Pleural effusions in non-Hodgkin's lymphoma. A cytomorphologic, cytochemical and immunologic study

Review of the records of 243 cases of cytologically diagnosed non-Hodgkin's lymphomas (NHL) revealed pleural effusions in 21 (8.6%). Cytologic examination of pleural fluid was done in 17 cases, of which 16 were reported as positive. Cytologic examination was supplemented with cytochemical staining (acid phosphatase, alpha naphthyl acetate esterase and periodic-acid-Schiff reactions) and E-rosetting studies in 12 cases. Of the 16 positive cases, 11 were malignant lymphomas consisting of convoluted lymphocytes. Acute lymphatic leukemia of the prothymocytic type (T-ALL) and chronic lymphocytic leukemia of the T-cell type (T-CLL) comprised one case each, and there were three cases of follicular center cell lymphomas, two of the cleaved-cell type and one of the Burkitt-type. Comparison of the cytomorphology of the tumor cells in the pleural effusion with those in fine needle aspiration smears from the solid tumors in 14 cases showed an identical appearance in 13 cases; in one, the Burkitt-type lymphoma, the cells were larger and more pleomorphic in the pleural effusion. This study indicates that the cytologic diagnosis and categorization of NHL of the convoluted-cell type is greatly enhanced by the study of neoplastic lymphocytes in a pleural effusion.     

Cell origins and diagnostic accuracy of interleukin 27 in pleural effusions

The objective of the present study was to investigate the presence of interleukin (IL)-27 in pleural effusions and to evaluate the diagnostic significance of pleural IL-27. The concentrations of IL-27 were determined in pleural fluids and sera from 68 patients with tuberculous pleural effusion, 63 malignant pleural effusion, 22 infectious pleural effusion, and 21 transudative pleural effusion. Flow cytometry was used to identify which pleural cell types expressed IL-27. It was found that the concentrations of pleural IL-27 in tuberculous group were significantly higher than those in malignant, infectious, and transudative groups, respectively. Pleural CD4(+) T cells, CD8(+) T cells, NK cells, NKT cells, B cells, monocytes, macrophages, and mesothelial cells might be the cell sources for IL-27. IL-27 levels could be used for diagnostic purpose for tuberculous pleural effusion, with the cut off value of 1,007 ng/L, IL-27 had a sensitivity of 92.7% and specificity of 99.1% for differential diagnosing tuberculous pleural effusion from non-tuberculous pleural effusions. Therefore, compared to non-tuberculous pleural effusions, IL-27 appeared to be increased in tuberculous pleural effusion. IL-27 in pleural fluid is a sensitive and specific biomarker for the differential diagnosing tuberculous pleural effusion from pleural effusions with the other causes.     

The differential diagnostic values of cytokine levels in pleural effusions

The aim is to examine whether the changes in pleural fluid interleukin (IL)-1beta, IL-2, IL-6, and IL-8 levels were significant in differential diagnosis of childhood pleural effusions. IL-1beta, IL-2, IL-6, and IL-8 levels in pleural fluids of all 36 patients were measured. The levels of IL-1beta, IL-2, IL-6, and IL-8 in pleural fluids were statistically significantly higher in the transudate group compared with those of the exudate group. The levels of IL-1 beta, IL-6, and IL-8 were also found to be statistically significantly higher in the empyema group compared with both the parapneumonic and the tuberculous pleural effusion groups. The levels of IL-2 and IL-6 were detected to be statistically significantly higher in the tuberculous pleural effusion group in comparison with those of the parapneumonic effusion group. The results showed that pleural fluids IL-1beta, IL-2, IL-6, and IL-8 could be used in pleural fluids exudate and transudate distinction.     

Carcinoembryonic antigen in effusions. A diagnostic adjunct to cytology

A total of 189 effusion specimens (100 benign and 89 malignant) submitted for cytologic examination were assayed for carcinoembryonic antigen (CEA) by an enzyme immunoassay to determine whether the addition of CEA evaluation to cytologic study would improve the diagnostic accuracy for the detection of malignancy. The sensitivity and specificity were 78% and 90%, respectively, for a cytologic diagnosis of malignancy and 68% and 99%, respectively, for a positive CEA (greater than 5 ng/mL). CEA assay was negative in the most common epithelial malignancies of the female genital tract (15 of 17 cases), mesotheliomas (5), lymphomas (7) and alveolar-cell carcinoma of lung (1). CEA assay was positive in 55 of 89 cases of malignancy, including 14 cases with cytologically negative malignant effusions. The CEA assay sensitivity for lung carcinoma (95% for adenocarcinoma, 100% for oat-cell carcinoma and 100% for carcinosarcoma), breast carcinoma (95%), and gastrointestinal carcinoma (100%) were all over 90%. No significant difference in the levels of CEA was noted between gastrointestinal and lung adenocarcinomas. Oat-cell carcinomas and squamous-cell carcinomas had lower values. In cases of an effusion with an unknown primary, an elevated CEA in the fluid is diagnostic of metastatic carcinoma arising from the breast, lung or gastrointestinal tract.     

Malignant melanoma metastatic to the liver. A cytomorphologic comparative study to identify reproducible diagnostic criteria

OBJECTIVE: To establish cytomorphologic criteria that might facilitate the identification of malignant melanoma (MM) cells with epithelioid (nevoid) morphology, in fine needle aspiration biopsy material from the liver. STUDY DESIGN: Aspirated material from 18 cases of MM with epithelioid features and 24 cases of benign liver lesions (BLL) were examined. The cases were selected based on the availability of corresponding tissue biopsies, adequate cell block material or sufficient number of direct smears to perform immunocytochemical staining. The presence or absence of 7 cytologic criteria were reviewed, and the results were evaluated by multivariate regression analysis. RESULTS: All evaluated criteria were significant for identifying MM cells and differentiating them from reactive hepatocytes (P < .001). Uniform atypia, cell dyscohesion, eccentric nuclei and irregular nuclear membranes supported MM, whereas, monolayered sheets or cordlike arrangement; coarse, granular cytoplasm; and occasional transgressing endothelium in true tissue fragments were evidence of BLL. CONCLUSION: A systematic evaluation of the cytomorphologic features described in this study, in conjunction with the clinical and radiologic findings, can be used to render an immediate, confident and accurate diagnosis of MM metastatic to the liver.     

Fine needle aspiration cytology of subareolar abscess of the breast. Spectrum of cytomorphologic findings and potential diagnostic pitfalls

The cytomorphologic findings of eight cases of subareolar abscess of the breast diagnosed by fine needle aspiration (FNA) biopsy are presented. A spectrum of cytologic findings was appreciated, including diagnostic anucleated squames associated with numerous neutrophils, keratinous debris, cholesterol crystals, parakeratosis and strips of squamous epithelium. A foreign-body reaction, with sheets of histiocytes and multinucleated foreign-body-type giant cells, was noted in some of the cases. Potential pitfalls for a false-positive diagnosis of malignancy included the presence of groups of atypical ductal cells, squamous atypia and fragments of exuberant granulation tissue. Seven of the eight cases had complete surgical excision of the lesion, which demonstrated the characteristic findings of dilated lactiferous ducts undergoing squamous metaplasia with rupture and surrounding extensive acute and chronic inflammation with foreign-body reaction. With recognition of the FNA cytologic findings of subareolar abscess of the breast, a specific diagnosis can be rendered, which then allows different treatment modalities depending on the stage of the lesion. Appreciation of the spectrum of cytologic features will enable the pathologist to make the correct diagnosis and thereby avoid potential pitfalls that can possibly lead to a false-positive diagnosis of malignancy.     

Immunocytochemical analysis and cytomorphologic diagnosis on fine needle aspirates of lymphoproliferative disease

Fine needle aspirates were used for the cytologic and immunologic analysis of 21 cases of lymphoproliferative disorders. Immunocytochemical studies performed on Cytospin preparations confirmed the cytomorphologic diagnosis in 19 cases. In one case, the morphology of both aspirates and surgically obtained material showed a reactive pattern while immunologic analyses were inconclusive on both types of material. Immunocytochemistry on tumor material obtained by fine needle aspirations was in agreement with immunohistochemistry on surgical biopsies in 15 of 16 patients with malignant lymphomas. We conclude that immunocytochemical studies performed on Cytospin material in conjunction with the cytologic diagnosis will lead to an increase in diagnostic accuracy as well as providing a means for subclassification of neoplastic lymphoid cells. Moreover, this technique appears to give results comparable to those obtained by histopathologic and immunohistochemical analysis on surgically removed lymph nodes.     

The diagnostic role of glycosaminoglycans in pleural effusions: A pilot study

Background

While modulation of the serotonin transporter (5HTT) has shown to be a risk factor for pulmonary arterial hypertension for almost 40 years, there is a lack of in vivo data about the broad molecular effects of pulmonary inhibition of 5HTT. Previous studies have suggested effects on inflammation, proliferation, and vasoconstriction. The goal of this study was to determine which of these were supported by alterations in gene expression in serotonin transporter knockout mice.

Methods

Eight week old normoxic mice with a 5-HTT knock-out (5HTT-/-) and their heterozygote(5HTT+/-) or wild-type(5HTT+/+) littermates had right ventricular systolic pressure(RVSP) assessed, lungs collected for RNA, pooled, and used in duplicate in Affymetrix array analysis. Representative genes were confirmed by quantitative RT-PCR and western blot.

Results

RVSP was normal in all groups. Only 124 genes were reliably changed between 5HTT-/- and 5HTT+/+ mice. More than half of these were either involved in inflammatory response or muscle function and organization; in addition, some matrix, heme oxygenase, developmental, and energy metabolism genes showed altered expression. Quantitative RT-PCR for examples from each major group confirmed changes seen by array, with an intermediate level in 5HTT +/- mice.

Conclusion

These results for the first time show the in vivo effects of 5HTT knockout in lungs, and show that many of the downstream mechanisms suggested by cell culture and ex vivo experiments are also operational in vivo. This suggests that the effect of 5HTT on pulmonary vascular function arises from its impact on several systems, including vasoreactivity, proliferation, and immune function.     

Causes of iron deficiency anemia in an adult inpatient population. Effect of diagnostic workup on etiologic distribution

The causes of iron deficiency anemia in a population of adults admitted to two Jerusalem hospitals within a period of 7 years were examined. About one half of the 262 patients with iron deficiency anemia were over 70 years old. The ratio of males to females exclusive of young females with menorrhagia was 1:1.8. Despite the combined use of various diagnostic procedures, no definite cause of iron deficiency anemia could be established in 34% of patients. Benign gastrointestinal lesions were found in about one half of the cases in both hospitals. The prevalence of GI neoplasms in hospital B with a more intensive use of endoscopic procedures was significantly higher than in hospital A (18% vs 5%, p less than 0.001). The relative usefulness of barium contrast vs endoscopic studies is illustrated by the fact that 22 diagnoses established by endoscopy were missed by barium studies, whereas only 2 of those established by barium studies were not visualized by endoscopy. A particularly high risk group were anemic males aged 50 to 69 years in whom the prevalence of GI neoplasms was 30%. These data indicate that reliance on traditional contrast radioscopy may result in misdiagnosis of a high proportion of gastrointestinal neoplasms.     

Tuberculosis of the breast. A cytomorphologic study

OBJECTIVE: Extrapulmonary tuberculosis occurring in the breast is rare despite the fact that 1-2 billion people worldwide suffer from tuberculosis. The aim of this study was to examine the cytomorphology of breast tuberculosis (breast TB) and to review the literature. STUDY DESIGN: Old records from the Cytopathology Laboratory, All India Institute of Medical Sciences, were reviewed from January 1980 to December 1998. Cases of breast TB where a cytologic diagnosis was rendered or a histologic diagnosis with prior fine needle aspiration cytology (FNAC) was available were selected. These slides were reviewed for determining the cytologic findings. RESULTS: One hundred sixty cases of breast TB were included in the study. Six males and 154 females with a clinical suspicion of carcinoma had undergone FNA that was reported as TB. The majority of the patients (111) were in the reproductive age group, 21-40 years. Of the 160 cases, 118 (73.75%) had cytomorphology diagnostic of tuberculosis--epithelioid cell granulomas with caseous necrosis. Eleven of the remaining 42 cases were positive for acid-fast bacilli (AFB) on Ziehl-Neelsen (ZN) staining, while 31 cases were confirmed to be tubercular on histology. ZN staining was done in 44 cases, and AFB were demonstrated in only 38.6% of cases. CONCLUSION: Up to 73% of breast TB can be confidently diagnosed when both epithelioid cell granulomas and necrosis are present. Also, the possibility that a woman in the reproductive age group who presents with a palpable lump in the breast may have tuberculosis must be kept in mind, especially as the incidence of breast TB may increase in the future with the global spread of AIDS.     

Early diagnosis of mesothelioma in serous effusions using AgNOR analysis

OBJECTIVE: To compare the results of conventional cytology, DNA image cytometry, immunocytochemistry and argyrophilic nucleolar organizer region (AgNOR) analysis for the diagnosis of malignant cells in serous effusions. STUDY DESIGN: One hundred twenty effusions, 40 with carcinoses, 40 with malignant mesotheliomas and 40 without tumor cells on follow-up were studied by conventional cytology and three adjunctive methods. RESULTS: Unequivocal tumor cells were detected in 92.5% of effusions due to carcinoses and in 45% due to mesotheliomas. Applying immunocytochemistry with BerEP-4 positivity and DNA image cytometry with aneuploidy as a marker revealed 100% of carcinoses and 71.7% of mesotheliomas. Applying the experimentally found thresholds of 2.5 AgNORs as "satellites" and 4.5 AgNORs as "satellites and clusters" together as mean values per nucleus resulted in a 95% correct rate of mesothelioma and 100% rate of carcinoma cell identification without false positive diagnoses. CONCLUSION: AgNOR analysis may be a useful adjunct to other methods in the routine diagnosis of malignant serous effusions. It seems to be the most sensitive method in early cytologic diagnosis of mesotheliomas in effusions. Seventy-three percent of malignant mesotheliomas were diagnosed cytologically at first on effusions. Forty-seven percent of patients with malignant mesotheliomas were identified at the early tumor stage T1 N0 M0.     

NK cell immunophenotypic and genotypic analysis of infants with severe respiratory syncytial virus infection

Respiratory syncytial virus (RSV) is the leading cause of severe lower respiratory tract infection in infants. Reduced numbers of NK cells have been reported in infants with severe RSV infection; however, the precise role of NK cells during acute RSV infection is unclear. In this study the NK and T cell phenotypes, LILRB1 gene polymorphisms and KIR genotypes of infants hospitalized with RSV infection were analyzed. Compared to controls, infants with acute RSV infection showed a higher proportion of LILRB1+ T cells; in addition, a subgroup of infants with RSV infection showed an increase in LILRB1+ NK cells. No differences in NKG2C, NKG2A, or CD161 expression between RSV infected infants and controls were observed. LILRB1 genotype distribution of the rs3760860 A>G, and rs3760861 A>G single nucleotide polymorphisms differed between infants with RSV infection and healthy donors, whereas no differences in any of the KIR genes were observed. Our results suggest that LILRB1 participates in the pathogenesis of RSV infection. Further studies are needed to define the role of LILRB1+ NK in response to RSV and to confirm an association between LILRB1 polymorphisms and the risk of severe RSV infection.     

High-performance capillary electrophoretic analysis of hyaluronan in effusions from human malignant mesothelioma

A procedure to quantify hyaluronan in effusions from human malignant mesothelioma using a highly sensitive and reproducible high-performance capillary electrophoresis (HPCE) method is presented. Following ethanol precipitation, hyaluronan and galactosaminoglycans were degraded to Δ^4.5-dissacharides with a mixture of chondroitinases ABC and AC. Heparan sulphate and proteins/glycoproteins were separated by ultrafiltration on a Centricon 3 membrane, and hyaluronan-derived disaccharides were analysed by direct injection of the filtrate into a HPCE system. Determination of hyaluronan in effusions from five healthy individuals and three patients with mesothelioma gave values comparable to those found using the HPLC method. One of the advantages of the HPCE method as compared to HPLC is the low solvent consumption. The much lower detection limit (attomole level) of the HPCE method may also allow the analysis of hyaluronan content in serum. The contribution of HPCE in diagnosis of a neoplasm, such as human malignant mesothelioma, illustrates the great potential of this technique in the field of life sciences.     

Castleman's disease: cytomorphologic and cytochemical features of a case

The cytomorphologic features of a case of giant lymph node hyperplasia, hyaline-vascular type, obtained by fine needle aspiration is described. Cytochemical studies of the vascular structures present in the aspirate suggest their origin as capillaries.     

Evaluation of proposed cytomorphologic criteria for the diagnosis of Chlamydia trachomatis in Papanicolaou smears

To evaluate the proposed cytomorphologic criteria for the cervical cytologic diagnosis of Chlamydia trachomatis infection, a study was made of 171 endocervical smears. All cytomorphologic elements that could be ascribed to Chlamydia trachomatis infection were correlated with the diagnostic confirmation of this microorganism by monoclonal antibody (MAb) staining. The presence of Chlamydia trachomatis was detected in 21 samples (12.28%) by MAb staining. Comparing the cytomorphologic results with the MAb results, the sensitivities and specificities of the Papanicolaou smear diagnoses were 19% and 86% using the cytologic criteria proposed by Gupta and coworkers, 38% and 87% using the criteria proposed by Kiviat and coworkers and 23% and 91% using the criteria proposed by Shiina. In view of (1) its low sensitivity, (2) the subjective elements and individual variations in the proposed cytologic criteria, (3) the similarity with Trichomonas vaginalis-produced exudates and (4) the implications of a misdiagnosis of a sexually transmitted disease, it is concluded that cervical cytology is not useful for ascertaining the presence of Chlamydia trachomatis.     

Heparanase expression: a potential ancillary diagnostic tool for distinguishing between malignant cells and reactive mesothelium in body cavity effusions

OBJECTIVE: Heparanase, an endoglycosidase that cleaves heparan sulphate, is frequently expressed in carcinomas and was suggested to play a role in cell invasion and metastasis. We investigated whether heparanase expression may serve as a reliable marker to discriminate benign mesothelial cells from malignant cells shed into body cavities. METHODS AND RESULTS: Cytological smears of effusions from 51 hospitalized patients were immunostained for heparanase. Strong immunoreactivity was noted in 35 of 40 (88%) carcinoma samples and in all three malignant mesothelioma cases. Only rare (<3%) reactive mesothelial cells were noted showing a faint negligible staining. Specificity was 100%, sensitivity 88%, and positive and negative predictive values were 100% and 89% respectively. CONCLUSIONS: Our results suggest that heparanase may be of value as a complementary component in a diagnostic panel of markers, contributing to its reliability and accuracy.