Spider Venoms Potentially Lethal to Humans

Spiders have one pair of venom glands, and only a few families have reduced them completely (Uloboridae, Holarchaeidae) or modified them to another function (Symphytognathidae or Scytodidae, see Suter and Stratton 2013). All other 42,000 known spider species (99%) utilize their venom to inject it into prey items, which subsequently become paralysed or are killed. Spider venom is a complex mixture of hundreds of components, many of them interacting with cell membranes or receptors located mainly in the nervous or muscular system (Herzig and King 2013). Spider venom, as it is today, has a 300-million-yearlong history of evolution and adaptation and can be considered as an optimized tool to subdue prey. In Mesothelae, the oldest spider group with less than 100 species, the venom glands lie in the anterior part of the cheliceral basal segment. They are very small and do not support the predation process very effectively. In Mygalomorphae, the venom glands are well developed and fill the basal cheliceral segment more or less completely. Many of these 3,000 species are medium- to large-/very large-sized spiders, and they have created the image of being dangerous beasts, attacking and killing a variety of animals, including humans. Although this picture is completely wrong, it is persistent and contributes considerably to human arachnophobia. The third group of spiders, Araneomorphae or “modern spiders”, comprises 93% of all spider species. The venom glands are enlarged and extend to the prosoma; the openings of the venom ducts are moved from the convex to the concave side of the cheliceral fangs and enlarged as well. These changes save the chelicerae from the necessity of being large, and hence, on the average, araneomorph spiders are much smaller than mygalomorphs. Nevertheless, they possess relatively large venom glands, situated mainly in the prosoma, and may also have rather potent venom.

Dicer is required for female reproductive tract development and fertility in the mouse.

Dicer encodes a riboendonuclease required for microRNA biosynthesis. Dicer was inactivated in Müllerian duct mesenchyme-derived tissues of the reproductive tract of the mouse, using an Amhr2-Cre allele. Although Amhr2-Cre; Dicer conditional mutant males appeared normal and were fertile, mutant females were infertile. In adult mutant females, there was a reduction in the size of the oviducts and uterine horns. The oviducts were less coiled compared to controls and cysts formed at the isthmus near the uterotubal junction. Unfertilized, degenerate oocytes were commonly found within these cysts, indicating a defect in embryo transit. Beads transferred into the mutant oviduct failed to migrate into the uterus. In addition, blastocysts transferred directly into the mutant uterus did not result in pregnancy. Histological analysis demonstrated that the mutant uterus contained less glandular tissue and often the few glands that remained were found within the myometrium, an abnormal condition known as adenomyosis. In adult mutants, there was ectopic expression of Wnt4 and Wnt5a in the luminal epithelium (LE) and glandular epithelium (GE) of the uterus, and Wnt11 was ectopically expressed in GE. These results demonstrate that Dicer is necessary for postnatal differentiation of Müllerian duct mesenchyme-derived tissues of the female reproductive tract, suggesting that microRNAs are important regulators of female reproductive tract development and fertility.

THE ROLE OF RECEPTOR TYROSINE KINASE AXL IN PANCREATIC DUCTAL ADENOCARCINOMA AND ITS REGULATION BY HEMATOPOIETIC PROGENITOR KINASE 1

Pancreatic ductal adenocarcinoma (PDA) is one of the most aggressive malignancies with less than 5% of five year survival rate. New molecular markers and new therapeutic targets are urgently needed for patients with PDA. Oncogenic receptor tyrosine kinase Axl has been reported to be overexpressed in many types of human malignancies, including diffuse glioma, melanoma, osteosarcoma, and carcinomas of lung, colon, prostate, breast, ovary, esophagus, stomach, and kidney. However, the expression and functions of Axl in PDA are unclear. We hypothesized that Axl contributes to the development and progression of PDA. We examined Axl expression in 54 human PDA samples and their paired benign pancreatic tissue by immunohistochemistry, we found that Axl was overexpressed in 70% of stage II PDAs, but only 22% of benign ducts (P=0.0001). Axl overexpression was associated with higher frequencies of distant metastasis and was an independent prognostic factor for both poor overall and recurrence-free survivals in patients with stage II PDA (p = 0.03 and 0.04). Axl silencing by shRNA in pancreatic cancer cell lines, panc-28 and Panc-1, decreased tumor cell migration and invasion and sensitized PDA cells to apoptosis stimuli such as γ-irradiation and serum starvation. In addition, we found that Axl-mediated Akt and NF-κB activation and up regulation of MMP2 were involved in the invasion, migration and survival of PDA cells. Thus, we demonstrate that Axl plays an important role in the development and progression of PDA. Targeting Axl signaling pathway may represent a new approach for the treatment of PDA. To understand the molecular mechanisms of Axl overexpression in PDA, we found that Axl expression was down-regulated by hematopoietic progenitor kinase 1 (HPK1), a newly identified tumor suppressor in PDA. HPK1 is lost in over 95% of PDAs. Restoration of HPK1 in PDA cells down-regulated Axl expression. HPK1-mediated Axl degradation was inhibited by leupeptin, baflomycin A1, and monensin, suggesting that HPK1-mediated Axl degradation was through endocytosis-lysosome pathway. HPK1 interacted with and phosphorylated dynamin, a critical component of endocytosis pathway. Overexpression of dominant negative form of dynamin blocked the HPK1-mediated Axl degradation. Therefore we concluded that HPK1-mediated Axl degradation was through endocytosis-lysosome pathway and loss of HPK1 expression may contribute to Axl overexpression in PDAs.

Vascular cell adhesion molecule 1 expression by biliary epithelium promotes persistence of inflammation by inhibiting effector T-cell apoptosis

Chronic hepatitis occurs when effector lymphocytes are recruited to the liver from blood and retained in tissue to interact with target cells, such as hepatocytes or bile ducts (BDs). Vascular cell adhesion molecule 1 (VCAM-1; CD106), a member of the immunoglobulin superfamily, supports leukocyte adhesion by binding a4b1 integrins and is critical for the recruitment of monocytes and lymphocytes during inflammation. We detected VCAM-1 on cholangiocytes in chronic liver disease (CLD) and hypothesized that biliary expression of VCAM-1 contributes to the persistence of liver inflammation. Hence, in this study, we examined whether cholangiocyte expression of VCAM-1 promotes the survival of intrahepatic a4b1 expressing effector T cells. We examined interactions between primary human cholangiocytes and isolated intrahepatic T cells ex vivo and in vivo using the Ova-bil antigen-driven murine model of biliary inflammation. VCAM-1 was detected on BDs in CLDs (primary biliary cirrhosis, primary sclerosing cholangitis, alcoholic liver disease, and chronic hepatitis C), and human cholangiocytes expressed VCAM-1 in response to tumor necrosis factor alpha alone or in combination with CD40L or interleukin-17. Liver-derived T cells adhered to cholangiocytes in vitro by a4b1, which resulted in signaling through nuclear factor kappa B p65, protein kinase B1, and p38 mitogen-activated protein kinase phosphorylation. This led to increased mitochondrial B-cell lymphoma 2 accumulation and decreased activation of caspase 3, causing increased cell survival. We confirmed our findings in a murine model of hepatobiliary inflammation where inhibition of VCAM-1 decreased liver inflammation by reducing lymphocyte recruitment and increasing CD8 and T helper 17 CD4 Tcell survival. Conclusions: VCAM-1 expression by cholangiocytes contributes to persistent inflammation by conferring a survival signal to a4b1 expressing proinflammatory T lymphocytes in CLD.

A numerical model for inert gas transport in the lung based on fractal airway morphology

Patients suffering from cystic fibrosis (CF) show thick secretions, mucus plugging and bronchiectasis in bronchial and alveolar ducts. This results in substantial structural changes of the airway morphology and heterogeneous ventilation. Disease progression and treatment effects are monitored by so-called gas washout tests, where the change in concentration of an inert gas is measured over a single or multiple breaths. The result of the tests based on the profile of the measured concentration is a marker for the severity of the ventilation inhomogeneity strongly affected by the airway morphology. However, it is hard to localize underlying obstructions to specific parts of the airways, especially if occurring in the lung periphery. In order to support the analysis of lung function tests (e.g. multi-breath washout), we developed a numerical model of the entire airway tree, coupling a lumped parameter model for the lung ventilation with a 4th-order accurate finite difference model of a 1D advection-diffusion equation for the transport of an inert gas. The boundary conditions for the flow problem comprise the pressure and flow profile at the mouth, which is typically known from clinical washout tests. The natural asymmetry of the lung morphology is approximated by a generic, fractal, asymmetric branching scheme which we applied for the conducting airways. A conducting airway ends when its dimension falls below a predefined limit. A model acinus is then connected to each terminal airway. The morphology of an acinus unit comprises a network of expandable cells. A regional, linear constitutive law describes the pressure-volume relation between the pleural gap and the acinus. The cyclic expansion (breathing) of each acinus unit depends on the resistance of the feeding airway and on the flow resistance and stiffness of the cells themselves. Special care was taken in the development of a conservative numerical scheme for the gas transport across bifurcations, handling spatially and temporally varying advective and diffusive fluxes over a wide range of scales. Implicit time integration was applied to account for the numerical stiffness resulting from the discretized transport equation. Local or regional modification of the airway dimension, resistance or tissue stiffness are introduced to mimic pathological airway restrictions typical for CF. This leads to a more heterogeneous ventilation of the model lung. As a result the concentration in some distal parts of the lung model remains increased for a longer duration. The inert gas concentration at the mouth towards the end of the expirations is composed of gas from regions with very different washout efficiency. This results in a steeper slope of the corresponding part of the washout profile.

The patent nasopalatine duct: a potential cause of unclear pain in the anterior maxilla

OBJECTIVE The aim of this report is to describe symptoms that can suggest the presence of a patent nasopalatine duct and to illustrate three cases. SUMMARY Patent nasopalatine ducts connecting the oral cavity with the nasal cavity are extremely rare. This malformation can be considered a developmental abnormality. Clinically, patent nasopalatine ducts appear as single or double spherical or oval apertures lateral or posterior to the incisive papilla. This type of anatomical malformation can be associated with an unclear pain sensation in the anterior maxillary region, which may be misinterpreted for example as toothache of endodontic origin. However, persisting nasopalatine ducts can also exist as an asymptomatic abnormality with no clinical sign of discomfort. Accordingly, understanding the differential diagnosis of a possible patent nasopalatine duct can prevent a general practitioner from performing unnecessary interventions, such as endodontic treatments, apical surgeries, or tooth extractions.

37,X/38,XY Mosaicism in a Cryptorchid Bengal Cat with Müllerian Duct Remnants

A 23-month-old tomcat was referred to our clinic because of male behavioral problems, cryptorchidism, and an undefined intra-abdominal organ resembling a uterus. Ultrasonography and computed tomography showed 2 fluid-filled tubular structures dorsolaterally to the bladder and connected to the pelvic urethra. The cat was castrated, and the tubular structures were surgically removed. Histology identified them as Müllerian duct remnants. The testes were hypoplastic, the epididymes and deferent ducts were normal. Cytogenetic analyses revealed the presence of a mosaic 37,X/38,XY karyotype which explains the clinical findings.

The comprehensiveness of the ESHRE/ESGE classification of female genital tract congenital anomalies: a systematic review of cases not classified by the AFS system.

STUDY QUESTION How comprehensive is the recently published European Society of Human Reproduction and Embryology (ESHRE)/European Society for Gynaecological Endoscopy (ESGE) classification system of female genital anomalies? SUMMARY ANSWER The ESHRE/ESGE classification provides a comprehensive description and categorization of almost all of the currently known anomalies that could not be classified properly with the American Fertility Society (AFS) system. WHAT IS KNOWN ALREADY Until now, the more accepted classification system, namely that of the AFS, is associated with serious limitations in effective categorization of female genital anomalies. Many cases published in the literature could not be properly classified using the AFS system, yet a clear and accurate classification is a prerequisite for treatment. STUDY DESIGN, SIZE AND DURATION The CONUTA (CONgenital UTerine Anomalies) ESHRE/ESGE group conducted a systematic review of the literature to examine if those types of anomalies that could not be properly classified with the AFS system could be effectively classified with the use of the new ESHRE/ESGE system. An electronic literature search through Medline, Embase and Cochrane library was carried out from January 1988 to January 2014. Three participants independently screened, selected articles of potential interest and finally extracted data from all the included studies. Any disagreement was discussed and resolved after consultation with a fourth reviewer and the results were assessed independently and approved by all members of the CONUTA group. PARTICIPANTS/MATERIALS, SETTING, METHODS Among the 143 articles assessed in detail, 120 were finally selected reporting 140 cases that could not properly fit into a specific class of the AFS system. Those 140 cases were clustered in 39 different types of anomalies. MAIN RESULTS AND THE ROLE OF CHANCE The congenital anomaly involved a single organ in 12 (30.8%) out of the 39 types of anomalies, while multiple organs and/or segments of Müllerian ducts (complex anomaly) were involved in 27 (69.2%) types. Uterus was the organ most frequently involved (30/39: 76.9%), followed by cervix (26/39: 66.7%) and vagina (23/39: 59%). In all 39 types, the ESHRE/ESGE classification system provided a comprehensive description of each single or complex anomaly. A precise categorization was reached in 38 out of 39 types studied. Only one case of a bizarre uterine anomaly, with no clear embryological defect, could not be categorized and thus was placed in Class 6 (un-classified) of the ESHRE/ESGE system. LIMITATIONS, REASONS FOR CAUTION The review of the literature was thorough but we cannot rule out the possibility that other defects exist which will also require testing in the new ESHRE/ESGE system. These anomalies, however, must be rare. WIDER IMPLICATIONS OF THE FINDINGS The comprehensiveness of the ESHRE/ESGE classification adds objective scientific validity to its use. This may, therefore, promote its further dissemination and acceptance, which will have a positive outcome in clinical care and research. STUDY FUNDING/COMPETING INTERESTS None.

In vivo definition of genetic and cellular aspects of mammalian sexual differentiation

The differentiation of the reproductive organs is an essential developmental process required for the proper transmission of the genetic material. Müllerian inhibiting substance (MIS) is produced by testes and is necessary for the regression of the Müllerian ducts: the anlagen of the uterus, fallopian tubes and cervix. In vitro and standard transgenic mouse studies indicate that the nuclear hormone receptor Steroidogenic factor 1 (SF-1) and the transcription factor SOX9 play an essential role in the regulation of Mis. To test this hypothesis, mutations in the endogenous SF-1 and SOX9 binding sites in the mouse Mis promoter were introduced by gene targeting in embryonic stem (ES) cells. In disagreement with cell culture and transgenic mouse studies, male mice homozygous for the mutant SF-1 binding site correctly initiated Mis transcription in the fetal testes, although at significantly reduced levels. Surprisingly, sufficient Mis was produced for complete elimination of the Müllerian duct system. However, when the SF-1 binding site mutation was combined with an Mis -null allele, the further decrease in Mis levels led to a partial retention of uterine tissue, but only at a distance from the testes. In contrast, males homozygous for the mutant SOX9 binding site did not initiate Mis transcription, resulting in pseudohermaphrodites with a uterus and oviducts. These studies suggest an essential role for SOX9 in the initiation of Mis transcription, whereas SF-1 appears to act as a quantitative regulator of Mis transcript levels perhaps for influencing non-Müllerian duct tissues. ^ The Mis type II receptor, a member of the TGF- b superfamily, is also required for the proper regression of the Müllerian ducts. Mis type II receptor-deficient human males and their murine counterparts develop as pseudohermaphrodites. A lacZ reporter cassette was introduced into the mouse Mis type II receptor gene, by homologous recombination in ES cells. Expression studies, based on b -galactosidase activity, show marked expression of the MIS type II receptor in the postnatal Sertoli cells of the testis as well as in the prenatal and postnatal granulosa cells of the ovary. Expression is also seen in the mesenchymal cells surrounding the Müllerian duct and in the longitudinal muscle layer of the uterus. ^

Life and death of the embryonic female reproductive tract

Regardless of genetic sex, amniotes develop two sets of genital ducts, the Wolffian and Müllerian ducts. Normal sexual development requires the differentiation of one duct and the regression of the other. I show that cells in the rostral most region of the coelomic epithelium (CE) are specified to a Müllerian duct fate beginning at Tail Somite Stage 19 (TS19). The Müllerian duct (MD) invaginates from the CE where it extends caudally to and reaches the Wolffian duct (WD) by TS22. Upon contact, the MD elongates to the urogenital sinus separating the WD from the CE and its formation is complete by TS34. During its elongation, the MD is associated with and dependent upon the WD and I have identified the mechanism for MD elongation. Using the Rosa26 reporter to fate map the WD, I show that the WD does not contribute cells to the MD. Using an in vitro recombinant explant culture assay I show that the entire length of the MD is derived from the CE. Furthermore, I analyzed cell proliferation and developed an in vitro assay to show that a small population of cells at the caudal tip proliferates, laying the foundation for the formation of the MD. I also show that during its formation, the MD has a distinctive mesoepithelial character. The MD in males regresses under the influence of Anti-Müllerian Hormone (AMH). Through tissue-specific gene inactivation I have identified that Acvr1 and Bmpr1a and Smad1, Smad5 and Smad8 function redundantly in transducing the AMH signal. In females the MD differentiates into an epithelial tube and eventually the female reproductive tract. However, the exact tissue into which the MD differentiates has not been determined. I therefore generated a MD specific Cre allele that will allow for the fate mapping of the MD in both females males. The MD utilizes a unique form of tubulogenesis during development and to my knowledge is the only tubule that relies upon a signal from and the presence of another distinct epithelial tube for its formation.^

Establishment of a complex inflammatory and protumorigenic microenvironment in mouse pancreas through cyclooxygenase-2 overexpression

Chronic inflammation is an established risk factor in the pathogenesis of many cancers. Pancreatic ductal adenocarcinoma, a malignancy with a particularly dismal prognosis, is no exception. Cyclooxygenase-2, a key enzyme induced by tissue injury, has a critical role in the generation of bioactive lipids known as prostaglandins. COX-2 overexpression is a frequent finding in pancreatic cancer, chronic pancreatitis and pancreatic intraepithelial neoplasias. To explore mechanisms through which chronic inflammation establishes and maintains a protumorigenic environment, we designed a mouse model overexpressing COX-2 in pancreatic parenchyma (BK5.COX-2 mice). We discovered that constitutive expression of COX-2 has a number of important sequelae, including upregulation of additional eicosanoid-generating enzymes and proinflammatory cytokines. Many of these molecular alterations precede the onset of significant histopathological changes. Increased levels of prostaglandins E2, D2, and F2α, 5-, 12-, and 15-hydroxyeiosatetraenoic acid (HETEs) were documented in tumors and pancreata of younger transgenic mice. Using a TaqMan™ Mouse Immune Panel, we detected elevated mRNAs for a number of proinflammatory cytokines (e.g., TNFα, IL-1β, IL-6). ^ Histological examination revealed early changes in the pancreas with similarities to human chronic pancreatitis, including loss of acinar cells, appearance of metaplastic ducts, and increased deposition of stroma. As the lesions progress, features typical of dysplastic and neoplastic cells emerged within the metaplastic ductal complexes, including cellular and nuclear atypia, crowding of cells, and loss of normal tissue architecture. The amount of fibroinflammatory stroma increased considerably; numerous small vessels were evident. A number of immunocytes from both the myeloid and lymphoid lineages were identified in transgenic pancreata. Neutrophils were the earliest to infiltrate, followed shortly by macrophages and mast cells. B and T cells generally began to appear by 8–12 weeks, and organized aggregates of lymphoid cells were often found in advanced lesions. ^ We tested the efficacy of several chemopreventive agents in this model, including celecoxib, a COX-2 selective inhibitor, pentoxifylline, a cytokine inhibitor, curcumin, a polyphenol with antioxidant and anti-inflammatory properties, and GW2974, a dual EGFR/ErbB2 inhibitor. Effects on lesion development were modest in the GW2974 and pentoxifylline treated groups, but significant prevention effects were observed with curcumin and celecoxib. ^

Distribution of the coal flow in the mill-duct system of the As Pontes Power Plant using CFD modeling

The efficiency of a Power Plant is affected by the distribution of the pulverized coal within the furnace. The coal, which is pulverized in the mills, is transported and distributed by the primary gas through the mill-ducts to the interior of the furnace. This is done with a double function: dry and enter the coal by different levels for optimizing the combustion in the sense that a complete combustion occurs with homogeneous heat fluxes to the walls. The mill-duct systems of a real Power Plant are very complex and they are not yet well understood. In particular, experimental data concerning the mass flows of coal to the different levels are very difficult to measure. CFD modeling can help to determine them. An Eulerian/Lagrangian approach is used due to the low solid–gas volume ratio.

Túneles en macizos calcáreos karstificados: impacto en las obras subterráneas de los fenómenos kársticos, su evaluación y tratamiento

En esta Tesis Doctoral se aborda un tema poco estudiado en el ámbito de los túneles y cuya problemática está basada en los riesgos e incertidumbres que representa el diseño y ejecución de túneles en macizos calizos karstificados. Mediante un estudio profundo del comportamiento de distintos casos reales de túneles en macizos kársticos calizos, aportando la realización de modelos y la experiencia constructiva del autor, se pretende proponer un procedimiento que permita sistematizar las actuaciones, investigación, evaluación y tratamiento en los túneles en karst, como herramienta básica para la toma de decisiones primarias correctas. Además, se proponen herramientas que pueden ayudar a mejorar el diseño y a decidir las medidas más eficientes para afrontar los problemas que genera el karst en los túneles, minimizando los riesgos para todos los actores, su probabilidad e impacto. Se profundiza en tres fases principales (que son referidas en la tesis en cuatro Partes): La INVESTIGACIÓN del macizo rocoso: La investigación engloba todas las actuaciones observacionales encaminadas a obtener el IK (Índice de Karstificación), así como las investigaciones necesarias mediante recopilación de datos superficiales, hidrogeológicos, climáticos, topográficos, así como los datos de investigaciones geofísicas, fotointerpretación, sondeos y ensayos geotécnicos que sean posibles. Mediante la misma, se debe alcanzar un conocimiento suficiente para llevar a cabo la determinación de la Caracterización geomecánica básica del macizo rocoso a distintas profundidades, la determinación del Modelo o modo de karstificación del macizo rocoso respecto al túnel y la Zonificación del índice de karstificación en el ámbito de actuación en el que se implantará el túnel. En esta primera fase es necesaria la correcta Definición geométrica y trazado de la obra subterránea: En función de las necesidades que plantee el proyecto y de los condicionantes externos de la infraestructura, se deben establecer los requisitos mínimos de gálibo necesarios, así como las condiciones de máximas pendientes para el trazado en alzado y los radios mínimos de las curvas en planta, en función del procedimiento constructivo y motivación de construcción del túnel (ferrocarril, carretera o hidráulico, etc...). Estas son decisiones estratégicas o primerias para las que se ha de contar con un criterio y datos adecuados. En esta fase, son importantes las decisiones en cuanto a las monteras o profundidades relativas a la karstificación dominante e investigación de las tensiones naturales del macizo, tectónica, así como las dimensiones del túnel en función de las cavidades previstas, tratamientos, proceso de excavación y explotación. En esta decisión se debe definir, conocida ya de forma parcial la geotecnia básica, el procedimiento de excavación en función de las longitudes del túnel y la clasificación geomecánica del terreno, así como sus monteras mínimas, accesos y condicionantes medioambientales, pero también en función de la hidrogeología. Se establecerá la afección esperable en el túnel, identificando en la sectorización del túnel, la afección esperable de forma general para las secciones pésimas del túnel. Con todos estos datos, en esta primera aproximación, es posible realizar el inventario de casos posibles a lo largo del trazado, para poder, posteriormente, minimizar el número de casos con mayores riesgos (técnicos, temporales o económicos) que requieran de tratamiento. Para la fase de EVALUACIÓN de la matriz de casos posibles en función del trazado inicial escogido (que puede estar ya impuesto por el proyecto, si no se ha podido intervenir en dicha fase), es necesario valorar el comportamiento teórico del túnel en toda su longitud, estudiando las secciones concretas según el tipo y el modo de afección (CASOS) y todo ello en función de los resultados de los estudios realizados en otros túneles. Se debe evaluar el riesgo para cada uno de los casos en función de las longitudes de túnel que se esperan que sean afectadas y el proceso constructivo y de excavación que se vaya a adoptar, a veces varios. Es importante tener en cuenta la existencia o no de agua o relleno arcilloso o incluso heterogéneo en las cavidades, ya que los riesgos se multiplican, así mismo se tendrá en cuenta la estabilidad del frente y del perímetro del túnel. En esta segunda fase se concluirá una nueva matriz con los resultados de los riesgos para cada uno de los casos que se presentarán en el túnel estudiado. El TRATAMIENTO, que se debe proponer al mismo tiempo que una serie de actuaciones para cada uno de los casos (combinación de tipos y modos de afección), debiendo evaluar la eficacia y eficiencia, es decir la relevancia técnica y económica, y como se pueden optimizar los tratamientos. Si la tabla de riesgos que se debe generar de nuevo introduciendo los factores técnicos y económicos no resulta aceptable, será necesaria la reconsideración de los parámetros determinados en fases anteriores. Todo el desarrollo de estas tres etapas se ha recogido en 4 partes, en la primera parte se establece un método de estudio e interpretativo de las investigaciones superficiales y geotécnicas, denominado índice de karstificación. En la segunda parte, se estudia la afección a las obras subterráneas, modelos y tipos de afección, desde un punto de vista teórico. La tercera parte trata de una recopilación de casos reales y las consecuencias extraídas de ellos. Y finalmente, la cuarta parte establece los tratamientos y actuaciones para el diseño y ejecución de túneles en macizos kársticos calizos. Las novedades más importantes que presenta este trabajo son: El estudio de los casos de túneles realizados en karst calizo. Propuesta de los tratamientos más efectivos en casos generales. La evaluación de riesgos en función de las tipologías de túnel y afecciones en casos generales. La propuesta de investigación superficial mediante el índice de karstificación observacional. La evaluación mediante modelos del comportamiento teórico de los túneles en karst para casos muy generales de la influencia de la forma, profundidad y desarrollo de la karstificación. In this doctoral thesis is studied the recommended investigation, evaluation and treatment when a tunnel is planed and constructed in karstic calcareous rock masses. Calcareous rock masses were selected only because the slow disolution produces stable conduct and caves instead the problems of sudden disolutions. Karstification index (IK) that encompasses various aspects of research karstic rock mass is presented. The karst rock masses are all unique and there are no similarities between them in size or density cavities ducts, but both their formation mechanisms, like, geological and hydrogeological geomorphological evidence allow us, through a geomechanical survey and geological-photo interpretation in the surface, establish a karst evaluation index specific for tunnelling, which allows us to set a ranking of the intensity of karstification and the associated stadistic to find caves and its risk in tunnelling. This index is estimated and useful for decision-making and evaluation of measures to be considered in the design of a tunnel and is set in degrees from 0 to 100, with similar to the RMR degrees. The sectorization of the tunnel section and the types of condition proposed in this thesis, are estimated also useful to understand the different effects that interception of ducts and cavities may have in the tunnel during construction and service life. Simplified calculations using two-dimensional models contained in the thesis, have been done to establish the relationship between the position of the cavities relative to the section of the tunnel and its size in relation to the safety factors for each ground conditions, cover and natural stresses. Study of over 100 cases of tunnels that have intercepted cavities or karst conduits have been used in this thesis and the list of risks found in these tunnels have been related as well. The most common and effective treatments are listed and finally associated to the models and type of affection generated to give the proper tool to help in the critical decision when the tunnels intercept cavities. Some existing studies from Marinos have been considered for this document. The structure of the thesis is mainly divided 4 parts included in 3 steps: The investigation, the evaluation and the treatments, which match with the main steps needed for the critical decisions to be made during the design and construction of tunnels in karstic rockmasses, very important to get a successfully project done.

Algoritmos genéticos en control activo de ruido

Este proyecto se centra en la implementación de un sistema de control activo de ruido mediante algoritmos genéticos. Para ello, se ha tenido en cuenta el tipo de ruido que se quiere cancelar y el diseño del controlador, parte fundamental del sistema de control. El control activo de ruido sólo es eficaz a bajas frecuencias, hasta los 250 Hz, justo para las cuales los elementos pasivos pierden efectividad, y en zonas o recintos de pequeñas dimensiones y conductos. El controlador ha de ser capaz de seguir todas las posibles variaciones del campo acústico que puedan producirse (variaciones de fase, de frecuencia, de amplitud, de funciones de transferencia electro-acústicas, etc.). Su funcionamiento está basado en algoritmos FIR e IIR adaptativos. La elección de un tipo de filtro u otro depende de características tales como linealidad, causalidad y número de coeficientes. Para que la función de transferencia del controlador siga las variaciones que surgen en el entorno acústico de cancelación, tiene que ir variando el valor de los coeficientes del filtro mediante un algoritmo adaptativo. En este proyecto se emplea como algoritmo adaptativo un algoritmo genético, basado en la selección biológica, es decir, simulando el comportamiento evolutivo de los sistemas biológicos. Las simulaciones se han realizado con dos tipos de señales: ruido de carácter aleatorio (banda ancha) y ruido periódico (banda estrecha). En la parte final del proyecto se muestran los resultados obtenidos y las conclusiones al respecto. Summary. This project is focused on the implementation of an active noise control system using genetic algorithms. For that, it has been taken into account the noise type wanted to be canceled and the controller design, a key part of the control system. The active noise control is only effective at low frequencies, up to 250 Hz, for which the passive elements lose effectiveness, and in small areas or enclosures and ducts. The controller must be able to follow all the possible variations of the acoustic field that might be produced (phase, frequency, amplitude, electro-acoustic transfer functions, etc.). It is based on adaptive FIR and IIR algorithms. The choice of a kind of filter or another depends on characteristics like linearity, causality and number of coefficients. Moreover, the transfer function of the controller has to be changing filter coefficients value thought an adaptive algorithm. In this project a genetic algorithm is used as adaptive algorithm, based on biological selection, simulating the evolutionary behavior of biological systems. The simulations have been implemented with two signal types: random noise (broadband) and periodic noise (narrowband). In the final part of the project the results and conclusions are shown.

Measurement of velocity in rotational flows using ultrasonic anemometry: the flowmeter

In this paper a previously developed theoretical model of the measurement process performed by a transit-time ultrasonic anemometer is applied to a fluid flowing through a circular section pipe. This model considers the influence of the shift of the acoustic pulse trajectory from straight propagation due to the flow on the measured speed. The aim of this work is to estimate the errors induced in the measured velocity by the shift of the acoustic pulse trajectory. Using different duct’s flow models, laminar and turbulent regimes have been analyzed. The results show that neglecting the effect of shift of the acoustic pulse trajectory leads to flow rate measurement underestimation.