Monocyte distribution width (MDW) as an infection indicator in severe patients attending in the Emergency Department: a pilot study.

OBJECTIVE: 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.

Colorectal carcinoma prognosis can be predicted by alterations in gene p53 exons 5 and 8.

BACKGROUND AND AIMS: Gene p53 alteration is a genetic event described in the progression from adenoma to colorectal carcinoma. Most of the p53 mutations occur in exons 5 to 8 in highly preserved regions and in the three main structural domains of the p53 protein. It is possible that mutations affecting different structural regions may present different effects on the p53 protein function and, due to this, they may have different prognostic meaning. MATERIALS AND METHODS: The study population consisted of 353 patients diagnosed with sporadic colorectal cancer. Mutations in 5-8 exons of p53 gene were detected by means of single strand conformation polymorphism (SSCP). All samples that showed different migration bands in SSCP were confirmed by sequencing. RESULTS: A total of 69 patients (19.7%) showed alterations of the gene p53. It was observed that mutation in codon 175 in exon 5 was related to tumors located in the colon (p = 0.01) and the mutation in the codon 288 in exon 8 was related to rectal tumors (p = 0.02). In the study of overall survival, mutation in codon 175 of exon 5 conferred a better prognosis and alterations of exon 8 were related to a worse prognosis in different population subgroups: in men, in patients younger than 71 years old, in the tumors located in the proximal colon, the ones moderately differentiated, and those that are mucinous. CONCLUSION: According to this study, mutations in different exons of p53 are related to different phenotypes in colorectal cancer. These phenotypes could mean differences in the clinical evolution of the patients.

Circulating tumor cells in colorectal cancer: correlation with clinical and pathological variables.

BACKGROUND: The CellSearch System is a technique to detect circulating tumor cells (CTCs) in patients with cancer. Few data have been published concerning the role of CTCs detection by this method in colorectal cancer. The aim of this study was to correlate the presence of CTCs with the commonest clinical and morphological variables. PATIENTS AND METHODS: Blood samples were collected from 97 patients and 30 healthy volunteers. Quantification of CTCs in 7.5 ml of blood was carried out with the CellSearch System. The results were expressed as number of CTCs/7.5 ml and the cut-off of >or=2 CTCs/7.5 ml was chosen to define the test as positive. RESULTS: Positive CTCs were detected in 34 of 94 patients (36.2%). Correlation was not found among positive CTCs and location of primary tumor, increased carcinoembryonic antigen level, increased lactate dehydrogenase level or grade of differentiation. Only stage correlated with positive CTCs (20.7% in stage II, 24.1% in stage III and 60.7% in stage IV, P = 0.005). CONCLUSIONS: CTCs detection by CellSearch is a highly reproducible method that correlates with stage but not with other clinical and morphological variables in patients with colorectal cancer. Colon cancer tumor cells are detectable in all stages. Further studies are warranted.

Telomerase activity in colorectal cancer, prognostic factor and implications in the microsatellite instability pathway.

AIM: To determine whether the telomerase activity is related to the Microsatellite instability (MSI) genetic pathway and whether it means a difference in the survival. METHODS: The population consisted of 97 colorectal cancer patients. MSI determination was performed in accordance with the NCI criteria using PCR and Genescan. Telomerase activity was determined by the TRAP-assay, an ELISA procedure based on the amplification of telomeric repeat sequences. RESULTS: 6.2% showed high MSI (MSI-H), 10.3% showed low MSI (MSI-L) and 83.5% did not show this alteration (MSS). Positive telomerase activity was detected in 92.8% of the patients. 83.3% of MSI-H tumors showed positive telomerase against 93.8% of MSS tumors. In the overall survival analysis the absence of telomerase activity conferred a better prognosis. CONCLUSION: Previous works have shown that tumors which develop via the MSI pathway present a better prognosis. No link between telomerase activity and MSI status is observed, although sample sizes are small. Patients with telomerase negative tumors had better overall survival than patients with telomerase positive tumors.

Role of the BRAF mutations in the microsatellite instability genetic pathway in sporadic colorectal cancer.

BACKGROUND AND AIMS: Between 10 and 15% of all cases of colorectal cancer are the result of microsatellite instability (MSI) in the genetic pathway due to an alteration in the DNA repair genes. Tumors with high MSI are characterized by a better prognosis. The BRAF oncogene has been linked to the MSI pathway in tumorogenesis. The objective of this study was to determine whether alterations in BRAF are related to MSI and whether they can result in differences in survival rates. METHODS: The study cohort comprised 351 patients diagnosed with sporadic colorectal cancer. MSI was determined in accordance with the National Cancer Institute's (NCI) recommendations by means of PCR and sequence analyses. Mutational analysis of the BRAF gene was performed by real-time PCR and subsequent sequencing of the altered samples. The methylation pattern of the hMLH1 gene was determined using methylation-specific PCR analyses of bisulfite-treated DNA and the results confirmed by sequencing. RESULTS: Of the patients tested, 6.9% showed high MSI and 3.7% showed a BRAF gene mutation. hMLH1 methylation was observed in 67.2% of the patients with MSI and/or the BRAF alteration. The BRAF mutation was related to the MSI genetic pathway (P < 0.0001) and with hMLH1 methylation. In the analysis of overall survival only MSI had an independent prognostic value for the risk of death. Patients with the BRAF mutation showed a higher risk of death, although this association was found not to be statistically significant. CONCLUSIONS: There is a subgroup of carcinomas which develop via the MSI pathway that carry an alteration of the BRAF gene. This alteration confers a poorer outcome on these patients within the total group of patients with MSI who have a better prognosis. This hypothesis should be further investigated in a larger study population due to the low incidence of BRAF mutations in colorectal 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: TC 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 (AU)

p16 Gene methylation in colorectal tumors: correlation with clinicopathological features and prognostic value.

BACKGROUND: Methylation of the CpG islands in the p16 gene promoter region is an important transcription repression mechanism that has been identified as an alternative mechanism for inactivating specific genes in cancer. Given that, DNA methylation is a common event in colorectal cancer. MATERIALS AND METHODS: The aim of this study was to establish the methylation status of the p16 gene in 104 patients with colorectal carcinoma and evaluate its prognostic value. DNA was bisulfite-modified and analyzed for p16 promoter methylation by methylation-specific PCR. RESULTS: Methylation of thep16 gene was determined in 18.3% of our patient population (19/104). The methylated state did not correlate with any clinicopathological factors. During a median follow-up period of 72 months, the overall survival rate for patients with gene methylation was 75% and without gene methylation it was 61% (p = 0.09). CONCLUSIONS: Although not statistically significant, this difference indicates a clinically valuable tendency.

Prognostic significance of the p185 protein in colorectal cancer.

The amplification and/or overexpression of the c-erbB-2/neu oncogene may play a role in tumor development and progression. The aim of this prospective study was to evaluate the prognostic value of p185 protein in colorectal cancer using immunohistochemical techniques. We analyzed 106 colorectal tumor tissue specimens from patients who had been operated on by the same surgeon and subjected to a median follow-up of 3 years. Thirty-three per cent of patients showed p185 overexpression related to an advanced stage of the disease. In patients with adenocarcinoma tumors of the colon without distant metastases, p185 detection was found to be of clinical prognostic relevance (p = 0.06).

Prevalencia de la enfermedad de Anderson-Fabry en varones con hipertrofia ventricular izquierda / Prevalence of Anderson-Fabry diseases in men with ventricular hypertrophy. AEBM reference 2003 award

La enfermedad de Anderson-Fabry es causa de hipertrofia ventricular izquierda en adultos. En este estudio se pretende analizar la incidencia de la enfermedad deAnderson-Fabry en varones con hipertrofia ventricular izquierda. Se trata de un estudio monocéntrico y prospectivo de 200 pacientes varones con hipertrofia ventricular izquierda. Se realiza la determinación de la actividad de alfa galactosidasa plasmática, confirmándose con la actividad enzimática en leucocitos. Se realizó el diagnóstico de enfermedad de Anderson-Fabry mediante análisis de actividad enzimática plasmática en tres pacientes, en dos de ellos se confirmó el diagnóstico mediante análisis de la actividad enzimática en leucocitos, descartándose dicho diagnóstico en el tercer paciente mediante esta técnica. La hipertrofia ventricular izquierda puede ser una manifestación de la enfermedad de Anderson-Fabry. Se debe tener en cuenta su diagnóstico en todos los pacientes varones con hipertrofia ventricular izquierda, especialmente si es moderada o severa, independientemente de la existencia de posibles causas de ésta (AU)

Importancia de la inestabilidad microsatélite en tumores borderline y carcinomas de ovario / Relevance of microsatellite instability in borderline tumors and ovarian carcinoma. AEBM reference 2003 award

El objetivo de nuestro estudio es determinar la inestabilidad de seis microsatélites (BAT25, BAT26, NME1, D17S250, D5S346 Y D2S123) y observar si está implicada en el desarrollo de los tumores esporádicos de ovario. El trabajo se ha realizado en 40 pacientes intervenidas quirúrgicamente por tumor de ovario esporádico. La comprobación de la inestabilidad se realizó por comparación del tejido sano y tumoral de cada una de las pacientes. Sólo se encontró inestabilidad en: inestabilidad en BAT26 en tumor borderline, inestabilidad en BAT25 en cistoadenocarcinoma de células claras e inestabilidad en NME1 encistoadenocarcinoma papilar seroso en diferentes pacientes. La inestabilidad microsatélite no parece estar implicada en la aparición o desarrollo de los tumores esporádicos de ovario (AU)

Loss of heterozygosity and methylation of p16 in renal cell carcinoma.

To investigate the possible role of genomic aberrations of chromosome 9p21 in the tumorigenesis of human renal cell carcinoma (RCC), 40 sporadic RCCs were studied using PCR analyses. The tumours were predominantly low stage and low grade. Loss of heterozygosity (LOH) was observed in nine of 39 informative cases, but no homozygous deletion was noticed. Hypermethylation of the promoter region of p16 occurred in eight of the 40 RCCs. No correlation was found between hypermethylation of the p16 gene and LOH on 9p21. A similar level of LOH and methylation was observed in the 40 RCCS regardless of histology, grade and stage. These results suggest that inactivation of p16 and the possibility of other unknown tumour suppressor genes located on other chromosomes could be involved in the pathogenesis of RCC.

Role of microsatellite instability in borderline ovarian tumors.

This study was designed to establish the role of microsatellite instability (MSI) in the development of sporadic tumors of the ovary. The instability of 6 microsatellites (BAT25, BAT26, NME1, D17S250, D5S346 and D2S123) was determined by comparing MSI in healthy and tumoral tissue in each of 40 patients undergoing surgery for a sporadic ovarian tumor. BAT26 and D2S123 instability was detected in borderline tumors, and ovarian carcinomas were found to present instability in the microsatellites BAT25, NME1 and D17S250. Our findings indicate that microsatellite instability lacks a significant role in the appearance or progression of sporadic ovarian tumors.

Metilación del gen p16 en el carcinoma broncogénico no microcítico. Implicación pronóstica / Methylation of gene p16 in non-small cell bronchogenic carcinoma. Prognostic implication

La importancia sociosanitaria del cáncer de pulmón radica en su elevada incidencia y mortalidad. 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. Los estudios genéticos han demostrado alteraciones en la región 21 del brazo corto del cromosoma 9 (9p21), que son frecuentemente identificadas en el cáncer humano. Esta región contiene un gen supresor llamado p16, que codifica las proteínas p16 y p19. Se ha observado que, aunque la pérdida de heterocigosidad (LOH) es un mecanismo frecuente de inactivación del gen, la metilación del promotor de p16 también es una alteración importante que suprime la función del gen. Se estudió una serie de 98 pacientes diagnosticados de carcinoma de pulmón no microcítico. Se analizó la LOH en 9p21 mediante el análisis de polimorfismos de microsatélites y el estado de metilación del gen p16. El 23,5 per cent de los pacientes presentaban LOH y/o inestabilidad en 9p21 y el 54,5 per cent metilación de p16. Los pacientes que no presentaban alteraciones genéticas en p16 tenían un riesgo relativo de fallecer 1,67 veces mayor (p = 0,1) que los que sí presentaban alguna de estas alteraciones (AU)

Valor pronóstico de la pérdida de heterozigosidad en la región 9p21 en el carcinoma broncogénico no microcítico / Prognostic value of loss of heterozygosity at chromosome 9p21 in nonsmall cell bronchogenic carcinoma

La importancia sociosanitaria del cáncer de pulmón radica en su elevada incidencia y mortalidad. En la actualidad se considera el cáncer como el resultado de una acumulación de alteraciones que afectan a diversos genes con distintas funciones celulares. Diversos estudios genéticos han demostrado una pérdida de material genético en la región 21 del brazo corto del cromosoma 9 (9p21), siendo una de las alteraciones genéticas más frecuentemente identificadas en el cáncer humano. Esta región contiene un gen supresor llamado p16, que codifica las proteínas p16 y p19.Se estudió una serie de 98 pacientes diagnosticados de carcinoma de pulmón no microcítico. Se analizó la pérdida de heterozigosidad (LOH) en 9p21 mediante el análisis de polimorfismos de microsatélites. El 23,5 por ciento de los pacientes presentaba LOH y/o inestabilidad en 9p21. Contrariamente a lo que se esperaba, los pacientes que no presentaban alteraciones genéticas en p16 tenían un riesgo relativo de fallecer 1,7 veces mayor (p = 0,1) que los que sí presentaban LOH y/o inestabilidad (AU)

Estudio del gen supresor p16 en el carcinoma broncogénico no microcítico. Valor pronóstico de sus alteraciones / Study of p16 supressor gene in the broncogenic in carcinoma non-small cell Pronostic Value

La importancia sociosanitaria del cáncer de pulmón radica en su elevada incidencia y mortalidad. 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 pérdida de material genético en la región 21 del brazo corto del cromosoma 9 (9p21), siendo una de las alteraciones genéticas más frecuentemente identificadas en cáncer humano. Esta región contiene un gen supresor llamado p16, que codifica las proteínas p16 y p19. Se estudió una serie de 98 pacientes diagnosticados de carcinoma de pulmón no microcítico. Se analizó la LOH en 9p21 mediante el análisis de polimorfismos de microsatélites. El 23,5 por ciento de los pacientes presentaba LOH y/o inestabilidad en 9p21.Contrariamente a lo que se esperaba, los pacientes que no presentaban alteraciones genéticas en p16 tenían un riesgo relativo de fallecer 1,7 veces mayor (p=0,1) que los que sí presentaban LOH y/o inestabilidad (AU)

[Loss of heterozygosity in the 9p21 region as an inactivation mechanism of the p16 suppressor gene]. / Pérdida de heterocigosidad en la región 9p21 como mecanismo de inactivación del gen supresor p16.

OBJECTIVE: To determine the loss of heterozygosity (LOH) on 9p21 (locus D9S1747) in patients with renal carcinoma by analysis of microsatellite polymorphisms. METHODS: 40 patients with sporadic renal cancer were studied. LOH on 9p21 was performed by analysis of microsatellite polymorphisms. RESULTS: 23.7% showed LOH on 9p21. No correlation was found between this genetic alteration and tumor features. CONCLUSIONS: LOH on 9p21 was found in 23.7% of the patients in this series. LOH was found in 26.9% of renal cell carcinomas, 25% of papillary carcinomas and 25% of Bellini duct carcinomas. LOH was not found in the other histological types.

Loss of heterozygosity on the short arm of chromosome 3 in renal cancer.

3% of human cancers are renal cell carcinomas (RCC). The most common chromosome abnormality found in this tumor is loss of heterozygosity (LOH) on the short arm of chromosome 3, which suggests that there must be one or more tumor suppressor genes between 3p14 and 3p21 near the VHL gene which play a relevant role in renal cancer development. DNA from normal and tumor tissue from 40 patients at various stages of RCC was analyzed for LOH at three microsatellites mapped to 3p (3p14.1-14.3; 3p21.2-21.3 and 3p25) by polymerase chain reaction). 42.5% of the tumors studied showed LOH on at least one locus. 30% showed LOH on only one locus; 5% on two loci and 7.5% on the three loci tested. LOH occurred only on nonpapillary tumors (p = 0.03). Interestingly, all the tumors with LOH on 3p21 were >/=25 mm (p = 0.04; relative risk 1.76, confidence interval: 1.3-2.3).

[5'CpG island methylation of the suppressor gene p16/CDKN2/MTS1 in renal carcinoma]. / Metilación de las islas 5'CpG del gen supresor p16/CDKN2/MTS1 en carcinoma renal.

OBJECTIVE: To determine the methylation status of the tumor suppressor gene p16 in patients with renal carcinoma by analysis of microsatellite polymorphisms. METHODS: 40 patients with sporadic renal cancer were studied. Tumor and normal tissue from each patient were tested for methylation status of the p16 gene by analysis of microsatellite polymorphisms. RESULTS: 20% showed methylation of p16. No correlation was found between this genetic alteration and tumor features. CONCLUSIONS: Methylation of p16 was found in 20% of the patients in this series. According to the analysis, 18.5% of renal cell carcinomas showed methylation, 50% of chromophobe cell carcinomas and 25% of carcinomas of Bellini's tubules. The rest of the histological types showed no methylation.

[P185 (Neu) oncoprotein in the prognosis of bladder carcinoma. Experience of 5 years]. / Oncoproteína P185 (Neu) en el pronóstico del carcinoma vesical. Experiencia a 5 años.

OBJECTIVE: To determine the utility of p185 oncogene in the biological characterization of transitional cell carcinoma and in the prediction of recurrence, and to analyze survival at 5 years mean follow-up. METHODS: A prospective clinical cohort study was conducted on 81 patients. Tissue specimens were obtained between November 1992 and November 1993. The study comprised two groups: nontumoral bladder tissue specimens from 20 patients (group I) and tissue specimens from 61 patients with bladder carcinoma (group II). p185 expression was determined by enzyme immunoanalysis (EIA). A statistical analysis of the results was performed. RESULTS: p185 oncoprotein levels were higher in patients with recurrence (1098.97 HNU/mg protein vs. 924.54 HNU/mg). Although higher levels of p185 were found in the patients that had died vs those who are alive, the differences were not statistically significant for overall survival or stratification by tumor grade or infiltration (p = 0.556; ns). CONCLUSIONS: Determination of p185 oncoprotein was found to be useful in the prediction of tumor recurrence at 5 years mean follow-up.

Expression of high p53 levels in colorectal cancer: a favourable prognostic factor.

The expression of p53 protein was examined in a series of 111 colorectal cancer adenocarcinomas with a long follow-up. A quantitative luminometric immunoassay (LIA) was used for the measurement of wild-type and mutant p53 protein in extracts from colorectal tumour cytosols, p53 being detected in 42% of the samples (range 0.0-52 ng (mg-1)). Using an arbitrary cut-off value of 2.7 ng mg(-1), 25% of the tumours were classified as manifesting high p53 levels. There was no association of p53 expression with patient age, sex, serum preoperative carcinoembryonic antigen (CEA) levels, tumour site and size, nodal status or TNM stage. Significant and independent correlation was found to exist between high p53 levels and prolonged disease-free survival (P = 0.05) at a median follow-up of 60 months. This survival advantage was most apparent among stage III cancer patients. The results from this study would suggest that expression of high p53 levels appear to be useful in selecting a group of colorectal cancer patients with a better prognosis.