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1.
J Clin Pathol ; 56(7): 532-6, 2003 Jul.
Article in English | MEDLINE | ID: mdl-12835300

ABSTRACT

AIMS: To investigate the physical status of human papillomavirus 16 (HPV-16) in low grade squamous intraepithelial lesions (LSILs) as a means of determining the percentage of viral integration. METHODS: Ninety two LSIL/HPV positive Thin Prep(TM) samples were initially tested for the E6 gene by the polymerase chain reaction (PCR) to identify the HPV-16 virus. To avoid false positive results, the specificity of the bands obtained from PCR was confirmed by Southern blot hybridisation with internal oligonucleotide probes. Next, a PCR screen for the E2 gene was performed to identify those samples in which the virus was integrated. Viral integration was detected in just over half of them. RESULTS: Twenty of the 92 samples were HPV-16 positive, as shown by PCR for the E6 gene. Southern blot analysis confirmed that 13 of these samples were positive for the viral E6 gene. Thus, viral integration was detected in just over a half of the samples positive for HPV-16. CONCLUSIONS: These data show that HPV-16 integration occurs in a subset of LSILs. The measurement of HPV-16 integration would be a helpful complementary tool for cytological evaluation in primary cervical screening to identify those patients at risk of developing high grade squamous intraepithelial lesions and cervical cancer.


Subject(s)
DNA, Viral/analysis , Papillomaviridae/genetics , Papillomavirus Infections/virology , Tumor Virus Infections/virology , Uterine Cervical Dysplasia/virology , Virus Integration , Adult , Carcinoma, Squamous Cell/virology , Female , Humans , Polymerase Chain Reaction/methods , Risk , Uterine Cervical Neoplasms/virology
2.
Acta Cytol ; 45(6): 995-8, 2001.
Article in English | MEDLINE | ID: mdl-11726131

ABSTRACT

OBJECTIVE: To develop a cytopreparation protocol suitable for satisfactory processing by the AutoCyte PREP System with the gynecologic specimens collected in PreservCyt fluid for the ThinPrep machine. STUDY DESIGN: The residue of a number of gynecologic specimens collected in PreservCyt and processed by ThinPrep were processed by AutoCyte PREP. Some modifications were made in the cytopreparatory protocol in order to obtain satisfactory specimens. RESULTS: The ThinPrep and AutoCyte PREP specimens were examined independently. The results were comparable, with a high degree of concordance between the two techniques. CONCLUSION: Gynecologic specimens collected in PreservCyt and following the ThinPrep specimen collection protocol can be processed using the AutoCyte PREP System. Minor protocol modifications provided comparable diagnostic material. Additional studies are needed to explore the feasibility of this approach and fulfill the various U.S. regulatory agency requirements for the liquid-based Pap test.


Subject(s)
Uterine Cervical Dysplasia/diagnosis , Uterine Cervical Neoplasms/diagnosis , Vaginal Smears/instrumentation , Cytological Techniques , Female , Hospitals, University , Humans , Laboratories, Hospital , Reproducibility of Results , Vaginal Smears/methods , Vaginitis/diagnosis , Vaginitis/microbiology
3.
Anal Quant Cytol Histol ; 23(5): 339-44, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11693559

ABSTRACT

OBJECTIVE: To explore the potential utility of immunostaining for CK20 and CD44 protein isoforms in evaluating cases of upper urinary tract transitional cell carcinoma (UTTCC). STUDY DESIGN: Of 105 consecutive patients diagnosed cytologically with UTTCC, 33 subsequently underwent open surgical procedures. Cytologic samples from these patients retrieved by aspiration and biopsy, and corresponding surgical specimens were graded and staged using World Health Organization/International Society of Urologic Pathologists criteria. Immunostaining for CK20, CD44 standard (CD44s) and CD44v6 isoform (CD44-v6) was performed on all available cytologic and surgical materials. Expression levels and distributions of these markers were correlated semiquantitatively with grade and stage. RESULTS: Cytologically assigned grade correlated with final histologic grade in 19 of 31 cases examined (61%). However, tumor invasion was not accurately assessable in cytologic samples from the majority of these cases. Statistically significant correlations of both increasing tumor grade and stage with abnormal CK20 expression were found. In addition, a significant relationship between focal CD44 isoform expression loss and tumor grade was identified. However, CD44 isoform expression loss did not significantly correlate with increasing tumor stage. CONCLUSION: Although cytologic tumor grading of UTTCC was accurate, invasion could not be adequately assessed. As an adjunct to morphologic analysis, immunostaining for CK20 and CD44 may aid in the clinical evaluation of UTTCC tumor stage and biologic behavior prior to definitive therapy.


Subject(s)
Carcinoma, Transitional Cell/diagnosis , Carcinoma, Transitional Cell/ultrastructure , Hyaluronan Receptors/metabolism , Intermediate Filament Proteins/metabolism , Neoplasm Staging/classification , Staining and Labeling/methods , Urologic Neoplasms/diagnosis , Urologic Neoplasms/ultrastructure , Humans , Hyaluronan Receptors/analysis , Intermediate Filament Proteins/analysis , Keratin-20 , Urothelium/pathology
4.
Cancer ; 93(4): 288-91, 2001 Aug 25.
Article in English | MEDLINE | ID: mdl-11507703

ABSTRACT

BACKGROUND: Diagnosing liver tumors by fine-needle aspiration biopsy is safe and accurate. However, there are cases that prove diagnostically difficult. Traditionally, immunostains for alpha-fetoprotein and polyclonal carcinoembryonic antigen have been used to distinguish adenocarcinomas from hepatocellular carcinomas (HCCs). In poorly differentiated tumors, these immunostains have limitations in both sensitivity and specificity. An hepatocyte-specific immunostain has been described in the surgical pathology literature. To the authors' knowledge, this hepatocyte antibody has not been studied in liver fine-needle aspiration biopsies. The authors examined the Hepatocyte Paraffin 1 (HP1) antibody for its diagnostic utility in this cytologic setting. METHODS: Cell-block material from 40 cases of HCC and 53 cases of metastatic adenocarcinoma were studied. Slides were stained for HP1 by the avidin-biotin complex method following antigen retrieval. The percentage of malignant cells that exhibited coarse granular staining in the cytoplasm was estimated for all cases of HCC, poorly differentiated HCC, and metastatic adenocarcinoma. RESULTS: HP1 was expressed in 83% of all HCCs but in only 56% of poorly differentiated HCCs. Only 2 of 53 (4%) of metastatic tumors expressed HP1. The overall sensitivity of HP1 was 79% and its specificity was 96%. CONCLUSION: HP1 was found to be a specific immunostain that may prove helpful in diagnosing all but the most undifferentiated liver tumors biopsied by fine-needle aspiration.


Subject(s)
Adenocarcinoma/pathology , Adenocarcinoma/secondary , Antibodies, Monoclonal , Carcinoma, Hepatocellular/pathology , Liver Neoplasms/pathology , Liver Neoplasms/secondary , Adenocarcinoma/immunology , Biopsy, Needle , Carcinoma, Hepatocellular/immunology , Diagnosis, Differential , Hepatocytes/immunology , Humans , Liver Neoplasms/immunology
7.
Diagn Cytopathol ; 23(3): 222-3, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10945916

ABSTRACT

Clear-cell features have been recognized in several different thyroid neoplasms. A case of thyroid follicular carcinoma with extensive clear- and Hurthle-cell features is described in a 37-yr-old white female, with cytochemical and immunohistochemical analysis. The tumor of the thyroid gland, with anterior neck soft-tissue extension, displayed clear cells on fine-needle aspiration, which were negative for thyroglobulin. The surgical specimen displayed predominately clear cells (80%), and only the nonclear-cell areas stained for thyroglobulin. Proper categorization of clear-cell lesions of the thyroid and soft tissues requires a multimodality approach, involving clinical/pathological correlation, morphological analysis, and ancillary tissue studies. Immunohistochemical stains for thyroglobulin are quite definitive in making the distinction between primary clear-cell thyroid tumors vs. metastatic clear-cell tumors. Cytologists should be aware, however, that the clear-cell areas of thyroid tumors might not stain for thyroglobulin.


Subject(s)
Adenocarcinoma, Follicular/diagnosis , Adenoma, Oxyphilic/diagnosis , Thyroid Neoplasms/diagnosis , Adenocarcinoma, Clear Cell/diagnosis , Adenocarcinoma, Clear Cell/secondary , Adenocarcinoma, Follicular/chemistry , Adenocarcinoma, Follicular/radiotherapy , Adenocarcinoma, Follicular/surgery , Adenoma, Oxyphilic/chemistry , Adenoma, Oxyphilic/radiotherapy , Adenoma, Oxyphilic/surgery , Adult , Diagnosis, Differential , Female , Humans , Immunohistochemistry , Thyroglobulin/analysis , Thyroid Neoplasms/chemistry , Thyroid Neoplasms/radiotherapy , Thyroid Neoplasms/surgery
8.
Acta Cytol ; 44(3): 301-4, 2000.
Article in English | MEDLINE | ID: mdl-10833982

ABSTRACT

OBJECTIVE: To investigate cyclin E expression as a possible marker for early cervical neoplasia using ThinPrep gynecologic specimens from premenopausal women. STUDY DESIGN: Archived ThinPrep liquid-based cervical/endocervical specimens (Cytyc Corporation, Boxborough, Massachusetts, U.S.A.) diagnosed as human papillomavirus infection (HPV) (20), atypical squamous cells of undetermined significance (ASCUS) (48) and within normal limits (WNL)/benign cellular changes (BCC) (21) were resampled in duplicate, fixed in 95% ethanol, subjected to immunocytochemical staining with the cyclin E antibody (clone 13A3, Novocastra Laboratories Ltd., Newcastle upon Tyne, U.K.) and HPV antibody (clone K1H8, Dako Corporation, Carpinteria, California, U.S.A.) and the expression scored by two pathologists and correlated with the cytologic diagnosis. A case was scored as positive if it contained > 10 abnormal squamous cells with nuclear immunocytochemical staining. RESULTS: The cylin E antibody assay was positive in 20 (100%) cases cytologically diagnosed as HPV. These cases were also anti-HPV antibody positive. Four cases (19%) cytologically diagnosed as WNL/BCC were cyclin E positive. Of these, two were anti-HPV antibody positive. Thirty-four (73%) cases cytologically diagnosed as ASCUS were positive for the cyclin E assay and for anti-HPV antibody staining. CONCLUSION: Cyclin E expression correlates strongly with morphologic features of HPV in ThinPrep specimens and may serve as a surrogate marker for HPV infection and early cervical preneoplastic lesions.


Subject(s)
Cyclin E/biosynthesis , Neoplasms, Squamous Cell/immunology , Papillomaviridae/immunology , Papillomavirus Infections/immunology , Tumor Virus Infections/immunology , Uterine Cervical Dysplasia/immunology , Uterine Cervical Neoplasms/immunology , Adult , Antibodies, Viral/analysis , Biomarkers , Cyclin E/immunology , Feasibility Studies , Female , Humans , Neoplasms, Squamous Cell/pathology , Papillomavirus Infections/pathology , Time Factors , Tumor Virus Infections/pathology , Uterine Cervical Neoplasms/pathology , Vaginal Smears , Uterine Cervical Dysplasia/pathology
9.
Diagn Cytopathol ; 22(6): 398-9, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10820537

ABSTRACT

Clear-cell features have been recognized in several different thyroid neoplasms. A case of thyroid follicular carcinoma with extensive clear- and Hurthle-cell features is described in a 37-yr-old white female, with cytochemical and immunohistochemical analysis. The tumor of the thyroid gland, with anterior neck soft-tissue extension, displayed clear cells on fine-needle aspiration, which were negative for thyroglobulin. The surgical specimen displayed predominately clear cells (80%), and only the nonclear-cell areas stained for thyroblobulin. Proper categorization of clear-cell lesions of the thyroid and soft tissues requires a multimodality approach, involving clinical/pathological correlation, morphological analysis, and ancillary tissue studies. Immunohistochemical stains for thyroglobulin are quite definitive in making the distinction between primary clear-cell thyroid tumors vs. metastatic clear-cell tumors. Cytologists should be aware, however, that the clear-cell areas of thyroid tumors might not stain for thyroglobulin.


Subject(s)
Adenocarcinoma, Follicular/diagnosis , Adenoma, Oxyphilic/diagnosis , Thyroid Neoplasms/diagnosis , Adenocarcinoma, Clear Cell/diagnosis , Adenocarcinoma, Clear Cell/secondary , Adenocarcinoma, Follicular/chemistry , Adenocarcinoma, Follicular/radiotherapy , Adenocarcinoma, Follicular/surgery , Adenoma, Oxyphilic/chemistry , Adenoma, Oxyphilic/radiotherapy , Adenoma, Oxyphilic/surgery , Adult , Diagnosis, Differential , Female , Humans , Immunohistochemistry , Thyroglobulin/analysis , Thyroid Neoplasms/chemistry , Thyroid Neoplasms/radiotherapy , Thyroid Neoplasms/surgery
10.
Acta Cytol ; 44(1): 31-6, 2000.
Article in English | MEDLINE | ID: mdl-10667156

ABSTRACT

OBJECTIVE: To determine the utility of immunohistochemical staining for p53 in cell block material for distinguishing reactive mesothelium from borderline or low grade ovarian carcinoma. STUDY DESIGN: Paraffin-embedded cell blocks from paracentesis and pelvic wash fluid of 44 cases of ovarian carcinoma and 20 cases containing only reactive mesothelium were immunostained for p53 using monoclonal antibody DO-7. Tumor grades ranged from borderline to high grade and were serous papillary (33), clear cell (3), mucinous (2), endometrioid (2), mixed serous papillary/clear cell (3) and undifferentiated (1). The three authors independently evaluated the staining, including estimation of the percentage and intensity of positive nuclear staining. RESULTS: A separation of positive from negative cases was seen when staining intensity was considered the critical parameter; moderate to strong staining was considered truly positive. Seventy-three percent (8/11) of borderline tumors, 80% (8/10) of low grade tumors and 65% (15/23) of intermediate to high grade tumors showed moderate to strong positivity. Percentage of staining was a less-reliable parameter as 25% of negative cases were positive by this assessment. CONCLUSION: p53 Immunohistochemistry, using monoclonal antibody DO-7 combined with standard morphologic evaluation, may be useful in distinguishing benign reactive mesothelium from borderline or low grade ovarian carcinoma.


Subject(s)
Ascitic Fluid/chemistry , Carcinoma/chemistry , Epithelial Cells/chemistry , Ovarian Neoplasms/chemistry , Tumor Suppressor Protein p53/analysis , Antibodies, Monoclonal , Carcinoma/pathology , Cell Nucleus/metabolism , Cell Nucleus/pathology , Cytodiagnosis/methods , Female , Humans , Immunoenzyme Techniques , Ovarian Neoplasms/pathology , Sensitivity and Specificity
11.
Acta Cytol ; 43(6): 1079-84, 1999.
Article in English | MEDLINE | ID: mdl-10578981

ABSTRACT

OBJECTIVE: Detecting malignant cells in the setting of reactive mesothelium can be difficult. Several techniques have been tried but without widespread acceptance. Sialosyl-Tn (STn) is an aberrantly glycosylated precursor of the MN blood group antigen frequently expressed in carcinomas and dysplastic epithelium. We investigated the STn monoclonal antibody for its clinical utility as an isolated stain to discriminate benign mesothelium from malignant cells. STUDY DESIGN: Cell block material from 72 cases of body cavity fluids were immunostained for STn using the avidin-biotin complex method without antigen retrieval. Slides were incubated overnight at 4 degrees C in a humidified chamber. RESULTS: Strong immunoreactivity was noted in 31/40 (77%) carcinomatous cases. Only moderate staining was noted in 1 of 28 (4%) benign effusions and weak staining in 5 (18%) additional benign cases. Specificity was 100%, sensitivity 78%, positive predictive value 100% and negative predictive value 76%. No staining was noted in four noncarcinomatous malignant effusions. CONCLUSION: STn may have diagnostic value in this cytologic setting as part of a diagnostic panel but not as an isolated stain.


Subject(s)
Antigens, Tumor-Associated, Carbohydrate , Ascitic Fluid/pathology , Biomarkers, Tumor , Carcinoma/immunology , Epithelial Cells , Pleural Effusion, Malignant/pathology , Diagnosis, Differential , Female , Humans , Immunohistochemistry
12.
Anal Quant Cytol Histol ; 21(2): 121-5, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10560479

ABSTRACT

OBJECTIVE: To determine whether various methods of fixation of surgical pathology specimens from breast carcinomas would influence the outcome of evaluation of the expression levels of c-erbB-2 (HER-2). For this, comparisons were made between (1) alcohol-fixed (95%) and air-dried smears from fresh surgical pathology specimens of breast carcinomas, and (2) formalin-fixed, paraffin-embedded tissue sections of the same specimens. STUDY DESIGN: Alcohol-fixed and air-dried smears or touch preparations were made from 30 fresh mastectomy/lumpectomy surgical pathology specimens from breast carcinomas. Immunohistochemistry was performed using the c-erbB-2 primary antibody against the extracellular domain of the c-erbB-2 gene product. Staining was simultaneously performed on formalin-fixed, paraffin-embedded tissue sections of the same specimens. A semiquantitative approach was used for evaluation of immunostaining by three independent investigators, and a consensus was reached. RESULTS: A total of 30 cases were reviewed. Tissue positivity was determined for c-erbB-2 in: 73% of alcohol-fixed specimens (n = 13 [3+] and n = 9 [2+]), 67% of air-dried smears (n = 9 [+3] and n = 11 [+2]) and 47% (n = 8 [+3]) and n = 6 [+2]) of formalin-fixed, paraffin-embedded tissue specimens. All formalin-fixed tissue specimens that were determined to positively express c-erbB-2 were also found to be positive on the alcohol-fixed smears. CONCLUSION: The incidence of c-erbB-2 expression in fresh cytologic material is significantly higher (P < .05) than in formalin-fixed, paraffin-embedded tissue. Alcohol-fixed smears demonstrate a slightly higher percentage of cell staining and stronger intensity of c-erbB-2 expression than the matched, air-dried smears. This is a sensitive and simple processing method that can be routinely applied in surgical pathology or fine needle aspiration biopsy specimens for the detection of c-erbB-2 (HER-2), with clinical implications.


Subject(s)
Adenocarcinoma/metabolism , Breast Neoplasms/metabolism , Receptor, ErbB-2/metabolism , Tissue Fixation/methods , Adenocarcinoma/pathology , Air , Biopsy, Needle , Breast Neoplasms/pathology , Ethanol , Female , Fixatives , Formaldehyde , Humans , Image Cytometry , Immunoenzyme Techniques
13.
Acta Cytol ; 43(4): 544-51, 1999.
Article in English | MEDLINE | ID: mdl-10432873

ABSTRACT

OBJECTIVE: To assess whether a light microscopic, semiquantitative approach could reliably distinguish between benign nonneoplastic, benign neoplastic and malignant oncocytic lesions of the breast and thyroid. STUDY DESIGN: Alcohol-fixed, Papanicolaou-stained fine needle aspiration smears of histologically proven goiter and chronic thyroiditis (18 cases), Hürthle cell adenomas (7 cases), Hürthle cell carcinomas (6 cases), fibrocystic disease (17 cases), papillomas and papillomatosis (7 cases) and apocrine carcinomas (6 cases) were rated by three independent observers using the following 10 cytologic criteria: cellularity, nuclear-cytoplasmic ratio, multinucleation, nuclear size, nuclear shape, anisonucleosis, multinucleolation, nucleolar-nuclear ratio, nucleolar size and nucleolar shape. Each of these 10 cytologic criteria was rated using a 1-3 scale. The scores for all 10 features were summed to give a total score for each case. The total scores were statistically analyzed to determine the validity and reproducibility of the summed criteria. RESULTS: The summed criteria of the total scores were reproducible between the three observers, with standard deviations ranging from 1.36 to 2.88 for thyroid and 1.72 to 2.00 for breast oncocytic lesions. The ability of the total scores to differentiate benign from malignant oncocytic lesions of the breast and thyroid was confirmed by a positive predictive value for malignancy of 67% for thyroid and 72% for breast and a negative predictive value for malignancy of 100% for nonneoplastic oncocytic lesions and > 90% for benign oncocytic neoplasms in both. The Kruskal-Wallis test revealed that the total scores were able to distinguish three diagnostic categories of nonneoplastic, benign neoplastic and malignant oncocytic breast and thyroid lesions, with P < .005. CONCLUSION: Without the expenditures of additional time, costs or materials, this semiquantitative approach compared favorably with contemporary morphometric studies involving the differential diagnosis of oncocytic cell pathology in fine needle aspiration cytology.


Subject(s)
Breast Neoplasms/pathology , Thyroid Neoplasms/pathology , Cytodiagnosis , Diagnosis, Differential , Double-Blind Method , Female , Fibrocystic Breast Disease/pathology , Humans , Microscopy/methods , Observer Variation
14.
Diagn Cytopathol ; 21(1): 27-9, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10405804

ABSTRACT

Our study evaluated the BTA (bladder tumor antigen) stat test kit as a primary screening device for the detection of transitional-cell carcinoma (TCC) of the bladder, with direct comparison by voided urine cytology (VUC) on the same specimens. The unfixed voided urine of 100 patients with no history of bladder cancer who had signs and symptoms of dysuria, incontinence, and gross hematuria and microhematuria were tested using the one-step BTA stat test kit before processing via the cytospin technique for fluid cytological evaluation. The patients in the study were followed for up to 12 mo with repeated urine cytological testing, cystoscopy, and bladder biopsy when clinically indicated. Nineteen cases tested positive, and 81 cases tested negative on the BTA stat test. VUC diagnosed three cases as unequivocally positive for TCC, 93 cases as negative, and four cases in which unqualified atypical urothelial cells were noted. TCC was confirmed by cystoscopy and bladder biopsy in three of three cases diagnosed by VUC and in three of 19 cases that tested positive by the BTA stat test. These findings resulted in an 84% false-positive rate for the BTA stat test and no false-positive cases for VUC during the 12-mo follow-up period. The results indicate that the sensitivity and specificity of BTA stat test are comparable to those of VUC; however, owing to a relatively high false-positive rate, it can at best act as an adjunct to urine cytological study for bladder cancer screening.


Subject(s)
Antigens, Neoplasm/urine , Carcinoma, Transitional Cell/diagnosis , Complement Factor H , Reagent Kits, Diagnostic , Urinary Bladder Neoplasms/diagnosis , Biomarkers, Tumor , Blood Proteins/urine , Carcinoma, Transitional Cell/immunology , Carcinoma, Transitional Cell/pathology , Carcinoma, Transitional Cell/urine , Evaluation Studies as Topic , Follow-Up Studies , Humans , Mass Screening , Neoplasm Invasiveness , Urinary Bladder Neoplasms/immunology , Urinary Bladder Neoplasms/pathology , Urinary Bladder Neoplasms/urine
15.
Oncol Rep ; 6(3): 695-8, 1999.
Article in English | MEDLINE | ID: mdl-10203617

ABSTRACT

Fluorescent in situ hybridization has shown promise in detecting malignant cells in body cavity effusions. However, this requires special preparatory techniques not used in many laboratories. We developed an in situ hybridization (ISH) procedure specifically for ethanol-fixed specimens, and using it tested the clinical utility of the chromosome 17alpha satellite probe (C17alpha). ISH with C17alpha was used in 12 malignant and 10 benign ethanol-fixed body cavity effusions. Cells were pretreated with protease K prior to ISH. The probe was detected by an anti-digoxigenin-horseradish peroxidase method. Signals were counted in 100 nuclei and the chromosome index (CI) and percent diploid cells calculated in each case. ISH was successfully performed in all cases. Malignant cells had an average CI of 2.23 with less than 44% diploid nuclei and 50% of specimens exhibited bizarre signals. Benign effusions had an average CI of 1.98 with over 84.6% diploid nuclei. Questionably bizarre signals were seen in two (20%) benign specimens. ISH can be performed on ethanol-fixed specimens. The C17alpha probe may prove valuable in detecting malignant cells in body cavity effusions.


Subject(s)
Chromosomes, Human, Pair 17 , DNA Probes , DNA, Satellite , Mesothelioma/genetics , Genes, erbB-2 , Genes, p53 , Humans , In Situ Hybridization , Mesothelioma/diagnosis , Mesothelioma/pathology , Pilot Projects
16.
Pathol Res Pract ; 195(2): 67-70, 1999.
Article in English | MEDLINE | ID: mdl-10093823

ABSTRACT

The retinoblastoma gene family is composed of three members: the retinoblastoma gene, one of the most studied tumor suppressor genes, and two related genes: p107 and pRb2/p130. These proteins are also known as the pocket proteins due to a unique structural and functional domain composed of subdomains A and B separated by a spacer region that is highly conserved among each of the proteins. These proteins exhibit unique growth suppressive properties that are cell type specific, suggesting that although the pocket proteins may complement each other, they are not fully functionally redundant. With the development of antibodies recognizing these three proteins it is now possible to detect expression in formalin-embedded specimens. Recent studies on 235 lung cancers, using immunohistochemical techniques, suggested an independent role for Rb2/p130 in the development and/or progression of human lung carcinoma. We found a statistically significant inverse relationship between the histological grading (degree of malignant potential) and the expression of pRb/p105, p107 and pRb2/p130 in squamous cell carcinomas, meaning that an increase in grading resulted in a significant decrease in protein expression. This phenomenon was particularly evident for pRb2/p130 (p < .0001) which had the highest percentage of undetectable levels in all the specimens examined and the tightest inverse correlation (p value) with both the histological grading and PCNA expression in the most aggressive tumor types, suggesting an important role for pRb2/p130 in the pathogenesis and progression of certain lung cancers. We further explored the expression of pRb2/p130 protein in routine archival FNAB cytological material from 30 Patients with lung cancer using immunocytochemical techniques, comparing protein expression with tumor type. Two pathologists evaluated the staining pattern and scored the percentage of positive cells. Of the 30 neoplasms, 27 displayed a positive staining for pRb2/p130. In particular, we detected pRb2/p130 in 9 (100%) squamous carcinomas, 11 (84%) adenocarcinomas, 5 (100%) BAC, and 2 (66%) SCC. The percentage of positive nuclei varied in different tumors with the highest expression level in adenocarcinomas. Immunocytochemistry represents a sensitive method for detection of pRb2/p130 expression in cytological or archival specimens, and the level of detection seems to be comparable to paraffin sections. Therefore, this methodology could be used in the preoperative evaluation of routine cytological specimens in order to improve the diagnostic and prognostic evaluation of lung cancer patients.


Subject(s)
Adenocarcinoma, Bronchiolo-Alveolar/metabolism , Carcinoma, Small Cell/metabolism , Carcinoma, Squamous Cell/metabolism , Lung Neoplasms/pathology , Phosphoproteins/metabolism , Proteins , Retinoblastoma Protein/metabolism , Adenocarcinoma, Bronchiolo-Alveolar/chemistry , Adenocarcinoma, Bronchiolo-Alveolar/diagnosis , Biopsy, Needle , Carcinoma, Small Cell/chemistry , Carcinoma, Small Cell/diagnosis , Carcinoma, Squamous Cell/chemistry , Carcinoma, Squamous Cell/diagnosis , Humans , Immunoenzyme Techniques , Lung Neoplasms/chemistry , Lung Neoplasms/metabolism , Phosphoproteins/analysis , Retinoblastoma Protein/analysis , Retinoblastoma-Like Protein p130
17.
Acta Cytol ; 43(1): 23-6, 1999.
Article in English | MEDLINE | ID: mdl-9987445

ABSTRACT

OBJECTIVE: To correlate the ranked review slides by the AutoPap Assisted Primary Screener with the final cytologic diagnosis and to assess whether the ranking of slides improves diagnostic accuracy. STUDY DESIGN: A total of 5,865 consecutive conventional and suitable cervical/vaginal smears, including high-risk (HR) cases, were processed by the AutoPap System. All slides designated Review were manually screened by two cytotechnologists using the Ranked Review Report. All abnormal findings and reactive/reparative changes were referred to two attending cytopathologists for a final diagnosis. After screening, the Review slides called Within Normal Limits (WNL) were selected according to rank for a further Quality Control (QC) review by the supervisor. All HR cases not selected by AutoPap for QC review were also rescreened. The final diagnoses of the possibly abnormal/reactive/reparative referred slides were recorded and distributed in five ranks according to the Ranked Review Report assignment. RESULTS: Of 5,865 slides, 5,120 (87%) qualified for scanning. The AutoPap System designated 3,840 (75%) slides for manual review. One thousand three hundred forty-five slides were assigned for QC review. After elimination of nonqualified slides, 763 remained. Rescreening detected 1 high grade squamous intraepithelial lesion (HSIL), 2 low grade squamous intraepithelial lesions (LSIL), 5 atypical squamous cells of undetermined significance (ASCUS) and 5 Benign Reactive Changes (BRC). QC of 364 HR cases revealed 1 LSIL, 2 ASCUS and 3 BRC. Ultimately, 313 (8.1%) smears were diagnosed as ASCUS or a more severe abnormality, 262 (6.8%) as reactive/reparative changes, 3,259 (84.8%) as WNL and 6 (0.1%) as unsatisfactory. Of 313 abnormal slides, 181 were ranked by the system in the 1st rank, 61 in the 2nd, 38 in the 3rd, 19 in the 4th and 14 in the 5th. No HSIL or more severe lesions occurred in the fourth and fifth ranks. CONCLUSION: Our study validated the claim by the manufacturer that a significant epithelial abnormality is more likely to be present if a high score is assigned to a slide. The preliminary results support use of the AutoPap Assisted Primary Screener to improve cytologic diagnosis.


Subject(s)
Mass Screening/standards , Pathology, Clinical/instrumentation , Pathology, Clinical/standards , Vaginal Smears/instrumentation , Vaginal Smears/standards , Female , Humans , Image Processing, Computer-Assisted/instrumentation , Image Processing, Computer-Assisted/standards , Laboratories, Hospital/standards , Mass Screening/instrumentation , Quality Control , Uterine Cervical Neoplasms/diagnosis , Uterine Cervical Neoplasms/pathology
18.
Acta Cytol ; 43(1): 27-9, 1999.
Article in English | MEDLINE | ID: mdl-9987446

ABSTRACT

OBJECTIVE: To evaluate the performance of the AutoPap System for primary screening of cervical/vaginal cytologic smears at the 25% no review rate at Jefferson University Hospital. STUDY DESIGN: A total of 5,865 consecutive conventional and suitable cervical/vaginal smears, including high-risk cases, were processed by the AutoPap System. Slides designated "no further review" (NFR) were manually screened by a cytopathologist (rapid screening) and a cytotechnologist (detailed screening) to detect epithelial abnormalities and assess the slide adequacy. The presence of Candida or Trichomonas was noted. Three cytopathologists determined the final diagnosis of all epithelial abnormalities by majority agreement. RESULTS: Of 5,865 slides, 5,120 (87%) qualified for scanning. The system classified 1,280 slides as NFR (25%) and 3,840 (75%) as review. Manual screening of 1,280 slides classified as NFR revealed 1,252 cases within normal limits, 10 atypical squamous cells of undetermined significance (4, favor low grade squamous intraepithelial lesion, 6 favor reactive) and 18 benign cellular changes. Adequacy assessment discrepancies were detected on 98 (19%) of 515 cases of satisfactory but limited by lack of endocervical component (SBLBLEC) and 15 (14%) of 105 cases on satisfactory but limited by inflammation/obscuration. Twenty-five percent of cases classified as SBLBLEC were vaginal smears. Trichomonas vaginalis was noted in 12 slides and Candida in 29. CONCLUSION: These preliminary results show that in our patient population, the AutoPap primary screener selected as NFR mostly slides within normal limits. The adequacy assessment discrepancies were comparable to those found in routine laboratory practice.


Subject(s)
Hospitals, University/standards , Laboratories, Hospital/standards , Mass Screening/standards , Vaginal Smears/standards , Candidiasis/diagnosis , Candidiasis/pathology , Epithelial Cells/pathology , Female , Humans , Image Processing, Computer-Assisted/instrumentation , Image Processing, Computer-Assisted/standards , Mass Screening/instrumentation , Trichomonas Vaginitis/diagnosis , Trichomonas Vaginitis/pathology , Uterine Cervical Neoplasms/diagnosis , Uterine Cervical Neoplasms/pathology , Vaginal Smears/instrumentation
19.
Diagn Cytopathol ; 20(2): 95-8, 1999 Feb.
Article in English | MEDLINE | ID: mdl-9951606

ABSTRACT

Recent work has shown CytoRich Red fixative system is effective in lysing red blood cells and reducing background in bloody fluid specimens. The scope of this study was to see whether CytoRich Red can lyse red blood cells in freshly prepared Pap and fine-needle aspiration smears. Paired smears from 20 bloody fine-needle aspirations were prepared. One slide was initially placed in CytoRich Red for up to 30 sec, removed, and then fixed in 95% alcohol. The other slide was placed directly into 95% alcohol. Ten paired Pap smears, one fixed with a commercial fixative and another immersed in a solution of CytoRich Red, were evaluated. All slides were stained with the Papanicolaou stain and analyzed for the amount of red blood cells, background material, and nuclear and cytoplasmic staining. In 100% of all smears utilizing CytoRich Red, red blood cells were significantly reduced without hindering staining. Significant loss of cells from the slides sent in CytoRich Red solution was not observed. CytoRich Red fixative can be effective in reducing red blood cells and background on freshly made smears. In both gynecological and nongynecological cases, diagnostic cells were well preserved and not compromised by blood.


Subject(s)
Blood Cells/cytology , Hemolysis , Papanicolaou Test , Reagent Kits, Diagnostic , Tissue Fixation/methods , Vaginal Smears/methods , Biopsy, Needle , Female , Humans , Specimen Handling
20.
Ear Nose Throat J ; 78(12): 905, 908-12, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10624054

ABSTRACT

Although fine-needle aspiration biopsy of salivary gland masses has been reported in the otolaryngology literature, the use of sonography to guide the biopsy of nonpalpable masses and masses seen on other cross-sectional imaging studies has not been described. Our goal was to evaluate sonographically guided biopsy of masses and lymph nodes related to the salivary glands. We analyzed the records of 18 patients who had undergone fine-needle aspiration biopsy of a salivary gland mass or lymph node with a 25-, 22-, or 20-gauge needle. A definitive cytologic diagnosis was made for 13 of the 18 patients (72%); cytology was suggestive but not definitive in three patients (17%) and insufficient in two (11%). Definitive diagnoses were made in three cases of reactive lymph node, in two cases each of lymph node metastasis and Warthin's tumor, and in one case each of pleomorphic adenoma, adenoid-cystic carcinoma, schwannoma-neurofibroma, parotid metastasis, parotid lymphoma, and Sjögren's-related lymphoid-epithelial lesion. Sonographically guided biopsy allows for confident needle placement in masses seen on computed tomography and magnetic resonance imaging. Sonography can usually distinguish a perisalivary lymph node from true intrasalivary masses, and it can help the surgeon avoid the pitfall of a nondiagnostic aspiration of the cystic component of masses. We conclude that sonographically guided biopsy of salivary gland masses can provide a tissue diagnosis that can have a direct impact on clinical decision making.


Subject(s)
Biopsy, Needle/methods , Lymphatic Metastasis/diagnostic imaging , Lymphatic Metastasis/pathology , Salivary Gland Neoplasms/diagnostic imaging , Salivary Gland Neoplasms/pathology , Adenoma, Pleomorphic/diagnostic imaging , Adenoma, Pleomorphic/pathology , Aged , Aged, 80 and over , Female , Humans , Lymph Nodes/diagnostic imaging , Lymph Nodes/pathology , Male , Middle Aged , Ultrasonography
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