Adriana Esther Rocher

Utilidad de la técnica de AgNOR en la interpretación de los derrames de cavidades serosas

Background: AgNOR technique detects, using silver salts, argyrophylic proteins of the nucleolar organizer region (NOR). The number and size of NOR reflect cell activity, proliferation and transformation and may help to differentiate benign from malignant cells. Aim: To assess the value of AgNOR assay to differentiate reactive mesothelial cells from malignant cells in serous effusions. Material and methods: Thirty one fluids obtained from 16 pleural, 14 peritoneal and one pericardial effusion, were studied. The fluids were processed with Giemsa and Papanicolau stains and with the AgNOR technique. The number of AgNOR dots were counted (only when it was possible to distinguish each individual dot) and the mean value per nucleus was calculated for each smear. Results: Mesothelial cells had a mean of 4,88 ± 1,5 dots compared with 13,78 ± 3,88 dots in the malignant cells (p<0,001). Conclusions: AgNOR assay can be useful for the differentiation of benign and malignant cells in serous effusions. (Rev Med Chile 2000; 128: 963-8)

Usefulness of ploidy, AgNOR and immunocytochemistry for differentiating benign and malignant cells in serous effusions.

Objective:  The objective of this study was to establish the value of different markers in differentiating reactive mesothelial cells from metastatic adenocarcinomatous cells in serous effusions (SE).

New Biochemical Parameters in the Differential Diagnosis of Ascitic Fluids

Background In the cases of ascitis, it is essential to determine their origin using the parameters obtained by the cytological and biochemical examinations. The aim of this study was to evaluate the usefulness of different biochemical markers and the number of cells in the differential diagnosis of ascitic fluid (AF). Methods One hundred ninety-one cases of AF were studied, who were admitted to the hospital from January 01, 2009 to December 31, 2014. One hundred fifty-two of them were included in the analysis, and the remaining 39 were excluded because they had more than one associated pathology, clotted or hemolyzed. Results The more frequent etiologies of AF were the cirrhosis (29%), the infections (22%) and the neoplasies (19%). Other pathologies reached 16%. Cutoff > 300 cells/mm3 detected the 78% of exudates. The AF/serum (S) of aspartate aminotransferase (AST) (> 0.5), lactate dehydrogenase (LDH) (> 0.6), proteins (PT) (> 0.5), cholesterol (COL) (> 0.4), and alanine aminotransferase (ALT) (> 0.5) correctly detected 80%, 78%, 72%, 70% and 70% of the exudates, respectively. Conclusion We proposed the utilization of a new cutoff of cellular counting, major of 300/mm3, since it would allow improving the detection of exudate ascites, without including the transudate ascites. AST AF/serum ratio (AF/S) showed the major usefulness in the differentiation and characterization of AF; LDH, proteins, cholesterol and ALT might be also acceptable in the above mentioned differentiation. The serum-ascites albumin gradient (SAAG) turned out to be a good marker of portal hypertension associated with cirrhotic processes. Creatine kinase (CK), alkaline phosphatase (ALP), amylase (AMI), total bilirubin (TB), triglycerides (TG) and glucose (GLU) did not allow differentiating exudates from transudates.

Symptomatic BK virus infection in an immunocompetent child diagnosed on urine cytology

Dear Editor, We report here a rare case of transient symptomatic BK virus infection in a 4-year-old immunocompetent child. Although not likely to be confused with neoplasia in this age group, the infection, when diagnosed on urine cytology, should be recognized as a sign of virus activation; as in this case, it is usually self-limiting. A 4-year-old male child presented with otitis media and increased nasopharyngeal secretion. After treatment with antibiotics for 1 week, his condition deteriorated and the patient showed dysuria and aesthenia. A renal echography was performed and showed a slight enlargement of the renal calyx. A study of the urinary sediment (without staining) showed large numbers of isolated cells with large nuclei; some of the cells had two or three nuclei. The urine did not show oval fat bodies, epithelial cylinders or renal tubular cells. Urine culture was negative. Serum creatinine levels fell within the normal range. Thirty millilitres of urine were centrifuged at 225 g for 10 minutes; several slides were prepared, fixed in 95% ethanol and stained with the Papanicolaou technique. The tests were repeated three more times during the week. The smears showed mononucleated cells, the majority with large nuclei of ground-glasslike appearance showing a large basophilic inclusion occupying almost all the nucleus (Decoy cells: Figure 1a). After destaining of the same slides, the presence of human polyomavirus antigen in the nuclei of infected exfoliated cells was demonstrated by immunocytochemical staining with mouse monoclonal antibody for the detection of polyomavirus capsid protein VP1 (Santa Cruz Biotechnology Inc, Santa Cruz, California, USA). After adding avidin–biotin–peroxidase complexes, the sections were developed with a diaminobenzidine substrate. Some cells showed brown (positive) nuclei (Figure 1b). One week after the acute episode, when the infection was receding, the cells presented cytoplasmatic eosinophilic inclusions (Figure 2). Although Decoy cells had been previously described in urine sediments, it was not until 1971 that they were identified as polyomavirus by Gardner et al. and Coleman et al. Two types of polyomavirus were discovered and they were named with the initials of the patients: BK (BKV) and JC (JCV). BKV was found in the urine of a patient with renal transplantation and JCV in the brain of a patient with progressive multifocal leukoencephalopathy; these types of polyomavirus differ in the size of the viral particles and in the serology. More than 80% of the adult population has had a primary infection, which remains in a latent state. The virus genome sequence can be detected by PCR in the kidneys and urinary tract (BKV and JCV), cerebral tissue (JCV) and very rarely in other organs. BKV infection is usually asymptomatic in children and is located in the kidneys and urinary tract. Occasionally the virus can be activated, either without apparent cause, or following immunosuppression due to chemotherapy, transplantation or AIDS. Cells containing activated virus are excreted in urine and must be differentiated from neoplastic cells or cells affected by chemotherapy. Malignant urothelial cells may show a large and dark nucleus and in general have irregular chromocentres with areas of irregular heterochromatin. In contrast to malignant cells, polyomavirus-infected cells show characteristic basophilic amorphous inclusions that occupy the whole nucleus, obscuring the chromatin structure (Decoy cells). In cases where differential diagnosis using only cytology is doubtful, PCR techniques in urine and immunocytochemistry with antibody anti-SV40has have been applied with good results. Decoy cells are found in the urine of transplant recipients, because of the association of BK virus with renal dysfunction following immunosuppression. Although BK-associated nephropathy develops in only 2–5% of renal transplant recipients, its prognosis when present is very poor, with irreversible graft failure developing in 45% of affected patients. Childhood cases of BKV are due to primary infection when there is no evidence of immunosuppression. The infected cells persist for 2 or 3 weeks and Correspondence: Professor Dr L. A. Palaoro, Avda Forest 1318, 4o B (1427), C.A.B.A., Argentina Tel.: 5411-4552-0449; Fax: 5411-45019008; E-mail: luispalaoro@yahoo.com.ar

Usefulness of cytology in the diagnosis of molluscum contagiosum on skin papules.

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Bronchoalveolar Wash: Proposal of a Cutting Point for Lactate Dehydrogenase in the Differential Diagnosis of Pulmonary Diseases

Background: The analysis of bronchoalveolar lavage (BAL) is of great clinical utility in the diagnosis of pulmonary diseases (PDs). The cytological study (total and differential cell count) performed routinely in these samples, has a high orientative power and is diagnostic in some cases. The study of soluble substances has provided little information. This study aimed to determine the cut-off point of lactate dehydrogenase (LDH) activity in the diagnosis of different lung diseases. Methods: Two hundred forty-three BALs were selected from a total of 306 patients with a single respiratory disease: acute pneumonia due to common germs (NCG, n = 126), tuberculosis (TB, n = 20), mycotic pneumonia (MN, n = 12, N = 37) and interstitial diseases (IDs, n = 48). Cytological study and measurement of LDH activity were performed. The mean, standard deviation, sensitivity (S), specificity (Sp) and Youden index (YI) of this enzyme were determined. Student’s parametric and non-parametric Mann-Whitney tests were done (P < 0.05: significant). Results: Comparing the means and standard deviations of LDH in the different PDs, a significant increase of this parameter was observed in the NCG compared to the other PDs: NCG vs. TB (P = 0.003); MN, MD and ID (P < 0.0001). Based on these significant differences observed, the cut-off point for LDH in NCG was evaluated. Different values were analyzed: LDH: ≥ 150 IU/L with S: 70%, Sp: 85% and YI: 0.55; ≥ 130 IU/L with S: 77%, Sp: 80% and YI: 0.57; ≥ 120 IU/L with S: 80%, Sp: 77% and YI: 0.57 and ≥ 100 IU/L with S: 86%, Sp: 74% and YI: 0.60. Conclusions: It is proposed to perform the measurement of LDH activity for the differential diagnosis of NCG in the PD, since its mean value was significantly higher than the rest of the PD, using a cut-off point of LDH ≥ 100 IU/L; it showed a higher S and YI for the diagnosis of NCG screening. Increased LDH activity in NCG could be associated with the high number of leukocytes present in this pathology, superior to the rest of the PD. Measurement of LDH activity along with cell count would contribute to the diagnosis of PD.

Urothelial cells in smears from cervix uteri.

Objectives: To establish the cytological criteria to identify the urothelial cells in cervical smears in order to avoid mistakes in the cytological diagnosis. Materials and Methods: Cervical smears from 34 post menopausal women with vesicovaginal fistulas, advanced bladder prolapse and genital erosive lichen planes (vulvar kraurosis) (Group 1) and transitional cell metaplasia of the cervix (TCM, Group 2) were stained with Papanicolaou technique. The cervical samples were taken during the routine annual examination for prevention of the uterine cancer. Results: The smears of cervix from Group 1 showed urothelial cells from the three layers of the transitional epithelium. The umbrella cells are the bigger ones with relatively large nuclei. Frequently, they are multinucleated with single or multiple nucleoli and a typical “frothy” cytoplasm (cytoplasmic vacuoles). The cells of the Group 2 showed nuclei with oval to spindled shapes, some tapered ends, less cytoplasm than squamous metaplastic cells, powdery chromatin, small nucleoli and nuclear grooves. Conclusions: The umbrella cells may be mistaken for dysplastic cells originating in low grade squamous intraepithelial lesions lesions (LSILs) due to their nuclear and cytoplasm sizes. Therefore, it is important to know the possibility of their appearance in the cervical smears, especially in post menopausal patients in order to avoid a false diagnosis of an intraepithelial lesion. It is unlikely that deeper cells of urothelium would be confused with high grade squamous intraepithelial lesion (HSIL) cells. However, their presence might be a reason of mistake in the diagnosis. TCM is an under-recognized metaplastic phenomenon of the cervix and vagina, which is a mimicker of high-grade squamous intraepithelial lesion. The differential characteristic between umbrella cells, cells from TCM and the deeper urothelial cells, and LSIL and HSIL are detailed in the present paper.