Agrobacterium-mediated transformation of the terpenoid indole alkaloid-producing plant species Tabernaemontana pandacaqui

Plants of the Apocynaceae family produce a wide range of terpenoid indole alkaloids (TIAs) which have important pharmaceutical applications. Studies of the molecular mechanisms controlling TIA biosynthesis may eventually provide possibilities to improve product yield by genetic modification of plants or cell cultures. However, these studies suffer from the lack of transformation/regeneration protocols for Apocynaceae plants. We chose to study the feasibility of Agrobacterium tumefaciens-mediated transformation of Tabernaemontana pandacaqui, because of the availability of an efficient regeneration procedure for this member of the Apocynaceae family. A procedure to produce transgenic T. pandacaqui plants was established, albeit with low efficiency. Transgenic expression was demonstrated of an intron-containing β-glucuronidase reporter gene and of a gene coding for the TIA biosynthetic enzyme strictosidine synthase from Catharanthus roseus, another Apocynaceae species.info:eu-repo/semantics/publishedVersio

Agrobacterium-mediated transformation of the terpenoid indole alkaloid-producing plant species Tabernaemontana pandacaqui

Plants of the Apocynaceae family produce a wide range of terpenoid indole alkaloids (TIAs) which have important pharmaceutical applications. Studies of the molecular mechanisms controlling TIA biosynthesis may eventually provide possibilities to improve product yield by genetic modification of plants or cell cultures. However, these studies suffer from the lack of transformation/regeneration protocols for Apocynaceae plants. We chose to study the feasibility of Agrobacterium tumefaciens-mediated transformation of Tabernaemontana pandacaqui, because of the availability of an efficient regeneration procedure for this member of the Apocynaceae family. A procedure to produce transgenic T. pandacaqui plants was established, albeit with low efficiency. Transgenic expression was demonstrated of an intron-containing β-glucuronidase reporter gene and of a gene coding for the TIA biosynthetic enzyme strictosidine synthase from Catharanthus roseus, another Apocynaceae species.info:eu-repo/semantics/publishedVersio

GEM operation in double-phase xenon

http://www.sciencedirect.com/science/article/B6TJM-4T7F5JF-5/2/897502f8049abd8c2dd3dffc3821f0f

A model of the reflection distribution in the vacuum ultra violet region

A reflection model with three components, a specular spike, a specular lobe and a diffuse lobe is discussed. This model was successfully applied to describe reflection of xenon scintillation light (175 nm) by PTFE and other fluoropolymers and can be used for Monte Carlo simulation and analysis of scintillation detectors. The measured data favors a Trowbridge-Reitz distribution function of ellipsoidal micro-surfaces. The intensity of the coherent reflection increases with increasing angle of incidence, as expected, since the surface appears smoother at grazing angles. The total reflectance obtained for PTFE is about 70% for VUV light at normal incidence in vacuum and estimated to be up to 100% in contact with liquid xenon

Temperature evaluation in ex-vivo bovine bones submitted to drilling processes

A necrose óssea induzida durante o processo de furação é um fenómeno frequente que contribui para problemas pós-operatórios. O calor de fricção gerado pelo contato entre a broca e a parede do furo é inevitável. No entanto, a utilização de técnicas avançadas para a obtenção do registo térmico durante a furação óssea é importante para a garantia da qualidade durante a furação. O objetivo deste estudo é apresentar dois métodos experimentais diferentes para analisar as condições de furação que geram as temperaturas mais baixas, evitando a ocorrência da necrose óssea. Foram utilizados ossos de bovino ex-vivo para simular o processo de furação considerando os efeitos do diâmetro da broca, velocidade de rotação e velocidade de avanço. Foram realizados diferentes testes experimentais para avaliar a sua repetibilidade. Os resultados identificaram o diâmetro da broca como o parâmetro mais crítico na indução de temperaturas mais elevadas durante a furação óssea.This research was supported by the Portuguese Foundation of Science and Technology under the research project UID/EMS/50022/2013. The third author acknowledges the funding of Project NORTE-01-0145-FEDER-000022-SciTech -Science and Technology for Competitive and Sustainable Industries, co-financed by Programa Operacional Regional do Norte (NORTE2020), through Fundo Europeu de Desenvolvimento Regional (FEDER).info:eu-repo/semantics/publishedVersio

A promoter region that controls basal and elicitor-inducible expression levels of the NADPH: cytochrome P450 reductase gene (Cpr) from Catharanthus roseus binds nuclear factor GT-1

NADPH:cytochrome P450 reductase (CPR) is essential for the activation of cytochrome P450 enzymes, which are involved in a wide variety of metabolic pathways in plants, including those related to defence responses. In the subtropical plant Catharanthus roseus several cytochrome P450 enzymes operate in the biosynthesis of defence-related terpenoid indole alkaloids (TIAs). In agreement with the importance of CPR in defence, Cpr mRNA levels in C. roseus were found to be enhanced by fungal elicitor preparations that also induce TIA biosynthesis and P450 gene expression. Here we describe the isolation of a C. roseus genomic DNA clone covering the 5¢ part of the Cpr gene and 1.6-kb of upstream sequences. Mapping of the transcription start site showed the untranslated leader sequence is approximately 280 bp long. To study the control of gene expression by the Cpr promoter, transcriptional fusions between Cpr promoter fragments and the gusA reporter gene were generated and their expression was analyzed in stably transformed tobacco plants. The Cpr promoter fragment extending from )1510 to )8, with respect to the ATG start codon, conferred basal and elicitor-inducible expression on the gusA reporter gene, strongly indicating that the Cpr gene of C. roseus is indeed controlled by this promoter region. Progressive deletion from the 5¢ end of the promoter to position )632 had little e ect on gusA expression. However, deletion to position )366 resulted in a complete loss of basal activity and largely eliminated elicitor-induced expression, indicating that the region from )632 to )366 contains the main transcription-enhancing cis-regulatory sequences. Electrophoretic mobility shift assays with tobacco nuclear extracts showed that binding sites for nuclear factor GT-1 are redundant in the Cpr promoter, but absent from the downstream part of the leader sequence. The presence of strong GT-1 binding sites in the main enhancer region ()632 to )366), is suggestive of a functional role for this factor in basal expression and elicitor responsiveness of the Cpr promoter.info:eu-repo/semantics/publishedVersio

A promoter region that controls basal and elicitor-inducible expression levels of the NADPH: cytochrome P450 reductase gene (Cpr) from Catharanthus roseus binds nuclear factor GT-1

NADPH:cytochrome P450 reductase (CPR) is essential for the activation of cytochrome P450 enzymes, which are involved in a wide variety of metabolic pathways in plants, including those related to defence responses. In the subtropical plant Catharanthus roseus several cytochrome P450 enzymes operate in the biosynthesis of defence-related terpenoid indole alkaloids (TIAs). In agreement with the importance of CPR in defence, Cpr mRNA levels in C. roseus were found to be enhanced by fungal elicitor preparations that also induce TIA biosynthesis and P450 gene expression. Here we describe the isolation of a C. roseus genomic DNA clone covering the 5¢ part of the Cpr gene and 1.6-kb of upstream sequences. Mapping of the transcription start site showed the untranslated leader sequence is approximately 280 bp long. To study the control of gene expression by the Cpr promoter, transcriptional fusions between Cpr promoter fragments and the gusA reporter gene were generated and their expression was analyzed in stably transformed tobacco plants. The Cpr promoter fragment extending from )1510 to )8, with respect to the ATG start codon, conferred basal and elicitor-inducible expression on the gusA reporter gene, strongly indicating that the Cpr gene of C. roseus is indeed controlled by this promoter region. Progressive deletion from the 5¢ end of the promoter to position )632 had little e ect on gusA expression. However, deletion to position )366 resulted in a complete loss of basal activity and largely eliminated elicitor-induced expression, indicating that the region from )632 to )366 contains the main transcription-enhancing cis-regulatory sequences. Electrophoretic mobility shift assays with tobacco nuclear extracts showed that binding sites for nuclear factor GT-1 are redundant in the Cpr promoter, but absent from the downstream part of the leader sequence. The presence of strong GT-1 binding sites in the main enhancer region ()632 to )366), is suggestive of a functional role for this factor in basal expression and elicitor responsiveness of the Cpr promoter.info:eu-repo/semantics/publishedVersio

Perspectives for Positron Emission Tomography with RPCs

In this study we address the feasibility and main properties of a positron emission tomograph (PET) based on RPCs. The concept, making use of the converter-plate principle, takes advantage of the intrinsic layered structure of RPCs and its simple and economic construction. The extremely good time and position resolutions of RPCs also allow the TOF-PET imaging technique to be considered. Monte-Carlo simulations, supported by experimental data, are presented and the main advantages and drawbacks for applications of potential interest are discussed.Comment: Presented at "RPC2001-VI Workshop on Resistive Plate Chambers and Related Detectors", Coimbra, Portugal, 26-27 November 2001 (5 pages

Detection of Scintillation Light of Liquid Xenon with a LAAPD

First observation of liquid xenon scintillation due to alpha-particles and gamma-rays with a large area avalanche photodiode immersed in the liquid is reported. An energy resolution of 10% (FWHM) and a coincidence time resolution of less then 1 ns (FWHM) were measured with 5.5 MeV alpha-particles and 511 keV gamma-rays, respectively. The quantum efficiency of the photodiode for xenon scintillation light (wavelength 178 nm) is estimated to be ~100%.Comment: 11 pages, 9 figures, to be published in Nuclear Instruments and Method

A survey of energy loss calculations for heavy ions between 1 and 100 keV

The original Lindhard-Scharff-Schi{\o}tt (LSS) theory and the more recent Tilinin theory for calculating the nuclear and electronic stopping powers of slow heavy ions are compared with predictions from the SRIM code by Ziegler. While little discrepancies are present for the nuclear contribution to the energy loss, large differences are found in the electronic one. When full ion recoil cascade simulations are tested against the elastic neutron scattering data available in the literature, it can be concluded that the LSS theory is the more accurate.Comment: Presented at the 10th International Symposium on Radiation Physics, 17-22 September, 2006, Coimbra, Portugal; style corrections, small change to fig.