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The best animal angioplasty model is the porcine model, which is expensive and not available in all laboratories. The aim of this study was to describe a new rat model of angioplasty. An injury was induced with the use of a standard percutaneous transluminal coronary angioplasty (PTCA) 1.5-mm balloon catheter. The neointimal tissue, arterial dimensions, and the injury index were assessed following angioplasty. Ki-67 expression was detected to evaluate cell turnover after balloon angioplasty. In contrast with the standard Clowes model, a significant neointimal formation was detected only in the presence of ruptured internal elastic lamina (IEL). A positive correlation between the percentage of ruptured IEL and the amount of neointimal tissue was also demonstrated. The percentage of IEL fracture correlates with the proliferation index by anti-Ki-67 immunolabeling 7 and 14 days after the angioplasty. Significant arterial negative remodeling was observed following PTCA balloon dilation. In conclusion, our inexpensive animal model of restenosis after angioplasty may have great relevance toward a better understanding of the mechanisms and toward assessment of new therapeutical strategies for this phenomenon.

Objective: To assess the possible contribution by a multiparametric quantitative approach to the cytologic diagnosis of oxyphilic cell (OC) thyroid lesions., Study Design: Ten cases of chronic lymphocytic (Hashimoto) thyroiditis and 10 nodular goiters containing oxyphilic cells plus 20 cases of tumors subsequently classified as oxyphilic cell adenomas (10 cases) or oxyphilic cell well-differentiated carcinomas (10 cases) were evaluated. The study was performed on May-Grünwald-Giemsa-stained smears for planimetric measurements. The same smears were destained and Feulgen restained for densitometric measurements. The latter were performed using static cytometry equipment measuring 100 and 20-30 lymphocytes per case for the determination of integrated optical density (IOD). The following parameters were considered: nuclear area, perimeter, maximum diameter, form ELL, form PE, IOD, 5c exceeding rate (5cER) and visual classification of histograms as euploid, polyploid and aneuploid., Results: Mean nuclear area of carcinomas was smaller than that of adenomas, goiter and thyroiditis. Nuclear area was larger in adenomas than in other benign lesions and carcinomas. All the other planimetric parameters were similar in the lesions examined. Four carcinomas and three adenomas were aneuploid, and all the rest were euploid. All the cases of thyroiditis and goiter were euploid or polyploid; four thyroiditis cases showed polyploid histograms and 5cER values > 1., Conclusion: Morphometric and densitometric procedures have a limited role in the discrimination of OC lesions, but small nuclear area values may be useful in distinguishing OC carcinoma from other lesions. The role of densitometry seems even more limited because aneuploid histograms may be found among adenomas and carcinomas. Further studies are needed to explain polyploidy and 5cER > 1 in Hashimoto thyroiditis.

cAMP-dependent protein kinase is anchored to discrete cellular compartments by a family of proteins, the A-kinase anchor proteins (AKAPs). We have investigated in vivo and in vitro the biological effects of the expression of a prototypic member of the family, AKAP75, on smooth muscle cells. In vitro expression of AKAP75 in smooth muscle cells stimulated cAMP-induced transcription, increased the levels of the cyclin-dependent kinase-2 inhibitor p27(kip1), and reduced cell proliferation. In vivo expression of exogenous AKAP75 in common carotid arteries, subjected to balloon injury, significantly increased the levels of p27(kip1) and inhibited neointimal hyperplasia. Both the effects in smooth muscle cells in vitro and in carotid arteries in vivo were specifically dependent on the amplification of cAMP-dependent protein kinase (PKA) signals by membrane-bound PKA, as indicated by selective loss of the AKAP75 biological effects in mutants defective in the PKA anchor domain or by suppression of AKAP effects by the PKA-specific protein kinase inhibitor. These data indicate that AKAP proteins selectively amplify cAMP-PKA signaling in vitro and in vivo and suggest a possible target for the inhibition of the neointimal hyperplasia after vascular injury.

Recent studies on paraffin-embedded tissue have shown that the cyclin-dependent kinase inhibitor p27(Kip1) is expressed in normal thyroid cells, whereas it is downregulated in neoplastic cells. This prospective study was undertaken to assess whether p27(Kip1) staining may also be applied to fine-needle aspiration biopsy (FNAB) samples of the thyroid. We present here our preliminary results on 100 FNABs examined for p27(Kip1) expression. p27(Kip1) expression was assessed by immunocytochemistry; the technique was optimized on smears prepared from a normal thyreocyte cell line (TL5), which conspicuously expresses p27(Kip1), and then applied to FNAB samples prospectively collected from 80 cases of nodular goiter and 20 cases of thyroid neoplasms (10 papillary carcinomas and 10 follicular neoplasms). The TL5 cell culture smears showed that methanol fixation, followed by heat-induced antigen retrieval, is the most suitable technique for p27(Kip1) staining on cytological samples. The FNAB smears similarly treated showed high p27(Kip1) expression (75%) in goiter and a significantly lower expression (35%) in neoplasms (P < 0.0001). Our preliminary results show that: 1) p27(Kip1) protein expression can be reliably assessed on cytological samples; and 2) p27(Kip1) stains nonneoplastic and neoplastic samples in a different fashion, and thus is a useful tool in thyroid cytology., (Copyright 2000 Wiley-Liss, Inc.)

cAMP signals are received and transmitted by multiple isoforms of cAMP-dependent protein kinases (PKAs), typically determined by their specific regulatory subunits. We describe changes in the cAMP signal transduction pathway during cell cycle progression in synchronized rat thyroid cells. Both PKA type II (PKAII) localization and nuclear cAMP signaling are significantly modified during G(0) and G(1)-S transitions. G(1) is characterized by PKA activation and amplified cAMP signal transduction. This is associated with a decrease in the concentration of RI and RII regulatory subunits and enhanced anchoring of PKAII to the Golgi-centrosome region. Just prior to S, the cAMP pathway is depressed. Up-regulation of the pathway by exogenous cAMP in G(1) inhibited the subsequent decay of the Cdk inhibitor p27 and delayed the onset of S phase. Forced translocation of endogenous PKAII to the cytosol down-regulated cAMP signaling, advancing the timing of p27 decay and inducing premature exit from G(1). These data indicate that membrane-bound PKA amplifies the transduction of cAMP signals in G(1) and that the length of G(1) is influenced by cAMP-PKA.

Aim: To investigate whether there is loss of the p27Kip1 protein in developing cervical cancer and whether p27Kip1 immunoreactivity has any relation to the proliferative indicator Ki-67., Methods: The expression of p27Kip1 and Ki-67 was assessed by immunohistochemistry in serial sections from normal epithelium (13), low grade (27) and high grade (19) squamous intraepithelial lesions (LSIL, HSIL), and invasive cervical cancer (23). In the SIL cases the presence of human papillomavirus (HPV) genomic sequences was assessed by in situ hybridisation. The results were evaluated by image analysis, and reported as mean score of the percentage of p27Kip1 and of Ki-67 positive cells in each histological group., Results: In general, p27Kip1 immunostaining was related to squamous differentation, and was intense in normal epithelium (47%), while it was reduced in SIL lesions as an effect of the decreased number of differentiating cells. However, decrease in the p27Kip1 expression was more evident in LSIL (36%) than in HSIL (39%); in the latter, p27Kip1 had a different intraepithelial distribution in that the staining extended to the basal cells. The average levels of p27Kip1 were similar in SIL lesions associated to low, intermediate, and high risk HPV types. Compared with normal epithelium and dysplasia, invasive cancer showed significantly lower p27Kip1 levels (23%). There was no relation between p27Kip1 and Ki-67 labelling indices in any of the histological groups examined., Conclusions: A reduction in p27Kip1 protein occurs in cervical cancer independently of the proliferative status. The changes in p27Kip1 expression may be related to the unregulated kinetics of developing cervical cancer.

DNA ploidy was evaluated by image cytometry in a series of 84 hepatocellular carcinomas diagnosed by fine-needle aspiration biopsy. In the series were included eight cases originally diagnosed as suspect and reclassified as well-differentiated hepatocarcinoma. The study was retrospectively performed on Papanicolaou-destained, Feulgen-restained smears. The 5c exceeding rate and the visual interpretation of the corresponding histograms were evaluated and compared with size of the tumors, serum alpha-fetoprotein values, hepatic functional staging, and patient survival. Sixty-eight cases were aneuploid and 16 euploid (9 diploid and 7 polyploid). Four of the eight cytologically suspect cases were aneuploid. Statistical analysis showed an association between size and cytologic grading, 5c exceeding rate and cytologic grading, and between aneuploidy and multiple tumors; in a Cox multivariate DNA content analysis, aneuploidy and multiple tumors were the two prognostically significant variables. DNA ploidy evaluation by static cytometry of hepatic tumors may be useful in the diagnosis on cytologic samples and could represent an independent prognostic parameter in predicting the survival outcome of patients with hepatocellular carcinoma.

Aim: To investigate the immunocytochemical staining pattern of mdm2 and p21WAF1 proteins in invasive cervical cancer and to determine its relation with the expression of p53 and with the high risk HPV infection., Methods: Immunocytochemistry for p53, mdm2, and p21WAF1 was performed in 31 paraffin embedded sections of invasive cervical cancer. The results were assessed by image analysis, evaluating for each protein the optical density of the immunostained area, scored as percentage of the total nuclear area. The presence of high risk human papillomavirus (HPV) infection was detected by using the polymerase chain reaction., Results: Immunostaining for both mdm2 and p21WAF1 was correlated with p53 expression; however, the correlation between p53 and mdm2 (R = 0.49; p < 0.01) was more significant than between p53 and p21WAF1 (R = 0.31; p < 0.05); the less stringent correlation between p53 and p21WAF1 might reflect the p53 independent mechanisms of p21WAF1 induction. Similar average levels of p53, mdm2, and p21WAF1 immunostaining were found in the presence or absence of high risk HPV-DNA, without significant differences between the two groups., Conclusions: These data suggest that mdm2 and p21WAF1 proteins are expressed in invasive cervical cancer and that their immunocytochemical staining pattern is not abrogated by the presence of high risk HPV genomic sequences.

FNA biopsy of 51 orbital masses is critically reviewed. Aspiration was performed with a 23 G needle inserted by an ophthalmologist; the smears were prepared by a cytologist. Forty-two cases (83%) were correctly diagnosed as benign or malignant either with (68%) or without (15%) correct specification of the histology. There were two false-negative and seven inadequate cases. Immunocytochemical stains were performed in five cases using the following antibodies: L26 (Pan B), UCHL1 (Pan T), kappa and lambda immunoglobulin light chains (three cases) in order to distinguish inflammatory pseudotumours from low-grade non-Hodgkin's lymphomas. In two cases we used CAM 5.2 (a monoclonal cytokeratin cocktail) and vimentin to ascertain the epithelial origin of two metastatic tumours. In five other cases cytospins were not adequately cellular for immunocytochemistry. Insufficient material and one false-negative sample were obtained from very fibrotic lesions or from posteriorly located lesions. The results are discussed and compared with other series reported in the literature. Orbital FNA biopsy may be considered a useful tool in the diagnostic approach to orbital masses in which the relatively high number of inadequate aspirations is offset by a low cost-benefit ratio.

Cell cycle progression is regulated by the combined action of cyclins, cyclin-dependent kinases (CDKs), and CDK inhibitors (CDKIs). p27KIP1, which has a high degree of similarity with p21WAF1, is a general CDKI thought to be involved in G1 arrest in response to agents that inhibit cell cycle progression. The aims of this study were 1) to establish the pattern of expression of p27KIP1 protein in nontumor lymphoid tissue, 2) to determine whether p27KIP1 is involved in lymphomagenesis, and 3) to address the possible relationship between p27KIP1 and p21WAF1 expression in reactive and tumor lymphoid tissue. p27KIP1 protein was found to be mainly present in quiescent lymphocytes in reactive lymphoid tissue as well as in peripheral blood lymphocytes, with an inverse expression for p27KIP1 and Ki-67 proteins. The same p27KIP1 expression pattern was observed in lymphomas, independently of histological type; small resting cells were p27KIP1 positive, and large proliferating cells were p27KIP1 negative. Therefore, tumors with a low proliferative index were mostly positive, whereas tumors characterized by a higher growth fraction bad low p27KIP1 protein levels. An unexpected finding was the existence of a group of six cases of high-grade lymphomas (three diffuse large B-cell lymphomas and three Burkitt's lymphomas) with homogeneously strong staining for p27KIP1 protein. All 6 of these cases belong to a group of 28 cases characterized by blockage of the p53 tumor suppressor pathway, as determined by genetic (p53 mutation) or immunophenotypic studies (p53+/p21-). p27KIP1 expression was not seen in any case of aggressive non-Hodgkin's lymphoma with an intact p53 pathway. The results indicate that p27KIP1 is down-regulated in lymphomas with a high proliferative index, although it is highly expressed in high-grade lymphomas with defects in the p53 pathway.

A case of osteomyelitis caused by Paracoccidioides brasiliensis primarily diagnosed by means of fine-needle aspiration biopsy is reported here in a 60-yr-old Italian patient who had lived in Venezuela for 40 yr. The cytologic and electron microscopic features of the exudate aspirated from a left femoral osteolytic area are described, and the differential diagnosis of this mycotic infection is discussed briefly.

Objective: To evaluate possible alterations of the mechanisms leading to apoptosis in hepatocellular carcinoma by studying bcl-2 expression on fine needle aspiration biopsy (FNAB) samples using immunocytochemistry., Study Design: The study was performed on a series of 84 hepatocarcinomas aspirated under ultrasound guidance. A Papanicolaou-stained smear for each case was destained and restained for bcl-2 by using the immunoperoxidase technique. bcl-2 Expression was then correlated with cytologic grading and the size of the tumor., Results: In 16 cases (19%), bcl-2 immunostaining gave a specific cytoplasmic signal. Fourteen of these positive cases were well differentiated, and two were pleomorphic tumors. Six positive cases were smaller than 5 cm, 3 were larger than 5 cm, and 7 were diffuse., Conclusion: A significant percentage of hepatocellular carcinomas produce and accumulate bcl-2 protein in cell cytoplasm. bcl-2 Expression can be detected on destained cytologic smears. bcl-2 Expression seems to correlate with the cytologic degree of differentiation but not with the size of the tumor.

Objective: To explore the possible advantages that plastic embedding offers to fine needle aspiration biopsy (FNAB)., Study Design: A series of 54 FNABs was fixed in Kryofix (30 cases) and acetone (24 cases), plastic embedded and cut using a rotative microtome. The 2-micron sections obtained were May-Grünwald-Giemsa and Papanicolaou stain in all cases; mucicarmine, Alcian Blue, periodic acid-Schiff (PAS), PAS-diastase and Feulgen stain for DNA cytometric evaluation were also applied in five cases. Immunocytochemical staining using the antibodies cytokeratin 1-8, epithelial membrane antigen, vimentin, Ki-67, lysozime, calcitonin and carcinoembryonic antigen was carried out with the immunoalkaline phosphatase method., Results: The fragments in the sections were perfectly two dimensional; the architectural features of the original tissues were preserved. Morphologic details of nuclear membranes, nucleoli and mitotic figures were excellent. Cytochemical and immunocytochemical stains were successful in all cases. Immunostains showed the precise location of the signal (nuclear, nucleolar or cytoplasmic), a clear background and preservation of morphology., Conclusion: Plastic embedding can be applied to FNAB to solve the problem of dense cellular groups, to study structural and cellular details, and to capitalize on the diagnostic material available for serial cytochemical, quantitative and immunocytochemical purposes.

Most of the data regarding the significance of c-erbB-2 oncogene expression as a prognostic marker in breast cancer have been generated in many large retrospective studies by retrieving the corresponding oncoprotein in archival paraffin embedded sections. Recently, employing fresh breast cancer cells obtained by means of fine-needle aspiration biopsy, we found a rate of c-erbB-2 positive breast tumors (58%) higher than that reported in paraffin-embedded tissue sections by others studies. The present analysis was undertaken to investigate the impact of routine tissue processing on the preservation of the c-erbB-2 immunoreactivity. This issue was addressed by assessing the relative rate of c-erbB-2 oncoprotein immunodetection on FNAB smears and matched surgical specimens of breast cancer. The expression of c-erbB-2 oncoprotein was evaluated using the alkaline phosphate-anti-alkaline phosphatase (APAAP) technique in 54 breast aspirates and corresponding surgical specimens of primary breast cancer. Twenty-six (48%) smears and 23 (43%) matched paraffin sections gave specific signal for c-erbB-2 oncoprotein. The slightly higher incidence of c-erbB-2 expression found on smears seems to be mainly due to the better antigen preservation in the fresh cytological preparations. We conclude that routine histological processing may affect c-erbB-2 immunoreactivity; therefore, in mounting prospective studies, it is advisable to assess c-erbB-2 status in fresh tissue. Moreover, the assessment of c-erbB-2 expression on aspirate samples may yield additional information to the pre-surgical prognostic evaluation of breast cancer diagnosed by FNAB.

The adrenal scintiscan with 123I-metaiodobenzylguanidine (MIBG), a reliable morphofunctional technique to evaluate catecholamine turnover in adrenal tumors, can be a useful method to investigate adrenal incidentalomas with arterial hypertension. A male patient, 44 yr old with diabetes, unstable arterial hypertension, and sudden paroxysms of tachycardia is described. The presence of a disomogeneous right juxta-adrenal neoplasm with calcifications was evidenced with ultrasound tomography and confirmed by computerized tomography (CT) scan. Adrenal 123I-MIBG scintiscan revealed a unilateral uptake at level of the right juxta-adrenal region, sized similarly to the neoplasm previously evidence by CT scan. Histological findings of the surgically removed neoplasm were consistent with an ancient schwannoma. Apart from pheochromocytomas, the MIBG uptake is commonly reported in neuroblastomas. In neuroblastoma, a bidirectional process of transdifferentiation has been previously reported in vitro between two coexistent cells: cells with specific uptake system for norepinephrine, with 123I-MIBG uptake capability, and cells oriented toward schwann/melanocytic line. The evidence of in vivo MIBG uptake in our schwannoma may be caused by the same possible phenotypic interconversion of above mentioned cell types. In conclusion, the presence of adrenal tumors with MIBG uptake capability, apart from pheochromocytomas, neuroblastomas, ganglioneuroblastomas, and ganglioneuromas, must be considered in the diagnosis of adrenal tumors.

The bcl-2 protein plays a role in the regulation of programmed cell death (PCD), overriding apoptosis. Its expression has been reported in breast ductal cells, where it is believed to be involved in the hormonal regulation of hyperplasia and involution. To date, bcl-2 gene product has not been investigated on breast cancer FNA. The expression of bcl-2 protein was evaluated using an immunoalkaline phosphatase technique in 54 pre-operative breast cancer aspirates and in paraffin-embedded sections from 20 matched surgical specimens. A high rate of bcl-2 protein expression was found on FNA samples (65%) and on the corresponding tissue sections (60%); there was a nearly absolute concordance in the two specimens, with 19/20 (95%) cases showing a concordant staining. These findings concur with the view that bcl-2 gene is frequently expressed in breast cancer, possibly through a hormonal-dependent pathway.

There is some evidence that a significant proportion (12%) of women, prospectively followed after negative cervical cytologic findings, develop tissue-proven high grade squamous intraepithelial neoplasia (HSIL) within a short period (up to 24 months). The present study was undertaken to address the validity of this statement since such a high prevalence of spontaneous HSIL, if true, may challenge the value of an annual cervicovaginal cytologic smear examination. Cytologic smears collected from 82 patients with tissue-proven HSIL and obtained in the preceding 24 months were evaluated. The vast majority (90%) of these smears revealed evidence of cervical epithelial abnormalities prior to the development of HSIL-88% at 24 months and 96% at 12 months. The cytomorphologic features of low grade squamous intraepithelial neoplasia were observed in 45% of the smears, while the remaining cases were not normal or negative but had cytologic evidence of human papillomavirus (HPV)-associated changes (described elsewhere), including binucleation and multinucleation, epithelial plaques and pearl formations, and hyperkeratosis, dyskeratosis and parakeratosis. Persistent negative cytology was found in only three cases. Although these HPV-related cellular changes may not have an independent predictive value for HSIL, our study demonstrated that their awareness and detection can help to identify women at risk of potentially more advanced lesions. Thus, an annual cytologic smear examination is valuable and accurately reflects the presence of HPV-associated epithelial changes and precursor lesions of HSIL.

Nucleolar Organiser Regions (NORs) and Proliferating Cell Nuclear Antigen (PCNA) were investigated on routine paraffin embedded histologic sections of 30 oral biopsy specimens (six cases of leukoplakia with low-degree of dysplasia, nine cases of leukoplakia with moderate-degree of dysplasia, nine cases of leukoplakia with severe-degree of dysplasia, six cases of squamous microinvasive carcinomas), tested for HPV-DNA by in situ hybridisation (ISH). The absolute number of NORs per nucleus and the percentage of nuclear positivity for PCNA were found to be different in each group of pathology, with further diversity due to the presence or absence of HPV-DNA. In the major part of HPV-positive lesions, the AgNOR number and percentage of cells positive for PCNA were found to be generally lower than in corresponding negative forms. Conversely, a few cases of HPV+ lesions showed significantly higher values both of AgNOR and PCNA, if compared to the other cases of HPV+ and HPV- lesions. These data suggested that high values of AgNOR and PCNA, in moderate and high grade oral dysplasia, could represent an "alarm signal" of a worse prognosis, and then a possible indication for a strict clinical management and/or a stronger treatment of some HPV-associated preneoplastic lesions.

A morphometric study was performed on the thyreocytes and Hürthle cells in 20 cases of nodular hyperplastic goiter (NHG) and in 8 cases of Hürthle cell-rich NHG. The study was carried out on cytologic samples obtained by fine needle aspiration twice from the same nodules in a selected series of patients before and after L-thyroxine therapy with the aim of evaluating the morphometric nuclear variations induced by that therapy. Thyreocytes and Hürthle cells were evaluated for nuclear Area, Perimeter, Maximum Diameter, Form Elliptic and Form Perimeter using a planimetric instrument. A statistically significant reduction in nuclear size was observed in 18 of 20 cases of NHG after L-thyroxine therapy, but the nuclear shape factors were modified only slightly or not at all. A slight reduction in the nuclear size of Hürthle cells was observed in five and an increase in the remaining three cases, but these differences were not statistically significant. Virtually no changes in shape factors were observed in this group of lesions before or after the therapy. These data are discussed and compared to the results of other series. Our results suggest that morphometry may be useful in the follow-up of NHG to evaluate the effects of L-thyroxine and that this therapy reduces the size of thyreocytes; the therapy, however, seems to have little or no effect on Hürthle cells.

This study analyzed a series of 140 consecutive fine needle aspiration biopsies of the spleen (sFNAB) collected over a period of nine years. sFNABs were performed by the authors using a 22-gauge spinal needle, and 23-gauge needle in pediatric patients, with a subdiaphragmatic approach. Ultrasound assistance was utilized in 35 cases. Four of the cytocentrifuge specimens were also prepared with part of the aspirated material and used for immunocytochemical staining. Study of these biopsy specimens revealed various benign conditions, such as white pulp hyperplasia, myeloid metaplasia, storage disease, and granulomatous processes and abscesses. Malignant neoplasms were represented by non-Hodgkin's and Hodgkin's lymphomas, leukemias, malignant histiocytoses and metastases. sFNAB was the only diagnostic procedure employed for the morphologic assessment of 60 benign and of 2 malignant cases. In all the other cases sFNAB gave the first diagnosis that was confirmed on surgical specimens. In 2 cases splenic bleeding occurred; one of them required splenectomy. Diagnostic and technical problems are discussed. The authors consider sFNAB a useful procedure, especially in the study of hematologic disorders and in all cases in which splenomegaly is not related or correlated with any clinical finding.

The aim of this study was to determine the relative rate of c-erbB-2 oncoprotein immunodetection on matched fine needle aspiration (FNA) smears and surgical specimens of breast cancer, and to correlate the c-erbB-2 expression with the assessment of the DNA ploidy status. The expression of c-erbB-2 oncoprotein was evaluated using an immunoalkaline phosphatase technique in 49 breast aspirates (four benign and 45 malignant lesions) and 21 matched surgical specimens. The DNA ploidy status was assessed by densitometric techniques on Feulgen-stained smears. Fifty-eight per cent of the smears obtained from 45 malignant lesions and 43% of the 21 corresponding paraffin sections contained cells that were stained by the antibody. The higher incidence of c-erbB-2 expression on smears seems to be due mainly to the better antigen preservation in the fresh cytological preparations. The correlation between c-erbB-2 oncogene expression and DNA ploidy assessment showed an increased incidence of oncogene expression in aneuploid tumours (71% vs 29%; P < 0.05).

Aims: To determine the relative diagnostic performance of non-isotopic in situ hybridisation (NISH) and a dot-blot assay for detecting human papillomavirus (HPV) on exfoliated cervical cells; and to correlate the results with cytopathological assessment., Methods: Cervical smears and cytological samples were obtained from 122 patients during the same clinical examination and the presence of HPV sequences determined by NISH and dot-blot analysis, respectively., Results: Dot-blot analysis gave an autoradiographic signal in 15 of 121 (12.4%) cases, while NISH detected viral genomes in 38 of 114 (33.3%) cases. Even in the presence of koilocytosis, where vegetative replication of the virus occurs, NISH was positive in over twice as many cases as dot-blot analysis (NISH 90%, dot-blot 40%), while in smears within normal cytological limits, where the viral copy number is likely to be considerably lower, the differences were more striking (NISH 31%, dot-blot 5%)., Conclusions: These data show that NISH on cytological smears is more sensitive than a standardised dot-blot hybridisation assay for detecting HPV infection in cytological material and is therefore a more appropriate screening tool.

Aim: To develop a methodology for direct detection of high risk human papillomavirus (HPV) infection in routine cervical smears by non-isotopic in situ hybridisation (NISH) which can be compared with cytopathological assessment of the same cells., Methods: The methodology was established using cultured cells and routine cervical smears hybridised with digoxigenin labelled probes for HPV, 16, 18, 31, and 33. The technique was applied to the analysis of 53 patients from a sexually transmitted disease clinic., Results: The optimal sensitivity achieved for single HPV detection in cultured cells was 1-2 copies of HPV 16 per cell and that for detection of a cocktail of HPV types in routine cervical smears was 2.5-12 copies per cell. Of parallel smears taken from patients with a normal Papinacolau-stained smear 33.3% (24) contained a HPV 16, 18, 31, and 33 signal indicating an occult HPV infection. The prevalence of these HPV types was similar in women in whom a cytopathological diagnosis of wart virus infection was made (64.7%, 17) and in patients with mild dyskaryosis (75%, 12)., Conclusions: The methodology evolved localises HPV sequences directly to epithelial cell nuclei, which can be morphologically assessed by haematoxylin counterstaining. Sample contamination with exogenous viral sequences can be distinguished from true infection. In this study, a HPV signal was not found in morphologically normal epithelial cells. The methods described will permit the detection of HPV sequences in routinely collected cervical smears and the evaluation of the natural history and potential clinical relevance of HPV infection without changes in clinical practice.

Aims: To determine the relative diagnostic sensitivity of non-isotopic in situ hybridisation (NISH) for the diagnosis of human papillomavirus (HPV) on matched smears and biopsy specimens; to compare the NISH signal type in the two samples; and to correlate the NISH data with the morphological diagnosis., Methods: HPV samples were assayed individually by NISH with digoxigenin labelled probes (HPV6, 11, 16, 18, and 33) on routinely collected paraffin wax embedded cervical biopsy specimens and for high risk HPVs with a cocktail of similarly labelled probes (HPV16, 18, 33) on matched smears. These were taken at the same colposcopic examination from 32 patients investigated for an abnormal cervical Papanicolaou (PAP) stained smear., Results: An HPV signal was present in 18 (56%) biopsy specimens and in 14 (44%) smears. There was higher concordance of sets of data in the presence of cytopathic wart virus changes. The superiority of biopsy over smear in detecting HPV was mainly the result of examining the entire cervical biopsy specimen rather than cells scraped from the cervical surface. The NISH signal type in both biopsy specimen and smear was similar; it has been shown that NISH type 1 signal correlates with episomal viral replication and type 2 and 3 signals with viral integration., Conclusions: These data show that NISH on cervical smears is a worthwhile primary screen for HPV infection. The NISH signal types in cervical smears are similar to those previously described in cervical biopsy specimens.

A series of 1153 fine-needle aspiration biopsies (FNAB) of breast masses obtained during the period 1985 to 1988 in one institution was examined. The following diagnostic accuracy values were found: sensitivity, 93.0%; specificity, 92.4%; predictive positive value, 93.7%; and efficiency, 92.8%. These results were compared with a previously published series for the period 1976 to 1984. The diagnostic accuracy values did not differ significantly between the two series.

Routine cervical smears (n = 262) from a Sexually Transmitted Diseases clinic were screened by non-isotopic in situ hybridization (NISH) stratifying human papillomavirus (HPV) infections into HPV6/11 (low risk) and HPV16/18/33 (high risk) categories. Of 188 patients with cytologically normal smears, HPV sequences were demonstrated in 41%. Of the 128 cases analysed by dual NISH, 16% contained low risk, 20% high risk and 5% both groups. In patients with cytological evidence of wart virus infection (WVI) only, 54% (n = 50) contained high-risk and 22% low-risk HPV types. The comparable incidences in CIN1/2 plus WVI (n = 24) were not significantly different: 54% and 17%, respectively. Cytological criteria underestimate the prevalence of HPV infection in patients with cytologically normal smears. This represents either 'occult' or 'latent' infection. The identical prevalence of HPVB16/18/33 in WVI only, and CIN1/2 plus WVI, suggests that the cytopathic effect induced by these HPVs may represent one end of a spectrum of morphological change which progresses to cervical intraepithelial neoplasia (CIN).

Leydig's cells tumor account for 1-2% of all testicular tumors. The authors report a rare case of benign Leydig's cells tumor in a young man with gynecomastia. They examine the embryogenesis, the hormonal modifications, the criteria of malignancy, the treatment and results. They present a review of the literature pertaining to this rare tumors.

The cytologic findings in nine cases of giant-cell tumor (osteoclastoma) of bone diagnosed by fine needle aspiration (FNA) biopsy are described. The aspirates contained a dual population of cells, consisting of mononucleated tumor cells and multinucleated giant cells (osteoclasts). The tumor cells were usually evident in the FNA smears as cohesive perivascular clusters; some were present as sheets or single cells. The most striking feature, which may best suggest the diagnosis of giant-cell tumor in an FNA smear, was the attachment of the osteoclasts to the cohesive groups of tumor cells. Even when the cells were more dissociated, there was a rather regular distribution of the two types of cells. This series further demonstrates that FNA biopsy permits an accurate cytopathologic diagnosis of giant-cell tumor and that its use can clarify the clinical and radiologic impressions of these bony lesions preoperatively. The differential diagnosis between osteoclastoma and other giant-cell containing bone lesions is discussed in detail.

A morphometric analysis was performed on histologic sections of 11 cases of aggressive basal cell carcinoma that recurred and/or metastasized after the first diagnosis (BCC2) and on 11 ordinary basal cell carcinomas (BCC1). The considered parameters were as follows: nuclear area, perimeter, maximum diameter, feret x, feret y, form perimeter, and form area. The results show numeric differences between the two groups for the first five parameters. A statistical analysis was performed between BCC1 and BCC2 for all the considered parameters using a two-sided t test for independent samples. The test showed significant differences between the first five parameters except for the two form factors. A multivariate analysis was performed using area and perimeter values of the compared groups that amplified the discrimination threshold between the two groups. The authors conclude that a morphometric assessment may bring significant contribution in the knowledge and in the outcome prediction of basal cell carcinoma.

A modified bronchial brush is presented as a new cytologic method to improve the value of fine-needle aspiration biopsy in the diagnosis of cystic lesions of bone. Four female and six male patients ranging in age from 10 to 44 years were studied. In all cases intralesional brushing was percutaneously performed under fluoroscopic control with local anesthesia. Fine-needle aspiration biopsy preceded intralesional brushing and was performed as described elsewhere. The brush was driven into the walls and septa of an osteolytic lesion through a 14-G needle; material obtained was fixed and processed for cytological and/or histological examinations.

A series of 1956 fine-needle aspiration biopsies (FNAB) of breast masses is described. The diagnostic accuracy of this series (sensitivity, 95.7%; specificity, 89.6%; predictive value of positivity, 95.9%; and efficiency, 94.0%) was compared with that published in the literature. Statistically significant differences were found between the level of diagnostic accuracy in series published by pathologists who diagnosed smears prepared by clinicians and surgeons (Group A), and those published by pathologists who performed palpation and aspiration, and made the cytologic diagnosis by themselves (Group B). These differences mainly consisted of a lower number of false-positives and "unsatisfactory" samples in Group B series.

We report one case of nodular sclerosing Hodgkin's disease with uncommon pseudosuppurative presentations in fine-needle aspiration biopsy (FNAB) samples and on histopathologic material. The criteria for cytologic diagnosis include atypical mononuclear cells and diagnostic SR cells in the purulent exudate. This lesion is considered a possible pitfall in FNAB cytology of Hodgkin's disease.

The literature on the subject is reviewed, special attention being paid to the complex problems of aetiological and pathogenetic classification involved by such lesions in the oral cavity. Stress in salso laid on topographical and histological aspects of pigmented lesions of melanocytic origin.