Ultrastructural analysis of fine needle aspirates from benign breast lesions.

Thirty six fine needle aspirates from various types of benign breast lesions were examined by electron microscopy and correlated with their cytological appearances. In all cases the parenchyma consisted of clumps of cohesive cells with the ultrastructural features of epithelial cells. In many cases, particularly from fibroadenomas, the parenchyma consisted of single layers of polarised epithelial cells showing lumen formation. Similar arrays of apocrine epithelial cells were observed in 60% of fibrocystic lesions. The more solid clumps from hyperplastic lesions consisted of epithelial cells of variable shape and electron density with disorganised lumen formation. Irrespective of the type of lesion, the epithelial cells were not normally subtended by myoepithelial cells or basal lamina. The extraction process seems to result in a shearing between the epithelium and basal lamina with lysis of the myoepithelial cells. Most naked nuclei probably result from lysed myoepithelial cells.     

Semiquantitative analyses of fine needle aspirates from benign and malignant breast lesions.

A scoring system for the assessment of fine needle aspirates of benign and malignant breast lesions was devised which showed a positive correlation (r = 0.67) between the scores obtained from the fine needle aspirates from ductal carcinomas and the Bloom and Richardson-type scores for their paired excision biopsy specimens. This system permitted grades II and III ductal breast carcinoma to be distinguished reliably from grade I tumours but no correlation with the lymph node state of patients with breast carcinoma was shown. Some overlap between the scores for grade I ductal carcinomas and some benign lesions was found, and this underlines a need for caution in the reporting of such equivocal aspirates. No cytological features that distinguished reliably ductal from lobular carcinoma were identified but the same spectrum of severity of cytological abnormality in the ductal and lobular carcinoma aspirates was seen. This system may be of prognostic value in the assessment of lobular carcinoma which has hitherto defied histological grading.     

False-positive reports in fine needle biopsy of breast lesions

A review of the literature reveals considerable variations in the diagnostic accuracy of fine needle biopsy (FNB) of breast lesions between series, partly due to different methods of calculation, different definitions, and insufficient numbers of cases with adequate follow-up to provide reliable statistics. The best larger series have a false-positive rate between 0.2 and 0.3%, slightly higher for non-palpable than for palpable lesions. The cytological patterns of a range of benign lesions which may cause diagnostic difficulties and may be misdiagnosed as malignant by FNB are described, and guidelines to reduce the risk of false-positive diagnoses are proposed.     

Iododeoxyuridine labelling of S-phase fraction in fine needle aspirates from breast carcinomas.

The suitability of measuring the S-phase fraction in human breast cancer by labelling tumour cells from fine needle aspirates (FNAs) in vitro with iododeoxyuridine (IdU) was studied in 11 patients. The S-phase fraction was measured both in preoperative FNAs labelled in vitro with IdU, and in FNAs taken from the same tumour when surgically removed after intravenous administration of IdU. Frozen sections were also immunostained for incorporated IdU. The mean S-phase fraction measured in FNAs after in vitro or in vivo labelling and in sections after in vivo labelling was 4.0, 3.6, and 3.1, respectively. Results of in vitro and in vivo labelling of FNAs with IdU were similar. However, as the S-phase fraction in breast cancer is generally low, the variation between the different measurements is too large; therefore, the S-phase fraction is not a suitable indicator of response to treatment.     

Interphase cytogenetics of chromosomes 11 and 17 in fine needle aspirates of breast cancer

The aims of this investigation were to compare quantitative with qualitative analysis of fluorescent in situ hybridization (FISH) centromere signals in interphase breast cancer cell nuclei and to evaluate the possible clinical utility of detecting numerical abnormalities of chromosomes 11 and 17 by FISH in the preoperative prediction of breast cancer histological grade. Commercial digoxigenin-labeled centromere probes to chromosomes 11 and 17 were hybridized to 69 malignant aspirates with histological follow-up. Aspirates were categorized as disomic or aneusomic for chromosomes 11 and 17 qualitatively; a subset of aspirates was also analyzed quantitatively. The quantitative and qualitative approaches resulted in almost identical categorisation. There was a significant association between the qualitative categorization of aspirates as aneusomic or disomic, the histological grade of the excised tumours (P = .0695, n = 69), and the cytological grade of the clinical aspirates (P = .006, n = 35). Although histological grade III tumors were almost invariably polysomic for one or both chromosomes, polysomy was also detected in grade I and II tumors. Qualitative FISH analysis was shown to be more sensitive than cytological grading in predicting histological grade III but was of lower specificity and was therefore not clinically useful.     

Mammography-guided stereotactic fine needle aspiration cytology of breast lesions

The purpose of the present study was to evaluate the efficacy of mammographic guided stereotactic fine needle aspiration cytology in 136 patients with breast lesions including mammographic features, cytomorphological findings and nuclear grading in malignant cases. Majority of the cases were predictable correctly by combination of the three modalities of diagnosis viz, clinical examination, mammography and cytology.     

Correlation of p53 mutations in ThinPrep-processed fine needle breast aspirates with surgically resected breast cancers.

Mutations of the p53 gene are one of the most common genetic changes found in cancer; their presence may be prognostic and even influence treatment for breast cancer. In this study, we investigated whether DNA could be extracted from the residual cells left in ThinPrep-processed breast fine-needle aspirates and whether p53 gene changes could be detected in the DNA. The results were then correlated with DNA extracted from the matched formalin-fixed, paraffin-embedded, surgically resected breast cancer when available. DNA was successfully extracted from 54 of 62 aspirates and all 31 surgical specimens. p53 gene mutations were detected in 10 of the 54 cytology specimens (18.5%) and consisted of base pair substitutions or deletions. Silent or intronic p53 changes were found in five additional aspirates. One of the aspirates had two gene alterations, resulting in a total of six gene changes. Five of these changes were located in introns 6 or 9 and the sixth was a silent (no amino acid change) change in exon 6. p53 Polymorphisms were detected in nine aspirates (16.3%) and were located in codon 47 (one aspirate), codon 72 (six aspirates), and codon 213 (two aspirates). All cases with surgical material available showed identical p53 mutations, alterations, and polymorphisms in the resected tumors compared with those detected in the corresponding aspirates. The results of this study show that DNA suitable for analysis of p53 gene sequence changes can be successfully extracted from ThinPrep-processed breast fine-needle aspirates, and that identical alterations are detected in both the cytology and surgical specimens.     

Fluorescence in situ hybridisation detection of erbB2 amplification in breast cancer fine needle aspirates.

AIM: To develop a method for the detection of amplification of the erbB2 oncogene in breast cancer fine needle aspirates using fluorescence in situ hybridisation (FISH) and to compare amplification with immunohistochemical detection of the erbB2 protein. METHODS: A digoxigenin labelled probe to the erbB2 gene was hybridised to 15 aspirates prepared from operative breast cancer specimens. A chromosome 17 centromere probe was also hybridised to the aspirates either separately or in combination with the erbB2 probe. The aspirates were scored for erbB2 amplification and chromosome 17 centromere number. Subsequently, paraffin wax embedded sections of the tumours were stained with the antibody CB11 and scored for the presence of membrane staining. RESULTS: Three of the 15 tumour aspirates showed high level amplification of erbB2 detected by FISH. These three tumours also showed chromosome 17 polysomy and diffuse membrane staining by immunohistochemistry. CONCLUSIONS: FISH can be used to detect erbB2 amplification in fine needle aspirates and results correlate with conventional immunohistochemical staining. Difficulties were encountered in the visualisation of the signals in non-amplified cases without the use of specialised digital imaging.     

Fluorescence in situ hybridisation detection of erbB2 amplification in breast cancer fine needle aspirates.

AIM: To develop a method for the detection of amplification of the erbB2 oncogene in breast cancer fine needle aspirates using fluorescence in situ hybridisation (FISH) and to compare amplification with immunohistochemical detection of the erbB2 protein. METHODS: A digoxigenin labelled probe to the erbB2 gene was hybridised to 15 aspirates prepared from operative breast cancer specimens. A chromosome 17 centromere probe was also hybridised to the aspirates either separately or in combination with the erbB2 probe. The aspirates were scored for erbB2 amplification and chromosome 17 centromere number. Subsequently, paraffin wax embedded sections of the tumours were stained with the antibody CB11 and scored for the presence of membrane staining. RESULTS: Three of the 15 tumour aspirates showed high level amplification of erbB2 detected by FISH. These three tumours also showed chromosome 17 polysomy and diffuse membrane staining by immunohistochemistry. CONCLUSIONS: FISH can be used to detect erbB2 amplification in fine needle aspirates and results correlate with conventional immunohistochemical staining. Difficulties were encountered in the visualisation of the signals in non-amplified cases without the use of specialised digital imaging.     

Optimal specimen processing of fine needle aspirates of non-hodgkin lymphoma

Fine needle aspiration (FNA) in conjunction with flow cytometry (FC) is a useful technique for non‐Hodgkin's lymphoma (NHL) diagnosis. We sought to investigate the effect of storage medium and time to processing on lymph node (LN) FNA viability. Benign LN FNAs were distributed among Roswell Park Memorial Institute (RPMI), Hanks' Balanced Salt Solution (HBSS) and Dulbecco's Modified Eagle's Medium (DMEM) storage media, and viability was compared at 0, 4.5 and 24 hours. FC survival analysis showed viable cells (%): at 0 hrs: HBSS 83.6%, RPMI 87.7%, DMEM 87.7%. At 4.5 hrs: HBSS 86.3%, RPMI 89.0%, DMEM 78.2%. At 24 hrs: HBSS 82.7%, RPMI 86.7%, DMEM 77.2%. FNA from a peri‐pancreatic LN involved by grade 2 follicular lymphoma was stored in RPMI at 4° C and analyzed at 1, 3, 5 and 7 days. Over 90% of follicular lymphoma cells were suitable for FC analysis at 1, 3, and 5 days after collection, decreasing to 76% at 7 days. In conclusion: RPMI appears to be the optimal storage medium compared to DMEM and HBSS.An NHL FNA sample stored at 4° C remains suitable for FC analysis for up to 5 days. Diagn. Cytopathol. 2012. © 2011 Wiley Periodicals, Inc.     

Microfilariae in fine needle aspirates: a report of 22 cases

The objective of the study is to document the value of fine needle aspiration cytology (FNAC) in the diagnosis of filariasis at all possible sites in both exfoliative cytologic material and fine needle aspirates. Both unguided and guided FNACs of all foci were studied over a period of two years between 1999 to 2000. Total 22 cases of filariasis were detected which included subcutaneous swellings(7), breast(3), thyroid(3), lymphnodes(3), effusions(3), cervical scrape(1), eyeball(1), sputum(1) and bronchial washing(1). In none of these cases was filariasis considered a diagnostic possibility. Cytologic smears showed eosinophils in 9 cases, oval ova and embryonated eggs in 2 cases. Microfilariae were associated with other diseases in 13 cases, including 6 cases of malignancy. Significant adherence of inflammatory cells and macrophages to microfilariae was present in 6 of the 22 cases. In endemic areas,filariasis should be considered one of the differential diagnosis of a swelling. Thus demonstration and identification of the parasite in cytologic smears played a significant role in the prompt recognition of the disease and institution of specific treatment, thus obviating the more severe manifestations of lymphatic frilariasis.     

Cytoplasmic vacuolation,intracytoplasmic lumina,and DPAS staining in ductal carcinoma of the breast

A retrospective study was carried out on 30 cases of histologically proven invasive ductal carcinoma of the breast with a prior fine-needle aspiration (FNA) cytology. On evaluating the May-Grünwald-Giemsa (MGG) FNA smears, cytoplasmic vacuolation was observed in 70% cases. Positivity with periodic acid Schiff-positive, diastase-resistant (DPAS) staining was observed in 90% of cases. The chi(2) value on a McNemar test was 4.16. Thus, DPAS staining was significantly superior to MGG staining for picking up cytoplasmic vacuoles (P < 0.05). In 56.67% cases, DPAS staining showed an improvement in score as compared to MGG smears. This was highly significant (P < 0.001) on Wilcoxon matched-pairs signed-ranks test. Applying the strict criteria of thick-walled cytoplasmic vacuoles with a central darkly stained dot, none of our cases revealed true intracytoplasmic lumina. Larger studies are required to establish a role for DPAS staining in separating borderline, in situ, and invasive breast lesions, and to see if such positvity can be incorporated into the grading systems for breast carcinoma.     

Nucleolar organizer regions distribution in fine needle aspiration cytological smears from breast lesions

Fine needle aspiration (FNA) cytology is now an integral part of the pre-operative investigation of breast lesions and the therapeutic protocol is today often planned on the basis of cytodiagnosis. However, from time to time the cytological picture may be equivocal or inconclusive. In recent years, nucleolar organizer region (NOR) scores have been explored for potential value in the diagnosis of malignancy as the scores in malignant nuclei are seen to be higher than in benign or reactive nuclei. With a view to applying NOR scoring in the evaluation of cytologically equivocal cases, we adopted the argyrophil technique for staining NOR s (AgNOR) in FNA cytological smears of 56 breast lesions, comprising 31 benign and 25 malignant lesions. Histological correlation was possible in 26 of these cases (17 malignant and 9 benign) and AgNOR scoring was done on paraffin sections of these as well. There was a significant difference between mean AgNOR scores in benign and malignant lesions in the cytological smears (P < 0.001). The AgNOR scores ranged from 2.5 to 5.0 per cell in benign lesions and 5.8 to 17.2 per cell in malignant lesions. None of the cases fell into the gray zone of overlap. One malignant lesion that was cytologically equivocal showed a mean AgNOR score of 6.08. The AgNOR scores on histological sections also showed a statistically significant difference (P < 0.001) between benign and malignant lesions with mean scores ranging from 1.34 to 2.58 dots per cell in benign lesions and scores of 2.42 to 5.28 dots per cell in malignant lesions. However, the scores overlapped in four cases and therefore it was considered unsuitable for routine diagnostic work. From this preliminary study, we conclude that an FNA AgNOR score of 5.0 and less strongly favours a benign lesion whereas a score above 5.0 would be in favour of a malignant lesion. A larger study would be needed to verify our impression that AgNOR scoring can be useful in cytologically equivocal cases.     

免疫组织化学标记物在乳腺细针吸取细胞学鉴别诊断中的意义

目的探讨免疫标记物对鉴别乳腺细针吸取细胞学（FNAC）良性病变和癌的意义。方法收集135例有随访资料、活检和组织病理学诊断对照的乳腺FNAC资料：良性病变88例,包括非增生性病变43例和增生性病变45例;乳腺癌47例,对其FNAC涂片和相应的石蜡切片作细胞周期蛋白（cyclin）D1、c—erbB-2、Ki-67、p21^CIP1/WAF1（简称p21）和34βE12的免疫组织化学APAAP和ABC法检测。利用SPSS11．5软件进行分析。结果（1）以上各标记物在良性非增生性和增生性病变中的标记差异无统计学意义。（2）以上各标记物在良、恶性病变中的标记差异均有统计学意义（P〈0．001）。多因素的logistic回归分析显示最有意义的组合标记物为cyclinD1（P〈0．001）、34βE12（P〈0．001）和c—erbB-2（P=0．003）．cyclinD1、c—erbB-2阳性和34βE12阴性提示为癌,其组合诊断的敏感性和特异性最高。组合标记物共同判断,cyclinD1和34βE12任一判断为癌,诊断的敏感性和特异性分别为95．7％和94．3％;这三个标记物任一判断为癌,诊断的敏感性进一步上升至97．9％,特异性下降至92．0％;这三个标记物任两个共同判断为癌,诊断的敏感性为72．3％,特异性为100．0％。（3）在癌组中,根据Robinson细胞学分级把癌分为3级,cyclinD1、34βE12和p21在各级癌中的表达差异不大,而c—erbB-2和Ki-67在1级癌的阳性表达率最低,仅为40．0％和33．3％,在3级癌中阳性表达率最高。组合cyclinD1和34βE12,cyclinD1和34βE12,任一判断为癌,1级和2级癌的检出率为93．3％和96．2％。结论所检测的免疫标记物对良、恶性病变的鉴别诊断价值较大,组合cyclinD1、34βE12和c—erbB-2可最有效地提高癌的诊断敏感性和特异性。对鉴别分化好的乳腺癌和乳腺良性病变,最有效的组合为cyclinD1和34βE12。     

The value of fine needle aspiration biopsy in the diagnosis and prognostic assessment of palpable breast lesions

In recent years there appears to be a growing movement toward the use of core needle biopsy (CNB) over fine needle aspiration biopsy (FNAB) for the detection of breast carcinoma. This tendency is caused in part by the idea that CNB can provide a more specific or definitive diagnosis as well as the belief that the assessment of prognostic/predictive factor is not possible or reliable on cytologic specimens. At our institution, FNAB of breast has been practiced for over 25 years with excellent cytologic-histological correlation. This practice has been beneficial not only for patients, but also for training physicians since nowadays a very small number of centers in the United States still routinely perform fine needle aspiration as a diagnostic tool in breast cases. To assess the diagnostic accuracy of FNAB in palpable breast lesions, we reviewed our experience during an 8-year-period and compared fine needle aspiration results with follow-up surgical specimens. From the cytology point of view, the lesions were divided as negative/benign, atypical/suspicious, positive, and insufficient. Only cases performed by pathologists were included. A total of 1,583 cases were retrieved from our archives. A definitive malignant diagnosis was reached in 357 cases. One-hundred and thirty-nine cases were classified as atypical/ suspicious, 135 cases had insufficient cells for establishing a diagnosis, and 952 were categorized as negative. A total of 408 follow-up surgical specimens were available for comparison with cytologic results. There were 19 false-negative, and no false-positive results were found. The majority of false-negative results were secondary to sampling errors. In 93% of the malignant cases, there was enough material obtained in cytological specimens to perform prognostic/predictive factors studies. Our data proves once again that FNAB is a reliable method for the initial evaluation and diagnosis of palpable masses of the breast. In addition, it also has the ability of providing necessary prognostic/predictive information, particularly for patients that may undergo neoadjuvant therapy.     

Cytologic diagnosis of papillary carcinoma of the breast in needle aspirates

Eleven cases of rare papillary carcinoma of the breast diagnosed by fine-needle aspiration cytology (FNAC) are reported. Five of these were pure papillary carcinomas and six were mixed papillary and ductal, lobular, or mucinous carcinomas. In each case, cytological material was collected by washing the needle and syringe contents into 30% alcohol in saline, and the Gelman cytosieve method was used for the cytological preparations. In this article, the cytological features of these tumors are described, including the presence of single papillae and papillary clusters, tall columnar cells, diathesis of blood with hemosiderin-laden macrophages, naked nuclei, and high cell recovery.     

The usefulness of cytogenetic analysis in fine needle aspirates for the histologic subtyping of sarcomas.

Conventional cytogenetic analysis performed from open biopsy tissue samples may be a useful adjunct for the histologic subtyping of bone and soft tissue sarcomas. However, its diagnostic utility in fine needle aspiration biopsy (FNAB) specimens is unclear. We retrospectively reviewed 24 consecutive FNAB bone and soft tissue sarcoma specimens, procured from 1995 to 2003, in which aspirated material was obtained for cytogenetic analysis. The study sample included eight Ewing sarcomas, six synovial sarcomas, five rhabdomyosarcomas, two myxoid liposarcomas, and one each of myxoid chondrosarcoma, osteosarcoma, and atypical lipoma. Cytogenetic analysis confirmed the t(X;18) in all six synovial sarcomas and the t(11;22) in three Ewing sarcomas. The t(2;13) was strongly suggested in one alveolar rhabdomyosarcoma. For two of these cases (both of which were synovial sarcomas), cytogenetic analysis was necessary for definitive diagnosis. While the positive cytogenetic results were supportive in the remainder, all were initially and accurately subtyped based on cytomorphology and/or immunohistochemistry. Cytogenetic analysis was noncontributory (eg no growth) in 14 sarcoma cases, but excluding the case of atypical lipoma, this did not preclude the rendering of an accurate diagnosis. Cytogenetic analysis can be performed on FNAB specimens from bone and soft tissue sarcomas and may be a useful diagnostic aid in difficult cases. However, when cell block material is available for immunohistochemistry, the majority of such cases are successfully subtyped.