Residence time distribution measurements in an external-loop airlift reactor: Study of the hydrodynamics of the liquid circulation induced by the hydrogen bubbles

A detailed study of the residence time distribution (RTD) analysis of liquid phase has been performed in an external-loop airlift reactor of 20 L nominal volume, regarded as a global unit and discriminating its different sections (riser, gas–liquid separator and downcomer) using the tracer response technique. The reactor was used as an electrochemical reactor in order to carry out the electrocoagulation/electroflotation (EC/EF). The gas phase created in the riser is the hydrogen produced by water electrolysis.In order to use this reactor for a continuous EC/EF, hydrodynamic studies were carried out to control the operating conditions and to help modelling the electrocoagulation. Current density, position of the electrodes in the riser and the volumetric liquid flow (inlet flow) are the key parameters for the hydrodynamics. The experimental results revealed that both in the downcomer and the riser–separator zones, the flow model is axial dispersion. Interesting results were obtained: –The superficial liquid velocity (ULd) at the downcomer, decreased when the volume inlet flow increased (0<QL<2 L/min). –The Peclet number obtained in the downcomer was correlated to the current density and the electrodes position. –In the riser–separator zone the Peclet number decreased with the superficial liquid velocity in the riser indicating that the dispersion increased with an increase of turbulence created in the separator by an increase of liquid velocity. –The percentage of flow that quits the reactor without reacting increased when the main flow increased and the current intensity decreased. The global RTD can be reconstituted by the signal resulting from the junction and that from riser–separator and downcomer zone by using the convolution technique. The experimental results confirm this reconstitution. The experiments confirm also that the liquid crosses the reactor without achieving loops in the case of the continuous flow

Defluoridation of drinking water by electrocoagulation/electroflotation in a stirred tank reactor with a comparative performance to an external-loop airlift reactor

Defluoridation using batch electrocoagulation/electroflotation (EC/EF) was carried out in two reactors for comparison purpose: a stirred tank reactor (STR) close to a conventional EC cell and an external-loop airlift reactor (ELAR) that was recently described as an innovative reactor for EC. The respective influences of current density, initial concentration and initial pH on the efficiency of defluoridation were investigated. The same trends were observed in both reactors, but the efficiency was higher in the STR at the beginning of the electrolysis, whereas similar values were usually achieved after 15 min operation. The influence of the initial pH was explained using the analyses of sludge composition and residual soluble aluminum species in the effluents, and it was related to the prevailing mechanisms of defluoridation. Fluoride removal and sludge reduction were both favored by an initial pH around 4, but this value required an additional pre-treatment for pH adjustment. Finally, electric energy consumption was similar in both reactors when current density was lower than 12 mA/cm2, but mixing and complete flotation of the pollutants were achieved without additional mechanical power in the ELAR, using only the overall liquid recirculation induced by H2 microbubbles generated by water electrolysis, which makes subsequent treatments easier to carry out

Kinetic study of defluoridation of drinking water by electrocoagulation/electroflotation in a stirred tank reactor and in an external-loop airlift reactor

A kinetic study of defluoridation of drinking water was carried out using the electrocoagulation/electroflotation process in two batch reactors of identical volume (20 L): a stirred tank reactor (STR) and an external-loop airlift reactor (ELALR). When the evolution of fluoride content was independent of stirring speed, experimental results showed that the kinetics of fluoride removal could be modelled using a variable-order-kinetic (VOK) approach coupled with a Langmuir–Freundlich adsorption model in the STR. Conversely, when mixing was less efficient, which is the case in the ELALR, experimental data could be fitted adequately only using a pseudo-first-order model. This constitutes however only an empirical approach based on a lumped parameter that accounts simultaneously for mass transfer, adsorption and electrochemical steps. In this case, only regression analysis could be used to establish a quantitative relationship between the kinetic constant and the operating conditions, such as current density and initial fluoride concentration

Defluoridation of Morocco drinking water by electrocoagulation/electroflottation in an electrochemical external-loop airlift reactor

An innovative application of external-loop airlift reactors as electrocoagulation/electroflotation cells with Al electrodes for defluoridation of drinking water was developed. Liquid overall recirculation and mixing were induced only by hydrogen microbubbles electrochemically generated from the cathode. This application was carried out in a 20 L external-loop air liftreactor both under semi-batch and continuous flow conditions. Results showed that liquid recirculation could be correlated to current density and gas–liquid dispersion height in the separator. Experimental data obtained at optimum conditions that favored simultaneously mixing and flotation confirmed that concentrations lower than 1.5 mg/L could be achieved when initial concentrations were between 10 and 20 mg/L. The effects of conductivity and pH agreed with the literature. Conversely, the low electrode surface vs. reactor volume ratio merged the formation of fluoroaluminum microflocs near the electrodes to fluoride adsorption on these particles in the riser and the separator sections, which differed from conventional EC cells. Consequently, defluoridation could be achieved at lower energy and electrode consumptions than in the literature. An optimum current density was defined at j = 6 mA/cm2 for pH 5, accounting simultaneously for mixing, reaction time, yield and operating costs. A promising attempt of transposition from batch to continuous process was also reported in this work, as flotation avoids the need for a downstream settling unit

Multiple roles for Plasmodium berghei phosphoinositide-specific phospholipase C in regulating gametocyte activation and differentiation

Critical events in the life cycle of malaria parasites are controlled by calcium-dependent signalling cascades, yet the molecular mechanisms of calcium release remain poorly understood. The synchronized development of Plasmodium berghei gametocytes relies on rapid calcium release from internal stores within 10 s of gametocytes being exposed to mosquito-derived xanthurenic acid (XA). Here we addressed the function of phosphoinositide-specific phospholipase C (PI-PLC) for regulating gametocyte activation. XA triggered the hydrolysis of PIP2 and the production of the secondary messenger IP3 in gametocytes. Both processes were selectively blocked by a PI-PLC inhibitor, which also reduced the early Ca2+ signal. However, microgametocyte differentiation into microgametes was blocked even when the inhibitor was added up to 5 min after activation, suggesting a requirement for PI-PLC beyond the early mobilization of calcium. In contrast, inhibitors of calcium release through ryanodine receptor channels were active only during the first minute of gametocyte activation. Biochemical determination of PI-PLC activity was confirmed using transgenic parasites expressing a fluorescent PIP2/IP3 probe that translocates from the parasite plasmalemma to the cytosol upon cell activation. Our study revealed a complex interdependency of Ca2+ and PI-PLC activity, with PI-PLC being essential throughout gamete formation, possibly explaining the irreversibility of this process

CRIMALDDI: platform technologies and novel anti-malarial drug targets

The Coordination, Rationalization, and Integration of antiMALarial drug Discovery & Development Initiatives (CRIMALDDI) Consortium, funded by the EU Framework Seven Programme, has attempted, through a series of interactive and facilitated workshops, to develop priorities for research to expedite the discovery of new anti-malarials. This paper outlines the recommendations for the development of enabling technologies and the identification of novel targets.Screening systems must be robust, validated, reproducible, and represent human malaria. They also need to be cost-effective. While such systems exist to screen for activity against blood stage Plasmodium falciparum, they are lacking for other Plasmodium spp. and other stages of the parasite's life cycle. Priority needs to be given to developing high-throughput screens that can identify activity against the liver and sexual stages. This in turn requires other enabling technologies to be developed to allow the study of these stages and to allow for the culture of liver cells and the parasite at all stages of its life cycle.As these enabling technologies become available, they will allow novel drug targets to be studied. Currently anti-malarials are mostly targeting the asexual blood stage of the parasite's life cycle. There are many other attractive targets that need to be investigated. The liver stages and the sexual stages will become more important as malaria control moves towards malaria elimination. Sexual development is a process offering multiple targets, even though the mechanisms of differentiation are still not fully understood. However, designing a drug whose effect is not curative but would be used in asymptomatic patients is difficult given current safety thresholds. Compounds active against the liver schizont would have a prophylactic effect and Plasmodium vivax elimination requires effectors against the dormant liver hypnozoites. It may be that drugs to be used in elimination campaigns will also need to have utility in the control phase. Compounds with activity against blood stages need to be screened for activity against other stages.Natural products should also be a valuable source of new compounds. They often occupy non-Lipinski chemical space and so may reveal valuable new chemotypes

Comparison of the cellular and biochemical properties of Plasmodium falciparum choline and ethanolamine kinases.

International audienceThe proliferation of the malaria-causing parasite Plasmodium falciparum within the erythrocyte is concomitant with massive phosphatidylcholine and phosphatidylethanolamine biosynthesis. Based on pharmacological and genetic data, de novo biosynthesis pathways of both phospholipids appear to be essential for parasite survival. The present study characterizes PfCK (P. falciparum choline kinase) and PfEK (P. falciparum ethanolamine kinase), which catalyse the first enzymatic steps of these essential metabolic pathways. Recombinant PfCK and PfEK were expressed as His6-tagged fusion proteins from overexpressing Escherichia coli strains, then purified to homogeneity and characterized. Using murine polyclonal antibodies against recombinant kinases, PfCK and PfEK were shown to be localized within the parasite cytoplasm. Protein expression levels increased during erythrocytic development. PfCK and PfEK appeared to be specific to their respective substrates and followed Michaelis-Menten kinetics. The Km value of PfCK for choline was 135.3+/-15.5 microM. PfCK was also able to phosphorylate ethanolamine with a very low affinity. PfEK was found to be an ethanolamine-specific kinase (Km=475.7+/-80.2 microM for ethanolamine). The quaternary ammonium compound hemicholinium-3 and an ethanolamine analogue, 2-amino-1-butanol, selectively inhibited PfCK or PfEK. In contrast, the bis-thiazolium compound T3, which was designed as a choline analogue and is currently in clinical trials for antimalarial treatment, affected PfCK and PfEK activities similarly. Inhibition exerted by T3 was competitive for both PfCK and PfEK and correlated with the impairment of cellular phosphatidylcholine biosynthesis. Comparative analyses of sequences and structures for both kinase types gave insights into their specific inhibition profiles and into the dual capacity of T3 to inhibit both PfCK and PfEK

Potent in vivo anti-malarial activity and representative snapshot pharmacokinetic evaluation of artemisinin-quinoline hybrids

BACKGROUND:Because Plasmodium falciparum displays increase tolerance against the recommended artemisinin combination therapies (ACT), new classes of anti-malarial drugs are urgently required. Previously synthesized artemisinin-aminoquinoline hybrids were evaluated to ascertain whether the potent low nanomolar in vitro anti-plasmodial activity would carry over in vivo against Plasmodium vinckei. A snapshot pharmacokinetic analysis was carried out on one of the hybrids to obtain an indication of the pharmacokinetic properties of this class of anti-malarial drugs. METHODS: In vitro activity of hybrids 2 and 3 were determined against the 3D7 strain of P. falciparum. Plasmodium vinckei-infected mice were treated with hybrids 1 - 3 for four days at a dosage of 0.8mg/kg, 2.5mg/kg, 7.5mg/kg or 15mg/kg intraperitoneally (ip), or orally (per os) with 2.7mg/kg, 8.3mg/kg, 25mg/kg or 50mg/kg. Artesunate was used as reference drug. A snapshot oral and IV pharmacokinetic study was performed on hybrid 2. RESULTS: Hybrids 1 - 3 displayed potent in vivo anti-malarial activity with ED50 of 1.1, 1.4 and <0.8mg/kg by the ip route and 12, 16 and 13mg/kg per os, respectively. Long-term monitoring of parasitaemia showed a complete cure of mice (without recrudescence) at 15mg/kg via ip route and at 50mg/kg by oral route for hybrid 1 and 2, whereas artesunate was only able to provide a complete cure at 30mg/kg ip and 80mg/kg per os. CONCLUSIONS: These compounds provide a new class of desperately needed anti-malarial drug. Despite a short half-life and moderate oral bioavailability, this class of compounds was able to cure malaria in mice at very low dosages. The optimum linker length for anti-malarial activity was found to be a diaminoalkyl chain consisting of two carbon atoms either methylated or unmethylated

Genetic and transcriptional analysis of phosphoinositide-specific phospholipase C in Plasmodium

Phosphoinositide-specific phospholipase C (PI-PLC) is a major regulator of calcium-dependent signal transduction, which has been shown to be important in various processes of the malaria parasite Plasmodium. PI-PLC is generally implicated in calcium liberation from intracellular stores through the action of its product, inositol-(1,4,5)-trisphosphate, and is itself dependent on calcium for its activation. Here we describe the plc genes from Plasmodium species. The encoded proteins contain all domains typically found in PI-PLCs of the δ class but are almost twice as long as their orthologues in mammals. Transcriptional analysis by qRT-PCR of plc during the erythrocytic cycle of P. falciparum revealed steady expression levels that increased at the late schizont stages. Genetic analysis in the P. berghei model revealed that the plc locus was targetable but that plc gene knock-outs could not be obtained, thereby strongly indicating that the gene is essential during blood stage development. Alternatively, we attempted to modify plc expression through a promoter exchange approach but found the gene to be refractory to over-expression indicating that plc expression levels might additionally be tightly controlled