The harnessing of peptide–monolith constructs for single step plasmid DNA purification

The availability of synthetic peptides has paved the way for their use in tailor-made interactions with biomolecules. In this study, a 16mer LacI-based peptide was used as an affinity ligand to examine the scale up feasibility for plasmid DNA purification. First, the peptide was designed and characterized for the affinity purification of lacO containing plasmid DNA, to be employed as a high affinity ligand for the potential capturing of plasmid DNA in a single unit operation. It was found there were no discernible interactions with a control plasmid that did not encode the lacO nucleotide sequence. The dissociation equilibrium constant of the binding between the 16mer peptide and target pUC19 was 5.0 ± 0.5 × 10⁻⁸ M as assessed by surface plasmon resonance. This selectivity and moderated affinity indicate that the 16mer is suitable for the adsorption and chromatographic purification of plasmid DNA. The suitability of this peptide was then evaluated using a chromatography system with the 16mer peptide immobilized to a customized monolith to purify plasmid DNA, obtaining preferential purification of supercoiled pUC19. The results demonstrate the applicability of peptide–monolith supports to scale up the purification process for plasmid DNA using designed ligands via a biomimetic approach.     

Development of a high resolution voxelised head phantom for medical physics applications

Computational anthropomorphic phantoms have become an important investigation tool for medical imaging and dosimetry for radiotherapy and radiation protection. The development of computational phantoms with realistic anatomical features contribute significantly to the development of novel methods in medical physics. For many applications, it is desirable that such computational phantoms have a real-world physical counterpart in order to verify the obtained results.In this work, we report the development of a voxelised phantom, the HIGH_RES_HEAD, modelling a paediatric head based on the commercial phantom 715-HN (CIRS). HIGH_RES_HEAD is unique for its anatomical details and high spatial resolution (0.18 × 0.18 mm² pixel size). The development of such a phantom was required to investigate the performance of a new proton computed tomography (pCT) system, in terms of detector technology and image reconstruction algorithms.The HIGH_RES_HEAD was used in an ad-hoc Geant4 simulation modelling the pCT system. The simulation application was previously validated with respect to experimental results. When compared to a standard spatial resolution voxelised phantom of the same paediatric head, it was shown that in pCT reconstruction studies, the use of the HIGH_RES_HEAD translates into a reduction from 2% to 0.7% of the average relative stopping power difference between experimental and simulated results thus improving the overall quality of the head phantom simulation.The HIGH_RES_HEAD can also be used for other medical physics applications such as treatment planning studies.A second version of the voxelised phantom was created that contains a prototypic base of skull tumour and surrounding organs at risk.     

Construction of small plasmid vectors for use in genetic improvement of the extremely acidophilic Acidithiobacillus caldus

The genetic improvement of biomining bacteria including Acidithiobacillus caldus could facilitate the bioleaching process of sulfur-containing minerals. However, the available vectors for use in A. caldus are very scanty and limited to relatively large broad-host-range IncQ plasmids. In this study, a set of small, mobilizable plasmid vectors (pBBR1MCS-6, pMSD1 and pMSD2) were constructed based on plasmid pBBR1MCS-2, which does not belong to the IncQ, IncW, or IncP groups. The function of the tac promoter on 5.8-kb pMSD2 was determined by inserting a kanamycin-resistant reporter gene. The resulting recombinant pMSD2-Km was successfully transferred by conjugation into A. caldus MTH-04 with transfer frequency of 1.38 ± 0.64 × 10^?5. The stability and plasmid copy number of pMSD2-Km in A. caldus MTH-04 were 75 ± 2.7% and 5–6 copies per cell, respectively. By inserting an arsABC operon into pMSD2, an arsenic-resistant recombinant pMSD2-As was constructed and transferred into A. caldus MTH-04 by conjugation. The arsenic tolerance of A. caldus MTH-04 containing pMSD2-As was obviously increased up to 45 mM of NaAsO₂. These vectors could be applied in genetic improvement of A. caldus as well as other bioleaching bacteria.     

Apport de la TEP/TDM au 18FDG dans la stadification initiale et l’évaluation précoce de la réponse thérapeutique des rhabdomyosarcomes pédiatriques

PurposeThe objective of this study was to retrospectively evaluate the impact of positron emission tomography/computed tomography (PET/CT) using fluorine-18-fluorodeoxyglucose (FDG), in comparison with conventional imaging modalities (CIM), for initial staging and early therapy assessment in paediatric rhabdomyosarcoma.Patients and methodsPrior to treatment, 18 patients (age range, 9 months to 18 years) with histologically proven rhabdomyosarcoma underwent FDG PET/CT in addition to CIM (magnetic resonance imaging of primary site, whole body CT and bone scintigraphy). After three courses of chemotherapy, 12 patients underwent FDG PET/CT in addition to CIM. RECIST criteria and visual analysis of FDG uptake were used for assessment of response. The standard of reference was determined by an interdisciplinary tumor board based on imaging material, histopathology and follow-up data (median = 5 years).ResultsPET/CT sensitivity was superior to CIM's concerning lymph node involvement (100% versus 83%, respectively) and metastases detection (100% versus 50%, respectively). PET/CT results changed therapeutic management in 11% of cases. After three courses of chemotherapy, the rate of complete response was 66% with PET/CT versus 8% with CIM. Five percent of patients relapsed during follow-up (median = 5 years).ConclusionThis study confirms that PET/CT depicts important additional information in initial staging of paediatric rhabdomyosarcomas and suggests a superior prognostic value of PET/CT in early response to chemotherapy assessment.     

Modelling and simulation of continuous L (+) lactic acid production from sugarcane juice in membrane integrated hybrid-reactor system

Modelling and simulation was done for a two-stage membrane-integrated hybrid reactor system for continuous production of L (+) lactic acid under non-neutralizing conditions. The model captures microbial conversion of sugar cane juice to lactic acid under substrate–product inhibitions with downstream purification by nanofiltration. All the major phenomena and the governing parameters like fluid flow, feed dilution, substrate–product inhibitions, Donnan and steric effects during micro and nanofiltration for cell recycle, product separation and purification have been reflected in the modelling. The model describes a green, integrated continuous process of direct lactic acid production starting with a cheap, renewable carbon source. The highest lactic acid concentration achieved after the final stage of nanofiltration was 66.97 g/L at 13 kg/cm² operating pressure when the overall productivity reached 12.40 g/(L h). The developed model could successfully predict production, purification and transport of lactic acid through two stage membrane modules. Performance of the model was very good as indicated in the high overall correlation coefficient (R² > 0.980) and the low relative error (RE < 0.1).     

High titer mevalonate fermentation and its feeding as a building block for isoprenoids (isoprene and sabinene) production in engineered Escherichia coli

Isoprenoids are important fine chemicals as material monomers, advanced fuels and pharmaceuticals. A variety of natural isoprenoids can be synthesized by engineered microbial strains. This work established a process by dividing the current isoprenoid pathway into the upstream fermentation process, from sugar to mevalonate (MVA), and the downstream process, from MVA to the target isoprenoids. The results showed that significant differences existed in the process conditions between the upstream and downstream fermentations. After individually optimizing the process conditions, the upstream MVA production (84.0 g/L, 34.0% and 1.8 g/ L/h) and downstream isoprene production (11.0 g/L and 0.23 g/L/h) were greatly improved in this two-step process. Flask fermentation experiments also confirmed that two-step route can significantly improve the sabinene titer to 150 mg/L (6.5-fold of the sabinene titer in an earlier flask study of our lab). Therefore, the two-step route proposed in this study may have potential benefits towards the current isoprenoids production directly from glucose. The high titer and yield of MVA indicate that MVA has great potential to be more broadly utilized as starting precursor in synthetic biology.     

Genetic stability of an Escherichia coli strain engineered to produce a novel therapeutic DNA vaccine encoding chicken type II collagen for rheumatoid arthritis

The development of therapeutic DNA vaccines capable of recovering immunological tolerance through the induction of both CD4 + CD25 + FoxP3 + regulatory and CD3 + CD8 + C28-suppressor T cells, and/or inhibition of both autoreactive CD4 + CD28+ type 1 T helper and autoantibody-producing B cells offers a promising new strategy for the treatment of rheumatoid arthritis. Previously, we developed pcDNA-CCOL2A1, a novel therapeutic DNA vaccine, which encodes the full-length chicken type II collagen sequence, and demonstrated that the efficacy of this vaccine for treating rheumatoid arthritis was comparable to that of the current “gold standard” treatment, methotrexate. In this study, we investigated the genetic stability of a strain engineered to produce the vaccine during continuous passage and long-term storage at different temperatures. By screening a panel of 12 strains, we identified a DH5α strain that exhibited high levels (12.30 ± 0.05 mg L⁻¹) of pcDNA-CCOL2A1 production after 15 h cultivation, and subsequently utilized this strain to establish a three-tier cells bank for future studies. Continuous passage of this strain for 100 inoculation times demonstrated that a higher percentage (>95%) of cells maintained the plasmid when cultivated under selective pressure (ampicillin) than under nonselective conditions, suggesting that the presence of antibiotics in the medium prevents the loss of the pcDNA-CCOL2A1 plasmid. Meanwhile, restriction digestion and gene sequencing analyses demonstrated that the pcDNA-CCOL2A1 vector remained stable, and that the plasmid sequence was conserved during this period. Lastly, the DH5α pcDNA-CCOL2A1 strain exhibited a high plasmid preservation (>90%) and high levels of plasmid production (9.05mg L⁻¹) after storage for 60 months at −80 °C. Furthermore the plasmid extracted from the DH5α pcDNA-CCOL2A1 strain after storage for 60 months at −80 °C was transfected to COS-7 cells, it can stably express the target protein chicken type II collagen. Conversely, this strain exhibited a complete loss of capability after 24 and 18 months storage at −20 °C and 4 °C, respectively. These findings will facilitate further pilot-scale testing, and even industrial-scale production, of the novel therapeutic vaccine pcDNA-CCOL2A1.     

The use of spatial ecological modelling as a tool for improving the assessment of geographic range size of threatened species

We analysed endemic threatened tree and reptile species of Socotra Island (Yemen), characterised by different ecological requirements and spatial distribution, in order to evaluate the usefulness of spatial ecological modelling in the estimation of species extent of occurrence (EOO) and area of occupancy (AOO). Point occurrences for the entire species range were used to model their spatial distribution by Random Forest (RF) and Generalised Linear Model (GLM). For each species the suitability area (SA) was obtained by applying the 0% omission error criterion on the probability map, and compared or integrated with EOO and AOO area obtained by topological methods such as the minimum convex polygon (MCP), α-hull and 2 km × 2 km grid.RF showed a lower prediction error than GLM. Higher accuracy was achieved for species with higher number of occurrences and narrower ecological niche. SA was always greater than AOO measured with the 2 km × 2 km grid method. SA was greater than EOO, measured by both MCP and α-hull methods, for species with localised distribution, while it was smaller for widely distributed species. EOO-α-hull area was equal or smaller than that calculated by MCP depending on the spatial distribution of species. AOO measured considering the SA within the EOO-MCP was greater than that measured using the standard 2 km × 2 km grid. Conversely, AOO calculated considering the suitable area within the EOO-α-hull showed variable results, being smaller or greater than the 2 km × 2 km grid AOO depending on the ecological niche and spatial distribution of species. According to our results, SEM does not provide an effective alternative to topological methods for the estimate of EOO and AOO. However, it may be considered a useful tool to estimate AOO within the boundaries of EOO measured by the α-hull method, because it reduces some potential sources of inconsistency and bias.     

Effect Factors for marine eutrophication in LCIA based on species sensitivity to hypoxia

Hypoxia is an important environmental stressor to marine species, especially in benthic coastal waters. Increasing anthropogenic emissions of nutrients and organic matter contribute to the depletion of dissolved oxygen (DO). Biotic sensitivity to low levels of DO is determined by the organisms’ ability to use DO as a respiratory gas, a process depending on oxygen partial pressure. A method is proposed to estimate an indicator of the intensity of the effects caused by hypoxia on exposed marine species. Sensitivity thresholds to hypoxia of an exposed ecological community, modelled as lowest-observed-effect-concentrations (LOEC), were compiled from literature for 91 benthic, demersal and benthopelagic species of fish, crustaceans, molluscs, echinoderms, annelids, and cnidarians, and converted to temperature-specific benthic (100 m depth) LOEC values. Species distribution and LOEC values were combined using a species sensitivity distribution (SSD) methodology to estimate the DO concentration at which the potentially affected fraction (PAF) of the community's species having their LOEC exceeded is 50% (HC50_LOEC). For the purpose of effect modelling in Life Cycle Impact Assessment (LCIA), Effect Factors (EF [(PAF) m³ kgO₂⁻¹]) were derived for five climate zones (CZ) to represent the change in effect due to a variation of the stressor intensity, or EF = ΔPAF/ΔDO = 0.5/HC50_LOEC. Results range from 218 (PAF) m³ kgO₂⁻¹ (polar CZ) to 306 (PAF) m³ kgO₂⁻¹ (tropical CZ). Variation between CZs was modest so a site-generic global EF of 264 (PAF) m³ kgO₂⁻¹ was also estimated and may be used to represent the average impact on a global ecological community of marine species exposed to hypoxia. The EF indicator is not significantly affected by the major sources of uncertainty in the underlying data suggesting valid applicability in characterisation modelling of marine eutrophication in LCIA.     

Strategies for high titre plasmid DNA production in Escherichia coli DH5α

The production of plasmid pEGFP-N1 in Escherichia coli DH5α was optimised. A strategy evaluating different media components separately was not successful (OD < 2.5, low plasmid titres), a statistical approach via a Plackett Burman design (11 parameters) allowed some improvement (7 mg/L plasmid, OD₆₀₀ 8.5). Generally, high biomass did not correlate with high plasmid titres. When conditions were transferred to the bioreactor (batch operation) little improvement in plasmid titres (10 mg/L plasmid, OD₆₀₀ 20) was observed. By switching to a fed-batch procedure with linear feeding these values increased to 20 mg/L plasmid (OD₆₀₀ 50). By using an adaptive feeding strategy, plasmid titres could be increased to 50 mg/L. Finally, by combining a growth controlled (reduced temperature (35 °C), low dO₂) initial batch phase with an adaptive feeding strategy in the fed-batch phase (37 °C, glucose-/dO₂-limitation) we were reproducibly able to produce up to 250 mg/L of plasmid DNA in cultures that reached a final OD₆₀₀ of 80.     

Kinetics of organic carbon removal by a mixed culture in a membrane bioreactor

The aim of this work was to model the biological activity and anticipate the kinetic behaviour of microorganisms and the overall performance of the process according to a specific model and running parameters. The bacterial inoculum used in these experiments was a mixture of cultures taken from the wastewater treatment plant in Montpellier. The fermentor, used in association with an ultrafiltration separation stage (with a filtration area of 0.2 m²) had a working volume of 15.8 l. For various working conditions (different solid retention times, different hydraulic retention times and substrate concentrations), the biomass concentration and the residual substrate concentration, expressed in terms of dry weight and chemical oxygen demand, respectively, were measured. The basic idea of modelling was related to the concept of maintenance. The coefficient of maintenance, E, and the theoretical conversion yield, y, were therefore calculated. The values of E and y, measured for total cell recycling experiments and for experiments with various solid retention times, remained similar and were found to equal 0.040 mgCOD mgVSS h⁻¹ and 0.36 mgVSS mgCOD⁻¹, respectively. Determining these two constants and modelling the treatment process made it possible to anticipate the optimal biomass concentration for a defined removal efficiency under different steady-state operating conditions.     

Enhanced production of valienamine by Stenotrophomonas maltrophilia with fed-batch culture in a stirred tank bioreactor

Valienamine is an important medicinal intermediate with broad use in the synthesis of some stronger α-glucosidase inhibitors. In order to improve valienamine concentration in the fermentation broth and make the downstream treatment easy, a fed-batch process for the enhanced production of valienamine by Stenotrophomonas maltrophilia in a stirred tank bioreactor was developed. Results showed that supplementation of validamycin A in the process of cultivation could increase the valienamine concentration. One-pulse feeding was observed to be the best strategy. The maximum valienamine concentration of 2.35 g L⁻¹ was obtained at 156 h when 86.4 g of validamycin A was added to a 15-L bioreactor containing 8 L fermentation medium with one-pulse feeding. The maximum valienamine concentration had a great improvement and was increased above 100% compared to batch fermentation in the stirred tank bioreactor. The pH-controlled experiments showed that controlling the pH in the process of one-pulse feeding fermentation had not obvious effect on the production of valienamine.     

Design,synthesis and molecular modeling of biquinoline–pyridine hybrids as a new class of potential EGFR and HER-2 kinase inhibitors

A new series of biquinoline–pyridine hybrids were designed and synthesized by a base-catalyzed cyclocondensation through one-pot multicomponent reaction. All compounds were tested for in vitro anticancer activities against two cancer cell lines A549 (adenocarcinomic human alveolar basal epithelial) and Hep G2 (liver cancer). Enzyme inhibitory activities of all compounds were carried out against EGFR and HER-2 kinase. Of the compounds studied, majority of the compounds showed effective anticancer activity against used cancer cell lines. Compound 9i (IC₅₀ = 0.09 μM) against EGFR and (IC₅₀ = 0.2 μM) against HER-2 kinase displayed the most potent inhibitory activity as compared to other member of the series. In the molecular modelling study, compound 9i was bound in to the active pocket of EGFR with four hydrogen bonds and two π–cation interactions having minimum binding energy ΔG_b = −54.4 kcal/mol.     

Transfection in perfused microfluidic cell culture devices: A case study

Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy.     

A targeted high-efficiency angiogenesis strategy as therapy for myocardial infarction

AimsThe aim of this study was to prove that an intramyocardial injection of a mixture of low-dose human growth factor (HGF) plasmid and microbubbles (MB) in combination with insonation was an effective therapy for myocardial infarction.Main methodsTwenty dogs with myocardial infarction were divided into 4 groups: (1) HGF, MB and ultrasound (HGF-US/MB), (2) HGF and US (HGF-US), (3) HGF alone and (4) surgery alone (control). In the HGF-US/MB group, HGF plasmid DNA (500 μg) mixed with 0.5 ml of MB solution was injected 5 min after coronary occlusion followed by insonation. With the exception of the control group, the other dogs were divided into two groups, one treated with the HGF gene and insonation and the other with the HGF gene only.Key findingsCompared to the HGF group, infarct size decreased from 32% ± 7% (control) to 23% ± 5% in the HGF-US/MB group 28 d later (P < 0.05). Capillary density increased from 21.7 ± 4.2/mm² (control) to 114.3 ± 28.9/mm² in the HGF-US/MB group (P < 0.01). Compared to the HGF group, there was a 14% decrease in the ratio of left ventricle weight/body weight and a 25% decrease in hydroxyproline content. We also observed a 29% and 20% decrease in collagen volume fraction of type I and type III collagen, respectively in the HGF-US/MB group.SignificanceIntramyocardial injection of HGF and MB in combination with insonation enhances neovascularization and reduces ventricular remodeling and infarct size.     

Design of new potent and selective secretory phospholipase A2 inhibitors. 6-Synthesis,structure–activity relationships and molecular modelling of 1-substituted-4-[4,5-dihydro-1,2,4-(4H)-oxadiazol-5-one-3-yl(methyl)]-functionalized aryl piperazin/one/dione derivatives

The group IIA human non-pancreatic secretory phospholipase A₂ (hnp-sPLA₂) is one of the enzymes implied in the inflammatory process. In the course of our work on inhibitors of this enzyme we investigated the influence of rigidity of the piperazine region on the biological activity. Several modifications were explored. Various linkers, such as amide, urea, carbamate, or alkoxyphenyl were inserted between the piperazine and the lipophilic chain. Also, modification of the piperazine core to incorporate carbonyl groups was studied. In an in vitro fluorimetric assay using the human GIIA (HPLA₂) and porcine pancreatic GIB enzymes, compound 60a (Y = phenoxy, R = C₁₈H₃₇, Z = CH₂) had the optimal activity with an IC₅₀ = 30 nM on HPLA₂. By means of molecular modelling we attempted to get informations towards comprehension of differences in activity.     

Optimization of integrated alkaline–extrusion pretreatment of barley straw for sugar production by enzymatic hydrolysis

In this work, an integrated one-step alkaline–extrusion process was tested as pretreatment for sugar production from barley straw (BS) biomass. The influence of extrusion temperature (T) and the ratio NaOH/BS dry matter (w/w) (R) into the extruder on pretreatment effectiveness was investigated in a twin-screw extruder at bench scale. A 2³ factorial design of experiments was used to analyze the effect of process conditions [T: 50–100 °C; R: 2.5–7.5% (w/w)] on composition and enzymatic digestibility of pretreated substrate (extrudate). The optimum conditions for a maximum glucan to glucose conversion were determined to be R = 6% and T = 68 °C. At these conditions, glucan yield reached close to 90% of theoretical, while xylan conversion was 71% of theoretical. These values are 5 and 9 times higher than that of the untreated material, which supports the great potential of this one-step combined pre-treatment technology for sugar production from lignocellulosic substrates. The absence of sugar degradation products is a relevant advantage over other traditional methods for a biomass to ethanol production process since inhibitory effect of such product on sugar fermentation would be prevented.     

Bioaugmentation of microbial communities in laboratory and pilot scale sequencing batch biofilm reactors using the TOL plasmid

The aim of this study was to investigate the effectiveness of bioaugmentation and transfer of plasmid pWWO (TOL plasmid) to mixed microbial populations in pilot and laboratory scale sequencing batch biofilm reactors (SBBRs) treating synthetic wastewater containing benzyl alcohol (BA) as a model xenobiotic. The plasmid donor was a Pseudomonas putida strain chromosomally tagged with the gene for the red fluorescent protein carrying a green fluorescent protein labeled TOL plasmid, which confers degradation capacity for several compounds including toluene and BA. In the pilot scale SBBR donor cells were disappeared 84 h after inoculation while transconjugants were not detected at all. In contrast, both donor and transconjugant cells were detected in the laboratory scale reactor where the ratio of transconjugants to donors fluctuated between 1.9 × 10^?1 and 8.9 × 10^?1 during an experimental period of 32 days. BA degradation rate was enhanced after donor inoculation from 0.98 mg BA/min prior to inoculation to 1.9 mg BA/min on the seventeenth day of operation. Survival of a bioaugmented strain, conjugative plasmid transfer and enhanced BA degradation was demonstrated in the laboratory scale SBBR but not in the pilot scale SBBR.     

Kinetic approach of aflatoxin B1-acetylcholinesterase interaction: a tool for developing surface plasmon resonance biosensors

This work presents a kinetic approach of the interaction between acetylcholinesterase (AChE) from electric eel and aflatoxin B1 (AFB1) or its protein conjugate (e.g., AFB1–HRP [horseradish peroxidase]) in order to develop a simple and sensitive detection method of these compounds. The dissociation constant K_d of the AChE/AFB1–HRP interaction (0.4 μM) obtained with the surface plasmon resonance (SPR) technique is very close to the inhibition constant reported in amperometric assay (K_i = 0.35 μM), proving that the conjugation of AFB1 to a carrier protein does not significantly influence the affinity of AFB1 for AChE. Thus, the AChE/AFB1–HRP couple can be used as mimic system for the binding of AChE to other AFB1–protein adducts and further used for developing biosensors for AFB1 bound to plasma proteins. The immobilization protocol was designed to minimize the nonspecific adsorption on the self-assembled monolayer (SAM) functionalized surface of the SPR chip without an additional hydrophilic linker, whereas the interaction protocol was designed to mark out the possible occurrence of mass transport limitation (MTL) effects. The detection limits (LODs) were 0.008 μM for AFB1–HRP (2.5 ng ml^?1 AFB1) and 0.94 ng ml^?1 for AFB1 itself, which is lower than recently reported values in spectrophotometric and amperometric assays.     

Performance of the Phytoplankton Index for Lakes (IPLAC): A multimetric phytoplankton index to assess the ecological status of water bodies in France

A new phytoplankton-based index was designed to respond to the Water Framework Directive (WFD) requirements concerning the assessment of lake ecological status. The “Indice Phytoplancton Lacustre” (IPLAC) is a multimetric index, taking into account biomass, abundance and species composition of communities. The first metric is based on the total phytoplankton biomass (MBA), the second on the abundance and taxonomic composition (MCS) of 165 indicator taxa. The IPLAC was developed on 2 independent databases, one for the calibration and the second for the validation of the metrics. The calibration dataset was composed of 255 “lake-years” from 214 distinct lakes sampled between 2005 and 2012. The validation dataset included 173 lake-years in order to confirm the response of the index to the trophic gradient and anthropogenic pressure.The results show that the IPLAC correctly highlights chemical pressure (eutrophication). Especially high Pearson correlations are shown with total phosphorus (r = −0.71, p-value <0.001), chlorophyll-a (r = −0.83, p-value <0.001) and water transparency (r = 0.73, p-value <0.001) which are the main proxies for the trophic level. Corine land cover was used as an indication of the anthropogenic pressure and good correlations are also found with the watershed land use, negatively correlated with agricultural area (r = −0.60, p-value <0.001), population density (r = −0.36, p-value <0.001) and positively with forest area (r = 0.57, p-value <0.001).The index is WFD-compliant and is dedicated to natural lakes and artificial water bodies in metropolitan France, and will be routinely used by the French Ministry of the Environment to assess lake ecological status through the phytoplankton community. However, the results must be carefully interpreted in two cases: reservoirs with large water level fluctuations, and samples that include less than 5 indicator species.