Acondicionamiento y mejora de la N-340a, travesía de Vinaròs. Tramo norte, alternativa 2

[EN] The main objective of this final degree project is the remodeling and improvement of the N 340a side street in their way to Vinaròs. This project responds to a necessity to adapt the road to a new uses around urban integration. The purpose of this Project is to become the road as a fully integrated street that nowadays is a road with absolute preference to vehicular traffic. We will made a full reconstruction and improvement of the N-340a. This document has been divided into two parts using different work methodologies. The first part is a teamwork, we will see a territorial characterization of the environment. We will propose different alternatives for urban crossing restructuration. The second part take¿s place as an individual job, we will develop the best alterative in a sub-section of the side street (northern section).[CA] L'objectiu fonamental d'aquest TFG és el condicionament i millora de la N-340a al seu pas per Vinaròs. Aquest avantprojecte respon a la necessitat d'adequació als seus nous usos i d'integració urbanística. La finalitat de l'actuació és que l'antiga travessia deixi de ser una via dissenyada amb preferència absoluta al trànsit rodat per convertir-se en una travessia urbana completament integrada. Aquest document s'ha dividit en dues parts emprant-hi metodologies de treball diferents; a la primera part, desenvolupada mitjançant un treball en equip, es realitza una caracterització territorial de l'entorn i es proposen i s'avaluen diferents alternatives per a la reconversió urbana de la travessia. A la segona part es desenvolupa, ja com a treball individual, l'alternativa òptima en un sub-tram de la travessia (Tram nord).[ES] El objetivo fundamental de este TFG es el acondicionamiento y mejora de la travesía de la N-340a a su paso por Vinaròs. Este anteproyecto responde a la necesidad de integración urbanística de la vía, adecuándola a sus nuevos usos. La finalidad de la actuación es que la antigua travesía deje de ser una vía diseñada con preferencia absoluta al tráfico rodado para convertirse en una vía urbana completamente integrada. Este documento se ha dividido en dos partes empleando en ellas metodologías de trabajo diferentes; en la primera parte, desarrollada mediante un trabajo en equipo, se realiza una caracterización territorial del entorno y se proponen y evalúan diferentes alternativas para la reconversión urbana de la travesía. En la segunda parte se desarrolla, ya como trabajo individual y a nivel de anteproyecto, la alternativa óptima en un sub-tramo de la travesía (Tramo norte).Pamies Catalán, D. (2016). Acondicionamiento y mejora de la N-340a, travesía de Vinaròs. Tramo norte, alternativa 2. http://hdl.handle.net/10251/68661.Archivo delegad

Mechanism-based models in reproductive and developmental toxicology

This chapter discusses the study of the currently available models for testing developmental toxicity (embryotoxicity and teratogenicity). The main alternative models for testing developmental toxicity are described. These models are divided between validated models (whole-embryo culture test (WEC), micromass test (MM) and embryonic stem cell test (EST)) and those that are not currently validated (although have proven scientific validity) as is the case of zebrafish, frog embryo teratogenesis assay (FETAX), in silico models for predicting embryotoxicity, in vitro cellular models different from the EST method, and methods using fragments of embryos. The non-validated alternative models for testing developmental toxicity are also explained here. To date, only three in vitro methods (MM, EST and WEC) have been validated by an international agency (ECVAM) in order to be used for testing the embryotoxicity potential of chemicals, although other models such as FETAX and zebrafish have also proved their validity for this purpose. Methods based on the employment of embryos allow the specific malformation expected after exposure to the chemical to be determined, while methods based on cellular systems are more relevant in order to determine the mechanism underlying the adverse observed effect and still display a wide field for improving their prediction capability

Chapter 7 - Alternative methods to animal experimentation for testing developmental toxicity

The available alternative methods for testing developmental toxicity comprise either cellular models or whole embryos of rodents, fish, or amphibian. The simplest cellular models consider the use of human or animal embryonic stem cells (embryonic or of induced pluripotency) under differentiation and one of the most widely used endpoints in these methods is the alterations in differentiated beating cardiomyocytes, although determination of other molecular markers is also extended. More complex cellular models consider the use of cocultures or 3D cultures or the so-called organoids. These models mimic the physiological environment in a much better way than the simple monolayer cultures. The use of whole embryos allows the determination of which teratogenic effects are expectable after exposure to developmental toxicants, which is one of the main disadvantages of the cellular methods. The appropriate assessment of chemical safety for development needs of a battery of alternative methods applied. Integrated Approaches to Testing and Assessment (IATA) would allow in the close future to perform safe and reliable assessment of developmental toxicity based on alternative methods

Reproductive toxicity: in vivo testing guidelines from OECD

The guidelines for testing the reproductive toxicity in vivo developed and validated by Organisation for Economic Cooperation and Development allow for a systematic and internationally accepted testing and assessment of chemicals. Within reproductive toxicity two main categories of guidelines are usually identified: one dedicated to testing developmental toxicity, starting before the gestation period, while the other guidelines test the reproductive toxicity as a whole, therefore including male and female fertility and development. In this chapter, we summarize the guidelines on in vivo reproductive toxicity, by describing the general principles of the studies, the main aspects of the procedure, the endpoints and the observations, data reporting, and the criteria needed for the interpretation of their results

OECD guidelines and validated methods for in vivo testing of reproductive toxicity

This chapter discusses the methods adapted by the OECD and some other protocols, including the general principles of the study, the main aspects of the procedure, the endpoints and the observations, data reporting and criteria for interpreting results, and summarizing the guidelines. OECD 414 provides general information concerning the effects of prenatal exposure on the pregnant test animal and on the developing organism. OECD 415 is designed to provide general information concerning the effects of the tested substance on male and female reproductive performance. OECD 416 test is designed to provide general information concerning the effects of a tested substance on the integrity and performance of the male and female reproductive systems, including gonadal function, the estrus cycle, mating behavior, conception, gestation, parturition, lactation and weaning, and the growth and development of the offspring. The study also provides information about the effects on the first generation (F1) including neonatal morbidity, mortality and preliminary data on prenatal and postnatal developmental toxicity. OECD 421 offers only limited means of detecting postnatal manifestations of prenatal exposure, or effects that may be induced during postnatal exposure. OECD 422 is intended for identification of possible health hazards likely to arise from repeated exposure over a relatively limited period of time. OECD 426 is designed to provide data, including dose–response characterization, on the potential functional and morphological effects on the developing nervous system of the offspring that may arise from exposure in uterus and during early life

Specific Effect of 5-Fluorouracil on α-Fetoprotein Gene Expression During the In Vitro Mouse Embryonic Stem Cell Differentiation

Embryonic stem (ES) cells are considered an important alternative to develop in vitro screening methods for embryotoxicity. Mouse ES cells can be cultured as cell suspension aggregates termed “embryoid bodies” (EBs) in which cells start to differentiate. We have studied the expression of several genes in the presence of a wide range of concentrations of 5-fluorouracil (5-FU). This well-established embryotoxic compound completely inhibited cell viability at 200 nmol/L in monolayer cultures. At lower concentrations, 5-FU led to decrease in the expression of the α-fetoprotein gene, a marker of the visceral endoderm, in the EBs. However, the expression of several mesodermal gene markers was not significantly affected at these concentrations. These results suggest a high sensitivity of the visceral endoderm differentiation to 5-FU. Therefore, the quantification of the α-fetoprotein gene after exposure to potential embryotoxicants should be considered an additional end point in future embryotoxicity assays in vitro with ES cells

Polyphenolic Extract of Barbary-Fig (Opuntia ficus-indica) Syrup: RP–HPLC–ESI–MS Analysis and Determination of Antioxidant, Antimicrobial and Cancer-Cells Cytotoxic Potentials

The traditionally derived syrup of Opuntia ficus-indica fruit is commonly used in homemade confectionery. Herein, the aqueous-acetone extract prepared from the Tunisian O. ficus-indica syrup was investigated. The qualitatively and quantitatively polyphenolic content was analysed using reversed-phase high-performance liquid chromatography–diode array detection (RP-HPLC–DAD) coupled to electrospray ionisation–mass spectrometry (ESI–MS). The extract contained 19.95 ± 2.01 mg phenolics per gram of fresh starting material with isorhamnetin 3-O-robinobioside as the major compound (22.76%). The syrup extract showed strong antioxidant potentials as assessed by both ABTS and DPPH functional methods. It exhibited effective antimicrobial activity, particularly against Staphylococcus aureus and Staphylococcus epidermidis with a minimal bactericide concentration (MBC) of 1.3 mg phenolics/ml. Furthermore, at final concentrations in the range of 41.38–186.25 μg polyphenols/ml, the extract decreased human SH-SY5Y neuroblastoma and 3T3 fibroblast in vitro cell viability in a dose- and time-dependent manner compared to non-treated control cells. The observed effects were significantly (P < 0.05) high against cancer lines. Extract concentrations higher than 106.43 μg/ml reduced cancer cells viability to 50–60% 1–3 h post-treatment. Further in vivo insight studies should emphasise and validate the herein obtained results

Genomic and Phenotypic Alterations of the Neuronal-Like Cells Derived from Human Embryonal Carcinoma Stem Cells (NT2) Caused by Exposure to Organophosphorus Compounds Paraoxon and Mipafox

Organophosphorus compounds (OPs) are pesticides of worldwide use due to the acute insecticidal effects mediated by the inhibition of esterases of the central nervous system (mainly acetylcholinesterase and neuropathy target esterase (NTE)). OPs need to inhibit acetylcholinesterase to be effective insecticides, but not NTE since its inhibition might cause progressive, irreversible delayed neuropathy in humans and other species. Additionally, other neurological and neurodevelopmental toxic effects with unknown targets have been reported in humans or animals chronically exposed to OPs. We used a mixed neuronal/glia culture derived from well-characterised human embryonal carcinoma stem cells (hNT2) to determine if neuropathic OP mipafox and non-neuropathic OP paraoxon are able to alter the neuronal differentiation process evaluated by gene expression studies, neuronal electrical activity measurements and neural cell morphology quantification. Exposure to paraoxon at non-cytotoxic concentrations altered the expression of different genes involved mainly in signalling pathways related to chromatin assembly and nucleosome integrity, generating cultures with a larger number of differentiated neurons-like cells and branching points than in the control. Moreover, these paraoxon-exposed neuronal-like cells displayed reduced electrical activity when compared with the control neurons as measured by Micro Electrode Array Chips. Similarly, exposure to mipafox, a known inhibitor of NTE activity, also reduced the electrical activity of hNT2 cultures differentiated into neurons-like cells, but no significant changes in cell morphology or gene expression were detected. Therefore, we conclude that paraoxon is able to strongly disturb in vitro neurodifferentiation, while the absence of morphological and transcriptional alterations did not allow us to conclude if the electrophysiological alterations detected in mipafox-exposed neurons are due to neurodevelopmental toxicity or to effects on mature neurons.JRC.I.2-Public Health Policy Suppor

A Human iPSC-derived 3D platform using primary brain cancer cells to study drug development and personalized medicine

Abstract A high throughput histology (microTMA) platform was applied for testing drugs against tumors in a novel 3D heterotypic glioblastoma brain sphere (gBS) model consisting of glioblastoma tumor cells, iPSC-derived neurons, glial cells and astrocytes grown in a spheroid. The differential responses of gBS tumors and normal neuronal cells to sustained treatments with anti-cancer drugs temozolomide (TMZ) and doxorubicin (DOX) were investigated. gBS were exposed to TMZ or DOX over a 7-day period. Untreated gBS tumors increased in size over a 4-week culture period, however, there was no increase in the number of normal neuronal cells. TMZ (100 uM) and DOX (0.3 uM) treatments caused ~30% (P~0.07) and ~80% (P < 0.001) decreases in the size of the tumors, respectively. Neither treatment altered the number of normal neuronal cells in the model. The anti-tumor effects of TMZ and DOX were mediated in part by selective induction of apoptosis. This platform provides a novel approach for screening new anti-glioblastoma agents and evaluating different treatment options for a given patient