Basophil Histamine Release Induced by Amoxicilloyl-poly-L-lysine Compared With Amoxicillin in Patients With IgE-Mediated Allergic Reactions to Amoxicillin.

BACKGROUND: Amoxicillin (AX) is the ß-lactam most often involved in IgE-mediated reactions. Diagnosis is based mainly on skin testing, although sensitivity is not optimal. We produced a new AX derivative, amoxicilloyl-poly-L-lysine (APL), and analyzed its recognition of IgE using the passive histamine release test (pHRT). METHODS: The study population comprised patients (n=19) with confirmed AX allergy and specific IgE to AX and controls (n=10) with good tolerance to AX. pHRT was performed using "IgE-stripped" blood from a single donor that was sensitized in vitro by patient sera and incubated with AX or APL. Histamine release was determined and expressed as nanograms of histamine released per milliliter of blood. RESULTS: The clinical symptoms were anaphylaxis (n=9), urticaria (n=7), erythema (n=2), and nondefined immediate reactions (n=1). The median (IQR) time interval between reaction and study was 90 (60-240) days and between drug intake and development of symptoms 24 (10-60) minutes. The median sIgE level was 3.37 (0.95-5.89) kUA/L. The sensitivity of pHRT to APL was 79% and the specificity 100%, which were higher than data obtained with pHRT to AX (63% sensitivity and 90% specificity). There was a positive correlation between maximal histamine release levels obtained with AX and APL (r=0.63). CONCLUSIONS: In patients with immediate hypersensitivity reactions to AX, APL showed higher sensitivity and specificity than the culprit drug, AX, when tested in vitro by pHRT. This indicates that APL can improve the in vitro diagnostic accuracy of allergic reactions to AX. Further assessment of skin testing is necessary.

Basophil histamine release induced by amoxicilloyl-poly-L-lysine compared with amoxicillin in patients with IgE-mediated allergic reactions to amoxicillin

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Bax expression in untreated breast cancer: an immunocytometric study of 255 cases.

BACKGROUND: Bax is one of the main effectors of apoptosis in breast cancer. However, in contrast with the antiapoptotic protein Bcl-2, which has been extensively studied in this tumor, there are relatively few clinical studies on the biological role of Bax in breast cancer. MATERIALS AND METHODS: The expression of the apoptosis-related Bax gene was studied in a series of 255 previously untreated breast cancers by means of immuno-flow cytometry. Additionally, and by the same method, the expression of the Bcl-2, VEGF and Nup88 genes were also studied. As variables of the study for the final statistical analysis, the histological variety of the tumors, histological and nuclear grade, the expression of hormone receptors, p53, Ki-67 or c-erb-B2, axillary node invasion, tumor size and DNA-ploidy were also included. RESULTS: The expression of the proapoptotic Bax protein was significantly associated with the expression of Nup88 (p<0.0001), VEGF (p=0.0014) and Bcl-2 (p=0.0063), all measured by the same method. An inverse correlation with c-erb-B2 expression, which almost attained statistical significance (p=0.058) was also registered. CONCLUSION: This study adds evidence to the little explored link between apoptosis and angiogenesis. Furthermore, it discloses a previously unreported relationship between Bax and Nup88 expression.

The small variant of the apoptosis-associated X-chromosome RBM10 gene is co-expressed with caspase-3 in breast cancer.

BACKGROUND: There are very few studies on the final triggers of apoptosis, the caspases, in breast cancer. MATERIALS AND METHODS: Caspase-3 expression was studied by means of reverse transcript polymerase chain reaction (RT-PCR) in a series of 108 previously untreated patients with breast cancer. Expression levels were correlated with those obtained using the same technique of the apoptosis-associated X-chromosome genes RBMX, RBM3, RBM10 small and RBM10 large variant; Bcl-2 and Bax; the angiogenesis-associated genes VEGF and CD105 (endoglin); hMAM and Nup88. The correlation with the expression of hormone receptors, c-erb-B2, mutant p53 and Ki-67, all measured by means of immunohistochemistry, was also studied, as well as that with standard clinical parameters such as histological type, tumor size, axillary metastasis and DNA-ploidy. RESULTS: The only statistically significant correlations observed between caspase-3 mRNA expression and the parameters tested were a direct one with both the Bax (p = 0.007) and the small variant of the X-chromosome RBM10 gene (p = 0.018), and an inverse one with the angiogenesis-associated CD105 (endoglin) gene (p = 0.044). CONCLUSION: These results indicate that very few genes are involved in the last steps of the apoptotic cascade in breast cancer, among them one of the X-chromosome RBM family. They also support the relatively unexplored link between apoptosis and angiogenesis.

Risk factors for ulcerative colitis: A population-based, case-control study in Spain.

BACKGROUND: Environmental factors seem to be very important in the aetiology of Ulcerative Colitis (UC), with smoking, contraceptive use, and hygiene being the factors most commonly linked to disease. AIM: To analyse the association between different risk factors and development of UC in our community. PATIENTS AND METHODS: This is a case-control, population-based study. The UC population consists of an inception-case population of all cases diagnosed, using Lennard-Jones criteria, in our community from 1st February 1992 to 31st January 1995 that were prospectively included. Controls were selected from healthy population and matched with patients for age, sex and rural/urban habitat. We used the SPSS/PC+ software, EpiInfo and Statistix for statistical analysis, giving the rates as point estimates and 95% confidence intervals (95%CI) or as mean±standard deviation in quantitative variables. For multivariate analysis we used conditional logistic regression. RESULTS: 205 patients were diagnosed of UC. 38 patients (18.5%) with UC were smokers, compared with 84 (40.8%) controls (p<0.001). Smoking behaved as a protector factor for UC (OR=0.55 (CI 95% 0.33-0.92) and ex-smoker acted as a risk factor (OR=1.94 (CI 95% 1.14-3.34). After the multivariate analysis, both associations were maintained. We did not detect statistical differences in the analysis of previous appendectomy, childhood hygiene or oral contraceptive use. Five of the 12 cases with family aggregation had first-degree relatives and 7 of them second-degree relatives. None of the controls had previous IBD history (p=0.0002). CONCLUSION: Ex-smoking and previous family history of inflammatory bowel disease appeared as risk factors for developing ulcerative colitis while current smoking behaved as a protective factor in this population.

Air pollution and cardiovascular admissions association in Spain: results within the EMECAS project.

OBJECTIVE: To evaluate the short term effect of air pollution on cardiovascular admissions in 14 Spanish cities METHODS: The period under study was from 1995 to 1999. Daily emergency admissions for all cardiovascular diseases (CVD) and heart diseases (HD) were obtained from hospital records, and the corresponding daily levels of particulates, SO2, NO2, CO, and ozone were recorded. The magnitude of association was estimated using Poisson generalised additive models controlling for confounding and overdispersion. For each cause, lagged effects, up to three days, of each pollutant were examined and combined estimates were obtained. For ozone the analyses were restricted to the warm period. One and two pollutant models were performed. RESULTS: Associations were more consistent in lag 0 (concurrent day) and 1 (lag 0-1), except in the case of ozone where there was a more delayed relation (lag 2-3). For combined estimates an increase of 10 microg/m3 in the PM10 levels in lag 0-1 was associated with an increase of 0.9% (95% CI: 0.4 to 1.5%) in the number of hospital admissions for CVD, and 1.6% (0.8 to 2.3%) for HD. For ozone the corresponding estimates for lag 2-3 were 0.7% (0.3 to 1.0) for CVD, and 0.7% (0.1 to 1.2) for HD. An increase of 1 mg/m3 in CO levels was associated with an increase of 2.1% (0.7 to 3.5%) in CVD admissions, and 4.2% (1.3 to 7.1%) in HD admissions. SO2 and NO2 estimates were more sensitive in two pollutant models CONCLUSIONS: A short term association between increases in daily levels of air pollutants and the number of daily admissions for cardiovascular diseases, with specificity for heart diseases, has been described in Spanish cities.

The S-phase fraction of the aneuploid cell subpopulation is the biologically relevant one in aneuploid breast cancers.

BACKGROUND: In the case of DNA-aneuploid tumors there are no clear guidelines as to which S-phase fraction is the more relevant one: that corresponding to either the diploid or the aneuploid population, or rather an average of both. MATERIALS AND METHODS: We studied 280 breast cancer specimens from previously untreated patients. Histologically, 231 were ductal infiltrating carcinomas, 30 lobular infiltrating carcinomas and 19 corresponded to other, less frequent varieties. Postsurgically, 164 cases (58.6%) were classified as T1, 87 (31.1%) as T2 and 7 as T3. The remaining 22 cases were multifocal, diffuse tumors. Flow cytometry was performed on fresh tumor tissue, and immunohistochemistry for hormone receptors, Ki67, c-erb-B2 and p53 on paraffin-embedded material. RESULTS: In diploid tumors, a high S-phase (above the 75th percentile) correlated significantly with Ki67 expression > or =20% (p<0.0001). In aneuploid tumors, however, this was only the case for the aneuploid fraction of tumor cells (p< 0.0001). A high S-phase of diploid tumors correlated directly and significantly with a high histologic grade (p=0.04), a high nuclear grade (p=0.01), tumor size (p=0.0008), and inversely with estrogen (p<0.0001) and progesterone (p<0.0001) receptor expression. In aneuploid tumors, the aneuploid tumor fraction showed a direct and significant correlation with a high histologic grade (p=0.005), a high nuclear grade (p=0.001), mutant p53 expression (p=0.0009), and inversely with estrogen (p<0.0001) and progesterone (p=0.0001) receptor expression. A high S-phase of the diploid cell fraction of aneuploid tumors, on the other hand, just showed an inverse correlation with high nuclear grade of the tumors (p=0.02), and none whatsoever with all other tested parameters.

[Surgery at follow-up in an incidence cohort of patients with Crohn's disease in Aragon (Spain): etiology, type of surgery and associated epidemiological factors]. / Cirugía de una cohorte incidente de pacientes con enfermedad de Crohn en Aragón: indicaciones, tipo de cirugía y factores de riesgo asociados.

INTRODUCTION: Few population-based studies have been published on predictive factors in the clinical course of Crohn's disease (CD). The only constant risk factor for postsurgical recurrence is smoking. The aim of this study is to describe surgical need, etiology and characteristics, and the distinct clinical variables that act as risk or protective factors for the indication of surgery in an incidence cohort of patients with a diagnosis of CD in Aragon. MATERIAL AND METHODS: Based on the results of a population-based, prospective study reporting the incidence of inflammatory bowel disease in Aragon, 88 patients with a diagnosis of CD were included in the present study. The patients medical records were reviewed and data on the following clinical variables from diagnosis (1992-1995) to February 2001 were gathered: surgery, type of surgery and etiology, anatomic distribution, number of relapses, remission, clinical course, death, smoking, oral contraceptive intake, and hospitalization. Descriptive and bivariate analyses were performed to investigate the association between these variables and surgery during follow-up. RESULTS: Eighty-eight patients with at least 6 months of follow-up were included (88/103; 85%), with a mean follow-up of 77 months (range, 6-110 months). Some kind of surgery during follow-up was required by 20.5% of our patients; in nearly 50% of these, surgery was indicated for intestinal obstruction. A second surgical intervention was required in 10.2% due to fistula and/or abscess or ileostomy for subsequent reconstruction of intestinal transit. Although ileal localization was more frequently associated with surgery, this association was not statistically significant. No association was found between surgery during follow-up and sex, age at diagnosis or oral contraceptive intake. Factors positively associated with surgery were a chronic clinical course and a greater number of hospitalizations. We found no positive or negative association with smoking, non-smoking or time free of smoking, but the total time of smoking was positively associated with surgery. CONCLUSIONS: Risk factors for surgery in patients with CD were a chronic clinical course, the number of hospitalizations and total time of smoking. Ileal localization was more frequently associated with surgery but this association was not statistically significant.

Cirugía de una cohorte incidente de pacientes con enfermedad de Crohn en Aragón: indicaciones, tipo de cirugía y factores de riesgo asociados / Surgery at follow-up in an incidence cohort of patients with crohn’s disease in Aragon (Spain): etiology, type of surgery and associated epidemiological factors

INTRODUCCIÓN: Con los estudios poblacionales disponibles conocemos poco sobre factores predictivos en el curso clínico de la enfermedad de Crohn (EC). El único factor consistente es el mayor índice de recurrencia quirúrgica de los pacientes fumadores. El objetivo de este estudio fue valorar la necesidad de cirugía, así como las distintas variables clínicas que actúan como factores de riesgo y/o protectores respecto a la indicación quirúrgica de una cohorte incidente de pacientes con diagnóstico de EC en Aragón. MATERIAL Y MÉTODOS: Basándonos en los resultados de un estudio poblacional y prospectivo incidente en Aragón, incluimos en el estudio a 88 pacientes diagnosticados de EC. Se analizaron las siguientes variables: cirugía, tipos de intervención quirúrgica y etiología, localización anatómica, número de brotes e ingresos, remisión, curso crónico, fallecimientos, exposición al tabaco y toma de anticonceptivos orales en el momento del ingreso. Se aplicaron análisis descriptivo y bivariante para investigar la asociación de dichas variables con la cirugía durante el seguimiento. RESULTADOS: De los 103 pacientes diagnosticados de EC, se obtuvo información con al menos 6 meses de seguimiento de 88, esto es, un 85%, con una media de 77 meses de seguimiento(mediana, 81; rango, 6-110). El 20,5% de los pacientes precisaron durante el seguimiento algún tipo de intervención quirúrgica, motivada en cerca del 50% por obstrucción intestinal; el 10,2% precisó una segunda cirugía por fístula y/o absceso, o la presencia de una ileostomía para posterior reconstrucción del tránsito intestinal. Aunque la afectación ileal se asoció más frecuentemente a la cirugía, esta diferencia no alcanzó significación estadística. Ni el sexo, ni la edad en el momento del diagnóstico, ni la toma de anticonceptivos se asociaron a la presencia de cirugía durante el seguimiento. Los pacientes que la precisaron habían presentado un curso clínico de más de un brote y un mayor número de ingresos. No observamos asociación positiva ni negativa con las variables «expuesto» al tabaco, no fumador ni tiempo libre de exposición, pero sí con el tiempo total de exposición al tabaco. CONCLUSIONES: La presencia de un curso crónico, el número de ingresos y el tiempo global de exposición al tabaco son factores de riesgo para la cirugía en los pacientes con EC

INTRODUCTION: Few population-based studies have been publishedon predictive factors in the clinical course of Crohn’s disease (CD). The only constant risk factor for postsurgical recurrence is smoking. The aim of this study is to describe surgical need, etiology and characteristics, and the distinct clinical variables that act as risk or protective factors for the indication of surgery in an incidence cohort of patients with a diagnosis of CD in Aragon. MATERIAL AND METHODS: Based on the results of a population-based, prospective study reporting the incidence of inflammatory bowel disease in Aragon, 88 patients with a diagnosis of CD were included in the present study. The patients’ medical records were reviewed and data on the following clinical variables from diagnosis (1992-1995) to February2001 were gathered: surgery, type of surgery and etiology, anatomic distribution, number of relapses, remission, clinical course, death, smoking, oral contraceptive intake, and hospitalization. Descriptive and bivariate analyses were performed to investigate the association between these variables and surgery during follow-up. RESULTS: Eighty-eight patients with at least 6 months of follow-up were included (88/103; 85%), with a mean follow-up of 77 months (range, 6-110 months). Some kind of surgery during follow-up was required by 20.5% of our patients; in nearly 50% of these, surgery was indicated for intestinal obstruction. A second surgical intervention was required in 10.2% due to fistula and/or abscess or ileostomy for subsequent reconstruction of intestinal transit. Although ileal localization was more frequently associated with surgery, this association was not statistically significant. No association was found between surgery during follow-up and sex, age at diagnosis or oral contraceptive intake. Factors positively associated with surgery were a chronic clinical course and a greater number of hospitalizations. We found no positive or negative association with smoking, non-smoking or time free of smoking, but the total time of smoking was positively associated with surgery. CONCLUSIONS: Risk factors for surgery in patients with CD were a chronic clinical course, the number of hospitalizations and total time of smoking. Ileal localization was more frequently associated with surgery but this association was not statistically significant

[Ulcerative pancolitis predicts the need for colectomy: study of an incident cohort of patients with ulcerative colitis in Aragón (Spain)]. / La afectación pancolónica inicial en la colitis ulcerosa predice la necesidad de colectomía: estudio poblacional basado en una cohorte incidente de pacientes en Aragón.

INTRODUCTION: Population-based studies show pancolitis as the only risk factor for colectomy in patients with ulcerative colitis (UC). AIM: To evaluate surgical need, its etiology and characteristics, and the distinct clinical variables that act as risk and/or protective factors for the indication for surgery in an incident cohort of patients with UC in Aragón. PATIENTS AND METHOD: Based on the results of a population-based, prospective study reporting the incidence of inflammatory bowel disease in Aragón, 168 patients with a diagnosis of UC were included in the present study. The patients' medical records were reviewed and the following clinical variables were collected from the time of diagnosis (1992-1995) to February 2001: surgery, type of surgical intervention and etiology, anatomic distribution, number of relapses, remissions, clinical course, death, smoking, oral contraceptives and hospitalizations. To investigate the association between these variables and surgery during follow-up, descriptive and bivariate analyses were performed. RESULTS: Of 204 patients diagnosed with UC, 168 (82.3%) with a follow-up of at least 6 months were included. The mean follow-up was 77 months (range: 6-110 months). Some type of surgery was required by 6.5% of our patients during follow-up and 3.6% needed a second surgical intervention. Surgery was indicated for various etiologies. Most patients underwent colectomy and ileostomy with subsequent surgical reconstruction of the intestinal tract. Pancolitis was a clear independent risk factor for colectomy in our patient cohort. No association was found between sex, age at diagnosis, and oral contraceptive intake with surgery during follow-up. Patients who underwent surgery had previously shown more than one relapse and several hospitalizations but did not necessarily show a chronic clinical course. We found no positive or negative association with any smoking-related variables: smoker, non-smoker, years of smoking or years free of smoking. CONCLUSION: Ulcerative pancolitis at diagnosis is a predictive factor for surgery in the short term in patients with a diagnosis of UC in Aragón. We found no other significant associations with the remaining epidemiological factors studied.

La afectación pancolónica inicial en la colitis ulcerosa predice la necesidad de colectomía: estudio poblacional basado en una cohorte incidente de pacientes en Aragón / Ulcerative pancolitis predicts the need for colectomy: study of an incident cohort of patients with ulcerative colitis in Aragón (Spain)

INTRODUCCIÓN: Sobre la base de estudios poblacionales, la única variable clínica consistente y predictiva de un determinado curso clínico que se ha descrito es el mayor riesgo decolectomía en los pacientes con pancolitis ulcerosa. OBJETIVO: Valorar la necesidad de cirugía, su etiología y sus características, así como las distintas variables clínicas que actúan como factores de riesgo o protectores respecto de la indicación quirúrgica de una cohorte incidente de pacientes con diagnóstico de colitis ulcerosa (CU) en Aragón. PACIENTES Y MÉTODO: Basados en los resultados de un estudio poblacional y prospectivo incidente en Aragón, incluimos en el estudio a 168 pacientes diagnosticados de CU; se revisaron sus historias clínicas y se recogieron las distintas variables clínicas desde su diagnóstico (1992-1995) hasta febrero de 2001: cirugía, tipos de intervención quirúrgica y etiología, localización anatómica, número de brotes e ingresos, remisión, curso crónico, fallecimiento, exposición al tabaco, anticonceptivos orales e ingresos. Se realizó un análisis descriptivo y un análisis bivariante para investigar la asociación de estas variables con la cirugía durante el seguimiento. RESULTADOS: Hemos obtenido información durante 6 meses de seguimiento de 168 pacientes (168/204) (82,3%), con un seguimiento medio de 77 meses (rango, 6-110 meses). Durante el seguimiento, el 6,5% de los pacientes precisó algún tipo de intervención quirúrgica y el 3,6%, una segunda cirugía. La indicación quirúrgica es variable; en la mayoría de los casos se realizó colectomía e ileostomía, y fue necesaria una segunda cirugía para la reconstrucción del tránsito intestinal. La pancolitis ulcerosa se comporta como un factor de riesgo independiente para la necesidad de cirugía. En nuestros pacientes, el sexo, la edad al diagnóstico y la toma de anticonceptivos no se asociaron a la presencia de cirugía. Los pacientes que precisaron cirugía durante el seguimiento habían presentado previamente un curso clínico de más de un brote y un mayor número de ingresos, pero no necesariamente un curso crónico. No objetivamos una asociación positiva ni negativa con las variables: expuesto, no fumador, tiempo libre de exposición al tabaco ni tiempo global de exposición. CONCLUSIÓN: La pancolitis ulcerosa al diagnóstico es un factor que se relaciona con la necesidad a corto plazo de la cirugía en los pacientes con diagnóstico inicial de CU en Aragón; no se objetivó ninguna otra asociación entre otros factores epidemiológicos estudiados

INTRODUCTION: Population-based studies show pancolitis as the only risk factor for colectomy in patients with ulcerative colitis (UC).AIM: To evaluate surgical need, its etiology and characteristics, and the distinct clinical variables that act as risk and/or protective factors for the indication for surgery in an incident cohort of patients with UC in Aragón. PATIENTS AND METHOD: Based on the results of a population based, prospective study reporting the incidence of inflammatory bowel disease in Aragón, 168 patients with a diagnosis of UC were included in the present study. The patients’ medical records were reviewed and the following clinical variables were collected from the time of diagnosis (1992-1995)to February 2001: surgery, type of surgical intervention andetiology, anatomic distribution, number of relapses, remissions, clinical course, death, smoking, oral contraceptives and hospitalizations. To investigate the association between these variables and surgery during follow-up, descriptive and bivariate analyses were performed. RESULTS: Of 204 patients diagnosed with UC, 168 (82.3%)with a follow-up of at least 6 months were included. Theme an follow-up was 77 months (range: 6-110 months). Some type of surgery was required by 6.5% of our patients during follow-up and 3.6% needed a second surgical intervention. Surgery was indicated for various etiologies. Most patients underwent colectomy and ileostomy with subsequent surgical re- construction of the intestinal tract. Pancolitis was a clear independent risk factor for colectomy in our patient cohort. No association was found between sex, age at diagnosis, and oral contraceptive intake with surgery during follow-up. Patients who underwent surgery had previously shown more than one relapse and several hospitalizations but did not necessarily show a chronic clinical course. We found no positive or negative association with any smoking related variables: smoker, non-smoker, years of smoking or years free of smoking. CONCLUSION: Ulcerative pancolitis at diagnosis is a predictive factor for surgery in the short

Down-regulation of BRMS1 mRNA Expression in Breast Cancer is not Related to the Presence of Axillary Node Metastasis.

BACKGROUND: BRMS1, recently identified in MDA-MB-435 breast cancer cells, seems to act as a potent antimetastatic gene in experimental tumor models. MATERIALS AND METHODS: To verify if BRMS1 exerts its action in a similar way in clinical tumors, BRMS1-mRNA expression was investigated in a series of 107 human breast carcinomas and correlated with the presence or not of axillary node metastases. Additionally, BRMS1 expression was correlated with all available clinical (histologic variety, histologic and nuclear grade) and biological parameters (ploidy, expression of Ki67, hormone receptors, c-erb-B2 and p53), as well as with the expression of hMAM- and Nup88-mRNA, previously shown by us to correlate with very low and very high aggressiveness of breast cancer, respectively. RESULTS: Down-regulation of BRMS1 expression in the tumors did not correlate with the presence of axillary node metastases. Furthermore, BRMS1 expression did not correlate with any other of the studied clinical or biological tumor parameters. CONCLUSION: In clinical breast cancers, down-regulation of BRMS1 expression seems not to mimic the very clear-cut antimetatstatic properties displayed in experimental tumor models.

The X-chromosome RBMX Gene is Expressed in Mammary Carcinoma.

BACKGROUND: A gene located on the q11.23 region of the male chromosome, RBMY, which plays a role in spermatogenesis, is down-regulated in testicular cancer. RBMY is a diverged X-Y shared gene. The corresponding X chromosome gene, RBMX, is located on Xq26. MATERIALS AND METHODS: We studied fresh tissues from 122 infiltrating breast cancers (99 ductal infiltrating, 19 lobular infiltrating and 4 tubular carcinomas) for the expression of RBMX by means of differential RT-PCR (reverse transcription-polymerase chain reaction), using beta-actin as an internal control and normalization standard. The obtained results were compared with all available clinical and molecular data of the studied tumors (estrogen and progesterone receptors (ER & PR), c-erb-B2, p53, Ki67, DNA-ploidy, Bcl-2, VEGF, CD105 (endoglin), histologic variety, histologic and nuclear grade and axillary node invasion). RESULTS: RBMX RT-PCR was successful in 120/122 instances. All 120 cases expressed RBMX. The only significant correlation found between RBMX expression and all the variables tested was an inverse one with CD105 (endoglin) mRNA-expression (r=-3063; p=0.0007). CONCLUSION: The X-chromosome RBMX gene is expressed in all breast cancers. The expression is inversely correlated with the expression of the angiogenesis-associated CD105 (endoglin) gene. The precise meaning of this association has still to be elucidated.

Elevated mammaglobin (h-MAM) expression in breast cancer is associated with clinical and biological features defining a less aggressive tumour phenotype.

BACKGROUND: Mammaglobin (h-MAM) is expressed mainly by breast epithelial cells, and this feature has been used to detect circulating breast cancer cells and occult metastases in sentinel axillary lymph nodes of breast cancer patients. However, the biological role of mammaglobin is completely unknown. METHODS: We studied 128 fresh-frozen breast cancer specimens by means of reverse transcriptase-polymerase chain reaction and quantified their h-MAM mRNA expression. This was then correlated with histological and nuclear grade, oestrogen and progesterone receptor expression, c-erb-B2 and mutant p53 expression, as well as with cellular proliferation measured by means of the Ki67 labelling index, DNA ploidy and S-phase, and finally with the presence or not of invaded axillary nodes in the mastectomy specimen. RESULTS: In the univariate analysis, high h-MAM expression (above the median for the whole group) correlated significantly (P < 0.05) with oestrogen and progesterone receptor expression, diploid DNA content, low Ki67 labelling index, low nuclear grade and almost significantly (P = 0.058) with the absence of axillary nodal invasion in the mastectomy specimen. In a final, multivariate model, only progesterone receptor expression, diploid DNA content and absence of nodal invasion were found to be independently associated with high h-MAM expression. CONCLUSION: All of the features associated with mammaglobin expression reflect, without exception, a less aggressive tumour phenotype. Further studies are needed to clarify whether this is attributable to h-MAM expression itself, or to another mechanism of which mammaglobin expression forms part.

Immunofluorometric study of Bcl-2 and Bax expression in clinical fresh tumor samples from breast cancer patients.

BACKGROUND: The balance between the expression of the antiapoptotic gene Bcl-2 and the proapoptotic gene Bax is considered a good indicator of the apoptotic activity of tumor cells. Bcl-2 and Bax expression seem also to individually play a prognostic role in breast cancer. Our aim was to study the expression of both genes in fresh breast cancer samples, and to correlate the obtained results with other available clinical and biological parameters of the tumors. MATERIALS AND METHODS: Fresh tumor specimens from 86 breast cancer patients were studied by means of immunofluorocytometry for the expression of the apoptosis-associated Bcl-2 and Bax genes. Additionally, DNA-ploidy was also measured. Paraffin blocks corresponding to the same tumors were used for immunohistochemistry, to study the expression of hormone receptors (ER and PR), p53, c-erb-B2 and the Ki67 labelling index. Fourteen patients had been treated with four cycles of induction chemotherapy (cyclo-phosphamide, adriamycin and 5-fluorouracil), and separate statistical analyses were carried out both for the whole group, and for the 62 patients not having received any treatment whatsoever, in order to exclude any potential influence of the chemotherapeutic treatment on the expression of the studied antigens. RESULTS: Bcl-2 expression correlated significantly with estrogen (p = 0.002) and progesterone (p = 0.012) receptor expression, as well as with c-erb-B2 (p = 0.045) expression in chemotherapy-naïve tumors, the correlation being completely lost if treated tumors were added to the study group. A high apoptotic index (Bcl2/Bax < 0.5) correlated significantly with progesterone receptor expression (p = 0.037) and c-erb-B2 expression (p = 0.018), and this correlation was maintained, whether previously treated tumors were included into the study or not (p = 0.038 and p = 0.027, respectively). Bax expression did not correlate with any other clinical or biological parameters of the tumors, including Bcl-2 expression. CONCLUSION: Bcl-2 and Bax-expression can be easily determined in clinical breast cancer specimens by means of immunofluorocytometry. Bcl-2-expression significantly correlates with hormone-receptor- and c-erb-B2-expression exclusively in previously untreated tumors. This, however, seems only to be the case when considering Bcl-2 expression in isolation, since a high apoptotic index, which considers the ratio of Bcl-2 versus Bax expression in the same tumor, seems not to be affected by the previous use of chemotherapy.

The EMECAM project: a multicentre study on air pollution and mortality in Spain: combined results for particulates and for sulfur dioxide.

OBJECTIVE: The EMECAM study is a collaborative effort to evaluate the impact of air pollution on mortality in Spain. In this paper the combined results are presented for the short term effects of particulates and sulfur dioxide on both daily mortality for all and for specific causes. METHODS: The relation between daily mortality for all causes, cardiovascular diseases, and respiratory diseases, and air pollution for particulates (daily concentrations) and SO(2) (24 and 1 hour concentrations) was assessed in 13 Spanish cities for the period 1990-6. With a standardised method, magnitude of association in each city was estimated by Poisson regression in a generalised additive model. Local estimates were obtained from both single and two pollutant analyses. Lastly, combined estimates for each cause and pollutant were obtained. RESULTS: For combined results, in single pollutant models a 10 microg/m(3) increase in the concentration of the mean of the concurrent and one day lag for black smoke was associated with a 0.8% (95% confidence interval (95% CI) 0.4 to 1.1%) increase in total mortality. The estimates for total suspended particles (TSPs) and particulate matter of aerodynamic diameter <10 microm (PM(10)) and total mortality were slightly lower. The same increase in concentrations of SO(2) was associated with a 0.5% increase in daily deaths. For groups of specific causes, higher estimations were found, specially for respiratory conditions. Peak concentrations of SO(2) showed significant associations with the three groups of mortality. When two pollutant analyses were performed, estimates for particulates, specially for black smoke, did not substantially change. The estimates for daily concentrations of SO(2) were greatly reduced, but, on the contrary, the association with peak concentrations of SO(2) did not show any change. CONCLUSIONS: There is an association between mortality and pollution through particulates among city populations in Spain. Peak rather than daily concentrations of SO(2) were related to mortality. Results suggest that populations in Spanish cities are exposed to health risks derived from air pollution.

Flow cytometry vs. Ki67 labelling index in breast cancer: a prospective evaluation of 181 cases.

BACKGROUND: Excessive proliferation is one of the first steps in oncogenic activation and one of the most important biological features defining the aggressiveness of tumors. Quantifying the proportion of tumor cells in S-phase by means of flow cytometry has shown, in the past, to be useful for defining high-risk subgroups in breast cancer. Several antigens closely associated with proliferation are also detectable by means of immunohistochemistry, offering in theory an easy to perform and cheap alternative to flow cytometry for measuring proliferation. To test this hypothesis, we compared both methods prospectively in a series of breast cancers. MATERIALS AND METHODS: We studied the proliferation rate of 181 breast cancers (152 ductal infiltrating, 17 lobular infiltrating, 12 other histological varieties), operated upon at our institution, by means of flow cytometry and the Ki67 labelling index, using the MIBI antibody. Ploidy (expressed as DNA content or DNA-index), S-phase fraction and the Ki67 labelling index were the variables of the study. The S-phase fraction was considered separately for diploid and aneuploid tumors, following the 1992 Maine Consensus guidelines and was judged abnormally elevated if higher than the 75th percentile for each group. The Ki67 labelling index was expressed as percent positive tumor cells, positive cells being those showing specific nuclear staining. RESULTS: DNA-ploidy and the Ki67 labelling index could be evaluated in all tumors. Of the total, 96 (53%) were diploid and 85 (47%) aneuploid. S-phase fraction could be measured in 172 out of the 181 tumors (95%). The 75th percentile cut-offs for diploid and aneuploid tumors were 9.9% and 15.8%, respectively. We found a significant correlation beween rising DNA content and increasing Ki67 index (r = 0.18; p = 0.022), as well as between the percentage of cells in S-phase of the whole tumor population and Ki67 (r = 0.22; p = 0.0055). A Ki67 cut-off of 50% or higher identified most aneuploid tumors, or a small group of diploid ones with a high S-phase fraction (specificity = 96.7%; positive predictive value 92.5%), however at the price of a very low sensitivity (62.6%). This was due to the presence of many aneuploid tumors with a low S-phase fraction. CONCLUSION: The Ki67 labelling index and S-phase fraction are significantly correlated. However, flow cytometry provides additional indirect information on tumor aggressiveness associated with DNA-ploidy. Further studies are needed to determine whether Ki67 alone is sufficient as a routinely applicable method.

Diferencias bioquímicas entre los quistes de mama benignos tipo II únicos y asociados a tipo I / Biochemical differences between single benign type ii breast cysts and those associated with type ii breast cysts

Introducción. Los quistes mamarios tipo II se caracterizan por presentar en el líquido altas concentraciones de Na+, albúmina, pH y cloruros (Cl-) y parecen corresponder al estadio más avanzado de la enfermedad macroquística. En el presente estudio hemos querido analizar la posible influencia de la coexistencia de quistes tipo I, reflejo de la actividad de la enfermedad, sobre las características bioquímicas de los quistes tipo II. Pacientes y métodos. El grupo estudio incluyó 124 líquidos de quistes tipo II (Na+/K+ > 1,5), de los cuales 72 fueron únicos y 52 asociados a quistes tipo I. En ellos determinamos las concentraciones de Na+, K+, Cl-, glucosa, albúmina, pH y volumen. Resultados. Los quistes tipo II únicos presentaron mayores valores de pH (p = 0,0306) e índice Na+K+ (p = 0,0205), así como menores de K+ (p = 0,0313) y volumen (p = 0,0014). No se constataron diferencias entre ambos grupos en las mujeres en fase folicular, pero sí en las de fase luteínica y menopáusicas. Cuando el dintel clasificador de los quistes fue establecido en una relación Na+/K+ > 3, observamos que los quistes tipo II únicos presentaban valores mayores de pH y menor volumen. Conclusiones. Los resultados anteriores indican que los quistes tipo II únicos presentan unas características bioquímicas distintas de cuando se asocian con quistes tipo I, de tal modo que en esta situación adquieren ciertas propiedades de estos últimos, lo cual puede ser el exponente de una fase más activa de la enfermedad (AU)