Importancia de los macrófagos cargados con hemosiderina en el diagnóstico de la hemorragia alveolar difusa / Importance of hemosiderin-loaded macrophages in diagnosis of diffuse alveolar hemorrhage / A importância dos macrófagos carregados com hemosiderina no diagnóstico de hemorragia alveolar difusa / Reconocimiento a la trayectoria del Prof. Dr. Juan Miguel Castagnino†

Resumen La hemorragia alveolar difusa (HAD) es un síndrome clínico con una alta mortalidad que compromete la función respiratoria. Su diagnóstico se basa en pruebas clínicas, radiológicas y citológicas. El objetivo del trabajo fue ratificar el valor de referencia de hemosiderófagos en lavados broncoalveolares (BAL) (hemosiderófagos ≥20%), correlacionar con la etiología y definir las condiciones preanalíticas para que la reacción de Perls alcance valores elevados de sensibilidad. De 109 muestras de pacientes con sospecha de HAD, se analizaron 90 por cumplir los criterios de inclusión; 36 resultaron positivas para HAD, 3 falsamente negativas y 51 resultaron negativas. La sensibilidad fue de 92% y la especificidad de 100%. La mediana de hemosiderófagos para muestras con diagnóstico de HAD fue de 70%. Se agruparon según la etiología: procesos infecciosos puros (PI), enfermedades autoinmunes puras (EA), enfermedades neoplásicas puras (EN), enfermedades autoinmunes más procesos infecciosos (EA+PI), enfermedades neoplásicas más procesos infecciosos (EN+PI), misceláneas (MI). La mediana de hemosiderófagos para cada grupo fue: PI (n=7) 50%, EA (n=15) 58%, EN (n=6) 73%, EA+PI (n=5) 80%, EN+PI (n=4) 80%, MI (n=2) 45% (p=0,57). El porcentaje de pacientes fallecidos fue de 49% (n=19), con una mediana de hemosiderófagos de 70%, en comparación con la de pacientes no fallecidos de 64% (p=0,25). Se ratificó el valor de referencia para establecer el diagnóstico de HAD en muestras de BAL obtenidas luego de las 36 h de comenzados los síntomas utilizando la reacción de Perls, la cual demuestra una alta sensibilidad y especificidad para dicho diagnóstico.

Abstract Difusse alveolar hemorrhage (DAH) is a clinical syndrome with high mortality. Its diagnosis is based on clinical, radiological and cytological tests. The objective of this study was to ratify the reference value of hemosiderophages in bronchoalveolar lavages (BAL) (hemosiderophagues ≥20%), to correlate with the etiology and define the pre-analytical conditions for the Perls reaction to reach high sensitivity values. Out of the 109 samples from patients with suspected ADH, 90 were analysed for meeting the inclusion criteria; 36 were positive for HAD, 3 were false negatives, and 51 were negative (sensitivity 92%; specificity 100%). The median number of hemosiderophagues for samples with a diagnosis of ADH was 70%; they were grouped according to etiology: pure infectious processes (PI), pure autoimmune diseases (AD), pure neoplastic diseases (ND), autoimmune diseases plus infectious processes (AD + PI), and miscellaneous (MI). The median number of hemosiderophagues for each group was: PI (n=7) 50%, AD (n=15) 58%, ND (n=6) 73%, AD + PI (n=5) 80%, ND + PI (n=4) 80%, MI (n=2) 45% (p=0.57). The percentage of deceased patients was 49% (n=19), with a median hemosiderophague of 70%, compared with 64% of non-deceased patients (p=0.25). The reference value to establish the diagnosis of ADH in BAL simples obtained 36 hours after the beginning of symptoms using the Perls reaction was ratified, which shows a high sensitivity and specificity to make the diagnosis of ADH.

Resumo A hemorragia alveolar difusa (HAD) é uma síndrome clínica com alta mortalidade que compromete a função respiratória. Seu diagnóstico se baseia em testes clínicos, radiológicos e citológicos. O objetivo do trabalho foi ratificar o valor de referência de hemossiderófagos em lavagens broncoalveolares (LBA) (hemossiderófagos ≥20%), relacioná-los com a etiologia e definir as condições pré-analíticas para que a reação de Perls alcance valores elevados de sensibilidade. De 109 amostras de pacientes com suspeita de HAD, 90 foram analisadas para cumprir com os critérios de inclusão; 36 resultaram positivas para HAD, 3 foram falsos negativos e 51 resultaram negativas. A sensibilidade foi de 92% e a especificidade de 100%. A média de hemossiderófagos para amostras com diagnóstico de HAD foi de 70%, eles foram agrupados de acordo com a etiologia: processos infecciosos puros (PI), doenças autoimunes puras (DA), doenças neoplásicas puras (DN), doenças autoimunes mais processos infecciosos (DA+PI), doenças neoplásicas mais processos infecciosos (DN+PI), miscelâneas (MI). A média de hemossiderófagos para cada grupo foi: PI (n=7) 50%, DA (n=15) 58%, DN (n=6) 73%, DA+PI (n=5) 80%, DN+PI (n=4) 80%, MI (n = 2) 45% (p= 0,57). A porcentagem de pacientes falecidos foi de 49% (n=19), com uma média de hemossiderófagos de 70%, em comparação com 64% de pacientes não falecidos (p=0,25). Foi ratificado o valor de referência para estabelecer o diagnóstico de HAD em amostras LBA obtidas 36 horas após o início dos sintomas através da reação de Perls, que apresenta alta sensibilidade e especificidade para esse diagnóstico.

Multiparametric Flow Cytometry versus Conventional Cytology in the Study of Leptomeningeal Involvement in Malignant Hematological Diseases.

BACKGROUND: CNS involvement is a complication in hematologic malignant neoplasms. The advantage of multiparametric flow cytometry (MFC) over conventional cytology (CC) in detecting occult leptomeningeal disease in CSF has been proven previously, as reported in the literature. In this study, we reviewed the experience of our laboratory in evaluating CSF specimens by MFC and CC after refinement of technical procedures. METHODS: MFC analysis was performed in 159 specimens. In 91 specimens, simultaneous CC and MFC analysis was requested and results compared. RESULTS: Neoplastic cells were identified in 27 (17.0%) of the total samples and in 17 (18.7%) of the paired specimens group by MFC, compared with 2 (2.2%) specimens with positive results as determined by CC. MFC enabled identification of malignant cells in low-cellularity specimens (<5 cells/µL) and all neoplasm categories. CONCLUSION: MFC allowed the detection of minimal numbers of tumor cells in CSF specimens from individuals with leukemia and lymphoma in whom CC had not been able to identify those tumor cells.

von Hansemann Cells from Fresh Urine Sediment Samples in the Diagnosis of Malakoplakia.

BACKGROUND: Malakoplakia is characterized by the presence of plaques with macrophages containing inclusion bodies. The diagnosis of this disease is carried out by biopsy of the lesion. The objective of this paper was to assess the value of fresh urine sediment in the diagnosis of malakoplakia. MATERIALS AND METHODS: Five suspected cases of malakoplakia that showed macrophages with inclusions called bodies of Michaelis-Gutmann (von Hansemann cells) in unstained urine sediment were processed with Papanicolaou, Giemsa, and periodic acid-Schiff (PAS) stains. Four of the five patients had a history of cystitis and had developed antibiotic resistance. The other patient had the characteristics cells in a routine urinalysis. RESULTS: Papanicolaou stain revealed intracytoplasmic eosinophilic or basophilic bodies, single or multiple in macrophages. Such bodies were stained deep red with PAS technique. Giemsa stain showed these bodies with a faint basophilic coloration, sometimes with a central core. Bladder biopsies established the definitive diagnosis, showing bodies within and outside macrophages, with a concentric "birds-eye" or "owl-eye" (targetoid) appearance. CONCLUSIONS: Finding of von Hansemann cells in fresh urine sediment of patients with cystitis and a history of resistance to antibiotic scan leads to the diagnosis of malakoplakia. Giemsa stain can show in some cases the characteristic central core of Michaelis-Gutmann bodies. Malakoplakia is probably the result of an acquired defect in macrophage function causing impairment of bactericidal activity. A correct diagnosis is important because the spread to ureters with bilateral stenosis and obstruction can lead to kidney failure.

Use of Sialic Acid as a Complementary Method in the Cytological Diagnosis: Preliminary Study.

INTRODUCTION: Cellular morphology does not allow, in many cases, to safely establish a diagnosis of malignancy or benignity. Sialic acid is found in the membranes of well-differentiated mature cells, normally located in the alpha-2,3 position. During tumor progression, changes occur in glycosylation of proteins and lipids, including alterations in the sialylation patterns of tumor cells. OBJECTIVE: To confirm the overexpression of alpha-2,6 sialinization in exfoliated cells of body fluids and bronchoalveolar lavage (BAL) as a malignant indicator mechanism, using glycan-binding lectins. MATERIALS AND METHODS: Thirty samples (20 effusion liquids and 10 BAL) diagnosed by Giemsa and Papanicolaou staining as negative and positive for malignancy, were studied. They were then stained with fluorescein-labeled Sambucus nigra lectin (Sigma Chemicals, USA), which specifically recognizes sialic acid in alpha-2,6 position. The fluorescence obtained at 515 nm evidenced the presence of sialic acid in the 2,6 position. RESULTS: Negative body fluids for malignancy showed a fine and homogeneous fluorescence pattern for reactive mesothelial cells. Neoplastic cells revealed a thick, heterogeneous pattern. In BAL, benign hyperplastic cells showed a homogeneous fine pattern while neoplastic cells showed a thick and heterogeneous fluorescence pattern. The pattern described was observed in all cases in the cell membrane. CONCLUSION: It was observed that the change in sialic acid conformation detected through Sambucus nigra Lectin could be used as a complementary method for the diagnosis of malignancy in different cytological samples.

[Quality of the exo-endocervical sampling in the prevention of uterine cervix cancer]. / Calidad de la toma exo-endocervical en la prevención del cáncer de cuello uterino.

Cervical and vaginal cytology, Papanicolaou test (PAP), is the most effective test for screening of preneoplastic lesions, and cervical cancer prevention. Its sensitivity goes from 50 to 98%, according to different statistics, and this variation is related to the sampling procedure. A satisfactory smear should be taken from the transformation zone, the junction between endocervix and exocervix. According to Bethesda, metaplastic and/or endocervical cells should be observed under the microscope. The traditional PAP smear includes an exo-endocervical sampling using the Ayre spatula; however, only near 50% of the smears are representative of the transformation zone. In this case report, we present the case of a 40-year-old woman who had negative cytology in five consecutive annual PAP smears, none of which showed metaplastic or endocervical cells. A new sample evidenced a carcinoma in situ (HSIL: high-grade squamous intraepithelial lesion). We emphasize the importance of performing a correct exo-endocervical sampling to allow prompt detection of all premalignant lesions and to prevent cervical cancer, which still persists with high mortality worldwide.

Calidad de la toma exo-endocervical en la prevención del cáncer de cuello uterino / Quality of the exo-endocervical sampling in the prevention of uterine cervix cancer

La citología cérvico-vaginal, test de Papanicolaou (PAP), es la técnica diagnóstica de cribado más efectiva para la detección de lesiones precancerosas y la prevención del cáncer de cuello uterino. La sensibilidad de la prueba varía en las diferentes estadísticas entre el 50% y el 98%; la causa de esta amplitud depende de la toma de muestra. Para que la toma se considere satisfactoria es necesario que se realice de la zona escamocolumnar, zona de transformación, y según el sistema Bethesda en el extendido se deben observar células metaplásicas y/o endocervicales. El PAP convencional incluye la toma exo-endocervical con espátula de Ayre; sin embargo, solo el 50% aproximadamente de las muestras son representativas de la zona de transformación. Para ejemplificar esta situación se presenta el caso de una mujer de 40 años que, a pesar de tener citologías negativas durante cinco años, ninguna con células endocervicales o metaplásicas, una toma adecuada mostró un carcinoma in situ (HSIL: lesión intraepitelial escamosa de alto grado). Recalcamos la importancia de la correcta toma exo-endocervical para poder detectar todas las lesiones premalignas y prevenir este tipo de cáncer que aún tiene alta tasa de mortalidad en todo el mundo.

Cervical and vaginal cytology, Papanicolaou test (PAP), is the most effective test for screening of preneoplastic lesions, and cervical cancer prevention. Its sensitivity goes from 50 to 98%, according to different statistics, and this variation is related to the sampling procedure. A satisfactory smear should be taken from the transformation zone, the junction between endocervix and exocervix. According to Bethesda, metaplastic and/or endocervical cells should be observed under the microscope. The traditional PAP smear includes an exo-endocervical sampling using the Ayre spatula; however, only near 50% of the smears are representative of the transformation zone. In this case report, we present the case of a 40-year-old woman who had negative cytology in five consecutive annual PAP smears, none of which showed metaplastic or endocervical cells. A new sample evidenced a carcinoma in situ (HSIL: high-grade squamous intraepithelial lesion). We emphasize the importance of performing a correct exo-endocervical sampling to allow prompt detection of all premalignant lesions and to prevent cervical cancer, which still persists with high mortality worldwide.

Comparación del recuento celular entre un método manual y un contador automatizado en líquidos de derrame / Comparison of manual microscope method and an automated cell count in serous fluids / Comparação da recontagem celular entre um método manual e um contador automatizado em líquidos de derrame

El objetivo del trabajo consistió en comparar el recuento celular total en los líquidos de derrame de cavidades serosas entre el método automatizado empleado en hematología y el método manual en hemocitómetro. Se procesaron 107 muestras: 45 líquidos ascíticos (LA) y 62 líquidos pleurales (LP) a los que se les realizó el recuento celular en cámara de Neubauer y en contador hematológico Sysmex XT 1800i. Se obtuvieron los siguientes resultados: 1) Regresión lineal: los coeficientes de correlación indicaron una alta correlación entre ambos métodos (LA r2: 0,999; p<0,0001 y LP r2: 0,997; p<0,0001). 2) Bland-Altman: El análisis de las figuras muestra una excelente concordancia entre ambos métodos. El error sistemático fue 51 para los LA y 97 para los LP, por lo que estos valores son despreciables dado el valor diagnóstico de los datos. Los resultados demuestran que los métodos son comparables entre sí y, por ende, se puede remplazar el recuento manual por el automatizado, de demostrada eficiencia y exactitud. Sin embargo, todos los líquidos requieren una observación al microscopio óptico previa al procesamiento por el contador hematológico, donde se apreciará la presencia de agrupamientos celulares como, por ejemplo, células neoplásicas en disposición glandular que dificultan el análisis por parte del equipo o la interpretación del resultado.

The purpose of this work was to compare the total cell count in liquids serous cavities between the automated method used in hematology and the manual method hemocytometer. A total of 107 samples were processed: 45 ascites fluids (LA for its name in Spanish) and 62 pleural fluids (LP for its name in Spanish). The cells were counted in improved Neubauer counting chamber and hematology analyzer Sysmex XT 1800i. The following results were obtained: 1) Linear Regression correlation coefficients indicated a high correlation between the two methods (LA r2: 0.999; p<0.0001 LP r2: 0.997; p<0.0001). 2) Bland-Altman analysis graphics showed excellent agreement between both methods. The systematic error was 51 for LA and 97 for LP; these values are insignificant considering the diagnostic value of the data. he results demonstrate that the methods are comparable and therefore can replace the manual counting by the automated method with proven efficiency and accuracy. However, all fluids require observation by optical microscope before being processed by the hematology analyzer, where the presence of cell clusters such as neoplastic cells in glandular disposition will be appreciated, which hinder the analysis by the equipment or interpretation of results.

O objetivo do trabalho consistiu em comparar a contagem total de células em líquidos de derrame de cavidades serosas entre o método automatizado utilizado em hematologia e o método manual em hemocitômetro. Foram processadas 107 amostras: 45 líquidos ascíticos (LA) e 62 líquidos pleurais (LP) nos quais se realizou a recontagem celular na câmara de Neubauer e no contador hematológico Sysmex XT 1800i. Foram obtidos os seguintes resultados: 1) Regressão linear: os coeficientes de correlação indicaram uma alta correlação entre ambos os métodos (LA r2: 0,999; p<0,0001 e LP r2: 0,997; p<0,0001). 2) Bland-Altman: A análise dos Figuras mostra uma excelente concordância entre ambos os métodos. O erro sistemático foi 51 para os LA e 97 para os LP, resultando estes valores desprezáveis dado o valor diagnóstico dos dados. Os resultados demonstram que os métodos são comparáveis entre si e, portanto, pode ser substituída a contagem manual pela automatizada, de eficiência e exatidão demonstradas. Entretanto, todos os líquidos requerem observação no microscópio óptico prévia ao processamento pelo contador hematológico. Nesse momento se apreciará a presença de agrupamentos celulares como, por exemplo, células neoplásicas em disposição glandular que dificultam a análise por parte da equipe ou a interpretação do resultado.

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.

Argentophilic nucleolus organizer region as a proliferation marker in cervical intraepithelial neoplasia grade 1 of the uterine cervix.

AIM: p16INK4a and argentophilic nucleolus organizer region (AgNOR) can be used as markers for progression of cervical intraepithelial neoplasia grade 1 (CIN1) of the uterine cervix. Our objective was to study the predictive value of the AgNOR technique as a progression marker of CIN1 and its correlation with p16INK4A. MATERIAL AND METHODS: One uterine cervix biopsy from each of 75 patients with diagnosis of CIN1 was selected. All of these patients underwent a second biopsy, and these were also used for the study. RESULTS: The second biopsies showed: regression (20 patients), persistent CIN1 (38 patients), progression to CIN2 (10 patients) and progression to CIN3 (seven patients). p16INK4A showed reactivity in 67 of the 75 first CIN1 biopsies: 12 of the 20 cases that cleared the lesions and the 55 cases with persistent or progressive lesions were positive for p16INK4a (specificity: 40%; sensitivity: 100%; positive predictive value [PPV]: 82%; negative predictive value [NPV]: 100%). Samples with AgNOR areas less than 3.0 µ(2) returned in all cases, but patients whose lesions persisted or progressed to CIN2/CIN3, showed AgNOR areas greater than 3.0 µ(2) in 50/55 cases (specificity: 100%; sensitivity: 91%; PPV: 100%; NPV: 80%). CONCLUSIONS: p16INK4a is expressed in a high percentage of returning lesions. AgNOR might be a better marker of proliferation of CIN1 than p16INK4a (PPV = 100%), which means that a value greater than 3.0 µ(2) indicates the persistence or progression of the lesion. As its NPV is 80%, a value of AgNOR area less than 3.0 µ(2) in CIN1 leaves a margin of doubt about the future behavior of the lesion.

Respuesta inflamatoria genital en la detección de alteraciones por virus del papiloma humano / The inflammatory genital reaction in the detection of alterations by human papilloma virus / Reagào inflamatoria genital na detecgào de alteragòes por Papilomavírus Humano

Disfunción vaginal (DV) (vaginosis/vaginitis) es el síndrome genérico de mayor prevalencia, alcanzando el 50% de todas las mujeres en edad fértil (sintomáticas y asintomáticas). El virus del Papiloma Humano (HPV) se detecta en 30 a 40% de mujeres en edad fértil (sintomáticas y asintomáticas) y se asocia a alteraciones pre-neoplásicas y a carcinoma invasor del cuello uterino. El diagnóstico sindrómico de DV y alteraciones inducidas por HPV es ineficiente y en la actualidad la morfología (macro y microscópica) es el gold standard, pero requiere ordenamiento. El Estudio del Contenido Vaginal es la prueba de laboratorio bacteriológico de mayor solicitud luego del urocultivo. BACOVA normatiza el diagnóstico de vaginosis/vaginitis y ERIGE aumenta el valor predictivo de células que alertan sobre alteraciones epiteliales. Desde 2007 al presente en los talleres BACOVA ERIGE (tinción de Giemsa) se evaluó la sensibilidad de la detección de células anormales exfoliadas. Un 99% de los participantes coincidió con la detección de koi-locitos. BACOVA/ERIGE no reemplaza al Papanicolaou de ninguna manera, pero puede y debe realizarse en laboratorios periféricos, con lo que además del diagnóstico de vaginosis/vaginitis con 100% de valor predictivo, aumentan la cobertura preventiva de estados proliferativos.

Vaginal dysfunction (DV) (vaginosis/vaginitis) is the generic syndrome of major prevalence, reaching 50% of all women in fertile age (symptomatic and asymptomatic). The Human Papillomavirus (HPV) is detected in 30-40% of women in fertile age (symptomatic and asymptomatic) and is associated to pre-neoplastic lesions and invading carcinoma of the uterine cervix. The diagnosis for the symptoms of DV and the alterations induced by HPV are inefficient and at present, the morphology (macroscopic and microscopic) is the standard gold, but it needs better classification. The Study of the Vaginal Content is the test of major request after urocultives in bacteriological laboratories. BACOVA establishes the procedure for the diagnosis of vaginosis/vaginitis and ERIGE increases the predictive value of cells that give the alarm on epithelial alterations. From 2007 to the present sensitivity in the detection of abnormal exfoliated cells from vagina and uterine cervix was evaluated during the BACOVA - ERIGE, (Giemsa's stain) workshops, 99% of the participants coincided with the detection of koilocytes. BACOVA/ERIGE does not replace the Papanicolaou by any means, but it can and must be performed in peripheral laboratories, where apart from the diagnosis of vaginosis/vaginitis with 100% of predictive value, it is possible to increase the detection of precocious proliferative changes of the squamous epithelium.

Disfungào vaginal (DV) (vaginose / vaginite) é a síndrome mais prevalente genérica, atingindo 50% de todas as mulheres em idade fértil (sintomáticas e assintomáticas). O Papilomavírus Humano (HPV) é detectado em 30-40% das mulheres em idade fértil (sintomáticas e assintomáticas) e está associado a alteragòes pré-neoplásicas e a carcinoma invasivo do colo do útero. O diagnóstico sindrómico de DV e alteragòes induzidas pelo HPV é ineficiente e atualmente a morfologia (macroscópica e microscópica) é o padrào ouro, mas precisa de ordenamento. O Estudo do Conteúdo Vaginal é o exame de laboratòrio bacteriológico mais solicitado, seguido da urocultura. BACOVA normatiza o diagnòstico de vaginose/ vaginite e ERIGE aumenta o valor preditivo de células que alertam a respeito de alteragòes epiteliais. Desde 2007 até hoje, nos workshops BACOVA/ERIGE (coloragào de Giemsa), foi avaliada a sensibili-dade da detecgào de células anormais esfoliadas. 99% dos participantes coincidiram com a detecgào de coilócitos. Bacova/Erige nào substitui o Papanicolaou de forma alguma, mas pode e deve ser feito em laboratórios periféricos, com o qual além do diagnóstico de vaginose / vaginite com 100% de valor preditivo, aumentam a cobertura preventiva de estados proliferativos.

Utilidad clínica de la citología espermática / Clinical usefulness of seminal cytology / Utilidade clínica da citologia espermática

En el examen del semen son importantes las determinaciones de la movilidad, el número y la morfología espermática, sin embargo no resultan suficientes. La observación de las células de la progenie espermática o de otras células epiteliales puede contribuir al mejoramiento del diagnóstico clínico. En el presente trabajo se presentan tres casos de estudios de muestras de sémenes donde la observación citológica fue el criterio más importante para la definición de la patología. Finalmente se discuten imágenes celulares publicadas en el Manual de OMS para el Examen y Procesamiento de Semen Humano, 5a edición, donde se confunden células germinales inmaduras con macrófagos. El propósito del trabajo es poner en evidencia la importancia del conocimiento citológico para una correcta interpretación de ciertas patologías seminales.

[Value of sputum analysis in the management of inflammatory airway diseases]. / Valor del estudio celular del esputo en el seguimiento de las enfermedades inflamatorias de la vía aérea.

Cellular analysis of sputum either spontaneous or by induced sputum technique, has become a widespread tool for the evaluation and guidance of treatment of inflammatory diseases of the airway, primarily asthma, COPD and eosinophilic bronchitis. Induced sputum method is a validated, standardized and non-invasive technique, useful in patients with difficulties to expectorate. Its implementation is simple and cost effective. Numerous investigations have shown the effectiveness of basing treatment decisions on the inflammatory component of the airway by counting cells in sputum. Several studies have demonstrated that in patients with asthma, results of this analysis can guide in defining strategies to reduce exacerbations and to improve lung function even in patients with severe asthma, as well as to decrease the remodeling; in addition, a reduction in exacerbations in COpD patients, monitored by this sputum examination, has also been described.

Valor del estudio celular del esputo en el seguimiento de las enfermedades inflamatorias de la vía aérea / Value of sputum analysis in the management of inflammatory airway diseases

El análisis celular del esputo, espontáneo u obtenido mediante la técnica de esputo inducido, se ha transformado en una herramienta ampliamente difundida para la evaluación y orientación del tratamiento de las enfermedades inflamatorias de la vía aérea, principalmente asma, enfermedad pulmonar obstructiva crónica y bronquitis eosinofílica. Se han aportado evidencias sobre la utilidad de la técnica del esputo inducido, validada y estandarizada, para ser empleada en pacientes con dificultades para expectorar. Numerosas investigaciones dieron cuenta de la efectividad de basar las decisiones terapéuticas en el componente inflamatorio de la vía aérea mediante el recuento de células en el esputo. Varios estudios mostraron que, en pacientes con asma el análisis celular de esputo guía en la determinación de estrategias para disminuir las exacerbaciones y para mejorar la función pulmonar, aun en pacientes con asma grave, para disminuir el remodelamiento; también se ha descrito su utilidad en pacientes con EPOC, para la disminución de las exacerbaciones.

Cellular analysis of sputum either spontaneous or by induced sputum technique, has become a widespread tool for the evaluation and guidance of treatment of inflammatory diseases of the airway, primarily asthma, COPD and eosinophilic bronchitis. Induced sputum method is a validated, standardized and non-invasive technique, useful in patients with difficulties to expectorate. Its implementation is simple and cost effective. Numerous investigations have shown the effectiveness of basing treatment decisions on the inflammatory component of the airway by counting cells in sputum. Several studies have demonstrated that in patients with asthma, results of this analysis can guide in defining strategies to reduce exacerbations and to improve lung function even in patients with severe asthma, as well as to decrease the remodeling; in addition, a reduction in exacerbations in COPD patients, monitored by this sputum examination, has also been described.

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.

Utilidad de los marcadores tumorales en líquidos de punción pleural / Usefulness of tumor markers in pleural fluids

El diagnóstico de la neoplasia pleural se basa en la demostración de células neoplásicas en fluido pleural (FP) o en biopsias de pleura. Sin embargo, aún en casos de malignidad hay un elevado porcentaje de informes falsos negativos (30-60%). Los FP neoplásicos presentan valores detectables de marcadores tumorales (MT) producidos por las células neoplásicas de la pleura. El objetivo de este trabajo ha sido evaluar la utilidad diagnóstica de algunos marcadores tumorales, de uso corriente en el laboratorio, en los fluidos de punción pleural. En 20 de ellos se analizaron: Antígeno cárcinoembrionario (CEA), fragmentos de la citoqueratina 19 (CYFRA 21-1) y Antígeno CA 125. Se efectuó el estudio fisicoquímico, recuento celular y examen citológico (Papanicolaou). En algunos casos se realizó diagnóstico anátomo-patológico. Los MT se dosaron por inmunoensayo de electroquimioluminiscencia. La comparación se efectuó por Kruskal-Wallis. Las muestras fueron clasificadas en 4 grupos y se determinó en cada uno mediana y rango para CEA (ng/mL), CYFRA (ng/mL) y CA 125 (UI/mL), respectivamente: 1) Citología positiva con diagnóstico previo de cáncer de pulmón (n: 5) 112 (2,3-1.610), 134,4 (45,8-600), 1.048 (498-2999); 2) Citología positiva con diagnóstico previo de cáncer de otro origen (n: 4) 15,28 (1,1-93,80), 108,1 (7,42-497,2), 1.827 (1.103-14.130); 3) Citología negativa con diagnóstico incierto (n: 4) 1,89 (0,91-2,96), 17,1 (1,5-29,6), 578,2 (27,8-12); 4) Citología no concluyente con diagnóstico incierto (n: 7) 31,8 (1,28-370,2), 96,3 (23,8-860), 585 (94,4-4.584). Se observó diferencia significativa entre los grupos. La combinación de citología y MT aumentó el diagnóstico de neoplasia pleural en 25%. Los resultados preliminares permiten concluir que un panel de marcadores tumorales en FP, sumado a los estudios tradicionales, representa una ayuda diagnóstica.

The diagnosis of pleural cancer is supported by the demonstration of neoplastic cells in pleural fluid or in pleural biopsies. However, even in malignancy there are a great number of false negatives results (30-60%). Tumor fluids. To establish the value of different tumor markers, frequently used in clinical laboratories, in the diagnosis of pleural fluids. 20 pleural fluids were processed for physical and chemical study, cellular counting, morphological examination (Papanicolaou stain) and electrochemiluminescense immunoassay for carcinoembryonic antigen (CEA), CYFRA 21-1 and CA 125. The results were analysed by Kruskal-Wallis. In some cases, biopsies were performed. The samples were classified in four groups, and the median and rank were calculated in each case (CEA, CYFRA and CA 125). 1) Positive cytology with previous diagnosis of lung cancer (n: 5) 112 (2.3-1610), 134.4 (45.8-600), 1048 (498-2999) 2) Positive cytology with previous diagnosis of not-lung cancer(n: 4) 15.28 (1.1-93.80), 108.1 (7.42- 497.2),1827 (1103-14130), 3) Negative cytology with uncertain diagnosis (n: 4)1.89 (0.91- 2.96), 17.1 (1.5-29.6), 578.2 (27.8-12, 4) Inconclusive cytology with uncertain diagnosis (n: 7) 31.8 (1.28-370.2), 96.3 (23.8-860), 585 (94.4 -4584). There were stastistic differences among the four groups. Joining the cytology to the assays for tumor markers raised sensitivity by 25%. The assay for tumor markers can be a complementary tool in the diagnosis of effusions.

Utility of AgNor, immunocytochemistry for CEA and tumor markers for diagnosis in serous fluids

Objective: The evaluation of serous fluids stained by morphological methods lacks, in many cases, the necessary accuracy to obtain the correct diagnostics. The objective of this work was to establish the value of complementary tools for the improvement of diagnosis in serous effusions. Methods: Fifty-six serous effusions were processed for morphological staining, immunocytochemistry of carcinoembryonic antigen (CEA), AgNOR counting and electrochemiluminescense immunoassay for tumor markers (TM): CEA, Ca125 and CYFRA 21-1. TM assays were also performed in sera from the same patients. The Sensitivity (Se) and Specificity (Sp) were evaluated for all the methods. Results: Cytology: Se 73 per cent, Sp 100 per cent, CEA by immunocytochemistry: Se 96 per cent, Sp 75 per cent, AgNOR:Se 86 per cent, Sp100 per cent, TM: a) in fluids: CEA, Ca125 and CYFRA 21-1, Se: 29 per cent, 66 per cent and 64 per cent respectively and Sp: 100 por cento, 87 por cento and 100 por cento respectively. b) in sera: CEA, Ca125 and CYFRA 21-1: Se: 27 per cent, 77 per cent and 47 per cent respectively and Sp: 100 per cent, 25 per cent and 75 per cent respectively. CEA (in cells) + TM (fluids): Se 100 per cent and Sp 75 per cent AgNOR + TM (fluids): Se 95 per cent and Sp 87 per cent TM Panel (CEA+Ca125+CYFRA 21-1): a) in fluids: Se 81 per cent and Sp 87 per cent b) in sera: Se 86 per cent and Sp 12 per cent Conclusion: AgNOR assay and immunocytochemistry for CEA were useful as complementary tools in the diagnosis using effusions, raising the Sensitivity of the Cytology from 73 per cent to 86 per cent and 96 per cent respectively. Sensitivity increased with the assays for a panel of TM in fluids, but the high costof these methods does not justify their use for non-conclusive smears.

Células Escamosas Atípicas de Significado Indeterminado: un citodiagnóstico subjetivo / Atypical Squamous Cells of Undetermined Significance: A subjective cytodiagnosis

El objetivo del presente trabajo es intentar la disminución de la subjetividad que acompaña al citodiagnóstico de células escamosas atípicas de significado incierto (ASCUS) por medio de un parámetro que pueda ser medido y sea útil como valor pronóstico. A 20 extendidos provenientes de cuello uterino coloreados con la técnica de Papanicolaou con diagnóstico de ASCUS (11 metaplasias atípicas, 6 casos de HPV con atipías y 3 casos de anomalías marcadas por hipoestrogenismo) se les aplicó la técnica de AgNOR (regiones del organizador nucleolar afines a Ag) y el área de AgNOR fue medida por citometría de imágenes. Los resultados fueron: 6,0 µ², s=1,60 µ² para metaplasia atípica, 0,90 µ², s=1,05 µ²para HPV con atipías y no detectable para extendidos hipoestrogénicos. Las diferencias entre áreas promedio fueron evaluadas con el test de Student. Se propone 4,4 µ² como valor de corte del área de AgNOR como índice de progresión de las metaplasias atípicas. Los casos de HPV con atipías son insuficientes para definir algún valor de área, y en las pacientes con anomalías marcadas por hipoestrogenismo se recomienda la repetición del extendido post-estímulo estrogénico.

The objective of this paper is to diminish the subjectivity of ASCUS (Atypical Squamous Cells of Undetermined Significance) diagnosis by means of a parameter useful in the prognosis of the pathology. AgNOR technique was applied to 20 cytologic smears from uterine cervix stained by Papanicolaou method and with ASCUS diagnosis: 11 atypical metaplasias, 6 with atypical keratinizing cells related to human papillomavirus (HPV) infection and 3 with regressive changes by estrogen deficiency. The AgNOR area was determined by image cytometry. The mean AgNOR areas were: 6.0 µ², s=1.60 µ² for atypical metaplastic cells, and 0.90 µ², s=1.05 µ² for smears with HPV with atypical keratinizing cells. AgNOR areas were not measurable in the cases of estrogen deficiency. Differences in the mean AgNOR protein areas were tested by Student's test. The cut-off value proposed for smears with atypical metaplastic cells is 4.4 µ² as a limit of progressive lesions. It was not possible to establish a value for cases with atypical keratinizing cells related to HPV because of the low number of smears. In estrogen deficiency, it is advisable to perform the estrogenic stimulus before taking any decision.

Usefulness of AgNOR technique and CEA expression in atypical metaplastic cells from cervical smears.

OBJECTIVE: To evaluate the proliferating capacity of atypical metaplastic cells in cervical smears by AgNOR technique and image analysis and investigate the probable relation with squamous intraepithelial lesions (SIL) using immunocytochemical assay for carcinoembryonic antigen (CEA). STUDY DESIGN: Eight cervical smears were stained with Papanicolaou stain for diagnosis of atypical metaplastic cells. After removal of the stain the smears were processed with a silver colloidal solution and the AgNOR area determined by image cytometry. Differences in the mean AgNOR protein area between reactive metaplastic cells and controls were tested by Student's t test. The CEA was investigated by immunocytochemical staining in smears with atypical metaplastic cells and smears from high-grade squamous intraepithelial lesions (HSIL). RESULTS: The mean AgNOR areas from atypical metaplastic cells (4.55, 6.66, 4.68, 5.30, 4.97, 6.20, 6.28, and 7.35) were significantly greater those that of intermediate metaplastic cells and cells from low-grade squamous intraepithelial lesions (LSIL) (0.77, 0.99, and 0.82, respectively). The atypical metaplastic cells showed values of mean AgNOR area intermediate between that of basal cells (3.28) and HSIL cells (7.73) or neoplastic cells (16.12). The CEA was strongly expressed by the atypical metaplastic cells. CONCLUSION: The expression of CEA in the atypical metaplastic cells underlies the probable relation to SIL. Although the organizer region areas raised high values, it would be necessary to use a greater number of cases to define whether the AgNOR area is indicative of the proliferative activity of the atypical metaplastic cells.

Técnica de detección de la región del organizador nucleolar en células exfoliadas de cuello uterino / Technique for the assay of nucleolus organizer region in exfoliated cells from uterine cervix

La técnica de detección de la región del organizador nucleolar (AgNOR) fue aplicada a 22 extendidos citológicos de exo y endocervix: 5 con células endocervicales, 2 hipotróficos (controles), 2 atróficos, 4 inflamatorios, 2 con células metaplásicas, 4 con células metaplásicas reactivas y 3 de lesiones intraepiteliales de bajo grado. Los promedios de partículas de AgNOR y el área coloreada aumentaron desde los controles hasta las células metaplásicas reactivas. Esta técnica permite identificar células exfoliadas del cuello uterino con mayor capacidad proliferativa, lo que resulta de utilidad en el citodiagnóstico de lesiones sospechosas en las coloraciones de rutina con el método de Papanicolaou

Células de Langerhans y lesiones intraepiteliales de cuello uterino / Langerhans cells and intraepithelial lesions of the uteri cervix

Las células de Langerhans fueron demostradas por un método inmunohistoquímico con el anticuerpo anti proteína S-100 en 22 cortes histológicos de cuello uterino: 6 controles, 8 lesiones intraepiteliales de bajo grado y 8 lesiones intraepiteliales de alto grado, en las cuales se tipificó virus de Papiloma Humano. El número de células de Langerhans decrece desde los epitelios normales hasta las lesiones de alto grado (controles: 186 ñ 5/mm², lesiones de bajo grado: 155 ñ 33/mm² y lesiones de alto grado: 39 ñ 17/mm²). Siendo dichas células importantes en el reconocimiento y procesamiento de antígenos exógenos, su disminución en casos de infecciones por virus de Papiloma Humano de alto riesgo se relacionaría con la progresión de la lesión