Ziehl-Neelsen stain in the pathology laboaratory: Performance and diagnostic aid for mycobacteria in bronchoalveolar lavage / Coloración de Ziehl-Neelsen en el laboratorio de patología: rendimiento y contribución al diagnóstico de micobacterias en el lavado broncoalveolar.

Introduction: With more than 100 years of use, the Ziehl-Neelsen stain is still currently used worldwide. Objective: To compare the performance of diagnostic tests used to determine mycobacteria in the clinic and pathology laboratory in bronchoalveolar lavage samples. Materials and methods: We retrospectively reviewed 737 bronchoalveolar lavage samples from 2019 to 2020 in the San Vicente Fundación hospital (Medellín, Colombia) comparing the performance of three tests done in parallel: mycobacteria and resistance PCR, culture, and Ziehl-Neelsen stain. Results: In total, 93/737 patients were classified as sick due to a positive result in any of the three tests. The culture, PCR, and Ziehl-Neelsen stain had a sensibility of 0.80, 0.76 y 0.51, respectively. However, only 5/75 (6.5%) of the positive cultures had results within the first four weeks and the rest in eight weeks. The PCR test combined with the Ziehl-Neelsen stain improved the sensibility of the PCR test alone from 0.76 a 0.88, a change that was statistically significant (p = 0.022). Conclusion: At least in bronchoalveolar lavage samples, culture is still the test with better sensibility. The use in parallel of the PCR test and the Ziehl-Neelsen stain improved in a statistically significant manner the performance of the PCR test alone, regardless of the higher turnaround time of the Ziehl-Neelsen stain.

Introducción. La coloración de Ziehl-Neelsen, con más de 100 años de uso, continúa vigente mundialmente. Objetivo. Comparar el rendimiento de las pruebas diagnósticas utilizadas para la determinación de micobacterias en el laboratorio clínico de patología en muestras de lavado broncoalveolar. Materiales y métodos. Se revisaron retrospectivamente 737 muestras de lavado broncoalveolar procesadas en el 2019 y el 2020 en el Hospital San Vicente Fundación (Medellín, Colombia) y se compararon las características de tres pruebas diagnósticas realizadas en paralelo: la reacción en cadena de la polimerasa (PCR) para micobacterias con detección de resistencia, el cultivo, y la coloración de Ziehl-Neelsen. Resultados. Se catalogaron como enfermos a 93 de los 737 pacientes a partir de los resultados positivos en alguna de las tres pruebas. El cultivo tuvo una sensibilidad de 0,80, la PCR una de 0,76 y la coloración de Ziehl-Neelsen una de 0,51. Sin embargo, solo 5 de 75 (6,5 %) cultivos fueron positivos a las cuatro semanas y el resto lo fue a las ocho semanas. La PCR combinada con la coloración de Ziehl-Neelsen mejoró la sensibilidad de la PCR por sí sola, de 0,76 a 0,88, diferencia que fue estadísticamente significativa (p=0,022). Conclusión. En las muestras de lavado broncoalveolar, el cultivo sigue siendo la prueba con mejor sensibilidad. El uso conjunto de la prueba de PCR y la coloración de Ziehl-Neelsen mejora significativamente la sensibilidad de la primera, lo que compensa la demora relativa en la entrega de los resultados debida al tiempo requerido para la tinción de Ziehl-Neelsen.

Coloración de Ziehl-Neelsen en el laboratorio de patología: rendimiento y contribución al diagnóstico de micobacterias en el lavado broncoalveolar / Ziehl-Neelsen stain in the pathology laboaratory: Performance and diagnostic aid for mycobacteria in bronchoalveolar lavage

Introducción. La coloración de Ziehl-Neelsen, con más de 100 años de uso, continúa vigente mundialmente. Objetivo. Comparar el rendimiento de las pruebas diagnósticas utilizadas para la determinación de micobacterias en el laboratorio clínico de patología en muestras de lavado broncoalveolar. Materiales y métodos. Se revisaron retrospectivamente 737 muestras de lavado broncoalveolar procesadas en el 2019 y el 2020 en el Hospital San Vicente Fundación (Medellín, Colombia) y se compararon las características de tres pruebas diagnósticas realizadas en paralelo: la reacción en cadena de la polimerasa (PCR) para micobacterias con detección de resistencia, el cultivo, y la coloración de Ziehl-Neelsen. Resultados. Se catalogaron como enfermos a 93 de los 737 pacientes a partir de los resultados positivos en alguna de las tres pruebas. El cultivo tuvo una sensibilidad de 0,80, la PCR una de 0,76 y la coloración de Ziehl-Neelsen una de 0,51. Sin embargo, solo 5 de 75 (6,5 %) cultivos fueron positivos a las cuatro semanas y el resto lo fue a las ocho semanas. La PCR combinada con la coloración de Ziehl-Neelsen mejoró la sensibilidad de la PCR por sí sola, de 0,76 a 0,88, diferencia que fue estadísticamente signifcativa (p=0,022). Conclusión. En las muestras de lavado broncoalveolar, el cultivo sigue siendo la prueba con mejor sensibilidad. El uso conjunto de la prueba de PCR y la coloración de ZiehlNeelsen mejora signifcativamente la sensibilidad de la primera, lo que compensa la demora relativa en la entrega de los resultados debida al tiempo requerido para la tinción de Ziehl-Neelsen.

Introduction: With more than 100 years of use, the Ziehl-Neelsen stain is still currently used worldwide. Objective: To compare the performance of diagnostic tests used to determine mycobacteria in the clinic and pathology laboratory in bronchoalveolar lavage samples. Materials and methods: We retrospectively reviewed 737 bronchoalveolar lavage samples from 2019 to 2020 in the San Vicente Fundación hospital (Medellín, Colombia) comparing the performance of three tests done in parallel: mycobacteria and resistance PCR, culture, and Ziehl-Neelsen stain. Results: In total, 93/737 patients were classifed as sick due to a positive result in any of the three tests. The culture, PCR, and Ziehl-Neelsen stain had a sensibility of 0.80, 0.76 y 0.51, respectively. However, only 5/75 (6.5%) of the positive cultures had results within the frst four weeks and the rest in eight weeks. The PCR test combined with the Ziehl-Neelsen stain improved the sensibility of the PCR test alone from 0.76 a 0.88, a change that was statistically signifcant (p = 0.022). Conclusion: At least in bronchoalveolar lavage samples, culture is still the test with better sensibility. The use in parallel of the PCR test and the Ziehl-Neelsen stain improved in a statistically signifcant manner the performance of the PCR test alone, regardless of the higher turnaround time of the Ziehl-Neelsen stain.

Biliary Tract Microbiota Similarities in Pancreatic Ductal Adenocarcinoma / Similitudes en la Microbiota de Tracto Biliar en Adenocarcinoma Ductal de Páncreas

Abstract Objective: to analyze microbiota profiles in the biliary tract, of pancreatic ductal adenocarcinoma (PDAC) patients and gallstones patients, in order to identify dif ferences, which may contribute to a better understanding of PDAC carcinogenesis. Methods: using microbiota analysis, a total of 25 samples from 14 patients were collected during surgery and compared. Samples were divided into three groups; one GS group (N = 3), and two PDAC groups; PDAC gallbladder group (N = 11) and PDAC brush group (N = 11). Results: upon comparison of bacterial communities' alpha and beta diversity indices and relative abundances by group (anatomic site) and condition (GS vs PDAC), we found no statistically significant results. However, we can highlight the high similarity of the compared parameters among the two different anatomic locations over the biliary tract in PDAC patients. Conclusion: to the best of our knowledge, this is the first study comparing two different anatomic locations over the biliary tract in PDAC patients. Among PDAC groups microbiota along the semi-closed duct system of the biliary tract showed substantial similarity, reflected in the alpha and beta diversity indices and relative abundances.

Resumen Objetivo: analizar los perfiles de microbiota en el tracto biliar de pacientes con adenocarcinoma ductal pancreático (PDAC) y pacientes con cálculos biliares (GS), con el fin de identificar diferencias, lo que puede contribuir a una mejor comprensión de la carcinogénesis de PDAC. Métodos: mediante análisis de microbiota, se recolectaron durante la cirugía un total de 25 muestras de 14 pacientes y se compararon. Las muestras se dividieron en tres grupos; Grupo GS (N = 3) y dos grupos PDAC; Grupo de vesícula biliar PDAC (N = 11) y grupo de cepillado PDAC (N = 11). Resultados: al comparar los índices de diversidad alfa y beta de las comunidades bacterianas y las abundancias relativas por grupo (sitio anatómico) y condición (GS vs PDAC), no encontramos diferencias estadísticamente significativas. Sin embargo, podemos destacar la gran similitud de los parámetros comparados entre las dos ubicaciones anatómicas diferentes en el tracto biliar en pacientes con PDAC. Conclusión: hasta donde sabemos, este es el primer estudio que compara dos ubicaciones anatómicas diferentes sobre el tracto biliar en pacientes con PDAC. Entre los dos grupos de PDAC, la microbiota del sistema de conductos semicerrados del tracto biliar, se encontró una similitud sustancial, reflejada en los índices de diversidad alfa y beta y en abundancias.

Cholangiocyte derived carcinomas and local microbiota.

Trillions of bacteria are present in the gastrointestinal tract as part of the local microbiota. Bacteria have been associated with a wide range of gastrointestinal diseases including malignant neoplasms. The association of bacteria in gastrointestinal and biliary tract carcinogenesis is supported in the paradigm of Helicobacter pylori and intestinal-type gastric cancer. However, the association of bacterial species to a specific carcinoma, different from intestinal-type gastric cancer is unresolved. The relationship of bacteria to a specific malignant neoplasm can drive clinical interventions. We review the classic bacteria risk factors identified using cultures and PCR (polymerase chain reaction) with new research regarding a microbiota approach through 16S rRNA (16S ribosomal ribonucleic acid gene) or metagenomic analysis for selected carcinomas in the biliary tract.

Biliary Tract Carcinogenesis Model Based on Bile Metaproteomics.

Purpose: To analyze human and bacteria proteomic profiles in bile, exposed to a tumor vs. non-tumor microenvironment, in order to identify differences between these conditions, which may contribute to a better understanding of pancreatic carcinogenesis. Patients and Methods: Using liquid chromatography and mass spectrometry, human and bacterial proteomic profiles of a total of 20 bile samples (7 from gallstone (GS) patients, and 13 from pancreatic head ductal adenocarcinoma (PDAC) patients) that were collected during surgery and taken directly from the gallbladder, were compared. g:Profiler and KEGG (Kyoto Encyclopedia of Genes and Genomes) Mapper Reconstruct Pathway were used as the main comparative platform focusing on over-represented biological pathways among human proteins and interaction pathways among bacterial proteins. Results: Three bacterial infection pathways were over-represented in the human PDAC group of proteins. IL-8 is the only human protein that coincides in the three pathways and this protein is only present in the PDAC group. Quantitative and qualitative differences in bacterial proteins suggest a dysbiotic microenvironment in the PDAC group, supported by significant participation of antibiotic biosynthesis enzymes. Prokaryotes interaction signaling pathways highlight the presence of zeatin in the GS group and surfactin in the PDAC group, the former in the metabolism of terpenoids and polyketides, and the latter in both metabolisms of terpenoids, polyketides and quorum sensing. Based on our findings, we propose a bacterial-induced carcinogenesis model for the biliary tract. Conclusion: To the best of our knowledge this is the first study with the aim of comparing human and bacterial bile proteins in a tumor vs. non-tumor microenvironment. We proposed a new carcinogenesis model for the biliary tract based on bile metaproteomic findings. Our results suggest that bacteria may be key players in biliary tract carcinogenesis, in a long-lasting dysbiotic and epithelially harmful microenvironment, in which specific bacterial species' biofilm formation is of utmost importance. Our finding should be further explored in future using in vitro and in vivo investigations.

Quilty effect areas are frequently associated with endocardial C4d deposition.

BACKGROUND: The meaning and clinical implications of the Quilty effect (QE) are not entirely clear. In some biopsies we have found complement split C4d deposition in QE areas, but we do fully comprehend the frequency or pathogenic relationships involved. The objective of this study was to gain insight into the immunologic events involved in the QE, and to understand if and how it relates to complement activation. METHODS: Protocol allograft biopsies (January to December 2005) with evidence of the QE, without cellular rejection or changes suspicious for antibody-mediated rejection, were selected for C4d, CD3, CD20 and CD68 immunohistochemistry. RESULTS: Among 128 allograft biopsies (42 patients), 17 (11 patients) fulfilled the inclusion criteria. Eleven of the 17 biopsies (64.7%), from 8 patients, showed C4d deposition in the endocardium; the positivity was interestingly linear in the endocardium and surrounded by the lymphocytes forming the Quilty lesion. In some cases, the linear C4d deposition extended to the endocardium surrounding the QE area. This pattern was not detected in any of 66 heart allograft biopsies without the QE. B cells were second to T cells in their contribution to the QE, comprising a median of 40% (range, 20% to 50%) of the cells. C4d deposition was not associated with clinical alterations. CONCLUSIONS: The QE is frequently associated with C4d deposition in the endocardium of patients without evidence of rejection. This event suggests a pathogenic relationship between the QE and complement activation. It is possible that the simultaneous presence of both features in an allograft heart biopsy, without evidence of rejection, indicates better adaptation of allograft to host ("accommodation"); however, the precise meaning and implications are not yet known.