A study of the SCN5A gene in a cohort of 76 patients with Brugada syndrome.

We aim to study the SCN5A gene in a cohort of Brugada syndrome (BS) patients and evaluate the genotype-phenotype correlation. BS is caused by mutations in up to 10 different genes, SCN5A being the most frequently involved. Large genomic rearrangements in SCN5A have been associated with conduction disease, but its prevalence in BS is unknown. Seventy-six non-related patients with BS were studied. Clinical characteristics and family risk profile were recorded. Direct sequencing and multiplex ligation-dependent probe amplification (MLPA) of the SCN5A gene for identification of mutations and larger rearrangements were performed, respectively. Eight patients (10.5%) had point mutations (R27H, E901K, G1743R (detected in three families), V728I, N1443S and E1152X). Patients with mutations had a trend toward a higher proportion of spontaneous type I Brugada electrocardiogram (ECG) (87.5% vs 52.9%, p = 0.06) and had evidence of familial disease (62.5%, vs 23.5%, p = 0.03). The symptoms and risk profile of the carriers were not different from wild-type probands. There were non-significant differences in the prevalence of type I ECG, syncope and history of arrhythmia in carriers of selected polymorphisms. None of the patients had any deletion/duplication in the SCN5A gene. In conclusion, 10.5% of our patients had mutations in the SCN5A gene. Patients with mutations seemed to have more spontaneous type I ECG, but no differences in syncope or arrhythmic events compared with patients without mutations. Larger studies are needed to evaluate the role of polymorphisms in the SCN5A in the expression of the phenotype and prognosis. Large rearrangements were not identified in the SCN5A gene using the MLPA technique.

[About a fatal infectious myositis: case report]. / A propos d'un cas de myosite infectieuse fatale.

Soft tissues' infections involve fascia and musculature which permit to define different anatomic entities as necrotizing fasciitis or myositis. We describe here the case of an infectious myositis whose initial clinical approach did not allow to get the right diagnosis. The tomodensitometry showed typical lesions which induced an accurate management, but could not avoid the death of the patient because of major wounds and a poor general background.

[The chondrosarcoma of the hand: review of the literature about a case and a proposition of a decision-making therapeutic tree]. / Le chondrosarcome de la main: revue de la littérature à propos d'un cas et proposition d'un arbre thérapeutique décisionnel.

We report a case of chondrosarcoma of the hand initially treated as a chondroma. Through a review of the literature, we review the clinical, radiological and histological diagnostic difficulties of this lesion. We propose a decision-making tree for the surgical coverage of the chondrosarcoma of the hand.

[Silicone or saline - filled implants for breast augmentation?]. / Critères intervenant dans le choix entre prothèse préremplie au gel de silicone et prothèse remplie au sérum physiologique en chirurgie mammaire d'augmentation.

The use is to oppose silicone and saline implants. Both are used in regular practice. What are the arguments to choose in between the two types? We separated three different criteria groups: prosthesis characteristics, surgical procedures and complications. For each criterion we made a review of literature looking for arguments in favour or against silicone or saline implants. The number of criteria to choose the correct type of prosthesis is very important. We conclude that surgeon experience; desires of the patient and evolution of prosthesis are the most important criteria.

Copper metabolism and biliary secretion in patients receiving orthotopic liver transplantation.

BACKGROUND: The quantitative aspects of biliary copper excretion in health and disease have not been fully defined yet. The aim of the study was to evaluate copper metabolism and biliary excretion of patients who have received an orthotopic liver transplant (OLT) during the immediate postoperative period. METHODS: We have studied retrospectively 16 patients undergoing primary OLT and eight undergoing cholecystectomy, and measured serum concentration of copper and its secretion in bile and urine by flame atomic absorption spectrometry (FAAS). RESULTS: We found a progressive increase of biliary copper secretion rates and a corresponding lowering of urinary copper during the postoperative period. Thus, in OLT patients, the mean of biliary copper secretion on day 1 is 0.7+/-0.2 micromol/day compared with 2.3+/-1.1 micromol/day on day 7 (p<0.01) and 6.1+/-2.5 micromol/day on day 15 (p<0.0001). The rate of copper output on day 5 after surgery is about one sixth of the value reported for patients who had undergone cholecystectomy. In patients suffering an acute rejection episode, there was an abrupt fall in bile flow (<15 ml/day) and excretion of biliary copper (<1 micromol/day), accompanied by an increase of urine copper excretion (>3 micromol/day), and both were recovered when the rejection episode was solved. We found an inverse relationship between the serum bilirubin (Bt), alkaline phosphatase (ALP) and the biliary copper excretion (p<0.01), and a direct relationship with urinary copper excretion (p<0.01). CONCLUSIONS: The copper measurements in urine and bile are non-invasive techniques, of low cost, rapid and easy to accomplish, and available in hospitals accredited for hepatic transplantation. These characteristics make these methods helpful in the monitoring of patients submitted to OLT for assessment of graft quality and subsequent outcome.

Radioterapia postmastectomía en pacientes con 1-3 ganglios positivos. Evidencia científica y práctica clínica / Postmastectomy radiotherapy in patients with 1-3 positive axillary nodes. Scientific evidence and clinical practice

Aún no ha sido establecido si las pacientes con cáncer de mama con un tumor primario menor o igual a 5 cm y con 1 a 3 ganglios axilares afectos en el momento del diagnóstico se beneficiarían de radioterapia postmastectomía. La radioterapia moderna ha demostrado su utilidad en pacientes con alto riesgo de recidiva local tanto en términos de mejor control loco-regional como en aumento de la supervivencia. Pero en el grupo de pacientes con riesgo moderado (pT1-2 N1 y 1 a 3 ganglios positivos) los ensayos aleatorizados están aún en marcha y no se espera obtener las primeras conclusiones en al menos diez años. La decisión de tratamiento se debería basar en el conocimiento del comportamiento biológico del cáncer de mama y en la evidencia científica acumulada. Revisamos extensamente ambos aspectos y concluimos que los pacientes en esta categoría tienen un beneficio en el control loco-regional que podría traducirse en mejor supervivencia en algunos subgrupos (AU)

Preoperative values of CA 15-3 and CEA as prognostic factors in breast cancer: a multivariate analysis.

The role of circulating tumor markers in providing prognostic information has not been widely studied. In the current study, serum levels of the carbohydrate antigen 15-3 (CA 15-3) and carcinoembryonic antigen (CEA) were determined preoperatively in 364 breast cancer patients with no clinical signs of metastasis. The prognostic relevance of these markers for recurrence (175/364) and death of disease (104/175) was determined by Cox multivariate analysis, including the comparison with classical prognostic factors. High levels of both tumor markers were associated with aneuploid tumors with high S-phase fraction and high ornithine decarboxylase activity. CA 15-3 was highly associated with the number of positive lymph nodes and peritumoral lymphatic or blood vessel invasion. No significant associations were found between CEA or CA 15-3 levels and histologic grade, necrosis and steroid receptor status. In univariate analysis, preoperative values, using optimum cutoff values of CA 15-3 (40 U/ml) and CEA (6 ng/ml), were statistically significant for relapse-free survival and overall survival. In multivariate analysis, only node status, DNA ploidy and ornithine decarboxylase activity were independent predictors for relapse-free survival; the estrogen receptor status was a predictor of overall survival. In node-negative patients, ornithine decarboxylase activity was the only factor selected for relapse-free survival. In node-positive patients, the number of lymph nodes and DNA ploidy were the only variables selected for relapse-free survival or overall survival. Estrogen receptor and ornithine decarboxylase activity were excluded for relapse-free survival, but were significant prognostic factors for overall survival.

[Temporary dependency of steroid-receptor prognostic value in breast cancer]. / Dependencia temporal del valor pronóstico de los receptores esteroides en el cáncer de mama.

BACKGROUND: The influence of prognostic factors in breast cancer may change during follow-up. Our aim was therefore to look for time-related changes in hazard ratios for steroid receptor status and the risk of relapse. PATIENTS AND METHOD: We studied 455 patients with operable primary breast cancer during a mean follow-up period of 72 months (range, 42-130 months). Clinical and histological characteristics were assessed and estrogen receptor (ER) and progesterone receptor (PgR) status were determined and compared to disease-free survival (DFS) and overall survival (OS). Analysis of DFS included fitting a multivariate Cox proportional hazards model, testing for non-proportionality, and examining diagnostic plots. RESULTS: A total of 66.8% of the tumors were ER positive and 54.7% PgR positive. Receptor status is highly associated with menopausal status, histological grade, peritumoral lymphatic or blood vessel invasion and mitotic index. No correlation was found between steroid receptor status and tumor size, node status or tumor stage. In univariate analysis, ER/PR vs ER+/PR+ display a hazard ratio of 2.15 (IC del 95%, 1.59-2.99) for DFS (p = 0.001) and 1.95 (IC del 95%, 1.38-2.59) for OS (p = 0.0043). In multivariate analysis, steroid receptor status, node status and mitotic index were independent prognostic factors for DFS and OS. By using Cox regression time-dependent covariates model, we show that the steroid receptor status hazard ratio is 5.6 at diagnosis, decreases after 4 years, and loses its significance after 10 years of follow-up. CONCLUSION: In patients with breast cancer the status steroid receptor have a relatively limited prognostic value and is lost within the first years.

Changes in elemental content during apoptotic cell death studied by electron probe X-ray microanalysis.

Recent data suggest that changes in ionic content, primarily potassium, play a pivotal role in the progression of apoptosis. However, the changes in total element content, i.e., sodium (Na), magnesium (Mg), phosphorous (P), chlorine (Cl), potassium (K), and calcium (Ca), during apoptosis have not been evaluated. Electron probe X-ray microanalysis (EPXMA) was used to measure total element content in U937 cells before and after the induction of apoptosis. As an experimental model we used U937 cells irradiated with ultraviolet (UV) light. Apoptosis was evaluated with phase-contrast microscopy, with scanning and transmission electron microscopy, and with the fluorescent dye bisbenzimide (Hoechst 33342). Plasma membrane permeability as a measure of cell death was determined by trypan blue dye exclusion. To investigate element content with EPXMA, cells were cryoprepared, i.e., cryofixed and freeze-dried, and analyzed as whole cells using a scanning electron microscope. We found that the UV irradiation induced rapid (within 2 h) morphological changes associated with apoptosis, such as plasma membrane blebbing, condensation of the chromatin, and the formation of membrane-bound apoptotic bodies. At this time, 95% of the apoptotic cells excluded trypan blue dye. EPXMA results demonstrated that UV light-irradiated apoptotic cells (cells with membrane-bound apoptotic bodies) had a lower Cl content (P < 0.001) and K content (P < 0.001) and a higher Na content (P < 0.001) in comparison with nonirradiated control cells. Also, P and Ca content was higher in apoptotic cells than in control cells, but this difference did not reach statistical significance. No differences were found in Mg. These data indicated that morphological changes characteristic of apoptotic cell death are related with significant changes in sodium, chlorine, and potassium content. In addition, we demonstrated that these changes in elemental composition were not associated with loss of cell membrane integrity.

A procedure to prepare cultured cells in suspension for electron probe X-ray microanalysis: application to scanning and transmission electron microscopy.

We describe a simple procedure to prepare cultured cells in suspension to analyse elemental content at the cellular level by electron probe X-ray microanalysis. Cells cultured in suspension were deposited onto polycarbonate tissue, culture plate well inserts, centrifuged at low g, washed to remove the extracellular medium, cryofixed and freeze-dried, and analysed in the scanning mode of a scanning electron microscope. We tested the effect of different washing solutions (150 mM ammonium acetate, 300 mM sucrose, and distilled water) on the elemental content of cultured cells in suspension. The results demonstrated that distilled water was the best washing solution to prepare cultured cells. In addition, the low Na content, high K content and high K/Na ratio of the cells indicated that this procedure, based on the centrifugation at low g followed by cryopreparation, constitutes a satisfactory method to prepare cultured cells in suspension. We also investigated the effects of different accelerating voltages on X-ray signal collection. The results showed that moderate accelerating voltages, i.e. 10-11 kV, should be used to analyse whole cells in the scanning mode of the scanning electron microscope. We show that this method of preparation makes it possible to prepare cryosections of the cultured cells, thus permitting analysis of the elemental content at the subcellular level, i.e. nucleus, cytoplasm and mitochondria, using a scanning transmission electron microscope.

Prognostic value of ornithine decarboxylase and polyamines in human breast cancer: correlation with clinicopathologic parameters.

The polyamines putrescine, spermidine, and spermine and ornithine decarboxylase (ODC), the rate-limiting enzyme in their biosynthetic pathway, play an important role in cell proliferation, differentiation, and transformation. In the present study, we have analyzed polyamine concentrations and ODC activity in samples from benign breast diseases (n = 36), benign breast tissue adjacent to the primary carcinoma (n = 19), and breast carcinoma (n = 104). ODC activity in primary carcinoma was significantly higher (2.42 +/- 0.22 nmol CO2/h g; P < 0.001) than that found in benign breast (0.62 +/- 0.15 nmol CO2/h g) or in breast tissue adjacent to the primary carcinoma (0.52 +/- 0.16 nmol CO2/h g). The total polyamine content of breast cancer tissues was higher than in benign breast diseases (704.3 +/- 38.3 nmol/g wet weight versus 295.8 +/- 27.4 nmol/g wet weight) and correlated well with ODC activity (Pearson, r = 0.42; P < 0.001). ODC activity correlated with histological grade, peritumoral lymphatic or blood vessel invasion, S-phase fraction, and cathepsin D. Total polyamine concentration increased with S-phase fraction, cathepsin D, and aneuploidy. No significant correlation was found between ODC or polyamines and tumor size, lymph node involvement, or steroid receptor status. A major finding in our study was that ODC activity was an independent prognostic factor for recurrence and death. The results indicate that the estimation of ODC activity and polyamines in human breast carcinoma might be useful to determine tumor aggressiveness and suggest that ODC may have a potential value as both a prognostic factor and a chemoprevention target in human breast cancer.

Backscattered electron imaging of cultured cells: application to electron probe X-ray microanalysis using a scanning electron microscope.

We report a simple method to study the elemental content in cultured human adherent cells by electron probe X-ray microanalysis with scanning electron microscopy. Cells were adapted to grow on polycarbonate tissue culture cell inserts, washed with distilled water, plunge-frozen with liquid nitrogen and freeze-dried. Unstained, freeze-dried cultured cells were visualized in the secondary and backscattered electron imaging modes of scanning electron microscopy. With backscattered electron imaging it was possible to identify unequivocally major subcellular compartments, i.e. the nucleus, nucleoli and cytoplasm. X-ray microanalysis was used simultaneously to determine the elemental content in cultured cells at the cellular level. In addition, we propose some improvements to optimize backscattered electron and X-ray signal collection. Our findings demonstrate that backscattered electron imaging offers a powerful method to examine whole, freeze-dried cultured cells for scanning electron probe X-ray microanalysis.