Rosario Granados

Bethesda classification of cervicovaginal smears: Reproducibility and viral correlates

Fifty-five cervicovaginal smears from women with squamous intraepithelial lesions (SILs) were independently evaluated on two separate occasions by four cytopathologists using a binary classification system (the Bethesda system). Smears were categorized as low-grade (LSIL) or high-grade (HSIL) using previously published criteria. All women had subsequent cervical biopsies containing human papillomavirus (HPV) DNA amplified with the polymerase chain reaction and typed by restriction fragment polymorphism analysis. Three or more observers agreed on classification in 49 of 55 cases (87%); unanimous diagnoses were rendered in 31 cases (56%). Interobserver and intraobserver reproducibility ranged from fair to near-excellent (kappa values 0.40 to 0.63; 0.63 to 0.74, respectively). HPV types included HPV 16 (27%), 18 (7%), 31 (9%), 35 (4%), 39 (4%), 6 (10%), 11 (2%), novel types (30%), and multiple types (4%). High-risk HPV types (16, 18, 31, 35, and 39) were significantly associated (P = .03) with consensus HSIL diagnoses (agreement of three or more observers). This was primarily because of the strong association of HPV 16 with HSIL (P = .001). After excluding HPV 16, the other high-risk HPV types (18, 31, 35, and 39) were no longer significantly associated with consensus HSIL diagnoses (P > .5). Conversely, LSIL diagnoses were significantly associated with non-high-risk HPV types (all HPV types except 16, 18, 31, 35, and 39; P = .006). Binary cytological classification of cervicovaginal SILs is reproducible among cytopathologists. Such classification correlates well with most low-risk HPV types and with the prototypic high-risk HPV 16 but not with other high-risk HPV types.

Classification of renal proteinuria: a simple algorithm.

Abstract Total protein, albumin, α1-microglobulin, and immunoglobulin G (IgG) were analyzed in 1622 urine samples without Bence-Jones proteinuria or gross hematuria. There was correlation with the histological picture obtained on renal biopsy in 61 patients. We established 24-h reference intervals for α1-microglobulin and IgG on 659 urine samples with total protein and albumin excretion rates below 100 mg/24 h and 30 mg/24 h, respectively, and creatinine clearance above 80 ml/min. The central 95% reference interval was found to be between 4 and 17 mg/24 h for α1-microglobulin and between 3 and 8.5 mg/24 h for IgG. In 80 urine samples with albumin excretion rate above 30 mg/24 h and α1-microglobulin and IgG within their reference intervals, we analyzed the 95% central interval of the distribution of the IgG/albumin ratios, and it was found to be within 0.01 and 0.20 (0.90 confidence interval: 0.17–0.24). Proteinuria was considered to be of the selective glomerular type if the albumin excretion rate was abnormal and the IgG/albumin ratio was under 0.20, even when the IgG excretion was within a pathological range. For the classification of proteinuria as predominantly tubular, we estimated the α1-microglobulin/albumin ratio in 173 urine samples with normal excretion rates of albumin and IgG and pathological excretion of α1-microglobulin. The discriminating value of 0.91 (0.90 confidence interval: 0.78–1.08) was accepted in order to define proteinuria of a tubular origin in the presence of a pathological albumin excretion rate. The association between albumin and IgG excretion rates and tubular reabsorption of the α1-microglobulin normally filtered by the glomerulus was studied in 33 urine samples from patients with no histologically significant tubulo-interstitial or vascular disease and a serum creatinine concentration below 141 μmol/l. The optimal curve-fitting function between albumin plus IgG and α1-microglobulin excretion rates was of the quadratic type (r=0.927). Mixed proteinuria was considered when both, albumin and α1-microglobulin excretion rates were pathological and could not be included in the previously described groups.

Applying the Paris System for Reporting Urine Cytology Increases the Rate of Atypical Urothelial Cells in Benign Cases: A Need for Patient Management Recommendations

The Paris System (TPS) for reporting urinary cytology attempts to unify the terminology in this field. Objectives: To analyze the impact of adopting TPS by measuring nomenclature agreement and cytohistological correlation. Materials and Methods: Voided urine liquid-based cytology samples corresponding to 149 biopsy-proven cases (76 high-grade carcinomas, 40 low-grade carcinomas, and 33 benign lesions), were reclassified by the same pathologist using TPS. Diagnostic agreement and sensitivity for both nomenclature systems was measured. Results: When using TPS, the rate of atypical samples increased 8 times (from 3 to 24.2%) in benign cases, 10 times (from 2.5 to 25%) in low-grade carcinomas, and 2.4 times (from 6.6 to 15.8%) in high-grade carcinomas. The false-positive rate (abnormal cytology in negative or low-grade carcinoma cases) increased from 11 to 34.2%. Sensitivity was higher (63 vs. 49%) with TPS at the expense of a lower specificity (73 vs. 91%). The agreement between both nomenclatures was moderate for negative and high-grade carcinoma cases (k = 0.42 and 0.56, respectively) and weak for low-grade tumors (k = 0.35). Conclusions: Adopting TPS for reporting urine cytology results in a considerable increase in atypical diagnoses, improving sensitivity but lowering specificity. Appropriate management recommendations for patients with an atypical cytological diagnosis are required.

Kidney histopathological findings in fatal pandemic 2009 influenza A (H1N1)

Dear Editor, A new pandemic was originated by a novel influenza A (H1N1) virus [1– 3]. Severe cases were characterized by acute respiratory distress syndrome (ARDS), shock, and acute kidney injury (AKI) [3]. Lung histopathological changes in fatal cases showed signs of diffuse alveolar damage, necrotizing bronchiolitis, and occasional alveolar hemorrhage [2]. However, histopathological changes in organs other than the lungs are not known. Here we report kidney histopathological findings and describe for the first time the specific kidney cell type targeted by pandemic 2009 influenza A (H1N1) virus infection. With the approval of our Ethics Committee and with closest relative informed consent, renal biopsies from four patients who died in the intensive care unit (ICU) with diagnosis of confirmed influenza A (H1N1) virus infection were studied by microscopy after hematoxylin and eosin (HE), Masson’s or periodic acid-Schiff (PAS) staining. Cell nuclei were revealed by staining with 4’,6-diamidino-2-phenylindole (DAPI). Localization of viral antigen and specific kidney cells was carried out by double immunofluorescence (IF) labeling [4] using antibodies (Santa Cruz) specific for either: (1) aquaporin 1, a marker of proximal tubular cells; (2) CD10, a marker of proximal tubular cells; (3) cytokeratin 7, a marker of distal tubular cells; or (4) CD34, a marker of endothelial cells, and a rabbit antiserum specific for influenza nucleoprotein (NP). This antibody was generated by immunization of rabbits with purified recombinant NP and validated by IF, Western blotting, and immunoprecipitation of control and influenzainfected human cells [5]. This antibody is cross-reactive with several influenza A virus subtypes (data not shown). Secondary antibodies were fluorescein isothiocyanate (FITC)labeled goat anti-mouse immunoglobulin G (IgG) (Santa Cruz) and Alexa 546-conjugated goat anti-rabbit IgG. Sections were studied under confocal microscopy (Leica SP5), and single optical sections are presented. Only cases 3 and 4 were diagnosed with AKI. Cases 3 and 4 had focal changes consistent with acute tubular necrosis (ATN) in the distal tubules (epithelial cell swelling, individual cell necrosis, and shedding of

Cytological Diagnosis of Bilateral Breast Implant‐Associated Lymphoma of the ALK‐Negative Anaplastic Large‐Cell Type. Clinical Implications of Peri‐Implant Breast Seroma Cytological Reporting

The cytological examination of peri‐prosthetic breast effusions allowed the diagnosis of bilateral breast‐implant ALK‐negative anaplastic large cell lymphoma (BI‐ALCL) in the case reported. Ten years after reconstructive surgery with bilateral breast implants, a large unilateral seroma developed and was cytologically analyzed. The presence of CD30 and CD4‐positive large‐sized atypical lymphoid cells exhibiting horseshoe‐shaped nuclei and a brisk mitotic activity rendered the diagnosis of BI‐ALCL. Similar cells were seen in the peri‐prosthetic fluid intraoperatively collected from the contralateral breast. Although initial histological analysis of the capsulectomy specimens showed unilateral tumor, the cytological findings prompted a more thorough tissue sampling, resulting in the diagnosis of bilateral disease. BI‐ALCL usually follows an indolent clinical course; however, there are reported cases with an aggressive behavior. While the presence of bilateral disease is a putative risk factor for a bad prognosis, the small number of cases reported precludes a definitive assessment of this risk. Since most BI‐ALCL present with late seromas, cytologic analysis of these effusions in women with breast implants should be mandatory. Cytology is a safe tool for diagnosis and follow‐up of patients with breast implant‐related late seromas, sometimes proven more sensitive than histological analysis. Complete bilateral capsulectomy and a detailed histological analysis should follow a cytological diagnosis of BI‐ALCL in a breast effusion in order to avoid false negative diagnoses. Our case constitutes the first published report of a bilateral BI‐ALCL diagnosed by cytology. Diagn. Cytopathol. 2016;44:623–627.

Flow cytometric DNA analysis on fine needle aspiration biopsies of liver lesions

Flow cytometric DNA analysis on fine needle aspiration biopsies of liver lesions

Increased risk of malignancy for non‐atypical urothelial cell groups compared to negative cytology in voided urine. Morphological changes with LBC

Liquid‐based cytology (LBC) has recently become the preferred method for urine cytology analysis, but differences with conventional cytology (CC) have been observed. The purpose of this study is to analyze these differences and the clinical relevance of non‐atypical urothelial cell groups (UCG) in voided urine specimens. Reporting terminology is discussed. Initially, diagnostic categories from 619 LBC and 474 CC samples, reviewed by five different pathologists, were compared (phase 1). Five years after LBC was implemented and applying strict cytologic criteria for UCG diagnosis, 760 samples were analyzed (phase 2) and compared to previous LBC specimens. Diagnostic differences, interobserver variability and clinicopathological correlation with a 6‐month follow‐up, were analyzed. UCG increased from 6.5% with CC to 20.7% (218%, 3.2 fold, P < 0.0001) with LBC. This difference was not related to interobserver variability. Five years later, the rate of UCG had decreased to 13 2%. While 6% of cases with a negative cytology had urothelial carcinoma (UC) within 6 months of diagnosis, this percentage increased to 15.7% with UCG. The sensitivity of the UCG category for UC was low (30.4%), but the specificity and the negative predictive value (NPV) were high (87.1% and 94%, respectively). LBC increases UCG when compared to CC. This can be corrected with observeŕs experience and using set cytological criteria. Due to its association with carcinoma, the presence of UCG in voided urine should be framed in a diagnostic category other than “negative for malignancy.” Diagn. Cytopathol. 2016;44:582–590.

Cervical cancer screening cotesting with cytology and MRNA HPV E6/E7 yields high rates of CIN2+ lesions in young women

European guidelines recommend primary HPV testing for cervical cancer screening. However, the starting age remains to be defined, with an undecided window between 30 and 35 years. This pilot study compares the effectiveness of primary HPV testing to that of cytology for the detection of high‐grade (CIN2+) lesions stratified by age.

0895. Identification and validation of a mirna as a diagnostic biomarker of diffuse alveolar damage in an animal model of acute lung injury and adult respiratory distress syndrome in mechanically ventilated patients

0895. Identification and validation of a mirna as a diagnostic biomarker of diffuse alveolar damage in an animal model of acute lung injury and adult respiratory distress syndrome in mechanically ventilated patients P Cardinal-Fernandez, A Ferruelo, N Rego, Y Rojas, A Ballen-Barragan, R Granados, C Jaramillo, E Lopez-Hernandez, L Martinez-Caro, N Nin, R Herrero, MA de la Cal, A Esteban, JA Lorente