Benign squamous cells with pleomorphic microvilli in urine or bladder washings

Exfoliated cells in catheterized urine or bladder washings from 40 patients were observed by light microscopy (LM) and by scanning electron microscopy (SEM). The specimens from seven of these patients (six postmenopausal females and one 85-year-old male) contained squamous cells with pleomorphic microvilli (PMV) on their surfaces. Four of these cases had no bladder lesions by cystoscopic examination. Three patients had recurrent papillary transitional-cell carcinoma of the bladder, and the cytologic specimens from two of them contained transitional cells with PMV. The distinction between squamous and transitional cell is readily made by SEM, based primarily on cell shape and thickness. The presence of PMV on otherwise-benign-appearing squamous cells in urine or bladder washing specimens may be a source of confusion in the interpretation of SEM findings. The presence of PMV on exfoliated squamous cells in cytologic material from the human urinary tract does not seem to have the same diagnostic and prognostic significance as the presence of PMV on transitional cells.     

Control specimens for immunocytochemistry in liquid‐based cytology

T. Hansen, H. Pedersen, V. Brauner and J. Hariri Control specimens for immunocytochemistry in liquid‐based cytology Objective Immunostaining necessitates the use of positive as well as negative controls, which is usually an easy procedure in immunohistochemistry (IHC). To find suitable control specimens for immunocytochemistry (ICC) is, on the other hand, a challenging task and to the best of our knowledge is not sufficiently dealt with in the English literature. The aim of this trial was to develop an applicable method to select, collect, process and store control specimens for ICC using liquid‐based cytology (LBC). Methods The study included 21 different antibodies, which were known to react with at least one of the cellular components from tonsils, serous fluids and bronchial washings. The LBC specimens from the tonsils were collected as SurePath? specimens (BD, Bencton, Dickinson and Company) by brushing the cut‐surface of a fresh tonsil and then immersing the brush head into the SurePath? vial. The serous fluids and bronchial washings were fixed in CytoRich Red? (BD). Some of the cellular suspensions from the tonsils and equal amounts of the serous fluid and the bronchial washings were also mixed as a cocktail. Unstained SurePath slides were then prepared on the PrepStain? (BD) Non‐GYN Program, and the unstained and dry slides were then stored at 5 °C to test the effect of storage on the preservation of the antigenicity. ICC was then performed on BenchMark‐XT?. Results Cellular components in unstained SurePath? slides reacted positively with relevant antibodies. Slides that were stored for up to 40 days did not loose staining intensity. Conclusion Specimens from body fluids and cell‐suspensions that are collected by brushing the cut‐surface from different types of fresh tissues or organs can be used as control specimens either separately or as mixtures. Dry and unstained slides can then be prepared and stored in a refrigerator for at least 40 days without loosing staining intensity.     

Immunocytochemical characterization of large-cell carcinomas of the lung. Role, limitations and technical considerations.

To clarify the use of cytologic preparations, particularly those previously stained by the Papanicolaou method, for the immunocytochemical evaluation of large-cell carcinomas (LCCs), 37 cytologic preparations from cases diagnosed as LCC were examined using a battery of immunocytochemical stains for keratin, chromogranin, common leukocyte antigen (CLA) and B72.3. Thirty-two specimens were from the thoracopulmonary region (12 fine needle aspirates of the lung, 7 bronchial brushings, 5 bronchial washings, 2 sputa and 6 pleural fluids); the remaining specimens were fine needle aspirates of 3 lymph nodes, 1 vertebral body and 1 liver. Of the specimens analyzed, 30 of 37 were positive for keratin and 7 of 35 were positive for B72.3 (6 were positive for both). Only 1 of 37 was positive for CLA while none of 37 was positive for chromogranin. Six specimens showed no reaction with either keratin, B72.3 or chromogranin. These results confirm that the majority of LCCs consist of epithelial cells of either a squamous or an adenocarcinomatous type. They also show that immunocytochemistry is a useful diagnostic adjunct that can be applied to cytologic preparations previously stained by the Papanicolaou method; this is important since immunostaining is often considered after undifferentiated malignant cells are encountered in a previously stained preparation. However, a thorough understanding of some technical limitations is critical in the evaluation of the results of this technique when it is applied to cytologic specimens.     

The recognition of Pneumocystis carinii in routine Papanicolaou-stained smears

Using definite criteria it is possible to accurately evaluate routine Papanicolaou-stained cytologic smears for the presence or absence of Pneumocystis carinii. Strict attention must be paid to the cellular environment and the background material intimately associated with the cells. In 133 cytology specimens evaluated from proximal and deep bronchial washings and brushings, 71 were considered positive for P. carinii and 62 were called negative. Ten of the latter were either unsatisfactory or equivocal. The 71 positives correlated in every instance with parallel Grocott methenamine silver-stained transbronchial biopsies or brushings. Fifty-one of the 52 satisfactory cytologic negatives also correlated with the biopsy and brushing findings. There was a single false negative. This high degree of correlation indicates that the Papanicolaou-stained specimen can be a valuable tool in the early diagnosis of pneumocystosis.     

Role of special stains in the diagnosis of Pneumocystis carinii infection from bronchial washing specimens in patients with the acquired immune deficiency syndrome

Fifty bronchial washing specimens from 36 patients with acquired immune deficiency syndrome (AIDS) were retrospectively reviewed to assess the sensitivity of the various special stains used to diagnose Pneumocystis carinii. In 76% of the cases, the Diff-Quik stain was positive; it was the easiest and most rapid of the special stains used. The sensitivity was increased to 92%, 96% and 100%, respectively, by also doing cresyl echt violet, Grocott's Gomori methenamine silver and both the cresyl violet and Grocott stains in addition to the Diff-Quik stain. We conclude that the Diff-Quik stain is a fairly reliable and rapid screening procedure for making the diagnosis of Pneumocystis infection in bronchial washings from AIDS patients. The routine Papanicolaou stain gave less sensitive results in the smears of the washing specimens, but does give a markedly improved yield in bronchoalveolar lavage specimens.     

Exfoliative cytology of multiple endobronchial granular cell tumor

A multicentric endobronchial granular cell tumor (GCT) in a 50-year-old man was diagnosed by the cytologic study of bronchial lavage specimens. The paraffin-embedded sections contained small clusters of medium-sized round tumor cells that had eccentric nuclei without nucleoli and eosinophilic finely granulated cytoplasm, which was positive with the periodic acid-Schiff stain. These cells were distinguishable from the macrophages and bronchial and squamous cells also found in the specimens. The excised tumors histologically mimicked a squamous cell carcinoma. Since 10% of all GCTs occur in the lung, where a multiple presentation can especially mimic a metastatic malignant process, it is important that the possibility of a granular cell tumor be considered in the screening of exfoliative cytologic specimens from the lung.     

Cytomorphology of granular-cell tumor of the bronchus. A case report

The cytologic features of a bronchial granular-cell tumor clinically mimicking a bronchogenic carcinoma are presented. Distinctive granular cells similar to the main cellular components of the surgical specimen were found in abundance in bronchial brushings and washings obtained during bronchoscopy. Our findings and conclusions confirm previously published studies. Granular-cell tumors of the bronchus can easily be diagnosed on cytologic examination if the entity is considered in the differential diagnosis of clinically suspected lung tumors.     

Cytology of bronchial benign granular-cell tumor

The cytologic features of a case with multiple bronchial benign granular-cell tumors are reported and compared with those of previously reported cases. Characteristic tumor cells were found in the bronchoscopic brushing smears and in cell block sections (but not smears) prepared from the washing fluid. These findings were confirmed by the bronchial biopsy and histologic study of the resected tumors. A cytologic diagnosis of bronchial granular-cell tumor should not be difficult because the cytologic appearance of the tumor cells is characteristic; however, the possibility of a concomitant tumor, such as adenocarcinoma or small-cell carcinoma, should be considered and excluded.     

The sensitivity of detection of asbestos bodies in sputa and bronchial washings

While asbestos bodies (ABs) in sputum and/or bronchial washings are highly specific markers for significant asbestos exposure, comparison of the sensitivity between sputum cytology and bronchial washing cytology for the detection of ABs had not been documented. Review of the files of the Cytopathology Laboratory, Methodist Hospital, Houston, Texas, for the period 1973 to 1984 identified 11 patients with slides available for review who (1) had been examined by both sputum cytology and bronchial washing cytology and (2) had at least one specimen positive for ABs. Of the 11 evaluable cases, all had ABs in the bronchial washings but ony 6 had ABs in the sputum. In addition, iron stain (e.g., the Prussian blue stain) was found to be more sensitive than the Papanicolaou stain for the detection of ABs in these cases. These findings indicate that iron-stained bronchial washing specimens should be preferred for the cytologic detection of asbestos exposure.     

Detection of adenocarcinoma in peritoneal washings by staining with monoclonal antibody B72.3

Peritoneal washings are routinely performed in the staging evaluation of carcinomas of the ovary and in "second-look" explorations; major problems in the evaluation of these specimens continue to be the distinction between atypical mesothelium and adenocarcinoma and the identification in an otherwise inflammatory specimen of rare cells of adenocarcinoma, which may be undetected by the most trained individual. Monoclonal antibody (MAb) B72.3, reactive with an oncofetal, tumor-associated glycoprotein (termed TAG-72; MW greater than 1000 kd) expressed in a variety of epithelial malignancies but not generally expressed in benign or malignant mesothelium, was reacted with sections from the paraffin-embedded cell blocks of 185 peritoneal washings from 180 patients with extant cancer or a prior history of malignancy. One hundred four of the washings were initially interpreted as atypical mesothelium, with no evidence of malignancy; when reacted with MAb B72.3, 6 of these specimens demonstrated groups of metastatic adenocarcinoma cells not appreciated by the usual cytologic criteria. Of the 81 washings interpreted as showing cells of adenocarcinoma, 73 demonstrated expression of TAG-72 from both gynecologic and nongynecologic malignancies. In the remaining 49 cases without an associated malignant process, MAb B72.3 did not stain atypical mesothelium, but did react, however, with benign endometrial cells and müllerian inclusions in two cases. MAb B72.3 may be used as a diagnostic adjunct to the routine cytologic evaluation of malignancy in peritoneal washings; reactivity with MAb B72.3 may indicate the need for further evaluation to define the presence of a malignancy or an advanced cancer.     

P-11THE IMPACT OF INTRODUCING LIQUID BASED CYTOLOGY INTO A ROUTINE SCREENING LABORATORY

Introduction:  A glacial acetic acid wash performed retrospectively or prospectively on visibly bloodstained cervical ThinPrep® specimens can reduce the unsatisfactory rate and increase the number of diagnostic epithelial cells. This study was undertaken to determine which specimens are most likely to benefit from a prospective glacial acetic acid wash.
Methods:  Bloodstained ThinPrep® specimens selected for routine lysing prior to processing were macroscopically assessed and scored based on the level of blood present (+ to +++). Both unlysed and lysed slides were prepared from each specimen and microscopically examined.
Results:  Fifty-eight specimens (32 scored +, 12 ++ and 14+++) were assessed. Three unlysed slides prepared from the ++ specimens and 12 from +++ specimens were evaluated as unsatisfactory due to excessive blood and inadequate numbers of squamous cells. In contrast, only one of the unlysed slides from the 32 + specimens was considered to be unsatisfactory. Almost all the lysed slides were satisfactory and generally more cellular than the unlysed slides. Abnormal cells were found in four cases (both unlysed and lysed paired slides).
Discussion:  Although the acetic acid wash increases cellularity of bloodstained ThinPrep® specimens, prospectively lysing all bloodstained specimens is an unnecessary procedure. Lysing only very heavily bloodstained specimens prior to processing will reduce laboratory workload, costs and the possibility of specimen mix up. Occasionally a retrospective wash may be required but screening staff should be aware that although blood may be present on an unlysed ThinPrep® slide, a lysed slide should not be requested unless there are insufficient numbers of squamous cells present.     

Oncocytic carcinoid tumor of the lung: a case report of diagnostic pitfall in filter membrane preparation of bronchial washings

BACKGROUND: Oncocytic carcinoid tumor of the lung is a rare variant of pulmonary carcinoid. This report describes the morphologic appearance of this rare tumor on filter membrane preparation along with potential pitfalls. CASE: A 49-year-old woman presented with cough and expectoration. On chest radiograph a mass lesion was seen in the upper zone of the right lung. Bronchial washings were sent for evaluation. On filter membrane (Millipore) preparation of bronchial washing the possibility of a non-small cell carcinoma, possibly squamous, was suggested. Right upper lobectomy was subsequently performed and a histologic diagnosis of oncocytic carcinoid given. The cytomorphologic features of this tumor on the Millipore preparation were reviewed. CONCLUSION: Differential diagnosis of oncocytic carcinoid should be kept in mind while assessing cytologic material when tumor cells show abundant granular cytoplasm and prominent nucleoli. Oncocytic carcinoid also must be differentiated from oncocytoma and granular cell tumor. Immunocytochemistry and electron microscopy are useful in confirming the diagnosis.     

P‐11THE IMPACT OF INTRODUCING LIQUID BASED CYTOLOGY INTO A ROUTINE SCREENING LABORATORY

Introduction: A glacial acetic acid wash performed retrospectively or prospectively on visibly bloodstained cervical ThinPrep® specimens can reduce the unsatisfactory rate and increase the number of diagnostic epithelial cells. This study was undertaken to determine which specimens are most likely to benefit from a prospective glacial acetic acid wash. Methods: Bloodstained ThinPrep® specimens selected for routine lysing prior to processing were macroscopically assessed and scored based on the level of blood present (+ to +++). Both unlysed and lysed slides were prepared from each specimen and microscopically examined. Results: Fifty‐eight specimens (32 scored +, 12 ++ and 14+++) were assessed. Three unlysed slides prepared from the ++ specimens and 12 from +++ specimens were evaluated as unsatisfactory due to excessive blood and inadequate numbers of squamous cells. In contrast, only one of the unlysed slides from the 32 + specimens was considered to be unsatisfactory. Almost all the lysed slides were satisfactory and generally more cellular than the unlysed slides. Abnormal cells were found in four cases (both unlysed and lysed paired slides). Discussion: Although the acetic acid wash increases cellularity of bloodstained ThinPrep® specimens, prospectively lysing all bloodstained specimens is an unnecessary procedure. Lysing only very heavily bloodstained specimens prior to processing will reduce laboratory workload, costs and the possibility of specimen mix up. Occasionally a retrospective wash may be required but screening staff should be aware that although blood may be present on an unlysed ThinPrep® slide, a lysed slide should not be requested unless there are insufficient numbers of squamous cells present.     

Appearanc of cytotoxic cells within the bronchus after local infection with herpes simplex virus.

Cells from rabbit spleens, bronchial washings (BW) and bronchus-associated lymphoid tissues (BALT) were examined for their ability to lyse cells infected with herpes simplex virus (HSV). Specific lysis of HSV-infected cells was mediated by BW cells as early as 4 days after intratracheal infection of the rabbits with the virus whereas lysis by spleen cells and BALT cells was not detected until 7 or more days after infection. Lysis by spleen cells was initially detected 7 days after intraperitoneal injection of the virus but lysis by BW and BALT cells was not observed until 14 days after infection. Although spleen, BW, and BALT cells could lyse antibody-coated target cells, antibodies detectable by antibody-dependent cellular cytotoxicity could not be detected in bronchial washings until 7 or more days after infection. The data suggest that cells capable of direct cytotoxicity of virus-infected cells appear within the bronchus after local infection by the virus.     

The nature of antigens common to both the metacestodes of Taenia crassiceps and its laboratory host

Metacestodes of Taenia crassiceps were cultured in laboratory rats and mice, washed in glycine/hydrochloric acid (Gly/Hcl) buffer pH 2.4 and phosphate buffered saline (PBS) pH 7.2 and a soluble extract of the metacestodes (SEM) was prepared from larvae washed in both buffers. The SEM and the original washings were examined for host IgG and a common host antigen (CHA). CHA was present in both the SEM and washings but IgG only in the washings and, in metacestodes from mice only in the Gly/HCl washings). Metacestodes washed in Gly/HCl were not viable. Metacestodes were also maintained in vitro in a peptide nutrient agar and 0.9% saline at 37 degrees C and a SEM was prepared from these; metacestodes from mice had neither host IgG nor CHA but those from rats had CHA but not IgG. SEM prepared from 'normal' metacestodes showed that the CHA consisted of at least two entities, only one having a common identity with the IgG. These results suggest that molecules of both host IgG and a common host antigen are present on the tegument of these parasites.     

Quantitation of Ki-67 expression in the differential diagnosis of reserve cell hyperplasia vs. small cell lung carcinoma

OBJECTIVE: To investigate whether the detection of proliferation-associated Ki-67 antigen may be of value in differentiating between reserve cell hyperplasia (RCH) and small cell lung cancer (SCLC). STUDY DESIGN: Retrospectively, 20 Papanicolaou-stained bronchial brushes or washings from 20 patients were selected. Ten were diagnosed as RCH (and had no SCLC in follow-up) and the other 10 as SCLC (histologically confirmed). All 20 Papanicolaou-stained slides were restained with the monoclonal antibody MIB1, directed against Ki-67 antigen; that simple and reliable procedure was described recently. In each specimen 5 coherent cell groups were identified, corresponding to RCH or SCLC, respectively; photographed; and studied for Ki-67 antigen expression after MIB1 staining of the slides. At least 3 cell groups remained in each specimen. The Ki-67 labeling index (LI) of the specimens was determined as the number of MIB1-positive cells divided by the total number of cells in the remaining cell groups. RESULTS: All cases of SCLC showed a mean Ki-67 LI of at least .415 (mean .684, SD .151), whereas in the cases with RCH the mean Ki-67 LI never was more than .158 (mean .048, SD .049). The difference was highly significant (P<.001, Student's t test). Linear discriminant analysis resulted in a classifier with which we were able to discriminate correctly between SCLC and RCH in 100% of the 20 bronchial brushings and washings. CONCLUSION: The results clearly demonstrate that measuring proliferative activity in Papanicolaou-stained bronchial brushings and washings by MIB1 restaining of the slides may be of great practical value in accurately discriminating RCH from SCLC. The method is simple and can be performed in any laboratory that is able to carry out immunocytochemical staining. However, an additional (prospective) study with a series of difficult cases is necessary to confirm these findings.     

Factors significant in the diagnostic accuracy of lung cytology in bronchial washing and sputum samples. I. Bronchial washings

Some factors influencing the diagnostic accuracy for primary lung cancer in bronchial washings were studied in 276 consecutive cases seen between 1959 and 1974. Diagnostic accuracy increased during the years under study; the reasons included increasing expertise of the laboratory staff, better documentation of cytologic criteria and improved collection techniques. The overall accuracy was 74%. Detection of malignant cells was highest for squamous-cell and adenosquamous carcinomas (81%), small-cell carcinoma, adenocarcinoma and large-cell carcinoma (70%) and lowest for bronchioloalveolar-cell carcinoma (47%). Accuracy was 84% for central tumors as compared to 30% for peripheral lesions. Tumors of less than 2 cm in diameter yielded very poor results (15%) while those greater than 2 cm yielded 82% accuracy. The specificity of diagnosis of cell type in those specimens with malignant cells was over 93% for squamous-cell carcinoma, small-cell carcinoma and adenocarcinoma, 77% for large-cell carcinoma and below 50% for adenosquamous carcinoma, bronchioloalveolar carcinoma and the uncommon tumors. Two bronchial washings per case gave an appreciably better result (92%) than one per case (68%). The percentage of unsatisfactory specimens from those with cancer was 13.5 and from a control group was 29.9. Reasons for unsatisfactory specimens included limited cellular material, excessive blood and/or leukocytes and drying artifacts.     

Cytologic evaluation of low grade transitional cell carcinoma and instrument artifact in bladder washings

OBJECTIVE: To identify key cytologic features for the separation of low grade transitional cell carcinomas (TCCs) from nonneoplastic lesions in bladder washings. STUDY DESIGN: The cytomorphologic features of 95 bladder washing specimens showing papillary fragments, which included 50 low grade TCCs and 45 nonneoplastic lesions, were reviewed retrospectively. RESULTS: Bladder washings from low grade TCCs showed papillary and irregular groups of cells with ragged borders, cytoplasmic homogeneity and subtle nuclear changes, such as increased nuclear/cytoplasmic ratio and irregular nuclear border. Bladder washings after instrumentation from nonneoplastic lesions of the bladder showed cellular specimens with cohesive, ball-shaped and papillary clusters with smooth borders lined with a denser-staining cytoplasmic collar. Reactive urothelial cells often displayed loose aggregates with irregular borders but no cytoplasmic collar. CONCLUSION: In bladder washing cytology, nuclear changes and cytoplasmic homogeneity play a major role in the diagnosis of carcinoma.     

Cytologic evaluation of bronchoalveolar lavage specimens in immunosuppressed patients with suspected opportunistic infections

The role of bronchoalveolar lavage cytology in the diagnostic evaluation of immunosuppressed patients with suspected opportunistic pulmonary infections was evaluated by comparing two groups of patients who underwent fiberoptic bronchoscopy. Bronchoalveolar lavage specimens were compared with other available diagnostic techniques, including bronchial washings, bronchial brushings, transbronchial lung biopsies and open lung biopsy. Prior to the initiation of a protocol for bronchoalveolar lavage, a specific etiology for the pulmonary infiltrate using the above combined modalities was identified in 23 of 47 cases, for an overall diagnostic rate of 49%. The combined bronchial washings and brushings (cytologic procedures) identified a specific etiology in 9 of 47 (19%) of the cases. There were ten cases in which a cytologically identifiable organism (Pneumocystis, virus or fungus) was not present in the bronchial washings and brushings and one missed case of malignancy, for a false-negative rate of 23%. With the addition of the lavage technique and better sampling of the distal airways, a specific etiology for the pulmonary infiltrate was identified in 32 of 48 (67%) of the cases. This is comparable to the values of 40% to 65% cited in the literature for diagnosis of infectious disease by open lung biopsy. The lavage cytologic procedure identified a specific etiology in 22 of 48 (46%) of the cases, and the false-negative rate was reduced to 6%. With the excellent sampling of the bronchoalveolar lavage and the improved cytology results, the need for transbronchial or open lung biopsy has been eliminated in immunosuppressed patients with suspected opportunistic pulmonary infections. This allows these patients to be studied on an outpatient basis.     

Cytologic features of nonfatal herpesvirus tracheobronchitis

Herpesvirus infections of the lower respiratory tract have most commonly been reported in patients with severe burns, immunosuppression or malignancies. Two patients without any of these underlying conditions developed severe herpetic tracheobronchitis, diagnosed by cytologic examination and confirmed by serologic studies. Serial examination of sputum, bronchial brushings and bronchial washings permitted observation of the evolution and progression of cellular changes found in herpesvirus infection of the lower respiratory tract. both patients recovered without specific antiviral therapy, but both developed superinfection with gram-negative organisms, requiring intensive antibiotic therapy. The distinctive features of herpesvirus infection in the tracheobronchial tree are similar to those recognized elsewhere in the body. Early findings include a variety of nonspecific changes in nuclear chromatin configurations; multinucleated cells may be common but do not often contain the central intranuclear inclusion bodies seen in later stages. These distinctive central intranuclear inclusions disappear in a few days, leaving only reparative changes in the surface epithelium. Herpesviruses are increasingly being reported in the literature as an etiologic agent of acute tracheobronchitis in otherwise healthy individuals.