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Grant
Agency: Cordis | Branch: FP7 | Program: CP-TP | Phase: KBBE.2012.3.5-03 | Award Amount: 7.70M | Year: 2012

Water4Crops provides a combination of technical improvements in the field of bio-treatment and agricultural water use within a transdisciplinary identification of novel agri-business opportunities. Water4Crops aims at: a) developing innovative biotechnological wastewater treatments for improved water recycling, b) initiating the co-creation of alternative combinations of bio-treatment, recycling of high value elements, and combinations for bioproducts leading to a better commercialization of biotechnology and agricultural products in Europe and India, c)improving water use efficiency at field level through agronomics, plant breeding and locally adapted new irrigation technologies and accurate crop water requirement measurements techniques. Water4Crops will boost bio-based economy by applying a double track approach. First a comprehensive set of key Green-Economy technologies for: 1) valorization of volatile fatty acids; 2) obtaining: natural antioxidants (polyphenols), biopolymers (PHAs), energy (biomethane); 3) new substances for selective recovery of valuable products from wastewater; 4) tailoring effluent properties from decentralized innovative bioreactors; 5) low bio-sludge production by SBBG Reactors; 6) removal of organopollutants by nanobiocatalysts; 7) reduced clogging of wetlands; 8) virus monitoring detection assays; 9) suitable precision irrigation systems for reclaimed water; 10) new monitoring for increase crop water productivity; 11) understanding the genetic mechanisms regulating drought-adaptive traits across maize, sorghum, millet and tomato; 12) optimized waste water related combinations of species/genotypes x environment x management. Second, new product market combinations will be identified. The co-creation process will be organized by two Mirror cases (Emilia Romagna area in Italy and Hyderabad region in India) within a specific Science-Practice Interface (INNOVA platforms). Developing the new applications and business opportunities with regional enterprises and stakeholder will move India and Europe towards a Green Economy.


Georgiev V.,Florida A&M University | Schumann A.,Vita 34 AG | Pavlov A.,University of Food Technologies | Pavlov A.,Bulgarian Academy of Science | Bley T.,TU Dresden
Engineering in Life Sciences | Year: 2014

Plant tissue and organ cultures in vitro usually face technological challenges. When submerged cultivation of plant cells in a controlled environment is desired, the characteristic growth morphology and physiology of differentiated organ cultures present a problem in process scale-up. Temporary immersion systems (TIS) were developed several decades ago. These systems are providing the most natural environment for in vitro culture of plant shoots and seedlings. Over the past few years, TIS have been recognized as a perspective technology for plant micropropagation, production of plant-derived secondary metabolites, expression of foreign proteins, and potential solutions in phytoremediation. Nowadays, several TIS, operating on similar or divergent technological principles, have been developed and successfully applied in the cultivation of various plant in vitro systems, including somatic embryos and transformed root cultures. In this article, the operational principle and technological design of the most popular TIS are reviewed. In addition, recent examples of the application of temporary immersion technology for in vitro cultivation of plant tissue and organ cultures at laboratory and pilot scales are discussed. Finally, future prospects and challenges to the industrial realization of that fast-developing technique are outlined. © 2014 WILEY-VCH Verlag GmbH & Co.


Grant
Agency: Cordis | Branch: FP7 | Program: MC-IAPP | Phase: FP7-PEOPLE-2013-IAPP | Award Amount: 921.62K | Year: 2014

The scientific and technological (S&T) goal of the project is to develop and validate an innovative approach combining phytoremediation and production of biomass on heavy metal contaminated (HMC) areas which could be used as local energy carrier. The S&T objectives of the project are as follows: Objective1) selection of optimal plant species suitable for phytoremediation driven energy crops production. Experimental plots will be established in Poland (arable land) and Germany (extreme HMC site) to test the pre-selected species Miscanthus gigantheus, Sida hermaphrodita, Spartina pectinata, Panicum virgatum. Guidelines on selection of the most appropriate species will be elaborated using brownfields management tools. The technical feasibility, economic viability and environmental benefits of the approach will be also analysed. Objective 2) is to develop a microbiological method stimulating the biomass yield and phytoremediation effect at HMC sites and. Interactions between plant and microbes will be investigated and tested as they play a crucial role at sites where plant growth is affected by HM contamination. Plant growth promoting rhizobacteria, bacterial endophytes and mycorhizal fungi enhancing growth of the selected species will be identified. They will help developing innovative inocula to be applied respectively to the brownfield management target (i.e. cleanup or use for biomass production). Objective 3) is to demonstrate an environmentally safe way of converting the HMC biomass into energy in a small scale local installation with special focus on gasification as a promising technology which may become a competitive niche alternative for handing HMC biomass. Cost effectiveness and environmental benefits of this method will be evaluated together with technology optimization guidelines for this type of fuel. The S&T objectives will be realized by transfer of knowledge: secondments and external recruitment between 6 consortium partners from industry and academia.


Giannopoulou E.Z.,Helmholtz Center Munich | Giannopoulou E.Z.,Forschergruppe Diabetes e.V. | Puff R.,Helmholtz Center Munich | Puff R.,Forschergruppe Diabetes e.V. | And 11 more authors.
Pediatric Diabetes | Year: 2014

Background: The application of autologous cord blood in children with type 1 diabetes has been found to be safe, but not to preserve beta-cell function in a previous study, which, however, had not included a control group. Objective: To compare the changes of metabolic and immune function over time between cord blood infused children and natural controls. Subjects and methods: Seven children with newly diagnosed type 1 diabetes underwent a single autologous cord blood infusion and 10 children were enrolled as natural controls in a non-randomized, controlled, open label intervention trial. Primary analyses were performed 1 year following cord blood infusion. Cases and controls were compared regarding metabolic [area under the curve (AUC) and peak C-peptide, insulin use, and HbA1c] and immune outcome (islet autoantibody titer and T-cell response), adjusted for age, gender, diabetes duration, and baseline levels. Results: There were no significant adverse events related to the infusion. Metabolic and immune outcomes were not significantly different at 12months follow-up between infused children and controls (e.g., adjusted p=0.244 for AUC C-peptide, adjusted p=0.820 for insulin use, adjusted p=0.772 for peripheral regulatory T cells). Six-month change of AUC C-peptide correlated significantly with the number of infused CD34+ cells (r=0.931, p=0.002). Conclusions: An autologous cord blood infusion does not change the natural course of metabolic and immune parameters after disease onset. However, the content of CD34+ cells in the stored blood sample might offer potential for improvement of future cell therapies. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.


Scholbach J.,University of Leipzig | Schulz A.,Fraunhofer Institute for Cell Therapy and Immunology | Westphal F.,University of Leipzig | Egger D.,Vita34 AG | And 5 more authors.
PLoS ONE | Year: 2012

To study the function and maturation of the human hematopoietic and immune system without endangering individuals, translational human-like animal models are needed. We compare the efficiency of CD34+ stem cells isolated from cryopreserved cord blood from a blood bank (CCB) and fresh cord blood (FCB) in generating highly engrafted humanized mice in NOD-SCID IL2Rγnull (NSG) rodents. Interestingly, the isolation of CD34+ cells from CCB results in a lower yield and purity compared to FCB. The purity of CD34+ isolation from CCB decreases with an increasing number of mononuclear cells that is not evident in FCB. Despite the lower yield and purity of CD34+ stem cell isolation from CCB compared to FCB, the overall reconstitution with human immune cells (CD45) and the differentiation of its subpopulations e.g., B cells, T cells or monocytes is comparable between both sources. In addition, independent of the cord blood origin, human B cells are able to produce high amounts of human IgM antibodies and human T cells are able to proliferate after stimulation with anti-CD3 antibodies. Nevertheless, T cells generated from FCB showed increased response to restimulation with anti-CD3. Our study reveals that the application of CCB samples for the engraftment of humanized mice does not result in less engraftment or a loss of differentiation and function of its subpopulations. Therefore, CCB is a reasonable alternative to FCB and allows the selection of specific genotypes (or any other criteria), which allows scientists to be independent from the daily changing birth rate. © 2012 Scholbach et al.


Boltze J.,Fraunhofer Institute for Cell Therapy and Immunology | Boltze J.,University of Leipzig | Reich D.M.,Fraunhofer Institute for Cell Therapy and Immunology | Hau S.,Fraunhofer Institute for Cell Therapy and Immunology | And 7 more authors.
Cell Transplantation | Year: 2012

Experimental transplantation of human umbilical cord blood (hUCB) mononuclear cells (MNCs) in rodent stroke models revealed the therapeutic potential of these cells. However, effective cells within the heterogeneous MNC population and their modes of action are still under discussion. MNCs and MNC fractions enriched (CD34+) or depleted (CD34-) for CD34-expressing stem/progenitor cells were isolated from hUCB. Cells were transplanted intravenously following middle cerebral artery occlusion in spontaneously hypertensive rats and directly or indirectly cocultivated with hippocampal slices previously subjected to oxygen and glucose deprivation. Application of saline solution or a human T-cell line served as controls. In vivo, MNCs, CD34+ and CD34- cells reduced neurofunctional deficits and diminished lesion volume as determined by magnetic resonance imaging. MNCs were superior to other fractions. However, human cells could not be identified in brain tissue 29 days after stroke induction. Following direct application on postischemic hippocampal slices, MNCs reduced neural damage throughout a 3-day observation period. CD34+ cells provided transient protection for 2 days. The CD34- fraction, in contrast to in vivo results, failed to reduce neural damage. Direct cocultivation of MNCs was superior to indirect cocultivation of equal cell numbers. Indirect application of up to 10-fold MNC concentrations enhanced neuroprotection to a level comparable to direct cocultivation. After direct application, MNCs migrated into the slices. Flow cytometric analysis of migrated cells revealed that the CD34+ cells within MNCs were preferably attracted by damaged hippocampal tissue. Our study suggests that MNCs provide the most prominent neuroprotective effect, with CD34+ cells seeming to be particularly involved in the protective action of MNCs. CD34+ cells preferentially home to neural tissue in vitro, but are not superior concerning the overall effect, implying that there is another, still undiscovered, protective cell population. Furthermore, MNCs did not survive in the ischemic brain for longer periods without immunosuppression. © 2012 Cognizant Comm. Corp.


Schumann A.,Vita 34 AG | Torras-Claveria L.,University of Barcelona | Berkov S.,University of Barcelona | Berkov S.,Agrobioinstitute Dragan Tzankov Blvd. | And 4 more authors.
Biotechnology Progress | Year: 2013

The influence of different elicitors (copper sulfate, silver nitrate, salicylic acid and methyl jasmonate), on both the growth and alkaloid production of Leucojum aestivum shoots grown in a temporary immersion system was studied. Seven Amaryllidaceae alkaloids and three protoalkaloids were quantitatively determined by GC-MS analysis in leaves and bulblets, separately. Methyl jasmonate was found to significantly improve the production of galanthamine (GAL) in both leaves and bulblets. The content of GAL released to the liquid nutrient medium was also measured. The release of GAL into the liquid medium took place mainly in the first 2 weeks determined by harvesting the liquid nutrient medium after 2 weeks and measuring the GAL content (1st subculturing step). © 2013 American Institute of Chemical Engineers.


Sygnecka K.,University of Leipzig | Heider A.,University of Leipzig | Scherf N.,Carl Gustav Carus Institute | Alt R.,University of Leipzig | And 3 more authors.
Stem Cells and Development | Year: 2015

Mesenchymal stem cells (MSCs) have been identified as promising candidates for neuroregenerative cell therapies. However, the impact of different isolation procedures on the functional and regenerative characteristics of MSC populations has not been studied thoroughly. To quantify these differences, we directly compared classically isolated bulk bone marrow-derived MSCs (bulk BM-MSCs) to the subpopulation Sca-1+Lin-CD45--derived MSCs- (SL45-MSCs), isolated by fluorescence-activated cell sorting from bulk BM-cell suspensions. Both populations were analyzed with respect to functional readouts, that are, frequency of fibroblast colony forming units (CFU-f), general morphology, and expression of stem cell markers. The SL45-MSC population is characterized by greater morphological homogeneity, higher CFU-f frequency, and significantly increased nestin expression compared with bulk BM-MSCs. We further quantified the potential of both cell populations to enhance neuronal fiber growth, using an ex vivo model of organotypic brain slice co-cultures of the mesocortical dopaminergic projection system. The MSC populations were cultivated underneath the slice co-cultures without direct contact using a transwell system. After cultivation, the fiber density in the border region between the two brain slices was quantified. While both populations significantly enhanced fiber outgrowth as compared with controls, purified SL45-MSCs stimulated fiber growth to a larger degree. Subsequently, we analyzed the expression of different growth factors in both cell populations. The results show a significantly higher expression of brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor in the SL45-MSCs population. Altogether, we conclude that MSC preparations enriched for primary MSCs promote neuronal regeneration and axonal regrowth, more effectively than bulk BM-MSCs, an effect that may be mediated by a higher BDNF secretion. © Mary Ann Liebert, Inc.


Danova-Alt R.,Vita 34 AG | Danova-Alt R.,University of Leipzig | Heider A.,University of Leipzig | Egger D.,Vita 34 AG | And 2 more authors.
PLoS ONE | Year: 2012

Very small embryonic-like (VSEL) cells have been described as putatively pluripotent stem cells present in murine bone marrow and human umbilical cord blood (hUCB) and as such are of high potential interest for regenerative medicine. However, there remain some questions concerning the precise identity and properties of VSEL cells, particularly those derived from hUCB. For this reason, we have carried out an extensive characterisation of purified populations of VSEL cells from a large number of UCB samples. Consistent with a previous report, we find that VSEL cells are CXCR4 +, have a high density, are indeed significantly smaller than HSC and have an extremely high nuclear/cytoplasmic ratio. Their nucleoplasm is unstructured and stains strongly with Hoechst 33342. A comprehensive FACS screen for surface markers characteristic of embryonic, mesenchymal, neuronal or hematopoietic stem cells revealed negligible expression on VSEL cells. These cells failed to expand in vitro under a wide range of culture conditions known to support embryonic or adult stem cell types and a microarray analysis revealed the transcriptional profile of VSEL cells to be clearly distinct both from well-defined populations of pluripotent and adult stem cells and from the mature hematopoietic lineages. Finally, we detected an aneuploid karyotype in the majority of purified VSEL cells by fluorescence in situ hybridisation. These data support neither an embryonic nor an adult stem cell like phenotype, suggesting rather that hUCB VSEL cells are an aberrant and inactive population that is not comparable to murine VSEL cells. © 2012 Danova-Alt et al.


PubMed | VITA 34 AG, University "Marta Abreu" of Las Villas and Vrije Universiteit Brussel
Type: | Journal: Methods in molecular biology (Clifton, N.J.) | Year: 2016

Digitalis purpurea L. is one of the main economically viable sources of cardenolides (cardiac glycosides) for the pharmaceutical industry. Nevertheless, production of cardenolides in plants grown by traditional agriculture is not always an efficient process and can be affected by biotic and abiotic factors. This chapter provides two biotechnology strategies for biomass and cardenolide production in D. purpurea. Firstly, we report biomass production using a temporary immersion system (TIS), combined with cardenolide extraction and quantification. Secondly, an efficient protocol for genetic transformation via Agrobacterium tumefaciens is provided. These strategies can be used independently or combined in order to increase the content of cardiac glycosides in D. purpurea and to unravel biosynthetic pathways associated to cardiac glycoside production.

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