Monocyte distribution width (MDW): study of reference values in blood donors.

OBJECTIVES: Monocyte distribution width (MDW) is a new biomarker used as an early indicator of sepsis (ESId). It is often aids in the identification of patients who may develop sepsis. This study aims to establish the MDW reference interval (RI) within the healthy population of blood donors using EDTA-K2 as anticoagulant. Many hospitals use this biomarker as a means of identifying patients who present to the hospital with sepsis. METHODS: A total of 274 samples obtained from healthy donors were analyzed. MDW measurements were taken within 2 h post-extraction. The RI was estimated using various statistical methodologies, including the recommended CLSI EP28-A3c guideline, non-parametric and robust methods, along with the Harrell-Davis bootstrap method applied to the entire sample. RESULTS: The RI estimated through non-parametric method was 14.77 CI90 % (14.36-14.97)-21.13 CI90 % (20.89-21.68); RI using the robust method was 15.64-19.05 and RI using the Harrell-Davis bootstrap method was 14.73 CI90 % (14.53-14.92)-21.14 CI90 % (20.88-21.40). CONCLUSIONS: Based on clinical applicability, we recommend utilizing the RI derived from the non-parametric method, aligning with the CLSI recommendations. Furthermore, we consider that our results can be taken as a reference in other laboratories that serve a population similar to our study cohort.

Monocyte distribution width (MDW) as an infection indicator in severe patients attending in the Emergency Department: a pilot study / Ancho de distribución de los monocitos como indicador de infección en pacientes graves atendidos en Urgencias: estudio piloto

Background. The aim of the present study was to evaluate the diagnostic performance of monocyte distribution width (MDW) as a biomarker for sepsis diagnosis in severe patients attended in the Emergency Department for different conditions and not only infections. Methods. We performed an observational study in a consecutive prospective cohort including severe patients attending the Emergency Department with different conditions. MDW and other biomarkers were determined from samples obtained during the first care of patients. The diagnostic performance of the different biomarkers was determined based on the final diagnosis at patient discharge. Results: One hundred two patients, with a mean age of 76.7 (SD 16.5) years were included, 53 being (51.9%) male. Among the patients included, 65 (63.7%) had an infectious disease while the remaining had other different conditions. A MDW cut-off of 20.115 provided the best accuracy to identify infected patients, with a sensitivity of 89.2 (95% CI 79.4-94.7), a specificity of 89.2 (95% CI 75.3-95.7), a positive predictive value of 93.5 (95% CI 84.6-97.5), a negative predictive value of 82.5% (95% CI 68.0-91.3), a positive likelihood ratio of 8.25 (3.26-20.91), and a negative likelihood ratio of 0.12 (0.06-0.24). The area under the receiver operating characteristic curve for infection according to MDW was 0.943 (95% CI 0.897-0.989; p<0.001). Conclusions: A MDW > 20.115 may be associated with infection and could help to distinguish between infected and non-infected patients in severe patients. These results must be confirmed in new studies due to the limited patient sample included. (AU)

Introducción: El objetivo del presente estudio fue evaluar el desempeño diagnóstico del ancho de distribución de monocitos (MDW) como biomarcador para el diagnóstico desepsis entre pacientes graves atendidos en el servicio de urgencias por diferentes afecciones y no solo por infecciones. Métodos: Realizamos un estudio observacional en una cohorte prospectiva consecutiva que incluyó pacientes graves desde el punto de vista clínico que acudían a urgencias con diferentes patologías. El MDW y otros biomarcadores se determinaron a partir de muestras obtenidas durante la primera atención de los pacientes. Se estudio la precisión de los diferentes biomarcadores para apoyar el diagnósticode infección, basándonos en el diagnóstico final al alta del paciente. Resultados: Se incluyeron 102 pacientes, con una edad media de 76,7 (DE 16,5) años, siendo 53 (51,9%) del sexo masculino. Entre los pacientes incluidos, 65 (63,7%) pacientes tenían una enfermedad infecciosa y el resto otras condiciones diferentes. Un punto de corte MDW de 20,115proporcionó la mejor precisión para identificar pacientes infectados, con un sensibilidad de 89,2 (IC 95 % 79,4-94,7), una especificidad de 89,2 (IC 95 % 75,3-95,7), un valor predictivo positivo de 93,5 (IC 95 % 84,6-97,5), un valor predictivo negativo de 82,5% (IC 95% 68,0-91,3), un coeficiente de probabilidad positivo de 8,25 (3,26-20,91), y uncoeficiente de probabilidad negativo de 0,12 (0,06-0,24). El área bajo la curva característica operativa del receptor para la infección del MDW fue de 0,943 (IC del 95 %: 0,897-0,989; p<0,001). Conclusiones: Un MDW > 20.115 se asocia a padecer una enfermedad infecciosa en un paciente grave y podría ayudar a distinguir entre pacientes infectados y no infectados. Estos resultados deben ser confirmados en nuevos estudios debido a la muestra limitada de pacientes incluidos. (AU)

Leucemia/linfoma de células T del adulto: a propósito de un caso / Adult T-cell Leukaemia/Lymphoma: a case report

La leucemia/linfoma de células T del adulto es una neoplasia de linfocitos T causada por el retrovirus humano HTLV-1, con manifestaciones sistémicas y cutáneas muy variables. El HTLV-1 afecta a más de 5-10 millones de personas en el mundo y su distribución geográfica en zonas endémicas se encuentra íntimamente ligada a la prevalencia de leucemia/linfoma de células T del adulto. Existen distintas variantes en función de su presentación clínica siendo las de peor pronóstico la aguda y el linfoma. El diagnóstico diferencial presenta dificultades debido a la inespecificidad de los síntomas. En el laboratorio de análisis clínicos la sospecha diagnóstica es mediante el estudio citológico en sangre periférica con la observación de células características del leucemia/linfoma de células T del adulto

Adult T-cell leukaemia/lymphoma is a mature peripheral T-cell leukaemia of adults associated with infection by the human retrovirus HTLV-1 that presents with highly variable systemic and cutaneous manifestations. HTLV-1 is estimated to affect at least 5-10 million people worldwide, and its geographic distribution in endemic regions is closely related to ATLL prevalence. Adult T-cell leukaemia/lymphoma is classified into different clinical types according to its clinical characteristics, with acute and lymphomatous types having the worst prognosis. Differential diagnosis of adult T-cell leukaemia/lymphoma can be difficult due to the non-specific symptoms in affected individuals. The Clinical Analysis laboratory can help initiate the diagnosis by visualising pathognomonic cells for adult T-cell leukaemia/lymphoma in a peripheral smear

[Detection and quantification of circulating tumor cells in peripheral blood in patients with colon cancer]. / Detección y cuantificación de células tumorales en sangre periférica en pacientes con cáncer de colon.

BACKGROUND AND OBJECTIVE: The aim of this study is the detection and quantification of circulating tumor cells (CTC) in patients diagnosed with colon cancer and to establish whether they are related to the main clinicopathologic variables for this type of carcinoma. PATIENTS AND METHOD: Twenty-five colon cancer patients and 30 healthy volunteers were analysed. The quantification was performed using the CellSpotter Analyzer (Veridex LLC), that allows immunomagnetic isolation and immunospecific labelling of the cells for their enumeration. RESULTS: 72% of the colon cancer patients showed CTC and the mean number of cells found was 5 CTC/7.5 ml of peripheral blood. 52% of the samples contained 2 or more cells. Considering 2 cells as the cut-off point, a significant relationship with lactate dehydrogenase was found. CONCLUSIONS: This new technology which allows isolation and quantification of CTC in peripheral blood has proven to be valid for the detection of epithelial cells in colon cancer patients in every tumor stage. The results shown in this work confirm that cytokeratin 8, 18 and 19 are detected in CTC in this tumor type and will allow us to develop a protocol for the study of the relationship of quantification of theses cells and the clinical parameters involved in colon cancer.

Detección y cuantificación de células tumorales en sangre periférica en pacientes con cáncer de colon / Detection and quantification of circulating tumor cells in peripheral blood in patients with colon cancer

Fundamento y objetivo: El objetivo de este estudio es cuantificar las células tumorales circulantes (CTC) en pacientes diagnosticados de cáncer de colon y determinar su relación con las variables clinicopatológicas de interés en este tipo de carcinomas. Pacientes y método: La población del estudio está formada por 25 pacientes con cáncer de colon y 30 personas voluntarias sanas. El análisis de las CTC se realiza con el CellSpotter Analyzer (Veridex LLC), que aísla y determina las células mediante técnicas inmunomagnéticas. Resultados: El 72% de los pacientes con cáncer de colon estudiados presentan CTC, con una media de 5 células/7,5 ml de sangre periférica. El 52% presenta 2 o más de 2 CTC. Con este punto de corte, se observa relación de la determinación de CTC con la lactatodeshidrogenasa. Conclusiones: Este nuevo sistema de aislamiento y cuantificación de CTC en sangre periférica permite la detección de células epiteliales en pacientes con adenocarcinoma de colon de todos los estadios tumorales. Los resultados de este estudio confirman que con las citoqueratinas 8, 18 y 19 se detectan las CTC en este tipo de tumores y nos permite establecer un protocolo para estudiar la relación de la cuantificación de estas células con los parámetros clínicos del cáncer de colon

Background and objective: The aim of this study is the detection and quantification of circulating tumor cells (CTC) in patients diagnosed with colon cancer and to establish whether they are related to the main clinicopathologic variables for this type of carcinoma. Patients and method: Twenty-five colon cancer patients and 30 healthy volunteers were analysed. The quantification was performed using the CellSpotter Analyzer (Veridex LLC), that allows immunomagnetic isolation and immunospecific labelling of the cells for their enumeration. Results: 72% of the colon cancer patients showed CTC and the mean number of cells found was 5 CTC/7.5 ml of peripheral blood. 52% of the samples contained 2 or more cells. Considering 2 cells as the cut-off point, a significant relationship with lactate dehydrogenase was found. Conclusions: This new technology which allows isolation and quantification of CTC in peripheral blood has proven to be valid for the detection of epithelial cells in colon cancer patients in every tumor stage. The results shown in this work confirm that cytokeratin 8, 18 and 19 are detected in CTC in this tumor type and will allow us to develop a protocol for the study of the relationship of quantification of theses cells and the clinical parameters involved in colon cancer

Circulating tumoral cells lack circadian-rhythm in hospitalized metastasic breast cancer patients.

BACKGROUND: The relationship between breast cancer and circadian rhythm variation has been extensively studied. Increased breast tumorigenesis has been reported in melatonin-suppressed experimental models and in observational studies. OBJECTIVES: Circulating Tumor Cells (CTC) circadian- rhythm may optimize the timing of therapies. This is a prospective experimental study to ascertain the day-time and night-time CTC levels in hospitalized metastasic breast cancer (MBC) patients. MATERIAL AND METHODS: CTC are isolated and enumerated from a 08:00 AM and 08:00 PM blood collections. 23 MBC and 23 healthy volunteers entered the study. 69 samples were collected (23 samples at 08:00 AM and 23 samples at 08:00 PM from MBC; 23 samples from healthy volunteers). Results from two patients were rejected due to sample processing errors. No CTC were isolated from healthy-volunteers. RESULTS AND DISCUSSION: No-differences between daytime and night-time CTC were observed. Therefore, we could not ascertain CTC circadian-rhythm in hospitalized metastasic breast cancer patients.

[Microsatellite instability predicts better outcome in colorectal cancer patients]. / La inestabilidad de microsatélites predice un mejor pronóstico en los pacientes intervenidos de carcinoma colorrectal.

BACKGROUND AND OBJECTIVE: Two different pathways for the development of tumor have been described in colorectal carcinoma: the chromosomic instability, raised by suppressor genes and proto-oncogene alterations, and the microsatellite instability (MSI), caused by alterations in DNA repairing genes. PATIENTS AND METHOD: The frequency and the clinical meaning of the microsatellites instability pathway were determined in a consecutive prospective cohort of 106 patients who underwent surgical resection of colorectal carcinoma by a single surgeon. Microsatellite instability determination was established according to the criteria proposed by the National Cancer Institute in 1998. RESULTS: 9.4% of patients had a high instability and it was low in 11.3%; both groups displayed different clinico-pathological characteristics (age, sex, tumor site and histologic type). In the multivariant analysis of overall survival and disease free survival, high instability exhibited prognostic value independent of the rest of variables evaluated (p < 0.0001). CONCLUSIONS: The genetic alterations giving rise to microsatellite instability lead to a better prognosis in patients with colorectal cancer.

La inestabilidad de microsatélites predice un mejor pronóstico en los pacientes intervenidos de carcinoma colorrectal / Microsatellite instability predicts better outcome in colorectal cancer patients

FUNDAMENTO Y OBJETIVO: En el carcinoma colorrectal se describen 2 vías genéticas diferentes implicadas en la génesis del tumor: la inestabilidad cromosómica, debida a la alteración de genes supresores o protooncogenes, y la inestabilidad de microsatélites, originada por alteraciones en los genes reparadores del ADN. PACIENTES Y MÉTODO: En este estudio se determina la frecuencia y el significado clínico de la vía de la inestabilidad de microsatélites en una cohorte prospectiva consecutiva de 106 pacientes intervenidos por carcinoma colorrectal por un mismo cirujano. Para la determinación de la inestabilidad de microsatélites se han seguido los criterios propuestos por el National Cancer Instituteen 1998. RESULTADOS: El 9,4% de los pacientes muestra una alta inestabilidad y el 11,3% una inestabilidad baja. Ambos grupos presentan diferentes características clínico patológicas (edad, sexo, localización del tumor y tipo histológico). En el análisis multivariante de la supervivencia global y de la supervivencia libre de enfermedad, la alta inestabilidad presenta un valor pronóstico independiente del resto de las variables clínico patológicas analizadas (p < 0,0001). CONCLUSIONES: La alteración genética que supone la alta inestabilidad de microsatélites confierea los pacientes con cáncer colorrectal un mejor pronóstico

BACKGROUND AND OBJECTIVE: Two different pathways for the development of tumor have been described in colorectal carcinoma: the chromosomic instability, raised by suppressor genes and protooncogene alterations, and the microsatellite instability (MSI), caused by alterations in DNA repairing genes. PATIENTS AND METHOD: The frequency and the clinical meaning of the Microsatellites instability pathway were determined in a consecutive prospective cohort of 106 patients who underwent surgical resection of colorectal carcinoma by a single surgeon. Microsatellite instability determination was established according to the criteria proposed by the National Cancer Institute in1998. RESULTS: 9.4% of patients had a high instability and it was low in 11.3%; both groups displayed different clinico-pathological characteristics (age, sex, tumor site and histologic type). In the multivariant analysis of overall survival and disease free survival, high instability exhibited prognostic value independent of the rest of variables evaluated (p < 0.0001). CONCLUSIONS: The genetic alterations giving rise to microsatellite instability lead to a better prognosis in patients with colorectal cancer

Prognostic significance of p16INK4a alterations and 9p21 loss of heterozygosity in locally advanced laryngeal squamous cell carcinoma.

The p16INK4a gene, localized within chromosome 9p21, has been identified as a cyclin-dependent kinase inhibitor and may negatively regulate the cell cycle acting as a tumor suppressor. Genetic alterations involving the 9p21 region are common in human cancers. A consecutive series of 64 untreated patients (median of follow up 53 months) undergoing surgical resection for locally advanced laryngeal squamous-cell carcinomas (LSCCs) has been studied prospectively. Our purpose was to investigate p16 alterations (9p21 allelic loss, hypermethylation and point mutations) and their possible association with clinico-pathological data and flow cytometric variables (DNA-ploidy and S-phase fraction (SPF)), and to determine the possible prognostic role of this gene in these tumors. PCR-based techniques were used for investigating 9p21 loss of heterozygosity (LOH) and methylation promoter status of the p16 gene. p16 mutations were detected by PCR-SSCP (single strand conformation polymorphism) and sequencing. 9p21 LOH was detected in 16/62 (26%) informative tumors, point mutations in 5% (3/64) and hypermethylation in 9% (6/64) of the cases. p16 alterations were significantly associated with high SPF and DNA-aneuploidy. By univariate analysis, poor histologic differentiation, stage IV, DNA-aneuploidy and p16 point mutations proved to be significantly related to quicker relapse, whereas these same factors, and in addition high SPF, 9p21 LOH and any p16 alterations were significantly related to shorter overall survival. By Cox proportional hazards analysis only histologic grade (G3) and p16 point mutations were independently related to both disease relapse and death. Our study has identified p16 point mutations as important biomolecular indicators in LSCCs.

Pérdida de heterocigosidad en la región 9p21 como mecanismo de inactivación del gen supresor p16 / Loss of heterozygosity (LOH) on 9p21 as mechanism of inactivation of the p16 tumor suppresor gene

OBJETIVO: El objetivo de este trabajo es estudiar la LOH en la región 9p21 (locus D9S1747) en una población de pacientes con carcinoma renal mediante el análisis de los polimorfismos de microsatélites. MÉTODO: Hemos estudiado una serie de 40 pacientes diagnosticados de carcinoma renal esporádico, analizando la LOH en 9p21 mediante el análisis de polimorfismos de microsatélites. RESULTADOS: El 23,7 por ciento de los pacientes presentaba LOH en 9p21, no relacionándose esta alteración genética con ninguna de las características tumorales estudiadas. CONCLUSIONES: - En nuestra serie de pacientes el 23,7 por ciento presentaba LOH en la región 9p21 - El 26,9 por ciento de los carcinomas de células renales convencionales presentaban LOH, el 25 por ciento de los carcinomas papilares y el 25 por ciento de los carcinomas de túbulos de Bellini, el resto de los tipos histológicos no presentaban LOH (AU)

Metilación de las islas 5'CpG del gen supresor p16/CDKN2/MTS1 en carcinoma renal / Methylation of 5' CpG island of the tumor suppresor gene p16/CDKN2/MTS-1 in renal carcinoma

OBJETIVO: El carcinoma renal constituye el 2 por ciento de todos los cánceres humanos. En la actualidad se considera el cáncer como el resultado de una acumulación de alteraciones genéticas que afectan a diversos genes con distintas funciones celulares. Estudios genéticos han demostrado la existencia de un gen supresor en el brazo corto del cromosoma 9, en la región 9p21. A este gen se la ha denominado p16 y codifica las proteínas p16 y p19. Una de las formas de inactivación de este gen sin pérdida de material genético es la metilación en secuencias específicas CpG, en la región promotora del gen. MÉTODO: Se estudió una serie de 40 pacientes diagnosticados de carcinoma renal esporádico. Se analizó el estado de metilación del gen p16 mediante el análisis de los polimorfismos de microsatélites en tejido tumoral y en tejido sano del mismo paciente. RESULTADOS: El porcentaje de pacientes con el gen p16 metilado fue del 20 por ciento. Se relacionó esta alteración genética con las características tumorales, y con ninguna de estas variables se demostró asociación, resultando variables independientes. CONCLUSIÓN: El proceso de tumorogénesis requiere la acumulación de alteraciones genéticas en protoncogenes y genes supresores de tumores. Nuestro estudio sugiere que la alteración del gen supresor p16 a través de la hipermetilación de la región promotora del gen, aunque su incidencia sea baja, puede contribuir al desarrollo del cáncer renal (AU)