of undetermined significance in Papanicolaou smears: a cytohistologic correlation.

The purposes of this study were to delineate morphologic criteria for the terms reactive/reparative change (RC) and atypical squamous cells of undetermined significance (ASCUS), which were proposed by the Bethesda System, and to evaluate the impact of the new terminology on patient management. Using criteria defined in this article, 88 cervical smears originally reported as "atypia" were reclassified as RC (57) and ASCUS (31). Correlation with the colposcopically directed biopsies was as follows: 3/57 RC showed condyloma and 19/31 ASCUS had squamous lesions (14 condyloma, four mild, and one moderate dysplasia). The remainder of the cases had either cervicitis or no significant abnormality histologically. Using the proposed criteria, 61.3% of ASCUS correlated with positive biopsies, compared to 5.2% of the RC cases. Our results indicate that patients with smears showing RC are significantly less likely to have a squamous lesion as compared to those showing ASCUS, and therefore may be managed more conservatively.

Should we report cervical smears lacking endocervical component as unsatisfactory?

The purpose of the study is to correlate the accuracy of diagnosis of cervical smears with the presence/absence of an endocervical component. The referral smears and the smears obtained during colposcopic examination of 84 patients with biopsy-proven squamous lesions were evaluated for the presence of an endocervical component, and the cytologic interpretations were compared with the histologic findings. Of the 136 smears containing an endocervical component, 111 (81.6 percent) had good correlation with histology and 25 (18.4%) showed a discrepancy. Of the 30 smears lacking an endocervical component, 24 (80%) had good histologic correlation and 6 (20%) were discrepant. Our data showed no significant difference in the detection of squamous lesions in those smears with an endocervical component from those without. We conclude that, although the presence or absence of an endocervical component should be documented in the cytology report, its absence should not be an indication to report the cervical smears as unsatisfactory.