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    Preliminary Measurements of Be-10/Be-7 Ratio in Rainwater for Atmospheric Transport Analysis

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    The meteoric cosmogenic beryllium has been used as an essential geophysical tracer in the analysis of atmospheric flows and erosion soils since 1960. The first measurements Be-7 and Be-10 concentrations in rainwater from Mexico, have been carried out by using gamma decay spectroscopy and AMS techniques, respectively for each isotope. With this it was possible to report a preliminar value for the Be-10/Be-7 isotopic ratio in such environmental samples. The present work described preliminary results related to rainwater collected at mountain and metropolitan areas. Results are compared with predictions and previous measurements for both radioisotopes, observing a very sensible behavior particularly for the case of Be-7 activities

    Variability of the transport of anthropogenic CO2 at the Greenland-Portugal OVIDE section:Controlling mechanisms

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    The interannual to decadal variability in the transport of anthropogenic CO2 (Cant) across the subpolar North Atlantic (SPNA) is investigated, using summer data of the FOUREX and OVIDE high-resolution transoceanic sections, from Greenland to Portugal, occupied six times from 1997 to 2010. The transport of Cant across this section, Tcant hereafter, is northward, with a mean value of 254 ± 29 kmol s-1 over the 1997-2010 period. We find that Tcant undergoes interannual variability, masking any trend different from 0 for this period. In order to understand the mechanisms controlling the variability of Tcant across the SPNA, we propose a new method that quantifies the transport of Cant caused by the diapycnal and isopycnal circulation. The diapycnal component yields a large northward transport of Cant (400 ± 29 kmol s-1) that is partially compensated by a southward transport of Cant caused by the isopycnal component (-171 ± 11 kmol s-1), mainly localized in the Irminger Sea. Most importantly, the diapycnal component is found to be the main driver of the variability of Tcant across the SPNA. Both the Meridional Overturning Circulation (computed in density coordinates, MOCσ) and the Cant increase in the water column have an important effect on the variability of the diapycnal component and of Tcant itself. Based on this analysis, we propose a simplified estimator for the variability of T cant based on the intensity of the MOCσ and on the difference of Cant between the upper and lower limb of the MOCσ (ΔCant). This estimator shows a good consistency with the diapycnal component of T cant, and help to disentangle the effect of the variability of both the circulation and the Cant increase on the Tcant variability. We find that ΔCant keeps increasing over the past decade, and it is very likely that the continuous Cant increase in the water masses will cause an increase in Tcant across the SPNA at long timescale. Nevertheless, at the timescale analyzed here (1997-2010), the MOCσ controls the T cant variability, blurring any Tcant trend. Extrapolating the observed ΔCant increase rate and considering the predicted slow-down of 25% of the MOCσ, Tcant across the SPNA is expected to increase by 430 kmol s-1 during the 21st century. Consequently, an increase in the storage rate of Cant in the SPNA could be envisaged

    The synergetic effect from the combination of different adsorption resins in batch and semi-continuous cultivations of S. Cerevisiae cell factories to produce acetylated Taxanes precursors of the anticancer drug Taxol

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    In situ product recovery is an efficient way to intensify bioprocesses as it can perform adsorption of the desired natural products in the cultivation. However, it is common to use only one adsorbent (liquid or solid) to perform the product recovery. For this study, the use of an in situ product recovery method with three combined commercial resins (HP-20, XAD7HP, and HP-2MG) with different chemical properties was performed. A new yeast strain of Saccharomyces cerevisiae was engineered using CRISPR Cas9 (strain EJ2) to deliver heterologous expression of oxygenated acetylated taxanes that are precursors of the anticancer drug Taxol ® (paclitaxel). Microscale cultivations using a definitive screening design (DSD) were set to get the best resin combinations and concentrations to retrieve high taxane titers. Once the best resin treatment was selected by the DSD, semi-continuous cultivation in high throughput microscale was performed to increase the total taxanes yield up to 783 ± 33 mg/L. The best T5α-yl Acetate yield obtained was up to 95 ± 4 mg/L, the highest titer of this compound ever reported by a heterologous expression. It was also observed that by using a combination of the resins in the cultivation, 8 additional uncharacterized taxanes were found in the gas chromatograms compared to the dodecane overlay method. Lastly, the cell-waste reactive oxygen species concentrations from the yeast were 1.5-fold lower in the resin's treatment compared to the control with no adsorbent aid. The possible future implications of this method could be critical for bioprocess intensification, allowing the transition to a semi-continuous flow bioprocess. Further, this new methodology broadens the use of different organisms for natural product synthesis/discovery benefiting from clear bioprocess intensification advantages

    In situ solid-liquid extraction enhances recovery of taxadiene from engineered Saccharomyces cerevisiae cell factories

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    Microbial cell factories express diverse heterologous pathways for the production of a wide range of valuable natural products. However, the recovery and purification of such compounds is a major bottleneck in commercialization. In this study, a novel in situ solid phase adsorption strategy was investigated for enhanced recovery of taxadiene, a precursor to the blockbuster anticancer drug, paclitaxel, from engineered Saccharomyces cerevisiae. A synthetic adsorbent resin (HP-20) was employed to efficiently sequester taxadiene as it was secreted during growth and a carefully optimized desorption solvent was applied following cultivation to maximize recovery of both secreted and intracellular taxadiene, across a range of scales (2 – 250 mL). Resin concentration was found to have an impact on cellular growth, with the high concentration of 12 % (w/v) resulting in fragmentation of the resin beads, which was detrimental to growth. The optimal resin concentration and desorption solvent combination elucidated at microscale (2 mL) resulted in a two-fold improvement in taxadiene titer to 61 ± 8 mg/L, compared to the traditional liquid-liquid extraction approach (dodecane overlay). Taxadiene was found to be distributed evenly between resin beads and biomass. Performance of the optimal process was subsequently investigated through scale-up using controlled mini-bioreactors (250 mL). Here, a comparable taxadiene titer of 76 ± 19 mg/L was achieved despite a 125-fold scale-up in cultivation volume. This represented a 1.4-fold improvement in taxadiene recovery compared to previous mini-bioreactor scale cultivations using the dodecane overlay extraction approach

    Increased paclitaxel recovery from Taxus baccata vascular stem cells using novel in situ product recovery approaches

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    In this study, several approaches were tested to optimise the production and recovery of the widely used anticancer drug Taxol® (paclitaxel) from culturable vascular stem cells (VSCs) of Taxus baccata, which is currently used as a successful cell line for paclitaxel production. An in situ product recovery (ISPR) technique was employed, which involved combining three commercial macro-porous resin beads (HP-20, XAD7HP and HP-2MG) with batch and semi-continuous cultivations of the T. baccata VSCs after adding methyl jasmonate (Me-JA) as an elicitor. The optimal resin combination resulted in 234 ± 23 mg of paclitaxel per kg of fresh-weight cells, indicating a 13-fold improved yield compared to the control (with no resins) in batch cultivation. This resin treatment was further studied to evaluate the resins’ removal capacity of reactive oxygen species (ROS), which can cause poor cell growth or reduce product synthesis. It was observed that the ISPR cultivations had fourfold less intracellular ROS concentration than that of the control; thus, a reduced ROS concentration established by the resin contributed to increased paclitaxel yield, contrary to previous studies. These paclitaxel yields are the highest reported to date using VSCs, and this scalable production method could be applied for a diverse range of similar compounds utilising plant cell culture. Graphical Abstract: [Figure not available: see fulltext.]

    Copy number variation mapping and genomic variation of autochthonous and commercial turkey populations

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    This study aims at investigating genomic diversity of several turkey populations using Copy Number Variants (CNVs). A total of 115 individuals from six Italian breeds (Colle Euganei, Bronzato Comune Italiano, Parma e Piacenza, Brianzolo, Nero d\u2019Italia, and Ermellinato di Rovigo), seven Narragansett, 38 commercial hybrids, and 30 Mexican turkeys, were genotyped with the Affymetrix 600K single nucleotide polymorphism (SNP) turkey array. The CNV calling was performed with the Hidden Markov Model of PennCNV software and with the Copy Number Analysis Module of SVS 8.4 by Golden Helix\uae. CNV were summarized into CNV regions (CNVRs) at population level using BEDTools. Variability among populations has been addressed by hierarchical clustering (pvclust R package) and by principal component analysis (PCA). A total of 2,987 CNVs were identified covering 4.65% of the autosomes of the Turkey_5.0/melGal5 assembly. The CNVRs identified in at least two individuals were 362\u2014189 gains, 116 losses, and 57 complexes. Among these regions the 51% contain annotated genes. This study is the first CNV mapping of turkey population using 600K chip. CNVs clustered the individuals according to population and their geographical origin. CNVs are known to be indicators also of adaptation, as some researches in different species are suggesting

    Diagnostic procedures for non-small-cell lung cancer (NSCLC): recommendations of the European Expert Group

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    Background There is currently no Europe-wide consensus on the appropriate preanalytical measures and workflow to optimise procedures for tissue-based molecular testing of non-small-cell lung cancer (NSCLC). To address this, a group of lung cancer experts (see list of authors) convened to discuss and propose standard operating procedures (SOPs) for NSCLC. Methods Based on earlier meetings and scientific expertise on lung cancer, a multidisciplinary group meeting was aligned. The aim was to include all relevant aspects concerning NSCLC diagnosis. After careful consideration, the following topics were selected and each was reviewed by the experts: surgical resection and sampling; biopsy procedures for analysis; preanalytical and other variables affecting quality of tissue; tissue conservation; testing procedures for epidermal growth factor receptor, anaplastic lymphoma kinase and ROS proto-oncogene 1, receptor tyrosine kinase (ROS1) in lung tissue and cytological specimens; as well as standardised reporting and quality control (QC). Finally, an optimal workflow was described. Results Suggested optimal procedures and workflows are discussed in detail. The broad consensus was that the complex workflow presented can only be executed effectively by an interdisciplinary approach using a well-trained team. Conclusions To optimise diagnosis and treatment of patients with NSCLC, it is essential to establish SOPs that are adaptable to the local situation. In addition, a continuous QC system and a local multidisciplinary tumour-type-oriented board are essential

    Two Galaxy Clusters: A3565 and A3560

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    We report 102 new redshifts and magnitudes for a sample of galaxies to RF ~ 15.5 mag in a 2.17 deg x 2.17 deg region centered on the galaxy IC 4296, the most luminous member of the A3565 cluster. Up to the limiting magnitude we find 29 cluster members, and measure a velocity dispersion of 228 km/s. The estimated total mass for this system is ~ 3.0 x h**-1 10**13 Msun (where h = H0/100 km/s/Mpc), and its dynamical properties are quite typical of poor clusters presenting X-ray emission. We also find that galaxies with absorption lines are more concentrated towards the center of the cluster, while systems with emission lines are mainly located in the outer parts. The small velocity dispersion of the cluster, coupled to the known presence of an interacting pair of galaxies, and the large extent of the brightest cluster galaxy, could indicate that galaxy formation through mergers may still be underway in this system. The surveyed region also contains galaxies belonging to the Shapley Concentration cluster A3560. Within 30 arc min of the cluster center, we detect 32 galaxies, for which we measure a velocity dispersion of 588 km/s and a mass of ~2 x h**-1 10**14 Msun. However, because our sample is restricted to galaxies brighter than M*, these values should be considered only as rough estimates.Comment: 33 pages, including 6 tables and 9 postscript figures. Uses AAS Latex macros. Postscript file and ASCII versions of Tables 4 and 6 are available at http://www.dan.on.br/other_surveys/a3565.html. Scheduled for September 1999 issue of The Astronomical Journa
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