Homogenizing the Teaching of Engaged Medical Examinations to Large Numbers of Students: the Experience of the Digital Rectal Examination.

This project aims to complement and homogenise the teaching of indications and technique of digital rectal examination (DRE) through the use of simulators, and subsequently analysed the level of satisfaction with the training and skills acquired. The students were distributed into small groups. One of the workshop's coordinators synthesised indications and procedures of DRE. A teaching video was made with all the contents and was distributed between the trainers. During the workshop, trainers explained the indications and the method of performing the DRE. Then, the selected clinical cases were presented, followed by the DRE by specific simulators. Once the students had completed each exploration, the trainers explained each case and discussed it with students. The following week, an anonymous questionnaire was given to participants to evaluate the workshop. Of the 232 participating students, 53 (23%) responded to the questionnaire. The overall level of satisfaction was higher than 98% (score 4-5), reaching 100% in the evaluation of the practical contents, and 93% of the students would recommend the continuity of the workshop in the next courses. The DRE workshop was well received among medical students, with a high degree of voluntary participation and response rate to the subsequent survey. With this project, we have achieved a greater homogenisation of teaching within the subject of Urology, and greater confidence for the students when facing their future clinical practice.

Genetic counseling in prostate cancer: How to implement it in daily clinical practice? / Asesoramiento genético en cáncer de próstata: ¿cómo implementarlo en la práctica clínica diaria?

Prostate cancer plays an undeniably prominent role in public health in our days and health systems. Its epidemiological impact is quantitatively very close to that of other tumors such as colon cancer and breast cancer, in which genetic counseling is part of their routine clinical practice, both in the initial evaluation and in the selection of therapeutic strategies. Hereditary cancer syndromes, breast/ovarian and Lynch syndrome are part of genetic counseling in these tumors. Currently, we also know that they can be associated to prostate cancer. The time has come to implement genetic counseling in prostate cancer from the earliest stages of its approach, from initial suspicion to the most advanced tumors. We present an updated review carried out by our interdisciplinary working group on scientific literature, clinical practice guidelines and consensus documents, aimed at the creation and drafting of a'Protocol for genetic counseling in prostate cancer' for the study of germline, with easy application in different healthcare settings. This protocol is currently being implemented in our routine practice and provides answers to 3 specific questions: Who should receive genetic counseling for prostate cancer? Which gene panel should be analyzed? How should counseling be done according to the results obtained? Other aspects about who should perform genetic counseling, ethical considerations and regulations are also collected.

A Preliminary Study of the Ability of the 4Kscore test, the Prostate Cancer Prevention Trial-Risk Calculator and the European Research Screening Prostate-Risk Calculator for Predicting High-Grade Prostate Cancer.

INTRODUCTION: To prevent the overdiagnosis and overtreatment of prostate cancer (PC), therapeutic strategies have been established such as active surveillance and focal therapy, as well as methods for clarifying the diagnosis of high-grade prostate cancer (HGPC) (defined as a Gleason score ≥7), such as multiparametric magnetic resonance imaging and new markers such as the 4Kscore test (4KsT). By means of a pilot study, we aim to test the ability of the 4KsT to identify HGPC in prostate biopsies (Bx) and compare the test with other multivariate prognostic models such as the Prostate Cancer Prevention Trial Risk Calculator 2.0 (PCPTRC 2.0) and the European Research Screening Prostate Cancer Risk Calculator 4 (ERSPC-RC 4). MATERIAL AND METHODS: Fifty-one patients underwent a prostate Bx according to standard clinical practice, with a minimum of 10 cores. The diagnosis of HGPC was agreed upon by 4 uropathologists. We compared the predictions from the various models by using the Mann-Whitney U test, area under the ROC curve (AUC) (DeLong test), probability density function (PDF), box plots and clinical utility curves. RESULTS: Forty-three percent of the patients had PC, and 23.5% had HGPC. The medians of probability for the 4KsT, PCPTRC 2.0 and ERSPC-RC 4 were significantly different between the patients with HGPC and those without HGPC (p≤.022) and were more differentiated in the case of 4KsT (51.5% for HGPC [25-75 percentile: 25-80.5%] vs. 16% [P 25-75: 8-26.5%] for non-HGPC; p=.002). All models presented AUCs above 0.7, with no significant differences between any of them and 4KsT (p≥.20). The PDF and box plots showed good discriminative ability, especially in the ERSPC-RC 4 and 4KsT models. The utility curves showed how a cutoff of 9% for 4KsT identified all cases of HGPC and provided a 22% savings in biopsies, which is similar to what occurs with the ERSPC-RC 4 models and a cutoff of 3%. CONCLUSIONS: The assessed predictive models offer good discriminative ability for HGPCs in Bx. The 4KsT is a good classification model as a whole, followed by ERSPC-RC 4 and PCPTRC 2.0. The clinical utility curves help suggest cutoff points for clinical decisions: 9% for 4KsT and 3% for ERSPC-RC 4. This preliminary study should be interpreted with caution due to its limited sample size.

Combined treatment with bexarotene and rosuvastatin reduces angiotensin-II-induced abdominal aortic aneurysm in apoE(-/-) mice and angiogenesis.

BACKGROUND AND PURPOSE: Abdominal aortic aneurysm (AAA) is a degenerative vascular disease associated with angiogenesis. Bexarotene is a retinoid X receptor (RXR) ligand with anti-angiogenic activity. Statins also exert anti-angiogenic activity and activate PPARs. Because RXR ligands form permissive heterodimers with PPARs and a single anti-angiogenic drug may not be sufficient to combat the wide array of angiogenic factors produced during AAA, we evaluated the effect of combined low doses of bexarotene and rosuvastatin in a mouse model of AAA. EXPERIMENTAL APPROACH: The effect of the combined treatment was investigated in a murine model of angiotensin II-induced AAA in apoE(-/-) mice. This combination therapy was also evaluated in in vivo (Matrigel plug assay) and in vitro (endothelial cell differentiation assay) models of angiogenesis as well as the underlying mechanisms involved. KEY RESULTS: Co-treatment with bexarotene plus rosuvastatin reduced aneurysm formation, inflammation and neovascularization compared with each single treatment. In HUVEC, the combination of suboptimal concentrations of bexarotene and rosuvastatin inhibited angiotensin II-induced morphogenesis, proliferation and migration. These effects were accompanied by diminished production of pro-angiogenic chemokines (CXCL1, CCL2 or CCL5) and VEGF, and seemed to be mediated by RXRα/PPARα and RXRα/PPARγ activation. This combined therapy reduced the activation of members of the downstream PI3K pathway (Akt/mTOR and p70S6K1) in vivo and in vitro. CONCLUSIONS AND IMPLICATIONS: The combination of RXR agonists with statins at low doses synergistically interferes with the signalling pathways that modulate inflammation and angiogenesis and may constitute a new and safer therapeutic treatment for the control of AAA.

PPARß activation restores the high glucose-induced impairment of insulin signalling in endothelial cells.

BACKGROUND AND PURPOSE: PPARß enhances insulin sensitivity in adipocytes and skeletal muscle cells, but its effects on insulin signalling in endothelial cells are not known. We analysed the effects of the PPARß/δ (PPARß) agonists, GW0742 and L165041, on impaired insulin signalling induced by high glucose in HUVECs and aortic and mesenteric arteries from diabetic rats. EXPERIMENTAL APPROACH: Insulin-stimulated NO production, Akt-Ser(473) and eNOS-Ser(1177) phosphorylation, and reactive oxygen species (ROS) production were studied in HUVECs incubated in low- or high-glucose medium. Insulin-stimulated relaxations and protein phosphorylation in vessels from streptozotocin (STZ)-induced diabetic rats were also analysed. KEY RESULTS: HUVECs incubated in high-glucose medium showed a significant reduction in insulin-stimulated production of NO. High glucose also reduced insulin-induced Akt-Ser(473) and eNOS-Ser(1177) phosphorylation, increased IRS-1-Ser(636) and ERK1/2-Thr(183) -Tyr(185) phosphorylation and increased ROS production. The co-incubation with the PPARß agonists GW0742 or L165041 prevented all these effects induced by high glucose. In turn, the effects induced by the agonists were suppressed when HUVEC were also incubated with the PPARß antagonist GSK0660, the pyruvate dehydrogenase kinase (PDK)4 inhibitor dichloroacetate or after knockdown of both PPARß and PDK4 with siRNA. The ERK1/2 inhibitor PD98059, ROS scavenger catalase, inhibitor of complex II thenoyltrifluoroacetone or uncoupler of oxidative phosphorylation, carbonyl cyanide m-chlorophenylhydrazone, also prevented glucose-induced insulin resistance. In STZ diabetic rats, oral GW0742 also improved insulin signalling and the impaired NO-mediated vascular relaxation. CONCLUSION AND IMPLICATIONS: PPARß activation in vitro and in vivo restores the endothelial function, preserving the insulin-Akt-eNOS pathway impaired by high glucose, at least in part, through PDK4 activation.

Rehabilitación física en el paciente gran quemado / Physical rehabilitation in extensively burned patients

La rehabilitación del paciente gran quemado tiene como objetivo prevenir y minimizar las secuelas derivadas de la propia lesión y del encamamiento. Se sustenta sobre tres pilares: 1) tratamiento postural para prevenir retracciones; 2) cinesiterapia en sus diferentes modalidades para mantener/recuperar rangos articulares y fuerza muscular; 3) recuperación funcional. Es fundamental que el médico rehabilitador y el fisioterapeuta formen parte del equipo multidisciplinar que atiende al paciente. Un trabajo conjunto y coordinado garantizará mejores resultados e influirá positivamente en la calidad de vida del paciente (AU)

The rehabilitation of the extensively burned patient aims to prevent and minimize consequences of the own lesion this is supported by three pillars: 1) postural treatment to prevent contractures; 2) kinesiotherapy to maintain/restore joint range of movement and muscle strength; 3) functional recovery. Physical therapy is essential in the multidisciplinary team. A coordinated team assures better results and positively influences the quality of life of the patients (AU)

Identification of RFLP and NBS/PK profiling markers for disease resistance loci in genetic maps of oats.

Two of the domains most widely shared among R genes are the nucleotide binding site (NBS) and protein kinase (PK) domains. The present study describes and maps a number of new oat resistance gene analogues (RGAs) with two purposes in mind: (1) to identify genetic regions that contain R genes and (2) to determine whether RGAs can be used as molecular markers for qualitative loci and for QTLs affording resistance to Puccinia coronata. Such genes have been mapped in the diploid A. strigosa × A. wiestii (Asw map) and the hexaploid MN841801-1 × Noble-2 (MN map). Genomic and cDNA NBS-RGA probes from oat, barley and wheat were used to produce RFLPs and to obtain markers by motif-directed profiling based on the NBS (NBS profiling) and PK (PK profiling) domains. The efficiency of primers used in NBS/PK profiling to amplify RGA fragments was assessed by sequencing individual marker bands derived from genomic and cDNA fragments. The positions of 184 markers were identified in the Asw map, while those for 99 were identified in the MN map. Large numbers of NBS and PK profiling markers were found in clusters across different linkage groups, with the PK profiling markers more evenly distributed. The location of markers throughout the genetic maps and the composition of marker clusters indicate that NBS- and PK-based markers cover partly complementary regions of oat genomes. Markers of the different classes obtained were found associated with the two resistance loci, PcA and R-284B-2, mapped on Asw, and with five out of eight QTLs for partial resistance in the MN map. 53 RGA-RFLPs and 187 NBS/PK profiling markers were also mapped on the hexaploid map A. byzantina cv. Kanota × A. sativa cv. Ogle. Significant co-localization was seen between the RGA markers in the KO map and other markers closely linked to resistance loci, such as those for P. coronata and barley yellow dwarf virus (Bydv) that were previously mapped in other segregating populations.

Use of tyramide-fluorescence in situ hybridization and chromosome microdissection for ascertaining homology relationships and chromosome linkage group associations in oats.

The physical mapping of single locus sequences by tyramide-fluorescence in situ hybridization (Tyr-FISH) and the analysis of sequences obtained from microdissected chromosomes were assayed as potential tools for (1) determining homology and homoeology among chromosome regions of Avena species, and (2) establishing associations between linkage groups and specific chromosomes. Low copy number probes, derived from resistance gene analogues (RGAs) and 2.8-4.5 kb long, successfully produced hybridization signals on specific chromosomes. Four sets of homoeologous chromosome regions were identified in the hexaploids using 3 probes that produced 4 single locus markers in A. strigosa and 2 in A. eriantha. Laser capture microdissection of metaphase I cells of A. sativa monosomic lines allowed the isolation of critical univalents. Sequences derived from 2 RGAs were successfully amplified in DNA extracted from univalents. In one instance, it was possible to map a nucleotide polymorphism specific for 1 chromosome. An association was established between this chromosome and its linkage groups in 2 hexaploid genetic maps. The results indicate that Tyr-FISH is useful in the characterization of homoeologous chromosome segments in hexaploids, whereas chromosome microdissection, as employed in this work, needs to be improved before it can routinely be used with meiotic chromosomes.

[GEITDAH consensus on attention deficit hyperactivity disorder]. / Consenso del GEITDAH sobre el trastorno por deficit de atencion/hiperactividad.

In this article, the GEITDAH -the Spanish abbreviation of the Special Interest Group on Attention Deficit Hyper-activity Disorder (ADHD)- presents a consensus reached by experts in the management of ADHD from all over Spain. The consensus concerns fundamental aspects that should be the starting point for future local or regional consensus guides. Another aim of this consensus is also to reduce the amount of variability that occurs in the health care offered to patients with ADHD in our country, as well as to act as a stimulus in educational matters. That fact that it is not very long will make it more popular among greater numbers of people and this will allow these goals to be reached more effectively. The conclusions in the consensus guide have been constructed around an introduction dealing with basic aspects and recommendations for diagnosis, treatment (both pharmacological and psychotherapeutic), patient flow and organisational aspects.

A new chromosome nomenclature system for oat (Avena sativa L. and A. byzantina C. Koch) based on FISH analysis of monosomic lines.

Fluorescent in situ hybridization (FISH) with multiple probes was used to analyze mitotic and meiotic chromosome spreads of Avena sativa cv 'Sun II' monosomic lines, and of A. byzantina cv 'Kanota' monosomic lines from spontaneous haploids. The probes used were A. strigosa pAs120a (a repetitive sequence abundant in A-genome chromatin), A. murphyi pAm1 (a repetitive sequence abundant in C-genome chromatin), A. strigosa pITS (internal transcribed spacer of rDNA) and the wheat rDNA probes pTa71 (nucleolus organizer region or NOR) and pTa794 (5S). Simultaneous and sequential FISH employing pairs of these probes allowed the identification and genome assignation of all chromosomes. FISH mapping using mitotic and meiotic metaphases facilitated the genomic and chromosomal identification of the monosome in each line. Of the 17 'Sun II' lines analyzed, 13 distinct monosomic lines were found, corresponding to four monosomes of the A-genome, five of the C-genome and four of the D-genome. In addition, 12 distinct monosomic lines were detected among the 20 'Kanota' lines examined, corresponding to six monosomes of the A-genome, three of the C-genome and three of the D-genome. The results show that 19 chromosomes out of 21 of the complement are represented by monosomes between the two genetic backgrounds. The identity of the remaining chromosomes can be deduced either from one intergenomic translocation detected on both 'Sun II' and 'Kanota' lines, or from the single reciprocal, intergenomic translocation detected among the 'Sun II' lines. These results permit a new system to be proposed for numbering the 21 chromosome pairs of the hexaploid oat complement. Accordingly, the A-genome contains chromosomes 8A, 11A, 13A, 15A, 16A, 17A and 19A; the C-genome contains chromosomes 1C, 2C, 3C, 4C, 5C, 6C and 7C; and the D-genome consists of chromosomes 9D, 10D, 12D, 14D, 18D, 20D and 21D. Moreover, the FISH patterns of 16 chromosomes in 'Sun II' and 15 in 'Kanota' suggest that these chromosomes could be involved in intergenomic translocations. By comparing the identities of individually translocated chromosomes in the two hexaploid species with those of other hexaploids, we detected different types of intergenomic translocations.

Un nuevo caso de histiocitoma fibroso maligno dependiente de cápsula renal / A new case of malignant fibrous histiocytoma arising from the renal capsule

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Taurine chloramine inhibits functional responses of human eosinophils in vitro.

BACKGROUND: Eosinophils are prominent effectors of allergic inflammation. Taurine-chloramine (TauCl), a derivative of the amino acid taurine, shows antioxidant properties in different cell systems but its effects on eosinophils have not been reported. OBJECTIVE: To study the effects of TauCl and taurine on functional responses of isolated human eosinophils activated by different stimuli. METHODS: Human eosinophils were purified from the blood of healthy donors by a magnetic bead separation system. The effects of TauCl and taurine (0.1-1 mM) were investigated on the generation of superoxide anion (ferricytochrome-c reduction microassay), calcium signal (fluorimetry), p47phox-p67phox translocation (Western blot), leukotriene C4 (LTC4) production (enzymeimmunoassay), eosinophil peroxidase (EPO) release (spectrophotometry), eosinophil cationic protein (ECP) release (radioimmunoassay), apoptosis (flow cytometry with annexin V-propidium iodide), and nuclear factor-kappaB (NF-kappaB) activation (Western blot). RESULTS: TauCl inhibited superoxide anion generation triggered by N-formyl-Met-Leu-Phe (fMLP; 30 nM), phorbol myristate acetate (1 nM) and serum opsonized zymosan (0.5 mg/mL) with similar potency (IC50 approximately 200 microM) for the three stimuli, while taurine (0.1-1 mM) was scarcely effective. TauCl but not taurine inhibited p47phox-p67phox translocation. TauCl (200 microM) and taurine (1 mM) did not modify the [Ca2+]i responses to fMLP. TauCl inhibited the release of EPO (IC50 approximately 200 microM) and reduced ECP and LTC4 production from fMLP-activated eosinophils while taurine was without significant effects. TauCl (1 mM) did not change constitutive apoptosis but significantly attenuated the ability of granulocyte-monocyte colony-stimulating factor (GM-CSF) and IL-5 to prevent apoptosis. The activation of eosinophil NF-kappaB induced by GM-CSF and IL-5 was suppressed by TauCl. CONCLUSION: Taurine is without significant in vitro effects on human eosinophil functions but its derivative TauCl inhibits oxidative burst and generation of inflammatory mediators, and reverses the survival effect produced by inflammatory cytokines. Therefore, endogenous TauCl may help to suppress excessive inflammatory response in eosinophils at inflammatory sites.

Analysis of molecular markers in three different tomato cultivars exposed to ozone stress.

Three differentially expressed cDNAs have been isolated from ozone treated tomato seedlings. Their level of expression after ozone exposure has been analysed in three tomato cultivars with different sensitivity to ozone (Nikita, Alisa Craig and Valenciano). These comparative analyses have been extended to a number of genes involved in antioxidative, wounding or pathogenesis responses, showing several differences among cultivars that could be related with their different sensitivity to ozone. Gene response to ozone was affected not only by the period and dose of ozone exposure (short time or chronic), but also by growth conditions (controlled growth chamber or field). Comparison of gene expression patterns puts on evidence the needing of validation in field of experiments performed with plants grown under controlled conditions. Our results suggest that changes in genes expression, observed after ozone treatment in field, are affected by additional factors related to environmental clues.

Expresion del receptor de esteroides y xenobioticos (SXR) y del gen de multirresistencia drogas (MDR1) y de los polimorfismos de las enzimas GSTs, SULTs y CYP en tumores vesicales profundos, analisis de su expresion y correlación con otros factores pronósticos / Steroid and xenobiotic receptor (SXR), multidrug resistance gene (MDR1) and GSTs, SULTs and CYP polymorphism expression in invasive bladder cancer, analysis of their expression and correlation with other prognostic factors

Introducción: El receptor de esteroides y xenobióticos SXR se ha demostrado su activación por parte de numerosos medicamentos, incluidos potentes inductores del citocromo P450, como la rifampicina y el cotrimazol. La función del SXR es bien conocida, y consiste en regular de manera positiva la trascripción del citocromo P450 3A4 (CYP3A4) y el gen de multirresistencia a drogas (multidrug resistance gene) MDR1, se considera una llave clave en el mecanismo regulador del metabolismo de los xenobióticos encontrándose involucrado en todas las fases de detoxificación Múltiples enzimas involucradas en el metabolismo y la degradación de hidrocarburos policíclicos aromáticos (PAH) son polimórficas en humanos, incluyendo la glutation S-transferasa (GSTs), N-acetiltransferasa (NATs), sulfotransferas (SULTs)1A1 y el citocromo p450 (CYP)1B1. Objetivos: Los objetivos que nos hemos planteado son los siguientes: 1. Analizar la expresión del factor de trascripción SXR y del MDR1 en vejiga mediante RT-PCR en tiempo real, tanto en vejiga tumoral como vejiga normal. 2. Analizar la relación de los factores clínicos y patológicos con la expresión del SXR y del MDR1. 3. Analizar la expresión de los polimorfismos de CYP1B1, GSTM1 GSTT1 y SULT1A1, y su correlación con distintos factores clínico patológicos y moleculares. Material y Métodos: De manera prospectiva se calculó un tamaño muestral necesario para este estudio. Se incluyeron 67 pacientes de dos instituciones distintas (Hospital Universitario Miguel Servet (49 HUMS) y Clínica Universitaria de Navarra (18 CUN)), diagnosticados de cáncer vesical infiltrante y tratados mediante cistectomía radical, se le realizó la determinación de la expresión de SXR y MDR1 mediante PCR cuantitativa en tiempo real, así como de los polimorfismos CYP1B1, GSTM1 GSTT1 y SULT1A1 mediante RFLP (restricción de la longitud del fragmento del polimorfismo). Se correlaciona mediante tablas de contingencia la correlación con el resto de los factores pronósticos. Resultados: La media de seguimiento de los pacientes fue de 23,7 meses, con una mediana de 28,26 meses. De los 67 pacientes estudiados, 31 pacientes (46,3%) presentaron progresión de la enfermedad, bien en forma de recidiva local, metástasis a distancia o ambos, con un tiempo medio a recidiva de 12,4 meses, mediana de 10 meses, con un rango de 1,1 mes a 31,9 meses. 36 pacientes (53,7%) no presentaron evidencia de progresión de la enfermedad. El receptor de esteroides y xenobióticos SXR así como el gen de multirresistenia a drogas (Multidrug resistance gene (MDR1)), se expresan en vejiga normal (0,94ΔCt y 0,94ΔCt) y en vejiga tumoral de la pieza de cistectomía (1,09 ΔCt y 0,45 ΔCt). Hemos analizado su expresión de manera cuantitativa y de manera cualitativa. La expresión de SXR se correlaciona con la presencia de carcinoma in situ (p=0,024), infiltración vasculo-linfática (p=0,05) mientras que MDR1 se correlaciona con la presencia de infiltración vasculo linfática (p=0,05) A su vez ambos la presencia de ambos factores se correlaciona entre ellos (p=0,011) Los polimorfismos: CYP1B1, GSTM1, GSTT1 y SULT1A1, se expresan en vejiga pero su expresión no guarda correlación con ningún factor pronóstico Conclusiones: El SXR y el MDR1 se expresan tanto en vejiga normal y tumoral. Y que dicha expresión guarda una correlación con factores pronósticos con influencia en la supervivencia descritas en la literatura

Introduction: Steroid and Xenobiotic Receptor (SXR) has demonstrated its activation by numerous drugs, including cytochrome P450 potent inducers like rifampicina or cotrimazol. The role of SXR is well known, and lies regulating in a positive manner cytochrome P450 3A4 (CYP3A4) transcription and the multidrug resistance gene (MDR1), it’s considered a key in the xenobiotic detoxification mechanism, being involved in all phases of the detoxification process. Enzymes involved in Policyclic Aromatic hidrocarbures (PAH) metabolism and degradation are polymorphic in humans, including glutation S-transferases (GSTs), N-acetiltransferases (NATs), sulfotransferases (SULTs)1A1 and cytochrome p450 (CYP)1B1. Objectives: The objectives we’ve planned are: 1. Analyze the expression of the transcription factor SXR and MDR1 in bladder by means of RT-PCR real time, both in normal bladder and in tumoral bladder. 2. Analyze the relation between clinical and pathological factors with the expression of SXR and MDR1. 3. Analyze the expression of the polymorphims CYP1B1, GSTM1 GSTT1 and SULT1A1 and their correlation with different clinic-pathological and molecular factors. Material and Methods: In a prospective way the size of the sample was estimated. In 67 patients from two institutions (Hospital Universitario Miguel Servet (49 HUMS) and Clinica Universitaria de Navarra (18 CUN)), diagnosed of invasive bladder cancer and treated by means of radical cystectomy, were determined the expression of both SXR and MDR1 by means of real time PCR, as well as the polymorphisms CYP1B1, GSTM1 GSTT1 y SULT1A1 by means of RFLP (Restriction fragment length polymorphism). Correlations with other prognostic factors by contingency tables were performed. Results: Average follow up was 23,7 months with a median of 28,26 months. Of the 67 patients studied, 31 patients (46,3) presented disease progression, in form of local recurrence or in distant metastasis or both. With a average time to progression of 12,4 months and a median of 10 months, with a range of 1,1 month to 31,9 month. 36 patients (53,7%) did not have any evidence of disease progression during follow up. The Steroid and Xenobiotic Receptor as well as the Multidrug Resistance Gene (MDR1) are expressed in both normal bladder (0,94ΔCt y 0,94ΔCt) and tumoral bladder in the cystectomy specimen(1,09 ΔCt y 0,45 ΔCt). We’ve analyzed their expression in a quantitative manner and in a qualitative manner. The expression of SXR correlates with the presence of ca. in situ (p=0,024), vasculo-lymphatic invasion (p=0,05) mean while MDR1 correlates with presence of vasculo-lymphatic invasion (p=0,05) Both factors are correlate between each others (p=0,011). Polymorphisms: CYP1B1, GSTM1, GSTT1 and SULT1A1, are expressed in these patients but their expression doesn’t correlates with any prognostic factor Conclusions: Both SXR and MDR1 are expressed in normal bladder as well as in tumoral bladder. And their expression correlates with different prognostic factors with influence in the survival described in the literature

[Are there any differences between the characteristics and the behavior of bladder cancer according to the age of presentation? Our experience]. / Existen diferencias en las caracteristicas y en el comportamiento del tumor vesical según edad de presentación? Nuestra experiencia.

INTRODUCTION: Owing to the different results from the series that evaluate the behavior of the bladder cancer according to the age at the moment of the diagnosis, our objective is based on valuing the characteristics and behaviour according to age of appearance. METHODS: A retrospective study of bladder cancer diagnosed in our area during decade 1993-2003, distributed in 3 intervals of age and some characteristics and behaviour are valued. RESULTS: Elderly patients present greater tumors, non differentiated and with greater rate of progression to infiltrated. Moreover the age, the pathological stage and the tumorlike degree appear as independent significant factors in the multivariant study. CONCLUSIONS: In our experience, the patients greater than 70 years present neoplasms of similar clinical characteristics, although pathologically more aggressive, with greater percentage of progression and worse survival.

¿Existen diferencias en las características y en el comportamiento del tumor vesical según edad de presentación? Nuestra experiencia / Are there any differences between the characteristics and the bahavior of bladder cancer according to the age of presentation? our experience

Objetivo: Dados los diferentes resultados de las series que evalúan el comportamiento de los tumores vesicales según la edad en la que debutan, nuestro objetivo se basa en valorar las características y comportamiento según edad de aparición. Método: Se realiza un estudio retrospectivo de los tumores vesicales de nuevo diagnóstico de nuestra área durante el decenio 1993-2003, distribuidos en 3 intervalos etarios y se evalúan diferentes características y comportamiento. Resultados: Se demuestra que los pacientes de más edad presentan tumores más grandes, indiferenciados y con mayor tasa de progresión hacia infiltrantes. Además aparecen como factores significativos independientes en el estudio multivariante: la edad, el estadio patológico y el grado tumoral. Conclusiones: En nuestra experiencia, los pacientes de edad mayor de 70 años presentan neoplasias de características clínicas similares, aunque patológicamente más agresivas, con mayor porcentaje de progresión y peor supervivencia

Introduction: Owing to the different results from the series that evaluate the behavior of the bladder cancer according to the age at the moment of the diagnosis, our objective is based on valuing the characteristics and behaviour according to age of appearance. Methods: A retrospective study of bladder cancer diagnosed in our area during decade 1993-2003, distributed in 3 intervals of age and some characteristics and behaviour are valued. Results: Elderly patients present greater tumors, non differentiated and with greater rate of progression to infiltrated. Moreover the age, the pathological stage and the tumorlike degree appear as independent significant factors in the multivariant study. Conclusions: In our experience, the patients greater than 70 years present neoplasms of similar clinical characteristics, although pathologically more aggressive, with greater percentage of progression and worse survival

Roflumilast inhibits leukocyte-endothelial cell interactions, expression of adhesion molecules and microvascular permeability.

BACKGROUND AND PURPOSE: The present study addressed the effects of the investigational PDE4 inhibitor roflumilast on leukocyte-endothelial cell interactions and endothelial permeability in vivo and in vitro. EXPERIMENTAL APPROACH: In vivo, intravital video-microscopy was used to determine effects of roflumilast p.o. on leukocyte-endothelial cell interactions and microvascular permeability in rat mesenteric venules. In vitro, the effects of roflumilast N-oxide, the active metabolite of roflumilast in humans, and other PDE4 inhibitors on neutrophil adhesion to tumour necrosis factor alpha (TNFalpha)-activated human umbilical vein endothelial cells (HUVEC), E-selectin expression and thrombin-induced endothelial permeability was evaluated. Flow cytometry was used to determine the effect of roflumilast on N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced CD11b upregulation on human neutrophils. KEY RESULTS: In vivo, roflumilast, given 1 h before lipopolysaccharide (LPS), dose-dependently reduced leukocyte-endothelial cell interactions in rat mesenteric postcapillary venules. It also diminished histamine-induced microvascular permeability. Immunohistochemical analyses revealed that roflumilast prevented LPS-induced endothelial P- and E-selectin expression. In vitro, roflumilast N-oxide concentration-dependently suppressed neutrophil adhesion to TNFalpha-activated HUVEC and CD11b expression on fMLP-stimulated neutrophils. It also reduced TNFalpha-induced E-selectin expression on HUVEC, when PDE3 activity was blocked. HUVEC permeability elicited by thrombin was concentration-dependently suppressed by roflumilast N-oxide. While roflumilast N-oxide was as potent as roflumilast at inhibiting stimulated endothelial cell and neutrophil functions, both compounds were significantly more potent than the structurally unrelated PDE4 inhibitors, rolipram or cilomilast. CONCLUSIONS AND IMPLICATIONS: These findings further support earlier observations on the inhibition of inflammatory cell influx and protein extravasation by roflumilast in vivo.

Modulatory effects of N-acetyl-L-cysteine on human eosinophil apoptosis.

Eosinophils are oxidant-sensitive cells considered relevant in allergic inflammation. The present study aimed to examine the effects of the antioxidant N-acetyl-L-cysteine (NAC) on constitutive and cytokine-delayed apoptosis in human isolated eosinophils. Human eosinophils were purified from the blood of healthy donors by a magnetic separation system. Apoptosis and cellular glutathione were assessed by cytofluorometric analysis and nuclear factor (NF)-kappaB binding activity assessed by electrophoresis mobility shift assay. The rate of spontaneous apoptosis of human eosinophils after 24 h culture, as assessed by annexin-V-positive staining, was mean+/-sem 48.2+/-1.4%, n = 5. Granulocyte-macrophage colony-stimulating factor (GM-CSF; 10 ng.mL(-1)) decreased apoptosis to 19.4+/-1.8%, n = 5. NAC (5 mM) inhibited spontaneous apoptosis (33.6+/-2.7%, n = 5) but augmented apoptosis in the presence of GM-CSF (30.9+/-1.5%, n = 5). NAC (5 mM) also increased the rate of apoptosis in the presence of tumour necrosis factor (TNF)-alpha (10 ng.mL(-1)) and interleukin-5 (5 ng.mL(-1)). NAC (5 mM) increased eosinophil glutathione content. The increase in eosinophil NF-kappaB binding activity induced by GM-CSF and TNF-alpha was suppressed by NAC. In conclusion, N-acetylcysteine modulates eosinophil apoptosis by inhibiting constitutive apoptosis but reversing the survival effect produced by inflammatory cytokines in human eosinophils.

Inhibitory effects of N-acetylcysteine on the functional responses of human eosinophils in vitro.

BACKGROUND: Oxidative stress appears to be relevant in the pathogenesis of inflammation in allergic diseases like bronchial asthma. Eosinophils are oxidant-sensitive cells considered as key effectors in allergic inflammation. OBJECTIVE: The aim of this work was to study the effects of the clinically used antioxidant N-acetyl-L-cysteine (NAC) on the functional responses of human-isolated eosinophils. METHODS: Human eosinophils were purified from the blood of healthy donors by a magnetic bead separation system. The effects of NAC were investigated on the generation of reactive oxygen species (chemiluminescence and flow cytometry), Ca(2+) signal (fluorimetry), intracellular glutathione (GSH; flow cytometry), p47(phox)-p67(phox) translocation (Western blot) and eosinophil cationic protein (ECP) release (radioimmunoassay). RESULTS: NAC (0.1-1 mm) inhibited the extracellular generation of oxygen species induced by N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) and eotaxin (in the presence of IL-5) with -logIC(50) values of 3.61+/-0.03 and 3.36+/-0.09, respectively. Also, the intracellular generation of hydrogen peroxide was virtually abolished by NAC (0.5-1 mm). NAC (1 mm) did not alter the fMLP-induced Ca(2+) signal but augmented the eosinophil content of reduced GSH and inhibited p47(phox)-p67(phox) translocation. NAC inhibited the release of ECP ( approximately 90% inhibition at 1 mm) from fMLP-activated eosinophils. CONCLUSION: Inhibition by NAC of human eosinophil functions in vitro is potentially useful in the treatment of allergic inflammation.