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.

Cell-in-cell phenomenon in urinary sediment: a case report.

The internalization of apoptotic cells by non-phagocytic cells has been observed in different tissues and could be an important mechanism for the elimination of dying cells. Here, we describe a probable event of phagocytosis of apoptotic cells mediated by urothelial cells in urinary sediment. A 90-years-old male patient was admitted unconscious to the hospital, visible signs included: pale skin and dry mucous membranes, presumptively diagnosed as dehydration. Blood test revealed anaemia (haemoglobin 130 g/L) and hyperglycaemia (glucose 7.8 mmol/L), urinalysis showed a picture of urinary tract infection (leukocyturia and bacteriuria). The microscopic analysis of urinary sediment revealed the presence of urothelial cells and leukocytes internalized in urothelial cells. Anti-CD68 (membrane marker of macrophages) was tested by immunocytochemistry and a negative result was observed. Based on the findings phagocytosis of apoptotic cells mediated by urothelial cells was identified. This phenomenon can be observed in urinary sediment and should not be confused with a neoplastic process since it is a physiological event of cell elimination.

Estudio multicéntrico de disfunción vaginal de la Red Nacional de Laboratorios BACOVA de la República Argentina: prevalencia, influencia de factores seleccionados, evaluación clínica y distribución de casos por región / Multicenter study on vaginal dysfunction of the National Network of BACOVA Laboratories in Argentina: prevalence, influence of selected factors, clinical evaluation and case distribution by region / Estudo multicêntrico de disfunção vaginal da Rede Nacional de Laboratórios BACOVA da República Argentina: prevalência, influência de fatores selecionados, avaliação clínica e distribuição dos casos por região

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Abstract The aims of this study were to update the prevalence of vaginal dysfunction (VD) in non-pregnant women of childbearing age, pregnant women and menopausal women, to analyse microbiological aspects, to evaluate the influence of parity and contraception in the vaginal microenvironment, to analyse case studies, and to compare the prevalence in the five regions of Argentina: Northeast, Northwest, Centre, Cuyo and South. Eight thousand three hundred twenty-four (8324) vaginal content samples of patients from 39 institutions, examined between May 2019 and June 2020, were prospectively and consecutively studied. The samples were analysed applying the standardized BAVACO-VIR methodology (study of the balance of vaginal content and genital inflammatory response). Of the 8324 samples, 5947 (71.5%) corresponded to women of childbearing age; 1627 (19.5%) to pregnant women and 750 (9.0%) to menopausal women. The most frequent basic vaginal state (BVS) in the three groups was BVS I with normal microbiota, accounting for 33.5% of the entire studied population. Moreover, 66.5% of vaginal dysfunction states were detected. In the pregnant women and menopausal women group there was a significant increase in the frequency of normal BVS I, in contrast with the group of women of childbearing age, where a significant increase of vaginosis and vaginitis was observed. In asymptomatic women, a predominance of BVS I was detected in the three groups. In symptomatic women, there was a predominance of BVS II and V in pregnant women, of BVS II, IV and V in women of childbearing age, and of BVS III and V in menopausal women. Asymptomatic VD was detected in 56.6% of pregnant women, 62.8% of women of childbearing age and 50.9% of menopausal women. A significantly positive association was detected between the presence of yeasts and BVS II and V and the presence of trichomonas and BVS V. The multiparity history variable showed a statistically significant positive association with BVS V and a negative association with BVS I. With regard to contraception, in women who used hormonal contraceptives it was observed that oral contraceptives increased the frequency of BVS I and II and decreased the frequency of BVS III, IV and V of VD; in those who used an intradermal device, there was a decrease in the frequency of BVS I, II and III and an increase in the frequency of BVS IV and V, whereas in women using injectable contraceptives, there was no association. The intrauterine device decreased the frequency of BVS I and II and increased the frequency of BVS V; the condom increased the frequency of BVS IV and the rhythm method decreased the frequency of BVS II. In BVS I, a decrease in the frequency of signs and symptoms was detected. The prevalence of VD in some regions accounted for values over 80%, a higher figure than that described at the national and international levels, which reflects the poor and insufficient action in Sexual and Reproductive Health. The high percentage of asymptomatic women with VD highlights the importance of studying the vaginal content, even in the absence of symptoms. A history of multiparity and contraception showed a link with vaginal function, reflecting the importance of considering this fact in a gynecological evaluation. Although the relationship of BVS with signs and symptoms does not constitute a diagnostic tool, it contributes to the understanding of pathogenic mechanisms.

Resumo Os objetivos deste estudo foram: atualizar a prevalência de disfunção vaginal (DV) em mulheres não grávidas em idade fértil (MEF), mulheres grávidas (EMB) e mulheres menopausadas (MPN), analisar aspectos microbiológicos, avaliar a influência da paridade e contracepção no microambiente vaginal, analisar quadros clínicos e comparar as prevalências das 5 regiões do nosso país: Nordeste, Noroeste, Centro, Cuyo e Sur. 8324 conteúdos vaginais de pacientes de 39 instituições, que compareceram entre maio de 2019 e junho de 2020, foram estudados prospectiva e facilitadoconsecutivamente. As amostras foram analisadas aplicando-se a metodologia padronizada BACOVA-ERIGE (estudo do equilíbrio do conteúdo vaginal e resposta inflamatória genital). Das 8324 amostras de conteúdo vaginal, 5947 (71,5%) corresponderam às MEF; 1627 (19,5%) às EMB e 750 (9,0%) às MNP. O estado vaginal básico (EVB) mais frequente nos três grupos foi EVB I de microbiota normal e representou 33,5% de toda a população global. Foram detectados 66,5% dos estados de disfunção vaginal. No grupo EMB e MNP, um aumento significativo na frequência de EVB I normal foi registrado, em contraste com o grupo MEF, onde foi reconhecido um aumento significativo de EVB de vaginose e vaginite. Em mulheres assintomáticas, foi detectado predomínio de EVB I nos três grupos. Em mulheres sintomáticas foi detetado: nas EMB, predomínio de EVB II e V; nas MEF, predominância de EVB II, IV e V, e nas MNP, predominância de EVB III e V. 56,6% das EMB, 62,8% das MEF e 50,9% das MNP apresentaram DV na ausência de sintomas. Foi detectada associação significativamente positiva entre a presença de leveduras e EVB II e V e a presença de tricomonas e EVB V. A variável antecedente de multiparidade apresentou associação positiva estatisticamente significativa com EVB V e associação negativa com EVB I. Em relação à contracepção, observou-se em mulheres que usavam anticoncepcionais hormonais, que os anticoncepcionais orais aumentaram a frequência de EVB I e II e diminuíram a frequência de EVB III, IV e V de DV; Em mulheres com dispositivo intradérmico, foi observada uma diminuição na frequência de EVB I, II e III e um aumento na frequência de EVB IV e V, e finalmente em mulheres com anticoncepcionais injetáveis, nenhuma associação foi demonstrada. O dispositivo intrauterino diminuiu a frequência de EVB I e II e aumentou a frequência de EVB V; o preservativo aumentou a frequência de EVB IV e o método do ritmo diminuiu a frequência de EVB II. Na EVB I, foi detectada diminuição da frequência de sinais e sintomas. A prevalência de DV em algumas regiões ultrapassou valores de 80%, valor superior ao descrito a nível nacional e internacional, o que reflete a atuação precária e insuficiente em Saúde Sexual e Reprodutiva. O alto percentual de mulheres assintomáticas com DV demonstra a importância da realização do estudo do conteúdo vaginal, mesmo na ausência de sintomas. A história de multiparidade e contracepção mostrou ligação com a função vaginal, refletindo a importância da sua consideração na avaliação ginecológica. Embora a relação da EVB com os sinais e sintomas não constitua uma ferramenta diagnóstica, ela contribui para o entendimento dos mecanismos patogênicos.

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.

Is the Wnt/ß catenin signalling pathway activated in Seminoma?: An immunohistochemical study.

BACKGROUND: The loss of reduction of some adhesion molecules is associated with the invasive phenotype of carcinomas. The aim of this paper was to study the expression of some proteins related to the Wnt/ß catenin signalling pathway in seminomas by immunohistochemical techniques in order to assess the contribution of this pathway to the tumoral development. MATERIALS AND METHODS: Immunohistochemistry for E-cadherin, ß-catenin, vimentin, C-Myc, cyclin D1 (CD1) and placental alkaline phosphatase. (PALP) was carried out in 24 archival tissue blocks of seminomas. Two cellular lines were used as E-cadherin and ß-catenin controls. (JKT-1 and TCam-2). RESULTS: E-cadherin was positive in two seminomas and in one carcinoma in situ. (CIS), showing a membranous pattern. ßcatenin was principally expressed in Sertoli cells, in some malignant gonocytes of CIS and in the membranes of seminomatous cells. No ß-catenin immunostaining was detectable in the cytoplasm and the nuclei of neoplastic cells. Seminomas were weakly possitive for vimentin in 11/24 cases. None of the tumors displayed expression of C-Myc or CD1. CONCLUSIONS: The results does not indicate the activation of the Wnt pathway, due to the lack of vimentin expression in 13/24 seminomas, the low expression in the rest of the cases, the lack of ß-catenin in nuclei and the absence of CD1 and C-Myc expression. Further work is needed to confirm these observations and to test other signaling pathways in seminomas.

Correlation between the cytology of urine sediment in fresh sample and smears stained by Papanicolaou and Giemsa methods.

BACKGROUND: Urine excreted by the body has a variable composition in different physiological and pathological conditions. The cells that come from the renal pelvis, ureters, bladder, and urethra are carried by the urine, and therefore, they can be observed in fresh samples and in smears with Giemsa and Papanicolaou stain. AIM: The aim of this study was to show that high correlation that exists between the cytological examination of fresh urine samples and smears stained with Papanicolaou and Giemsa methods. MATERIALS AND METHODS: A total of 45 cases with no tumor of the urinary tract and 36 patients with lower urinary tract neoplasms were included in the study (20: Low-grade urothelial tumors; 16: High-grade urothelial tumors, squamous carcinomas, and adenocarcinomas). The sediments in the urine samples were observed in fresh specimen and in smears stained with Papanicolaou method. RESULTS: The meticulous observation of fresh urinary sediments allowed identification of diverse cellular types associated with varied pathologies. CONCLUSIONS: The cytological examination of urinary samples in fresh smears, and its later diagnostic confirmation with the Papanicolaou stain is important not only as a diagnostic procedure of tumoral or non-tumoral pathologies, but also as a method for the 'screening' of pre-cancerous lesions or carcinoma in situ, especially in high-risk populations.

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.

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 / ReagOo inflamatoria genital na detecgOo 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.(AU)

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, (Giemsas 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.(AU)

DisfungOo 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 padrOo 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 (coloragOo de Giemsa), foi avaliada a sensibili-dade da detecgOo de células anormais esfoliadas. 99% dos participantes coincidiram com a detecgOo de coilócitos. Bacova/Erige nOo 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.(AU)

A rare case of respiratory disorders associated with two autosomal recessive diseases and male infertility.

The study of nasal ciliary beat frequency (CBF) and ultrastructure may contribute to the understanding of pathognomonic cases of male infertility associated with defects in sperm motility. This study was designed to report a particular case of male infertility, characterized by the association of two respiratory autosomal recessive genetic diseases (alpha-1-antitrypsin deficiency [AAT-D] and primary ciliary dyskinesia [PCD]). A 39-year-old patient with complete sperm immotility, AAT-D, and bronchiectasis was studied in the Laboratory of Male Fertility, the Department of Urology, the Respiratory Center of a Pediatric Hospital, and in the Department of Clinical Medicine of a Rehabilitation Respiratory Hospital. Family history, physical examination, hormonal analysis, microbial assays, semen analysis, nasal ciliary function, and structure study by digital high-speed video photography and transmission electron microscopy are described. A noninvasive nasal biopsy to retrieve ciliated epithelium lining the inferior surface of the inferior nasal turbinates was performed and CBF was determined. Beat pattern was slightly curved and rigid, not wide, and metacronic in all the observed fields analyzed. CBF was 8.2 Hz in average (reference value, 10-15 Hz) Ultrastructural assay revealed absence of the inner dynein arms in 97% of the cilia observed. The final infertility accurate diagnosis was achieved by the study of nasal CBF and ultrastructure contributing to the patient health management and genetic counseling while deciding fatherhood. Beyond this particular case, the present report may open a new field of studies in male infertility, mainly in cases of asthenozoospermia.

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.

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.(AU)

Motility, number and morphology of sperms are very important in semen examination. Nevertheless, observation of germ cells or other epithelial cells can contribute to the improvement of the clinical diagnosis. In the present paper three clinical cases are presented where the cytological characteristic were the most important criteria to define the pathologies. Finally, some images published by the WHO in the last edition of the "Laboratory Manual for the Examination and Processing of Human Semen" , Fifth Edition, 2010 are discussed; in these images, germ cells are erroneously diagnosed as macrophages. The objective of the present work is to underline the importance of the cytology for a correct interpretation of certain seminal pathologies.(AU)

No exame de sÛmen sÒo importantes as determinaþ§es da motilidade, o número e a morfologia espermática, mas tudo isso nÒo é suficiente. A observaþÒo das células da progÛnie espermática ou de outras células epiteliais pode contribuir para a melhoria do diagnóstico clínico. No trabalho em questÒo sÒo apresentados trÛs casos de estudo das amostras dos sÛmens onde a observaþÒo citológica foi o critério mais importante para a definiþÒo da patologia. Finalmente sÒo discutidas imagens celulares publicadas no Manual da OMS para o Exame e Processamento do SÛmen Humano, 5¬ ediþÒo, onde as células germinativas imaturas sÒo confundidas com macrófagos. O objetivo deste trabalho é destacar a importÔncia do conhecimento citológico para a correta interpretaþÒo de certas patologias seminais.(AU)

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

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.(AU)

In vivo antigenotoxic activity of watercress juice (Nasturtium officinale) against induced DNA damage.

The present study was carried out to investigate the genotoxicity as well as possible protective activity against damage induced by cyclophosphamide (CP) of the aqueous juice of watercress (Nasturtium officinale, W.T. Aiton) in vivo. Male and female Swiss mice 7-8 weeks old (N = 48) were treated by gavage with 1 g kg(-1) body weight and 0.5 g kg(-1) body weight of watercress juice during 15 consecutive days. Genotoxicity and its possible protective effect were tested by the comet assay in peripheral blood cells and the micronucleus test in bone marrow. In addition, biopsies of the bladder, epididymis and testicles of mice were performed to extend the experimental design. Watercress juice per se did not induce genetic damage according to the comet assay and micronucleus study, exhibiting a protective activity against CP (P < 0.05 and P < 0.001, respectively). The comparative analysis of bladder histological changes obtained in the watercress plus CP group against those treated with CP alone suggests a probable protective effect. Further studies are needed in order to establish the protective role of watercress juice against DNA damage.

[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.