Chiral Analysis of Pesticides and Drugs of Environmental Concern: Biodegradation and Enantiomeric Fraction

The importance of stereochemistry for medicinal chemistry and pharmacology is well recognized and the dissimilar behavior of enantiomers is fully documented. Regarding the environment, the significance is equivalent since enantiomers of chiral organic pollutants can also differ in biodegradation processes and fate, as well as in ecotoxicity. This review comprises designed biodegradation studies of several chiral drugs and pesticides followed by enantioselective analytical methodologies to accurately measure the enantiomeric fraction (EF). The enantioselective monitoring of microcosms and laboratory-scale experiments with different environmental matrices is herein reported. Thus, this review focuses on the importance of evaluating the EF variation during biodegradation studies of chiral pharmaceuticals, drugs of abuse, and agrochemicals and has implications for the understanding of the environmental fate of chiral pollutants.The authors wish to thank Fundação para a Ciência e Tecnologia—FCT for financial support under the project Fluoropharma PTDC/EBB-EBI/111699/2009, the PhD grant attributed to Alexandra S. Maia SFRH/BD/86939/2012, QREN-POPH, European Social Fund, MCTES, PEst FCOMP-01-0124-FEDER-022718; PEst-OE/EQB/LA0016/2011, PEst-OE/SAU/UI4040/2014. This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT—Fundação para a Ciência e Tecnologia and European Regional Development Fund (ERDF), in the framework of the program PT2020 and by CESPU (ChiralDrugs_CESPU_2017

Albumin and mammalian cell culture: implications for biotechnology applications

Albumin has a long historical involvement in design of media for the successful culture of mammalian cells, in both the research and commercial fields. The potential application of albumins, bovine or human serum albumin, for cell culture is a by-product of the physico-chemical, biochemical and cell-specific properties of the molecule. In this review an analysis of these features of albumin leads to a consideration of the extracellular and intracellular actions of the molecule, and importantly the role of its interactions with numerous ligands or bioactive factors that influence the growth of cells in culture: these include hormones, growth factors, lipids, amino acids, metal ions, reactive oxygen and nitrogen species to name a few. The interaction of albumin with the cell in relation to these co-factors has a potential impact on metabolic and biosynthetic activity, cell proliferation and survival. Application of this knowledge to improve the performance in manufacturing biotechnology and in the emerging uses of cell culture for tissue engineering and stem cell derived therapies is an important prospect

Assessment of culturable bacterial endophytic communities colonizing Canna flaccida inhabiting a wastewater treatment constructed wetland

Microorganisms, including the bacterial populations living inside plant tissues (endophytes), and vegetation are components of constructed wetland (CW) systems playing pivotal roles in the water treatment process and in the ecosystem establishment. The present study focuses on the assessment of the culturable endophytic bacterial communities of Canna flaccida plants, the dominant plant species in a polycultured CW treating wastewater from a tourism facility. Bacterial endophytes were grouped by random amplified polymorphic DNA and identified by 16S ribosomal RNA gene sequencing. From the bacterial isolates, 103 were considered for phylogenetic analysis, falling in 25 genera within the gamma-, beta- alpha-Proteobacteria, Flavobacteria, Sphingobacteria, Actinobacteria and Bacilli classes. Forty-nine percent of the isolates belonged to the Enterobacteriaceae family, suggesting that the plants in CW systems may act as a sink of potential human pathogenic microorganisms; nevertheless their abundance on the plant tissues was reduced from the inlet (62%) to the outlet zone (38%). High diversity in terms of genera was found in C. flaccida tissues, although the number of genera common to both sampling zones was low, which suggests that the processes occurring within the CW, including the water characteristics gradient from the inlet to the outlet (e.g. total suspended solids, organics and fecal loads), had the ability to shape the diversity of the endophytic communities. (C) 2016 Elsevier All rights reserved.C.S.C. Calheiros and S.I.A. Pereira thank the grants SFRH/BPD/63204/2009 and SFRH/BPD/65134/2009, respectively, from Fundação para a Ciência e Tecnologia (FCT, Portugal), Fundo Social Europeu and Fundos Nacionais do MEC through the program QREN−POPH−Tipologia 4.1-Formação Avançada. This work was supported by National Funds from FCT−Fundação para a Ciência e a Tecnologia through project UID/Multi/50016/2013. The authors are thankful for the collaboration of Paço de Calheiros and Saint-Gobain Weber Portugal, S.A. The authors wish to thank Dr. Pedro Carvalho for reviewing the manuscript

Assessment of mycorrhizal colonisation and soil nutrients in unmanaged fire-impacted soils from two target restoration sites

International audienceThe mycorrhizal colonisation of plants grown in unmanaged soils from two restoration sites with a fire history in Northern Portugal was evaluated from the perspective of supporting restoration programmes. To promote restoration of original tree stands, Quercus ilex L. and Pinus pinaster Ait. were used as target species on two sites, denoted Site 1 and 2 respectively. The aim of the study was to assess whether mycorrhizal propagules that survived fire episodes could serve as in situ inoculum sources, and to analyse the spatial distribution of soil nutrients and mycorrhizal parameters. In a laboratory bioassay, P. pinaster and Q. ilex seedlings were grown on soils from the target sites and root colonisation by ectomycorrhizal (ECM) and arbuscular mycorrhizal (AM) fungi was determined. The ECM root colonisation levels found indicated that soil from Site 2 contained sufficient ECM propagules to serve as a primary source of inoculum for P pinaster. The low levels of ECM and AM colonisation obtained on the roots of plants grown in soil from Site 1 indicated that the existing mycorrhizal propagules might be insufficient for effective root colonisation of Q. ilex. Different ECM morphotypes were found in plants grown in soil from the two sites. At Site 2 mycorrhizal parameters were found to be spatially structured, with significant differences in ECM colonisation and soil P concentrations between regions of either side of an existing watercourse. The spatial distribution of mycorrhizal propagules was related to edaphic parameters (total C and extractable P), and correlations between soil nutrients and mycorrhizal parameters were found

Assessment of mycorrhizal colonisation and soil nutrients in unmanaged fire-impacted soils from two target restoration sites

International audienceThe mycorrhizal colonisation of plants grown in unmanaged soils from two restoration sites with a fire history in Northern Portugal was evaluated from the perspective of supporting restoration programmes. To promote restoration of original tree stands, Quercus ilex L. and Pinus pinaster Ait. were used as target species on two sites, denoted Site 1 and 2 respectively. The aim of the study was to assess whether mycorrhizal propagules that survived fire episodes could serve as in situ inoculum sources, and to analyse the spatial distribution of soil nutrients and mycorrhizal parameters. In a laboratory bioassay, P. pinaster and Q. ilex seedlings were grown on soils from the target sites and root colonisation by ectomycorrhizal (ECM) and arbuscular mycorrhizal (AM) fungi was determined. The ECM root colonisation levels found indicated that soil from Site 2 contained sufficient ECM propagules to serve as a primary source of inoculum for P pinaster. The low levels of ECM and AM colonisation obtained on the roots of plants grown in soil from Site 1 indicated that the existing mycorrhizal propagules might be insufficient for effective root colonisation of Q. ilex. Different ECM morphotypes were found in plants grown in soil from the two sites. At Site 2 mycorrhizal parameters were found to be spatially structured, with significant differences in ECM colonisation and soil P concentrations between regions of either side of an existing watercourse. The spatial distribution of mycorrhizal propagules was related to edaphic parameters (total C and extractable P), and correlations between soil nutrients and mycorrhizal parameters were found

Assessment of mycorrhizal colonisation and soil nutrients in unmanaged fire-impacted soils from two target restoration sites

The mycorrhizal colonisation of plants grown in unmanaged soils from two restoration sites with a fire history in Northern Portugal was evaluated from the perspective of supporting restoration programmes. To promote restoration of original tree stands, Quercus ilex L. and Pinus pinaster Ait. were used as target species on two sites, denoted Site 1 and 2 respectively. The aim of the study was to assess whether mycorrhizal propagules that survived fire episodes could serve as in situ inoculum sources, and to analyse the spatial distribution of soil nutrients and mycorrhizal parameters. In a laboratory bioassay, P. pinaster and Q. ilex seedlings were grown on soils from the target sites and root colonisation by ectomycorrhizal (ECM) and arbuscular mycorrhizal (AM) fungi was determined. The ECM root colonisation levels found indicated that soil from Site 2 contained sufficient ECM propagules to serve as a primary source of inoculum for P pinaster. The low levels of ECM and AM colonisation obtained on the roots of plants grown in soil from Site 1 indicated that the existing mycorrhizal propagules might be insufficient for effective root colonisation of Q. ilex. Different ECM morphotypes were found in plants grown in soil from the two sites. At Site 2 mycorrhizal parameters were found to be spatially structured, with significant differences in ECM colonisation and soil P concentrations between regions of either side of an existing watercourse. The spatial distribution of mycorrhizal propagules was related to edaphic parameters (total C and extractable P), and correlations between soil nutrients and mycorrhizal parameters were found

Fertilizer potential of slurry from intensive dairy cattle farms in Intensive production forestry systems

Intensive dairy cattle breeding has a relevant social and economic impact in Portugal, particularly in the northern region. This activity generates a high flow of livestock effluents (slurry), rich in important nutrients for plant growth, which can be introduced into intensive production forestry systems. These effluents can provide a good alternative to mineral fertilizers, not only from an economic perspective but, particularly, from the point of view of environmental protection. In the present study, the effect of increasing doses of slurry on tree growth, either with or without mycorrhizal arbuscular fungi (AMF) and plant growth-promoting bacteria (PGPB) inoculation, was evaluated in clones of Paulownia CoT2 and Populus i214, as they are genotypes that have a high efficiency in the mobilization of soil nutrients (namely N) and in the capture of CO2 from the atmosphere, as well as high biomass calorific value. For this purpose, a demonstration field trial was installed, occupying an area of 14,607 m2, where the trees were planted with the compasses of: 2.5 x 1.5 m and 2.5 x 0.75 m, respectively for Paulownia and for Poplar. Prior to transplantation to the field, some plants were inoculated with AMF and PGPB. In the field, the following treatments were performed: T0 - no fertilization, either mineral or organic; T1 - amount of slurry equivalent to 85 kg of N ha-1; T2 - amount of slurry equivalent to 170 kg of N ha-1; T3 - amount of slurry equivalent to 340 kg of N ha-1, both with and without inoculation. Results revealed a significant and positive effect of the slurry application, both in the diameter at breast height (DBH) and total stand height (TH), showing its high fertilizing potential, and, on the other hand, there was no increased contamination by nitrates and by pathogenic microorganism in the leachates for the experimental doses of slurry. Therefore, we can conclude that, under the experimental conditions, the slurry resulting from the intensive exploitation of cattle constitutes an alternative to exclusively mineral fertilization in intensive production forestry systems, either by increasing the production of biomass or by the absence of contamination of aquifers by nitrates and pathogens.</jats:p

Cell culture processes for monoclonal antibody production

Animal cell culture technology has advanced significantly over the last few decades and is now generally considered a reliable, robust and relatively mature technology. A range of biotherapeutics are currently synthesized using cell culture methods in large scale manufacturing facilities that produce products for both commercial use and clinical studies. The robust implementation of this technology requires optimization of a number of variables, including (1) cell lines capable of synthesizing the required molecules at high productivities that ensure low operating cost; (2) culture media and bioreactor culture conditions that achieve both the requisite productivity and meet product quality specifications; (3) appropriate on-line and off-line sensors capable of providing information that enhances process control; and (4) good understanding of culture performance at different scales to ensure smooth scale-up. Successful implementation also requires appropriate strategies for process development, scale-up and process characterization and validation that enable robust operation and ensure compliance with current regulations. This review provides an overview of the state-of-the art technology in key aspects of cell culture, e.g., generation of highly productive cell lines and optimization of cell culture process conditions. We also summarize the current thinking on appropriate process development strategies and process advances that might affect process development

Efficient optimization of process strategies with model-assisted design of experiments

Conventional design of experiments (DoE) methods require expert knowledge about the investigated factors and their boundary values and mostly lead to multiple rounds of time-consuming and costly experiments. The combination of DoE with mathematical process modeling in model-assisted DoE (mDoE) can be used to increase the mechanistic understanding of the process. Furthermore, it is aimed to optimize the processes with respect to a target (e.g., amount of cells, product titer), which also provides new insights into the process. In this chapter, the workflow of mDoE is explained stepwise including corresponding protocols. Firstly, a mathematical process model is adapted to cultivation data of first experimental data or existing knowledge. Secondly, model-assisted simulations are treated in the same way as experimentally derived data and included as responses in statistical DoEs. The DoEs are then evaluated based on the simulated data, and a constrained-based optimization of the experimental space can be conducted. This loop can be repeated several times and significantly reduces the number of experiments in process development