Simulation of a Multistage Compressor at Low Load Operation with Additional Bleed Air Extraction for Minimum Environmental Load Reduction

The need for more flexible operation of gas-fired power plants has led manufacturers to exploit possibilities to retrofit existing systems and increase turndown capabilities, lowering the minimum environmental load (MEL) - the lowest output at which the unit can operate and still meet environmental emissions limits. A possible measure for the compressor to enable a reduced MEL is to extract significant mass flow rates through the bleed ports in operation with a closed IGV to lower the mass flow entering the combustor, enabling a further load reduction while maintaining emissions. For this measure to be implemented, a stable operation of the compressor has to be ensured at the reduced MEL conditions. It is well known that bleed-air offtake at full speed shifts the loading towards the rear stages of the compressor. At MEL, the rear stages already operate at an increased loading compared to base-load. Therefore, to confirm the viability of bleed offtake as a turndown strategy, the effects on performance and stability have to be quantified for operation at MEL. Of particular interest is how an increase in air extraction through the bleed ports influences the stability of the compressor at MEL, especially when the offtake is from low pressure. Information on the degradation of the stability margin due to the additional bleed-air extraction at MEL is gained through numerical simulations. Full-compressor CFD simulations of an F-class gas turbine at reduced MEL conditions are performed. The influence of several geometrical and numerical modeling details is studied, and the model is validated against a comprehensive set of experimental data. The numerical results show good agreement with the experimental data, even with increased bleed air extraction. It can be concluded that bleed-air extraction is a capable method of reducing the compressor discharge mass flow rate significantly. The extraction rate is limited by the stability of the last compressor stages. To verify the integrity of the whole GT at operation under such conditions, e.g., the thermal state of hot parts, further analysis should be performed

High-level HIV-1 Nef transient expression in Nicotiana benthamiana using the P19 gene silencing suppressor protein of Artichoke Mottled Crinckle Virus

<p>Abstract</p> <p>Background</p> <p>In recent years, different HIV antigens have been successfully expressed in plants by either stable transformation or transient expression systems. Among HIV proteins, Nef is considered a promising target for the formulation of a multi-component vaccine due to its implication in the first steps of viral infection. Attempts to express Nef as a single protein product (not fused to a stabilizing protein) in transgenic plants resulted in disappointingly low yields (about 0.5% of total soluble protein). In this work we describe a transient expression system based on co-agroinfiltration of plant virus gene silencing suppressor proteins in <it>Nicotiana benthamiana</it>, followed by a two-step affinity purification protocol of plant-derived Nef.</p> <p>Results</p> <p>The effect of three gene silencing viral suppressor proteins (P25 of Potato Virus X, P19 of either Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus) on Nef transient expression yield was evaluated. The P19 protein of Artichoke Mottled Crinckle virus (AMCV-P19) gave the highest expression yield in vacuum co-agroinfiltration experiments reaching 1.3% of total soluble protein, a level almost three times higher than that previously reported in stable transgenic plants. The high yield observed in the co-agroinfiltrated plants was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs) indicating an effective modulation of RNA silencing mechanisms by AMCV-P19. Interestingly, we also showed that expression levels in top leaves of vacuum co-agroinfiltrated plants were noticeably reduced compared to bottom leaves. Moreover, purification of Nef from agroinfiltrated tissue was achieved by a two-step immobilized metal ion affinity chromatography protocol with yields of 250 ng/g of fresh tissue.</p> <p>Conclusion</p> <p>We demonstrated that expression level of HIV-1 Nef in plant can be improved using a transient expression system enhanced by the AMCV-P19 gene silencing suppressor protein. Moreover, plant-derived Nef was purified, with enhanced yield, exploiting a two-step purification protocol. These results represent a first step towards the development of a plant-derived HIV vaccine.</p

Plant antibodies for human antifungal therapy

There is an increasing interest in the development of therapeutic antibodies (Ab) to improve the control of fungal pathogens, but none of these reagents is available for clinical use. We previously described a murine monoclonal antibody (mAb 2G8) targeting β-glucan, a cell wall polysaccharide common to most pathogenic fungi, which conferred significant protection against Candida albicans, Aspergillus fumigatus and Cryptococcus neoformans in animal models. Transfer of this wide-spectrum, antifungal mAb into the clinical setting would allow the control of most frequent fungal infections in many different categories of patients. To this aim, two chimeric mouse-human Ab derivatives from mAb 2G8, in the format of complete IgG or scFv-Fc, were generated, transiently expressed in Nicotiana benthamiana plants and purified from leaves with high yields (approximately 50 mg Ab/kg of plant tissues). Both recombinant Abs fully retained the β-glucan-binding specificity and the antifungal activities of the cognate murine mAb against C. albicans. In fact, they recognized preferentially β1,3-linked glucan molecules present at the fungal cell surface and directly inhibited the growth of C. albicans and its adhesion to human epithelial cells in vitro. In addition, both the IgG and the scFv-Fc promoted C. albicans killing by isolated, human polymorphonuclear neutrophils in ex vivo assays and conferred significant antifungal protection in animal models of systemic or vulvovaginal C. albicans infection. These recombinant Abs represent valuable molecules for developing novel, plant-derived immunotherapeutics against candidiasis and, possibly, other fungal diseases

Predicting the Impact of Compressor Flexibility Improvements on Heavy-Duty Gas Turbines for Minimum and Base Load Conditions

The increasing importance of renewable energy capacity in the power generation scenario, together with the fluctuating consumer energy demand, forces conventional fossil fuel power generation systems to promptly respond to relevant and rapid load variations and to operate under off-design conditions during a major fraction of their lives. In order to improve existing power plants’ flexibility in facing energy surplus or deficit, retrofittable solutions for gas turbine compressors are proposed. In this paper, two different operation strategies, variable inlet guide vanes (IGVs) and blow-off extraction (BO), are considered for enabling partial load and minimum environmental load operation, and thus to identify implementation opportunities in existing thermal power plants. A typical 15-stage F-class gas turbine compressor is chosen as a test case and some energy demand scenarios are selected to validate the adopted solutions. The results of an extensive 3D, steady, CFD analysis are compared with the measurements coming from an experimental campaign carried out in the framework of the European Turbo-Reflex project. It will be shown how the combined strategies can reduce gas turbine mass flow rate and power plant output, without significantly penalizing efficiency, and how such off-design performance figures can be reliably predicted by employing state-of-the-art CFD models

Predicting the Impact of Compressor Flexibility Improvements on Heavy-Duty Gas Turbines for Minimum and Base Load Conditions

The increasing importance of renewable energy capacity in the power generation scenario, together with the fluctuating consumer energy demand, forces conventional fossil fuel power generation systems to promptly respond to relevant and rapid load variations and to operate under off-design conditions during a major fraction of their lives. In order to improve existing power plants&rsquo; flexibility in facing energy surplus or deficit, retrofittable solutions for gas turbine compressors are proposed. In this paper, two different operation strategies, variable inlet guide vanes (IGVs) and blow-off extraction (BO), are considered for enabling partial load and minimum environmental load operation, and thus to identify implementation opportunities in existing thermal power plants. A typical 15-stage F-class gas turbine compressor is chosen as a test case and some energy demand scenarios are selected to validate the adopted solutions. The results of an extensive 3D, steady, CFD analysis are compared with the measurements coming from an experimental campaign carried out in the framework of the European Turbo-Reflex project. It will be shown how the combined strategies can reduce gas turbine mass flow rate and power plant output, without significantly penalizing efficiency, and how such off-design performance figures can be reliably predicted by employing state-of-the-art CFD models

Retrofittable Solutions Capability for Gas Turbine Compressors

The increasing introduction of renewable energy capacity has changed the perspective on the operation of conventional power plants, introducing the necessity of reaching extreme off-design conditions. There is a strong interest in the development and optimization of technologies that can be retrofitted to an existing power plant to enhance flexibility as well as increase performance and lower emissions. Under the framework of the European project TURBO-REFLEX, a typical F-class gas turbine compressor designed and manufactured by Ansaldo Energia has been studied. Numerical analyses were performed using the TRAF code, which is a state-of-the-art 3D CFD RANS/URANS flow solver. In order to assess the feasibility of lower minimum environmental load operation, by utilizing a reduction in the compressor outlet mass-flow rate, with a safe stability margin, two different solutions have been analyzed: blow-off extractions and extra-closure of Variable Inlet Guide Vanes. The numerical steady-state results are compared and discussed in relation to an experimental campaign, which was performed by Ansaldo Energia. The purpose is to identify the feasibility of the technologies and implementation opportunity in the existing thermal power plant fleet

Production of an Engineered Killer Peptide in Nicotiana benthamiana by Using a Potato virus X Expression System

The decapeptide killer peptide (KP) derived from the sequence of a single-chain, anti-idiotypic antibody acting as a functional internal image of a microbicidal, broad-spectrum yeast killer toxin (KT) was shown to exert a strong microbicidal activity against human pathogens. With the aim to exploit this peptide to confer resistance to plant pathogens, we assayed its antimicrobial activity against a broad spectrum of phytopathogenic bacteria and fungi. Synthetic KP exhibited antimicrobial activity in vitro towards Pseudomonas syringae, Erwinia carotovora, Botrytis cinerea, and Fusarium oxysporum. KP was also expressed in plants by using a Potato virus X (PVX)-derived vector as a fusion to the viral coat protein, yielding chimeric virus particles (CVPs) displaying the heterologous peptide. Purified CVPs showed enhanced antimicrobial activity against the above-mentioned plant pathogens and human pathogens such as Staphylococcus aureus and Candida albicans. Moreover, in vivo assays designed to challenge KP-expressing plants (as CVPs) with Pseudomonas syringae pv. tabaci showed enhanced resistance to bacterial attack. The results indicate that the PVX-based display system is a high-yield, rapid, and efficient method to produce and evaluate antimicrobial peptides in plants, representing a milestone for the large-scale production of high-added-value peptides through molecular farming. Moreover, KP is a promising molecule to be stably engineered in plants to confer broad-spectrum resistance to phytopathogens

Humanization of a highly stable single-chain antibody by structure-based antigen-binding site grafting

The murine single-chain variable fragment F8 (scFv(F8)) is endowed with high intrinsic thermodynamic stability and can be functionally expressed in the reducing environment of both prokaryotic and eukaryotic cytoplasm. The stability and intracellular functionality of this molecule can be ascribed mostly to its framework regions and are essentially independent of the specific sequence and structure of the supported antigen-binding site. Therefore, the scFv(F8) represents a suitable scaffold to construct stable scFv chimeric molecules against different antigens by in vitro evolution or antigen-binding site grafting. Thanks to the favourable pharmacokinetic properties associated to a high thermodynamic stability of antibody fragments, such scFv(F8) variants may be exploited for a wide range of biomedical applications, from in vivo diagnosis to therapy, as well as to interfere with the function of intracellular proteins and pathogens, and for functional genomics studies. However, the potential immunogenicity of the murine framework regions represents a limitation for their exploitation in therapeutic applications. To overcome this limitation, we humanized a derivative of the scFv(FS), the anti-lysozyme scFv(11E), which is endowed with even higher thermodynamic stability than the parent antibody. The humanization was carried out by substituting the framework residues differing from closely related V-H and V-L domains of human origin with their human counterparts. Site-directed mutagenesis generated the fully humanized product and four intermediate scFvs, which were analyzed for protein expression and antigen binding. We found that the substitution Tyr 90 -> Phe in the V-H domain dramatically reduced the bacterial expression of all mutants. The back-mutation of Phe H90 to Tyr led to the final humanized variant named scFv(H5)H90Tyr. This molecule comprises humanized V-H and V-L framework regions and is endowed with HEL-binding affinity, stability in human serum and functionality under reducing conditions comparable to the murine cognate antibody. Consequently, the humanized scFv(H5)H90Tyr represents a suitable scaffold onto which new specificities towards antigens of therapeutic interest can be engineered for biomedical applications. (C) 2008 Elsevier Ltd. All rights reserved