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Quantification of 11 clinical strains of Mycobacterium avium subsp. paratuberculosis isolated from domestic (cattle, sheep, and goat) and wildlife (fallow deer, deer, wild boar, and bison) animal species in an automatic liquid culture system (Bactec MGIT 960) was accomplished. The strains were previously isolated and typed using IS 1311 PCR followed by restriction endonuclease analysis (PCR-REA) into type C, S, or B. A strain-specific quantification curve was generated for each M. avium subsp. paratuberculosis strain by relating the time to detection in the liquid culture system to the estimated log 10 CFU in each inoculum. According to their growth curves, the tested M. avium subsp. paratuberculosis strains were classified into two distinct groups. The first group included the S-type strain isolated from goat and all the sheep strains with C, S, and B genotypes. A second group contained the C- and B-type strains isolated from cattle, goat, and wildlife animals with the exception of the fallow deer strain. The strains isolated from cattle or sheep showed similar strain-specific standard curves irrespective of their genotype. In contrast, the strains isolated from goat or from wildlife animal species varied in their rates of growth in liquid culture. Universal-standard curves and algorithms for the quantification of each group of strains were generated. In addition, the liquid culture system was compared with a real-time quantitative PCR system for the quantification of the 11 M. avium subsp. paratuberculosis strains. Correlations between the estimated log 10 CFU and M. avium subsp. paratuberculosis DNA copy numbers were very high for all the tested strains ( R ≥ 0.9).

Tumor invasion is paradigmatic of the complex interactions connecting a carcinoma with its environment, and a reflex of the cellular and molecular heterogeneity that defines the initiation of dissemination and metastasis. The hostile situation generated by a growing carcinoma and a reactive stroma is at the basis of the promotion of carcinoma invasion and metastasis, with oxidative stress emerging as a main player in the acquisition of an aggressive tumor phenotype. In this review, we present this complex scenario with a focus on the contribution of the reactive environment and the oxidative stress to the cellular and molecular events associated with carcinoma invasion and metastasis. We also discuss the potential of oxidative stress as a source of biomarkers of advance disease, and as supplier of a therapeutic armamentarium against the initial steps of metastatic dissemination.

Resumen Introduccion y objetivos La literatura sobre la participacion de la inestabilidad de microsatelites en el carcinoma de celulas escamosas de cabeza y cuello muestra una gran variabilidad, probablemente debido a las diferencias en la metodologia de las pruebas. Utilizando un sistema de deteccion consensuado, nos planteamos como objetivo llegar a una estimacion fiable de la prevalencia de la inestabilidad de microsatelites en un subconjunto de carcinomas de celulas escamosas de cabeza y cuello. Metodos Se analizo el estado de inestabilidad de microsatelites en 43 pacientes no tratados previamente y diagnosticados de un carcinoma primario de celulas escamosas de laringe mediante una prueba de PCR multiple, incluyendo 5 marcadores repetidos de mononucleotidos. Resultados En 36 casos se observo un fenotipo estable o microsatelites estables (83,7%), y en 7 casos (16,3%) se mostro un fenotipo positivo de inestabilidad de microsatelites. Uno de los casos mostro inestabilidad en 3 de los 5 marcadores, otro mostro inestabilidad en 2 marcadores y 5 casos en un marcador. Entre los casos de inestabilidad de microsatelites positiva y los casos estables no hubo diferencias con respecto a la edad, el estadio del tumor, la afectacion de los ganglios linfaticos o las metastasis a distancia. Conclusiones Nuestros datos muestran que una parte de los carcinomas de celulas escamosas de laringe presentan inestabilidad de microsatelites. El conocimiento sobre el estado de inestabilidad de microsatelites de los pacientes permitira el ajuste de la terapia anti-cancerigena a nivel individual.

We developed a suicidal DNA vaccine (pIRF1A-G-pMT-M) for salmonid fish susceptible to Infectious Hematopoietic Necrosis Virus (IHNV). The suicidal vaccine consists of two operons: i) an inducible fish promoter, the interferon regulatory factor 1A promoter (pIRF1A), driving the expression of the IHNV viral glycoprotein (G) gene that induces protection, and ii) a ZnCl(2) inducible fish promoter, the metallothionein promoter (pMT), driving the expression of the IHNV matrix (M) protein that induces apoptosis. The vaccine induces an immune response to the G protein and then induces the cell to undergo apoptosis to eliminate the DNA vaccine-containing cell. Also developed is another suicidal construct (pCMV-luc-pMT-M) for monitoring the persistence of luciferase (luc) expression after induction of apoptosis. In this study, we evaluated the inducibility of the MT promoter with ZnCl(2) and the capacity of cells transfected with the suicidal vector pCMV-luc-pMT-M to undergo apoptosis after ZnCl(2) addition. We also demonstrated the protective immunity elicited by the suicidal DNA vaccine pIRF1A-G-pMT-M, the survival of fish after treatment with ZnCl(2), and the elimination of the suicidal vector in fish after ZnCl(2) treatment.

This paper attempts to clarify a design process that is being used by Kazuyo Sejima and Sou Fujimoto based on the use of scale models. Two typical cases are studied and represented graphically in order to map the workflow. The results reveal that the mutual influence between team members, the continuous process of production and selection are closer to an "editing process" rather than the conventional linear design process. The architectural quality and character of the work produced by Sejima and Fujimoto can be seen as a consequence of the process itself. The process based on the use of scale models becomes an object of design, and its advantages and disadvantages are discussed in this article. This systematical study is expected to offer new ideas to practitioners on how to integrate scale models in the design process and how to enhance creativity and collaborative teamwork.

Names within Chionoloma tenuirostre are evaluated for proper taxonomic position. A total of 20 names are treated as synonyms, among them, 10 are here synonymised for the first time ( Hyophila acutiuscula , Leptodontium subintegrum , Oxystegus cylindricus var. filifer , Trichostomum angustifolium , T. fallax , T. fallax f. minutum , T. fragilifolium , T. prionodon , T. siamense and T. uncinifolium ) and three names are excluded from the synonymy ( Barbula wollei , Tortula cylindrotheca and Trichostomum burmense ). In order to maintain nomenclatural stability, the typification of 12 names is discussed and lectotypes are designated.

El bloqueo del plexo cervical superficial ecoguiado es una técnica sencilla y que, complementada con una adecuada sedación, puede proporcionar las condiciones adecuadas para la realización de la exéresis de la glándula paratiroides sin necesidad de anestesia general. Dicha técnica anestésica favorece el alta temprana de los pacientes sometidos a este tipo de cirugía, que pueden de este modo verse más fácilmente incluidos en un régimen de Cirugía Mayor Ambulatoria. Es una técnica indicada para cirugías de corta duración, poco complejas, en pacientes colaboradores; aunque también estaría indicada en pacientes de alto riesgo evitando la necesidad de una anestesia general y asegurando una mayor estabilidad hemodinámica. ABSTRACT The ultrasound-guided superficial cervical plexus block is a simple technique and, complemented with adequate sedation, may provide adequate conditions for performing the exeresis of the parathyroid gland without the need for general anesthesia. This anesthetic technique favors the early discharge of patients undergoing this type of surgery, who can thus be more easily included in a Major Ambulatory Surgery regimen. It is a technique indicated for short-term, lesscomplex surgeries in collaborating patients; although it would also be indicated in high-riskpatients avoiding the need for general anesthesia and ensuring greater hemodynamic stability.

The tumor microenvironment (TME) is gaining increasing attention in oncology, as it is recognized to be functionally important during tumor development and progression. Tumors are heterogeneous tissues that, in addition to tumor cells, contain tumor-associated cell types such as immune cells, fibroblasts, and endothelial cells. These other cells, together with the specific extracellular matrix (ECM), create a permissive environment for tumor growth. While the influence of tumor-infiltrating cells and mechanical properties of the ECM in tumor invasion and progression have been studied separately, their interaction within the complex TME and the epithelial -to-mesenchymal transition (EMT) is still unclear. In this work, we develop a 3D co-culture model of lung adenocarcinoma cells and macrophages in an interpenetrating network hydrogel, to investigate the influence of the macrophage phenotype and ECM stiffness in the induction of EMT. Rising ECM stiffness increases both tumor cell proliferation and invasiveness. The presence of tumor-associated macrophages and the ECM stiffness jointly contribute to an invasive phenotype, and modulate the expression of key EMT-related markers. Overall, these findings support the utility of in vitro 3D cancer models that allow one to study interactions among key components of the TME.

[EN] It is possible that the architectural exhibition of greatest influence in diffusing the Modern Movement was the show organized by Philip Johnson and Henry-Russell Hitchcock at the Museum of Modern Art of New York in 1932. It was initially intended to offer the public a sample of ten scale models by the ten European and American architects most involved in the avant-garde, requesting each to submit the model of a project designed specifically for the exhibition. In the end, they were given somewhat greater freedom, as the exhibition was fleshed out with a selection of photographs of the work that each of them had built. Nonetheless, the most important items were the scale models, the most highly thought of being the one provided by Mies van der Rohe. This text considers the models displayed by all of the architects in the light of MoMA archives and other sources. It also looks at the relevance they had as a system of representation suited to spreading what the organizers eventually called International Style., [ES] Es posible que la exposición de arquitectura más relevante para la difusión del Movimiento Moderno fuera la organizada por Philip Johnson y Henry-Russell Hitchcock en el Museum of Modern Art de Nueva York, en 1932. Inicialmente se había pensado en ofrecer al público una muestra de diez maquetas de los diez arquitectos europeos y estadounidenses más implicados con las vanguardias, encargándoles a cada uno la maqueta de un proyecto diseñado ex profeso para la exposición. Finalmente se les concedió mayor libertad, ampliando la exposición con una selección fotográfica de la obra construida de cada uno de ellos. No obstante, las piezas más importantes fueron las maquetas, siendo la más apreciada la de Mies van der Rohe. En este texto se estudian, acudiendo al archivo del MoMA y a otras fuentes, los modelos expuestos por cada arquitecto y la relevancia que se les concedió en cuanto sistema de representación apto para difundir lo que los organizadores acabarían denominando el International Style.

This paper studies the visits of wild fauna, the influence of vegetal cover and fencing at water site election, and consumption tendency in water troughs designed for small game species distributed in an agricultural Mediterranean area during the summers from 2002 to 2005. Red-legged partridges (Alectoris rufa), lagomorphs (Oryctolagus cuniculus, Lepus granatensis), and other autochthonous species (birds and wild canids) visited water sources throughout the study. The number of visits by target species was higher than non-game. Lagomorphs preferred protected troughs (with surrounding vegetal cover) while partridges opted for open ones (without cover). Fencing had no effect on the visits of birds and lagomorphs, but it did reduce the visits of wild canids. Harsh climatic conditions determined an increase in water consumption in summer. Before designing a water trough device for this fauna, managers should consider that (1) target and non-target species could try to make use of it, (2) water points should be adapted to target species self protection, (3) fencing can prevent undesirable visitors (particularly predators), and (4) water requirements increase in the harsh moments of the dry season.

Resumen Son extremadamente infrecuentes los tumores no neoplasicos que aparece durante el embarazo o el puerperio y originan complicaciones obstetricas, la mayor parte de las veces debido al tamano que pueden llegar a alcanzar. El hallazgo de una masa anexial es, en general, casual ya que las no complicadas producen escasa sintomatologia. El uso generalizado de la ecografia durante el embarazo y el puerperio permite su diagnostico. El manejo de un tumor anexial durante la gestacion o el puerperio debe ser individualizado, dependiendo de los sintomas de la paciente, la edad gestacional y el tamano y las caracteristicas de la tumoracion. El quiste folicular gigante solitario luteinizado del puerperio es una lesion poco frecuente del ovario que puede surgir en el embarazo y aumentar de tamano durante el puerperio hasta que es detectado. Presentamos el caso de un quiste folicular gigante luteinizado del puerperio diagnosticado en una paciente en la 6.a semana del puerperio. Realizamos asimismo una revision de las masas ovaricas no neoplasicas mas frecuentes.

Introduccion y objetivo Los adenocarcinomas nasosinusales (ACNS) son tumores epiteliales raros, primarios de las fosas nasales y los senos paranasales, que se caracterizan por presentar estructuras glandulares. Los objetivos de este trabajo son: determinar las alteraciones genicas en los ACNS mediante MLPA (multiplex ligation-dependent probe amplification) y establecer la relacion entre los cambios genicos del tumor y el comportamiento clinico-evolutivo de los pacientes. Material y metodo Realizamos un estudio longitudinal prospectivo a 20 pacientes con ACNS controlados en nuestro servicio entre 1998 y 2004. A las muestras tumorales se les extrajo el ADN para realizar la MLPA. Resultados La categoria T de los pacientes fue para T2: 4 (20 %), T3: 6 (30%), T4a: 3 (15 %) y T4b: 7 (35 %). Todos eran inicialmente N0 y M0. El 85 % habia estado expuesto, en el medio laboral, al polvo de madera. El tratamiento indicado fue quirurgico (100 %) con radioterapia complementaria (70 %). La supervivencia global a los 5 y 10 anos fue del 42 y el 22 %, respectivamente. Las ganancias genicas mas frecuentes fueron: PTP4A3 y PDCD8 (65 %), TNRFSF7 (50 %), RECQL4 y LMO2 (45 %); las perdidas: BCL2 (70 %), IL13 (55 %), ABCB1 y RB1 (50 %), PIK3CA y CDH1 (45 %). Conclusiones La aparicion de metastasis se correlaciono de forma significativa con perdidas en F3, MIF y BRCA1. La peor supervivencia con perdidas en F3, BCRA1 y MIF y ganancias en PIK3CA, UTY y RELA. En el analisis multivariable alcanzaron significacion estadistica las perdidas en BRCA1 y F3.

Introduction and aim Intestinal-type sinonasal adenocarcinomas (ITACs) are rare epithelial tumours, primarily originating in the nasal cavities and paranasal sinuses, and characterized by glandular structures. The aims of this study are: to determine the genetic alterations in ITACs by MLPA (Multiplex ligation-dependent probe amplification) and to correlate the findings to the clinical behaviour and follow-up information of the patients. Material and method We performed a longitudinal prospective study on 20 patients with ITAC, seen in our department between 1998 and 2004. DNA was extracted from primary tumour samples and analyzed by MLPA. Results The T stage of our series was T2: 4 (20%), T3: 6 (30%), T4a: 3 (15%), and T4b: 7 (35%). All cases initially were N0 and M0. Seventeen patients (85%) had professional exposure to wood dust. All patients underwent surgical intervention and 70% received complementary radiotherapy. Overall 5 and 10 year survival was 42% and 22%, respectively. Gains were found most frequently for PTP4A3 and PDCD8 (65%), TNRFSF7 (50%), RECQL4 and LMO2 (45%), and losses for BCL2 (70%), IL13 (55%), ABCB1 and RB1 (50%), PIK3CA and CDH1 (45%). Conclusions Losses of F3, MIF, and BRCA1 significantly correlated with the posterior development of metastases and with worse survival. Also gains of PIK3CA, UTY, and RELAcorrelated with poor clinical outcome. Losses of BRCA1 and F3 were significant in multivariate analysis.

Introduccion Las recidivas tumorales (RT) causan elevada mortalidad en los pacientes con cancer escamoso de cabeza y cuello (CECC) y complican mucho la actitud terapeutica. Objetivo Describir los aspectos clinicos de las RT y su implicacion en la supervivencia. Pacientes y metodo Se revisa a 652 pacientes con CECC y se recoge las caracteristicas clinicas de las RT observadas. Resultados Las RT se presentaron en el 19,9 % (130 pacientes) de nuestra serie de CECC. Las localizaciones mas frecuentes son mesofaringe (32%), hipofaringe (24 %) y cavidad oral (21 %). El 50 % de las RT son de caracter locorregional; el 43 %, locales, y el 7 %, estomales. La aparicion de una RT reduce al 15 % la supervivencia general de los pacientes con CECC, es mayor en la laringe glotica (38 %) y la supraglotica (27 %), y baja al 2-4% en mesofaringe e hipofaringe. Conclusiones Las RT son mas frecuentes en tumores faringeos y de tipo locorregional. El pronostico en esta localizacion es muy malo y el tratamiento de rescate apenas modifica la supervivencia.

Background: Lymph node metastasis is the mayor cause of mortality in patients with head and neck squamous cell carcinomas (45%). The genetic changes underlying metastasis are still largely unknown and genetic markers to predict lymph node positivity still need to be found. The aim of this study was to search such markers by using Multiplex Ligation-dependent Probe Amplification (MLPA), a semi-quantitative PCR technique to detect gene copy number alterations. Methods: Thirty-seven genes were analysed by MLPA in 34 larynx and 22 pharynx carcinomas. Results: Losses of CDKN2A (9p21) and MLH1 (3p22) and gains of CCND1, EMS1 (both at 11q13), RECQL4 and PTP4A3 (both at 8q24) were the most frequent aberrations in both larynx and pharynx carcinomas. Amplifications were detected at EMS1, CCND1 and ERBB2 (17q21). A correlation between loss of N33 (8p22) and poor survival was found (p=0.02). Gain of EMS1 had the same relation with survival but not significant (p=0.08). Lymph node positive tumors presented a specific pattern of genetic alterations, with losses of N33, STK11 (19p13) and TP53 (17p13), the latter especially in larynx tumors. Conclusion: We propose that these 3 genes might play a role in the development of metastasis in larynx and pharynx squamous cell carcinomas.

Introduction For most patients with squamous head and neck cancer (HN-SCC), locoregional tumour recurrence (TR) carries an extremely poor prognosis and is therapeutically challenging. Objective To define the clinical aspects of TR and their implication on the survival in patients with HN-SCC. Patients and method The clinical management and the outcome of 652 patients with HN-SCC were reviewed. Results The overall incidence of TR in this series of HN-SCC was 19.9% (n=130). The most frequent locations of the primary cancers were oropharynx (32%), hypopharynx (24%), and oral cavity (21%). The rates of recurrence were locoregional 50%, local 43% and stomal recurrence 7%. The appearance of a TR reduces the overall survival of patients with HN-SCC to 15%. Survival is better in glottic (38%) and supraglottic (27%), carcinomas, and worse in oro-hypopharynx tumours (2-4%). Conclusions RT are more frequent in pharyngeal tumours, especially locoregional recurrences. Patients with recurrence in pharynx were definitely associated with poor prognosis and in these cases salvage surgery seems not to improve survival rates.

The mitochondrial genome of mosses is characterized by a highly conserved structure and genic content. This is confirmed here through the assembly and annotation of the mt genome of the moss Oxystegus tenuirostris, family Pottiaceae, for which it is assembled. A phylogenetic tree is inferred from the whole genome of 16 species of mosses to validate the sequence of O. tenuirostris by confirming its shared ancestry with Syntrichia. The genome is 105 001 bp long, with a GC content of 39.2%, comprising 40 protein coding, 24 tRNA, and 3 rRNA genes. All introns reported from the mt genome of all but one peristomate moss are present, whereas no region of 50 bp or more is repeated within the genome. The genic content and order is identical to that of most mosses, highlighting that the mt genome is static not only across the phylogenetic depth but also breadth of the moss tree of life.

International audience; Nouakchott est une capitale où le processus d’urbanisation et les différents projets de planification et « modernisation » adoptés ont produit de très fortes disparités et ségrégations. La mobilité est considérée par les pouvoirs publics comme un besoin et non pas comme un droit et on ne réfléchit pas au fait que ce droit peut être garanti aussi par une diminution de la nécessité de se déplacer. L’article se propose de montrer comment la mobilité est prise en charge à Nouakchott et analyse les facteurs qui tout en amplifient la congestion de la ville, fabriquent les inégalités territoriales.

Administered by intramuscular injection, a DNA vaccine (pIRF1A-G) containing the promoter regions upstream of the rainbow trout interferon regulatory factor 1A gene (IRF1A) driven the expression of the infectious hematopoietic necrosis virus (IHNV) glycoprotein (G) elicited protective immune responses in rainbow trout (Oncorhynchus mykiss). However, less laborious and cost-effective routes of DNA vaccine delivery are required to vaccinate large numbers of susceptible farmed fish. In this study, the pIRF1A-G vaccine was encapsulated into alginate microspheres and orally administered to rainbow trout. At 1, 3, 5, and 7d post-vaccination, IHNV G transcripts were detected by quantitative real-time PCR in gills, spleen, kidney and intestinal tissues of vaccinated fish. This result suggested that the encapsulation of pIRF1A-G in alginate microparticles protected the DNA vaccine from degradation in the fish stomach and ensured vaccine early delivery to the hindgut, vaccine passage through the intestinal mucosa and its distribution thought internal and external organs of vaccinated fish. We also observed that the oral route required approximately 20-fold more plasmid DNA than the injection route to induce the expression of significant levels of IHNV G transcripts in kidney and spleen of vaccinated fish. Despite this limitation, increased IFN-1, TLR-7 and IgM gene expression was detected by qRT-PCR in kidney of vaccinated fish when a 10μg dose of the oral pIRF1A-G vaccine was administered. In contrast, significant Mx-1, Vig-1, Vig-2, TLR-3 and TLR-8 gene expression was only detected when higher doses of pIRF1A-G (50 and 100μg) were orally administered. The pIRF1A-G vaccine also induced the expression of several markers of the adaptive immune response (CD4, CD8, IgM and IgT) in kidney and spleen of immunized fish in a dose-dependent manner. When vaccinated fish were challenged by immersion with live IHNV, evidence of a dose-response effect of the oral vaccine could also be observed. Although the protective effects of the oral pIRF1A-G vaccine after a challenge with IHNV were partial, significant differences in cumulative percent mortalities among the orally vaccinated fish and the unvaccinated or empty-plasmid vaccinated fish were observed. Similar levels of protection were obtained after the intramuscular administration of 5μg of pIRF1A-G or after the oral administration of a high dose of pIRF1A-G vaccine (100μg); with 70 and 56 relative percent survival values, respectively. When fish were vaccinated with alginate microspheres containing high doses of the pIRF1A-G vaccine (50 or 100μg), a significant increase in the production of anti-IHNV antibodies was detected in serum samples of the vaccinated fish compared with that in unvaccinated fish. At 10 days post-challenge, IHNV N gene expression was nearly undetectable in kidney and spleen of orally vaccinated fish which suggested that the vaccine effectively reduced the amount of virus in tissues of vaccinated fish that survived the challenge. In conclusion, our results demonstrated a significant increase in fish immune responses and resistance to an IHNV infection after the oral administration of increasing concentrations of a DNA vaccine against IHNV encapsulated into alginate microspheres., This work was supported by grants from the Ministerio de Economía y Competitividad (MINECO) and the Consejo Superior de Investigaciones Científicas (Spain) (AGL2010-18454 and 2010–20E084, respectively). N. Ballesteros is grateful to the MINECO for the award of a PhD student fellowship.

[ES] Durante los últimos años la concienciación de los consumidores sobre la importancia del bienestar de los animales destinados a la producción de alimentos ha ido en creciente aumento. Así, se han desarrollado estudios y sistemas para evaluar y controlar la calidad del bienestar principalmente en las especies explotadas en sistemas intensivos y se han publicado protocolos con diversos parámetros a determinar. Sin embargo, todos ellos presentan problemas no existiendo una metodología universalmente aceptada. Por ello, partiendo de criterios y parámetros basados en el animal y en los alojamientos y manejo utilizados en protocolos existentes y en otros considerados importantes para la salud y el de bienestar de las vacas de leche por profesores de los Dpts. de Producción Animal y de Medicina, Cirugía y Anatomía Veterinaria de la Facultad de Veterinaria de León, se ha desarrollado un programa de software libre (Happy Cow) por parte de alumnos de la Escu, [EN] During the past years the consumer’s awareness about the importance of welfare of animals used for food production is increasing. In this way, studies and systems to evaluate and monitor the quality of the welfare, mainly in intensive systems productions, have been developed and guidelines and protocols with parameters to be determined have been published. However, they all have various problems and there is no universally accepted methodology. Therefore, based on criteria and parameters based on the animal and on facilities and handling recommended on the aforementioned project and other variables considered important for dairy cows health and welfare by professors from the Departments of Animal Production and Medicine, Surgery and Veterinary Anatomy, Faculty of Veterinary Medicine of León, a free software program (Happy Cow) has developed by students of the School of Industrial Engineering and Computer ULE that achieves two objectives: to be used

Mycobacterium avium subsp. paratuberculosis is the cause of Johne's disease in cattle and other ruminants. M. avium subsp. paratuberculosis infection of the bovine host is not well understood; however, it is assumed that crossing the bovine intestinal mucosa is important in order for M. avium subsp. paratuberculosis to establish infection. To examine the ability of M. avium subsp. paratuberculosis to infect bovine epithelial cells in vitro, Madin-Darby bovine kidney (MDBK) epithelial cells were exposed to M. avium subsp. paratuberculosis . It was observed that bacteria can establish infection and replicate within MDBK cells. M. avium subsp. paratuberculosis also has been reported to infect mammary tissue and milk, and we showed that M. avium subsp. paratuberculosis infects bovine mammary epithelial cells (MAC-T cell line). Using polarized MAC-T cell monolayers, it was also determined that M. avium subsp. paratuberculosis crosses apical and basolateral surfaces with approximately the same degree of efficiency. Because M. avium subsp. paratuberculosis can be delivered to the naïve host by milk, it was investigated whether incubation of M. avium subsp. paratuberculosis with milk has an effect on invasion of MDBK cells. M. avium subsp. paratuberculosis exposed to milk entered epithelial cells with greater efficiency than M. avium subsp. paratuberculosis exposed to broth medium or water ( P < 0.01). Growth of M. avium subsp. paratuberculosis within MAC-T cells also resulted in augmented ability to subsequently infect bovine MDBK cells ( P < 0.001). Microarray analysis of intracellular M. avium subsp. paratuberculosis RNA indicates the increased transcription of genes which might be associated with an invasive phenotype.

Readily available p-tolylsulfinimines undergo highly stereoselective [3 + 2] cycloadditions with azomethine ylides generated from α -iminoesters and LDA to produce N-sulfinylimidazolidines. In the presence of Lewis acids, p-tolylsulfinimines react with glycine iminoester enolates to produce N-sulfinylimidazolidines, after cyclization of open chain intermediates. These mechanistically diverse processes take place with excellent regio-, stereo-, and facial selectivities, and the latter is opposite to most known reactions involving sulfinimines. Some of the resulting imidazolidines have been transformed into examples of a novel class of nonsymmetrical vicinal diamines using reductive and/or hydrolytic protocols.

Aquabirnaviruses, such as the infectious pancreatic necrosis virus (IPNV), Novirhabdo- viruses, such as the infectious hematopoiteic necrosis virus (IHNV) and the viral hemorrhagic septicemia virus (VHSV), cause considerable losses to the salmonid industry worldwide. Coinfections of 2 viruses have been described, but the interactions between rhabdoviruses and birnaviruses have not been examined closely. Using virus titration, flow cytometry and RT-PCR assays, we compared the effect of IPNV on the replication of IHNV and VHSV in tissue culture cells. RT-PCR assays indi- cated that simultaneous infection of IPNV with VHSV does not affect the replication of the rhab- dovirus either in the first or successive passages; the infective titers were similar in single and double infections. In contrast, coinfection of IPNV with IHNV induced a fall in infectivity, with reduced expression of IHNV viral antigens in BF-2 cells from Lepomis macrochirus and a loss of 4.5 log10 units of the infective titer after 3 successive passages. It was possible to stimulate BF-2 cells to produce significant interferon-like activity against IHNV but not against VHSV.

Invasive head and neck squamous carcinomas are among the cytogenetically most complex tumors. Perhaps for this reason, there is little consensus on the prognostic value of specific chromosomal aberrations. Here we present Results of CGH analysis of 56 clinically well-characterized set of head and neck cancers, consisting of larynx and pharynx only. The aim was to find possible associations with clinical outcome. The major chromosome arms showing gains were (in decreasing order): 3q, 7q, 8q, 5p, 11q13, 17q and 18p, and losses occurred at 3p, 11qter, 4p, 18q, and 5q. The segments most frequently amplified were 3q26-qter, 11q13, 11q22, 3q12–13, 18p11.3, 18q11.2 and 8q24.3. Tumors with stages III and IV, and lymph node positive tumors had a worse clinical outcome. Surprisingly, no specific chromosomal abnormality correlated with disease-free survival. The only aberration that correlated to one of the clinico-pathological parameters was amplification 11q13, that occurred solely in lymph node positive, stage IV tumors. However 11q13 amplification did not correlate with disease-free survival. These Results seem to indicate that genetic alterations at the level of chromosomes have limited prognostic value in patients with invasive larynx and pharynx squamous cell carcinomas.

Squamous cell carcinoma of the pharynx and larynx (SCCPL) is a genetically complex disease and is frequently associated with nonrandom chromosomal alterations. Fifty primary SCC of the pharynx (oropharynx, n=11, and hypopharynx, n=11) and larynx (n=28) were examined for numerical aberrations of chromosomes 8, 9, 11, and 17 with a panel of chromosome-specific repetitive DNA probes by fluorescence in situ hybridization (FISH). DNA ploidy analysis was also performed by flow cytometry (FCM). Aneusomic copy numbers of chromosomes 8, 9, 11, and 17 were discovered in 66%, 68%, 68% and 78% of tumors, respectively. FCM showed abnormal DNA content in 74% of cases (mean DNA index=1.69). Polysomy was the main finding in both DNA-aneuploid and DNA-diploid tumors (64.5% of cases). Numerical aberrations of chromosomes 8 and 11 correlated to DNA ploidy by FCM (P

A survey of routes for aminal cleavage of N-sulfinylimidazolidines has been carried out, and selective conditions to cleave the aminal moiety while preserving the sulfinamide group unaltered have been found. Thus, the treatment of enantiopure N-sulfinylimidazolidines with aqueous H(3)PO(4) in THF affords enantiopure N-sulfinyldiamino esters in excellent yields, while the presence of MeOH as cosolvent allows for the simultaneous removal of the sulfinamide group and aminal cleavage. The behavior of these substrates in a variety of chemical transformations has been explored.

0.0001). Cox regression revealed that the 3q gain was the strongest prognostic factor; other features did not have additional prognostic value. In conclusion, loss of 1p is associated with a high MAI. A gain of 3q is a stronger predictor of recurrence than grade, MAI, and other features in invasive breast cancers.

A new procedure for the asymmetric synthesis of imidazolidines and vicinal diamines is reported. The 1,3-dipolar cycloaddition between nonracemic p-tolylsulfinimines and azomethine ylides generated in situ from alpha-iminoesters and LDA produces N-sulfinylimidazolidines with a high degree of stereocontrol. In contrast, the presence of Lewis acids promotes formation of the cycloadducts through a highly diastereoselective process with opposite stereochemistry. Subsequent transformations of the imidazolidines including oxidative, reductive, and hydrolytic processes that provide easy access to vicinal diaminoalcohols have been explored. Among these, reductive cleavage of the aminal with LiAlH4 is an extremely efficient and general reaction for the synthesis of enantiopure N-sulfinyl-N'-benzyldiaminoalcohols

DNA plasmid vectors were constructed with inducible rainbow trout promoters to take the place of the cytomegalovirus immediate early promoter in DNA vaccines for fish. DNA fragments containing the promoter regions upstream of the rainbow trout Mx1 and interferon regulatory factor 1A (IRF1A) genes were obtained and their nucleotide sequences were determined. Vectors containing the G gene of IHNV linked to the trout promoters were found to be very effective as vaccines in rainbow trout. In addition, we demonstrated that the IRF1A promoter is transcriptionally active in mouse cells NIH-3T3 suggesting that this fish promoter might be used for mammalian DNA vaccine development as well.

The objectives of this work were to recover high-calorific value/low-ash content coals from coal fines cleaning wastes by agglomeration with colza oil and to decrease the risk of spontaneous combustion of the dump where the coal wastes are disposed of. Three Spanish coal fines wastes from different coal cleaning plants were agglomerated with refined colza oil over a wide range of oil concentration. The results were evaluated by the percentages of organic matter recovery and ash rejection as well as the ash content of the recovered coal and the amount of organic matter remaining in the dump after oil agglomeration. It was concluded that the agglomeration of coal fines wastes with colza oil allows the recovery of significant amounts of energy that is currently wasted and reduces the risk of spontaneous combustion of coal dumps as well as the amount of coal fines wastes to be handled and disposed of.

[EN] The drawings and historic cartography are an exceptional value for the knowledge of any city, being able to see through these documents, traces or remains of missing or changed locations in the course of time. Although graphical documents that remain from the city of Oviedo is rather low until mid nineteenth century, stands out because of his interest some plans and drawings that allow us graphically how to restore the city and its architectural heritage, [ES] Los dibujos y la cartografía histórica tienen un valor excepcional para el conocimiento de cualquier ciudad, pudiendo llegar a apreciar, a través de estos documentos, las huellas o vestigios de lugares desaparecidos o transformados con el paso del tiempo. Aunque los documentos gráficos que se conservan de la ciudad de Oviedo son más bien escasos hasta mediados del siglo XIX, cabe destacar por su interés algunos planos y dibujos que nos permiten restituir gráficamente la forma de la ciudad y su patrimonio edilicio

The Asian moss Pseudosymblepharis perlongifolia, previously known only from Malaysia, is reported for the first time for America on the basis of several collections from Maricao (Puerto Rico). The principal distinctive characters that separate it from the nearest species of Pseudosymblepharis and related genera are discussed. The species is described and illustrated.

The enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies againstMycobacterium aviumsubsp.paratuberculosisin wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deerM. aviumsubsp.paratuberculosisisolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detectM. aviumsubsp.paratuberculosisantibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations.

Sinonasal squamous cell carcinomas (SCC) are rare tumors, etiologically related to occupational exposure to wood and leather dust. In spite of surgical and radiotherapeutic advances, the 5 year survival is still 30–50%. Therefore, alternative treatment options are needed. We report the establishment and characterization of six unique human sinonasal SCC cell lines, named SCCNC1, 2, 4, 5, 6 and 7. In vitro growth and invasion characteristics were evaluated and genetic profiles were compared to those of the original primary tumors. The population doubling times ranged from 21 to 34 hours. Cell lines SCCNC2 and 7 were highly invasive in matrigel. Five cell lines carried a high number of copy number alterations, including amplifications and homozygous deletions, while one showed only three abnormalities. Sequence analysis revealed three cell lines with TP53 mutation and none with KRAS or BRAF. Overexpression of p53 was observed in five, and of EGFR in four cell lines. None of the cell lines showed strong immunopositivity of p16 or presence of human papilloma virus. In conclusion, we have created six new cell lines that are clinically and genetically representative of sinonasal SCC and that will be a useful tool for the preclinical testing of new therapeutic agents.

Choroidal neovascularization (CNV) is an important cause of severe central vision loss that may appear as a complication of infectious and noninfectious posterior uveitis. Inflammatory CNV is the third cause of CNV after age-related macular degeneration (AMD) and pathologic myopia [1]. The prevalence of inflammatory CNV in eyes with posterior uveitis is about 2 % at the time of initial presentation [2, 3]. Although most cases of inflammatory CNVrespond well to the intravitreal vascular endothelial growth factor (VEGF) inhibitors, bevacizumab (Avastin; Genentech, Inc., South San Francisco, CA, USA) [4-6] and ranibizumab (Lucentis; Genentech, Inc.) [7], there is a proportion of cases that might be suboptimally responders [8]. Several authors have reported the outcomes of switching these VEGF inhibitors to aflibercept (VEGF-TrapEye, Eylea; Regeneron, Tarrytown, NY, USA) for CNV secondary to age-related macular degeneration (AMD) [9, 10]. However, little is known of this switch in indications other than neovascular AMD. Herein, we report the short-term efficacy of intravitreal aflibercept injections in three cases diagnosed with inflammatory CNV and persistent retinal fluid in spite of intravitreal ranibizumab injections.

The analysis of the early macrophage responses, including bacterial growth within macrophages, represents a powerful tool to characterize the virulence of clinical isolates of Mycobcaterium avium susbp. paratuberculosis (Map). The present study represents the first assessment of the intracellular behaviour in ovine monocyte-derived macrophages (MDMs) of Map isolates representing distinct genotypes (C, S and B), and isolated from cattle, sheep, goat, fallow deer, deer, and wild boar. Intracellular growth and survival of the selected isolates in ovine MDMs was assessed by quantification of CFUs inside of the host cells at 2 h p.i. (day 0) and 7 d p. i. using an automatic liquid culture system (Bactec MGIT 960). Variations in bacterial counts over 7 days from the baseline were small, in a range between 1.63 to 1.05-fold. After 7 d of infection, variations in the estimated log10 CFUs between all the tested isolates were not statistically significant. In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S) or with two C-type isolates from distinct hosts (cattle and sheep); which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas. Upon 10 d p.i., both Map isolates were able to induce the formation of granulomas comparable to the granulomas observed in clinical specimens with respect to the cellular components involved. In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.

Blood and kidney leukocytes were purified from two-year-old rainbow trout carrying infectious pancreatic necrosis virus (IPNV) and the cells were processed to detect the virus by two methods: 1) indirect immunofluorescence stain followed by flow cytometry analysis, and 2) RT-PCR or nested-PCR amplification. These methods were compared with separate homogenization of visceral samples, followed by inoculation on cell cultures and seroneutralization, a procedure routinely employed by most disease diagnostic laboratories. IPNV was isolated by homogenization of samples in all seven specimens examined. The assay took between 7 and 28 days required for the appearance of cytopathic effects, which is usual in subclinical samples. From purified leukocytes, IPNV was detected by RT-PCR in five out of the seven specimens. The nested-PCR improved the sensitivity of the assay and gave positive results for all the fish. In addition, flow cytometry demonstrated the presence of IPNV in all the fish in 8-24 h. Examination of leukocytes is a practical way of detecting IPNV and much less time-consuming than the homogenization technique., This study was supported by grants from the Comision Interministerial de Ciencia y Tecnologla, project AGF 98-0837 and Comunidad Autonoma de Madrid, project 079/23/98.

The Forum of European National Highway Research Laboratories (FEHRL) Investigation on Longitudinal and Transverse Evenness of Roads (the FILTER project) is a research project on evenness developed by FEHRL. The laboratories involved in this project are from Belgium, Denmark, France, Germany, the Netherlands, Slovenia, Spain, Sweden, and the United Kingdom. Guy Descornet (of the Belgium Road Research Center) has coordinated the project. The FILTER project is divided into some tasks that deal with theoretical and experimental analysis of data on evenness. One of these tasks is to analyze the European data obtained from an experiment that was conducted by Technical Committee C1, Surface Characteristics, of the Permanent International Association of Road Congresses—World Road Association in 1998. This experiment was accomplished in the United States, Japan, and Europe (the FILTER experiment). The FILTER experiment was carried out in Germany and the Netherlands in September 1998. The participants (who used the profilometers commonly used in Europe) should report their results on the profiles and indices of both transversal evenness and longitudinal evenness. The Road Research Center of the Center of Research and Experimentation of Public Works (Madrid, Spain) was in charge of analyzing the results regarding longitudinal indices from the FILTER experiment. The method used to analyze these data and some of the conclusions obtained are explained. The work contributes to the achievement of the FILTER project, one aspect of which is to try to harmonize the different methods of measuring evenness in Europe.

La qualité des espaces publics, leurs formes, la spatialisation et le fonctionnement de leurs équipements et dispositifs économiques, leurs ambiances, leurs modes d’intégration dans le système des centralités intra-urbaines sont au cœur de la réflexion sur les devenirs de la ville contemporaine. L’offre commerciale peut participer à la création d’un espace urbain attractif, intense et habitable dans les centres comme dans les zones périphériques. Quelle place doivent prendre les structures du commerce dans une nouvelle programmation urbaine permettant d’assurer une attractivité redéfinie des espaces urbains ? Et plus exactement, comment intégrer les acteurs du système commercial aux politiques de renouvellement des espaces publics dans une perspective d’urbanisme durable ? Le commerce est un des éléments structurant et modulant l’intensité urbaine, les pratiques de mobilité, le lien social, l’animation et l’identité des quartiers lausannois. On explore ici quelques pistes de réflexion relatives au modèle de régulation de l’urbanisme commercial lausannois et à la manière dont les activités commerciales peuvent participer à la mise en place de stratégies de qualification de l’espace public et d’organisation des centralités urbaines. The quality of urban space, its morphology, spatial planning and ambiences, as well as the way that these centralities are integrated within the urban network and the hierarchy of centrality nodes are at the core of the creation of and thought about the contemporary city. The breakthrough of retail policies in Lausanne could become an extremely helpful planning tool, oriented towards making proximate neighborhoods both residentially and economically attractive. How does vicinity of retail enhance urbanity, and how does it influence in urban intensity, and the livability of public space? More precisely, how can we integrate retail related role players in the enhancement of the public realm’s urbanity within a context of sustainable urban design? The present article carries out an enquiry of how retail activities take part in strategies to improve the quality of the public realm and the organization of urban centralities. It reveals some hints that could help urban practitioners when making decisions regarding retail oriented developments. Die Qualität der öffentlichen Räume, ihre Morphologie, ihre Raumordnung und die Organisation von ihren wirtschaftlichen Einrichtungen und Massnahmen, die Umgebung sowie die Art dieser Zentralitäten innerhalb des urbanischen Netzwerkes integriert wurden und die Hierarchie der Zentralitätsknoten sind im Mittelpunkt der Betrachtung bezüglich der Weiterentwicklung der zeitgenossischen Stadt. Der Handel ist einer der Bereiche, der die urbane Intensität, das Mobilitätsystem, den sozialen Kontakt, das Leben und die Identität der Viertels von Lausanne struckturiert und moduliert. Kaufmännliche Richtlinien in Lausanne könnten extrem hilfreiches urbanisches Mittel zur Entwicklung eines attraktiven und wirtschaftlichen urbanischen Raumes und Wohngebietes in Stadtmitten und an Stadträndern werden. Wie kann Handel im Nah Bereich die Urbanisierung aufwerten/verbessern und wie kann er die urbane Intensität und die Lebensqualität der öffentlichen Räume einwirken? Genauer gesagt, wie können wir einzelhandelbezogenen Rollenspieler in der Aufwertung der Urbanisierung der öffentlichen Räume im Zusammenhang mit einer nachhaltigen urbanen Perspektive einführen? Der vorliegende Artikel führt eine Untersuchung durch, die die Einwirkung der kommerziellen Urbaniserung auf die Srategien zur Qualitätsverbesserungen von dem öffentlichen Raum und zur Organisation/Ordnung von den Stadtzentralitäten betont. Es zeigt Anleitungen auf, die Stadtpläner weiterhelfen können, wenn sie Entscheidungen im Bereich einzelhandelbezogener Entwicklungen treffen müssen.

P. 263-270 The activity of 2 populations of wild rabbits (Oryctolagus cuniculus, L. 1758), consisting of 14 adults (>9 mo of age) each (4 males and 10 females), was analysed over 2 consecutive years. Rabbits were captured in the wild and kept in 2 separate enclosures of 0.5 ha, with each enclosure divided into 2 zones: a smaller area where warrens were located (breeding area) and a larger area where food and water were provided (feeding area). Seven rabbits in each enclosure were individually tagged with a microchip (2 males and 5 females) and, after installing 2 detection devices, it was possible to identify which of the 2 areas they were located in and record the length of time spent in each. To regulate the size of the breeding population, young rabbits produced in the enclosures were captured and removed regularly. Considering the number of movements between areas and the time spent in the feeding area, a circadian activity pattern was found, reporting 2 maximum activity peaks coinciding with twilight (18.35% of the total movements, 6-8 a.m.) and daybreak (22.95%, 7-10 p.m.) while activity was dramatically decreased during the midday hours (1.86%, 10 a.m.-4 p.m.). Rabbits displayed a seasonal pattern throughout the year, with maximum activity levels during winter (45.76% of the total movements, January-March) and spring (42.91%, April-June), which could be related to higher reproductive activity at this time of the year as a higher breeding output was reported in June and September. The levels of activity exhibited by males (13.44% daily activity rate) were significantly higher than those displayed by females (9.80%). No significant differences were found regarding time spent on the feeding area in relation to season or gender. The average duration of each foray to the feeding area was higher during the summer, higher for females than males and higher during the middle of the night than the rest of the day. SI

The aim of this work was to obtain high calorific value products from coal fines cleaning wastes by agglomeration with vegetable oils. These residues are mainly being disposed of in dumps, causing important economic and environmental problems. Three Spanish coal fines wastes from different coal cleaning plants were agglomerated with crude and refined sunflower and soybean oils over a wide range of oil concentrations. The response of these fines wastes to agglomeration with the oils, was evaluated by the percentages of coal matter recovery, ash rejection and efficiency index. Speaking in terms of products quality, the best results were attained at the lowest oil concentrations, especially when the refined ones were used. In these cases, the agglomeration with vegetable oils allowed the recovery from coal fines wastes of a ready to burn fine coal fuel.

Mycobacterium tuberculosis,Mycobacterium leprae,Mycobacterium bovis,andMycobacterium aviumsubsp.paratuberculosiscan survive within host macrophages in a dormant state, encased within an organized aggregate of immune host cells called granuloma. Granulomas consist of uninfected macrophages, foamy macrophages, epithelioid cells, and T lymphocytes accumulated around infected macrophages. Within granulomas, activated macrophages can fuse to form multinucleated giant cells, also called giant Langhans cells. A rim of T lymphocytes surrounds the core, and a tight coat of fibroblast closes the structure. Severalin vivomodels have been used to study granuloma’s structure and function, but recently developedin vitromodels of granuloma show potential for closer observation of the early stages of host’s responses to live mycobacteria. This paper reviews culture conditions that resulted in three-dimensional granulomas, formed by the adhesion of cell populations in peripheral blood mononuclear cells infected with mycobacteria. The similarities of these models to granulomas encountered in clinical specimens include cellular composition, granulomas’ cytokine production, and cell surface antigens. A reliablein vitrodormancy model may serve as a useful platform to test whether drug candidates can kill dormant mycobacteria. Novel drugs that target dormancy-specific pathways may shorten the current long, difficult treatments necessary to cure mycobacterial diseases.