Elissa Ephrem

Amal Najjar

Catherine Charcosset

Hélène Greige-Gerges
email: greigegeorges@yahoo.com

Selection of nerolidol among a series of terpenic and phenolic compounds for its potent activity against Lactobacillus fermentum ATCC 9338

Keywords: Cyclodextrins, Lactobacillus fermentum, nerolidol, phenolic compounds, terpenes CFU: colony forming unit, DMSO: dimethylsulfoxide, HP-β-CD: hydroxypropyl-β-cyclodextrin, HPLC: high performance liquid chromatography, LAB: lactic acid bacteria, MBC: minimal bactericidal concentration, MIC: minimal inhibitory concentration, Ner: nerolidol

Essential oil components are widely used for their antibacterial activity against spoilage microorganisms in food. Lactobacillus fermentum is a Gram-positive lactic acid bacteria responsible for the deterioration of various food products, including beverages and dairy products. In this study, 17 terpenic and 11 phenolic compounds were screened against the food spoilage microorganism Lactobacillus fermentum ATCC 9338. The antibacterial activity of the tested compounds was dependent on hydrophobicity and particular chemical features. Nerolidol solubilized in dimethylsulfoxide exhibited the highest antibacterial activity and showed low minimal inhibitory (MIC: 25 μM) and minimal bactericidal (MBC: 50 μM) concentrations.

Moreover, no viable cells were detected within 16 h of incubation at 50 μM. The important antibacterial activity of nerolidol against L. fermentum is probably related to the high hydrophobicity, the aliphatic chain length, and the presence of the hydroxyl group.

Hydroxypropyl-β-cyclodextrin/nerolidol inclusion complex showed MIC and MBC values of 100 and 200 μM, respectively. The total bacterial kill was observed after 12 h of incubation. The results obtained with the inclusion complex are probably due to the time required to allow nerolidol to be released from the inclusion complex.

Introduction

Lactobacillus fermentum is an ubiquitous Gram-positive, rod-shaped, anaerobic, thermoacidophilic lactic acid bacteria (LAB), which can originate from plants, animals, meat, dairies, fruits, and cereals. L. fermentum is an obligate heterofermentative bacteria which ferments various types of sugars (lactose, fructose, maltose, sucrose) under anaerobic conditions. In addition to lactic acid, L. fermentum produces acetic acid, ethanol, carbon dioxide, and minor byproducts, such as diacetyl, hydrogen peroxide, and different alcohols (e.g. 3-methylbutanol), esters (e.g. ethyl acetate), and carbonyls (e.g. nonanal) compounds. L. fermentum is not harmful to humans and its presence is necessary for the fermentation of different food products, including sourdough, cocoa, and certain beverages. Certain L. fermentum strains are probiotic agents and express outstanding health-promoting characteristics when consumed [START_REF] Lacerda | Strain-specific probiotics properties of Lactobacillus fermentum , Lactobacillus plantarum and Lactobacillus brevis isolates from Brazilian food products[END_REF]. Nonstarter lactic acid bacteria, including L. fermentum, cause defects in certain food products, such as slits or cracks in hard cheeses, lack of flavors, or bloated packaging in dairy products [START_REF] Ledenbach | Microbiological spoilage of dairy products[END_REF][START_REF] Nath | Etiology of white spot defect in swiss cheese made from pasteurized milk[END_REF][START_REF] Kurmann | Studies of the defect excessive development of eyes in Gruyere cheese[END_REF]. L. fermentum also spoils various types of beverages and can grow in fruit juice, leading to the production of undesirable compounds [START_REF] Juven | Identification of chemical constituents of tomato juice which affect the heat resistance of Lactobacillus fermenturn[END_REF][START_REF] Parish | lsolattion and identification of lactic acid bacteria from samples of citrus Molasses and unpasteurized orange juice[END_REF][START_REF] Shearer | Heat resistance of juice spoilage microorganisms[END_REF][START_REF] Ray | Fundamental food microbiology[END_REF][START_REF] Robinson | Encyclopedia of Food Microbiology[END_REF]. It is one of the LABs responsible for the desired malolactic fermentation in wine, resulting in the conversion of malic acid into lactic acid, acetate, succinate, and carbon dioxide [START_REF] Matthews | Lactic acid bacteria as a potential source of enzymes for use in vinification[END_REF]. However, the presence of LABs could lead to wine spoilage as the control of bacterial growth and malolactic fermentation is difficult to achieve, thus altering the wine organoleptic properties [START_REF] Matthews | Lactic acid bacteria as a potential source of enzymes for use in vinification[END_REF][START_REF] Edwards | Lactobacillus kunkeei sp . nov. : a spoilage organism associated with grape juice fermentations[END_REF][START_REF] Bartowsky | Bacterial spoilage of wine and approaches to minimize it[END_REF]. The growth of heterofermentative LABs (such as L. fermentum) in wine causes an increased acidity, cloudiness, and mousy odor [START_REF] Erkmen | Food Microbiology: Principles Into Practice[END_REF]. On the other hand, L. fermentum can grow in beer, as it is resistant to hop-compounds [START_REF] Bokulich | The microbiology of malting and brewing[END_REF], leading to beer spoilage [START_REF] Bokulich | The microbiology of malting and brewing[END_REF][START_REF] Dolezil | Variations amongst beers and lactic acid bacteria relating to beer spoilage[END_REF] and aroma alteration [START_REF] Bokulich | The microbiology of malting and brewing[END_REF].

Different strategies have been adopted to overcome the microbial spoilage of food, including the addition of chemical additives and physical treatments. Physical treatments include various preservation techniques, such as thermal, ultraviolet light, ultrasound, pulsed electric field, and high hydrostatic pressure technologies [START_REF] Aneja | Emerging preservation techniques for controlling spoilage and pathogenic microorganisms in fruit juices[END_REF]. However, the application of these treatments is limited due to changes in the organoleptic properties of the food product [START_REF] Mayer | Phonoloxidase and their significance in fruit and vegetables[END_REF][START_REF] Messens | The use of high pressure to modify the functionality of food proteins[END_REF][START_REF] Ross | Combining nonthermal technologies to control foodborne microorganisms[END_REF] and sometimes due to high cost [START_REF] Raso | Influence of temperature andpressure on the lethality of ultrasound[END_REF]. On the other hand, chemical preservatives used against food microbial spoilage, including benzoates, sorbates, propionates, nitrates, and nitrites, can cause allergic responses and could be converted to potential carcinogens [START_REF] Anand | Artificial preservatives and their harmful effects: looking toward nature for safer alternatives[END_REF]. The high demand of fresh and "safe" food, free of synthetic additives and contaminants, have increased the interest of using natural products for food preservation. Natural antimicrobials may derive from plants, animals, and microorganisms. Plant essential oils are largely exploited due to their wide spectrum of antimicrobial activity against spoilage bacteria and food-borne pathogens [START_REF] Gutierrez | Efficacy of plant essential oils against foodborne pathogens and spoilage bacteria associated with ready-to-eat vegetables: antimicrobial and sensory screening[END_REF]. Plant antimicrobials include different chemical classes, among which are saponins, tannins, flavonoids, terpenes, simple phenols, and phenolic acids [START_REF] Cowan | Plant products as antimicrobial agents[END_REF][START_REF] Gutiérrez-Larraínzar | Evaluation of antimicrobial and antioxidant activities of natural phenolic compounds against foodborne pathogens and spoilage bacteria[END_REF].

The majority of natural antimicrobials are hydrophobic and poorly stable, which limit their use in aqueous media. Solvents, such as dimethylsulfoxide (DMSO) [START_REF] Lee | Antifungal effect of eugenol and nerolidol against Microsporum gypseum in a guinea pig model[END_REF], dimethylformamide [START_REF] Kubo | Antimicrobial activity of green tea flavor components and their combination effects[END_REF], and ethanol [START_REF] Brehm-Stecher | Sensitization of Staphylococcus aureus and Escherichia coli to antibiotics by the sesquiterpenoids nerolidol, farnesol, bisabolol, and apritone[END_REF][START_REF] Zhao | Identification of cyclodextrin inclusion complex of chlorogenic acid and its antimicrobial activity[END_REF], are used to dissolve hydrophobic compounds in aqueous solutions. However, the use of organic solvents in food products is not desirable. Various encapsulation systems (nanoemulsions, liposomes, nanoparticles, solid lipid nanoparticles, cyclodextrins, etc.) were introduced to the food industry as a novel strategy to overcome the poor water solubility of food antimicrobials and to enhance their stability in food matrices [START_REF] Blanco-Padilla | Food antimicrobials nanocarriers[END_REF][START_REF] Weiss | Nanostructured Encapsulation Systems: Food Antimicrobials[END_REF]. Cyclodextrins are natural oligosaccharides widely used in food products for their safety and their ability to deliver hydrophobic compounds (e.g. antioxidants, antimicrobials, etc.) [START_REF] Astray | A review on the use of cyclodextrins in foods[END_REF]. Moreover, different derivatives of cyclodextrin have been synthetized to enhance the aqueous solubility of native cyclodextrins [START_REF] Del | Cyclodextrins and their uses: a review[END_REF].

The objective of this study is to select a natural potent antibacterial agent against L. fermentum.

The selected antibacterial is then complexed with hydroxypropyl-β-cyclodextrin (HP-β-CD) and the obtained inclusion complex is tested against the chosen bacterium in culture medium.

Therefore, a wide range of terpenic and phenolic phytochemicals was screened against the bacterium, under its optimal growth conditions. The tested compounds belong to different subclasses and possess variable structures. Seventeen terpenes were tested, among which eight monoterpene hydrocarbons (camphene, p-cymene, limonene, α-phellandrene, α-pinene, βpinene, α-terpinene, γ-terpinene), a sesquiterpene hydrocarbon (β-caryophyllene), four monoterpene alcohols (borneol, geraniol, linalool, menthol), a sesquiterpene alcohol (nerolidol), two monoterpene esters (bornyl acetate, linalyl acetate), and a monoterpene ketone (camphor) (Fig. 1). In addition, eleven phenols were tested, among which four phenylpropenes (transanethole, eugenol, isoeugenol, estragole), a phenylpropene ester (eugenyl acetate), two hydroxycinnamic acids (p-coumaric acid, trans-ferulic acid), a phenol ether (anisole), a methylphenol (o-cresol), a flavonoid (quercetin), and a stilbenoid (resveratrol) (Fig. 2). Nerolidol (Ner) showed the most potent antibacterial activity against L. fermentum among the 28 tested compounds. The potency of nerolidol solubilized in DMSO and hydroxypropyl-βcyclodextrin/nerolidol (HP-β-CD/Ner) complex was assessed against L. fermentum by determining the minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC) values under the optimal conditions for L. fermentum growth. The time required for both forms of nerolidol to cause a total bacterial kill was determined by time-kill analysis.

Materials and methods

Materials

β-Caryophyllene, geraniol, isoeugenol (98% mixture cis and trans), nerolidol (98%, mixture of cis (40%) and trans (60%) isomers), (-)-β-pinene, ethanol, and DMSO were purchased from Sigma-Aldrich (Missouri, United States). trans-Anethole (99%) and linalyl acetate were purchased from Sigma-Aldrich (Madrid, Spain). Anisole, camphene, o-cresol, eugenol, eugenyl acetate, (-)-menthol, and α-phellandrene were purchased from Sigma-Aldrich (Schnelldorf, Germany). (-)-Borneol, (-)-bornyl acetate, p-cymene, (R)-(t)-limonene, linalool, (+)-α-pinene, αterpinene, and γ-terpinene were purchased from Sigma-Aldrich (Buchs, Switzerland). (±)-Camphor and trans-ferulic acid were purchased from Sigma-Aldrich (Hong Kong, China).

Quercetin was purchased from Sigma-Aldrich (Haryana, India). p-Coumaric acid was purchased from Sigma-Aldrich (Irvine, United kingdom). Hydroxypropyl-β-cyclodextrin was purchased from Wacker-Chemie (Lyon, France). De Man, Rogosa, and Sharpe (MRS) broth and MRS agar were purchased from Laboratorios Conda (Madrid, Spain).

Bacterial strain and culture

L. fermentum ATCC 9338 was purchased from American Type Culture Collection (Manassas, Virginia, USA). L. fermentum cultures were routinely maintained at 4 °C on MRS agar. Before each antimicrobial assay, fresh cultures were prepared in sterile MRS broth and incubated at 37 °C for 22 h under anaerobic conditions. A bacterial suspension was prepared by diluting the bacterial culture in MRS broth to a final concentration of 25 x 10 5 colony forming unit (CFU)/ml.

Screening of natural terpenic and phenolic compounds for antibacterial activity 2.3.1. Screening rounds

Twenty-eight natural terpenic and phenolic compounds were screened for their antibacterial activity against L. fermentum at 3500, 500, 250, and 100 μM. The tested molecules were dissolved in DMSO, except β-caryophyllene and camphene which were dissolved in ethanol, and the obtained solutions were homogenized by hand agitation prior to each test. Antibacterial agent solutions (25 µl) were mixed with MRS broth (4.8 ml) in glass culture tubes (20 x 100 mm). The tubes were then inoculated with 200 μl of a diluted L. fermentum suspension (25 x 10 5 CFU/ml) to yield a bacterial concentration of 10 5 CFU/ml at baseline. Bacterial cultures (5 ml) exempt of any agent, or containing 25 µl of DMSO or ethanol, served as controls. All cultures were incubated at 37 °C for 22 h under anaerobic conditions. Screening rounds were conducted with bioactive compounds at 3500, 500, and 250 μM, successively. After each screening round, molecules demonstrating an anti-proliferative activity against L. fermentum at a given concentration were identified. Whereas, compounds demonstrating a bactericidal effect against L. fermentum were selected for another screening round at a lower concentration. The compounds exhibiting a total bactericidal activity or a total anti-proliferative activity against L. fermentum at 250 µM were screened at 100 µM. Each test was performed in triplicate and under sterile conditions.

Determination of the anti-proliferative activity of molecules

The anti-proliferative activity of the molecules against L. fermentum was assessed by UV-visible spectroscopy at 660 nm using Uviline 9100-9400 spectrophotometer (GmbH, Germany). The optical density of each tube was measured and compared to the control. The percentage of bacterial proliferation inhibition was calculated as follows:

Bacterial proliferation inhibition (%) = ,

where OD 660m and OD 660c are the optical densities of the tubes containing the molecule and the control tube, respectively. Each test was done in triplicate.

Preparation of HP-β-CD/Ner inclusion complex

HP-β-CD/Ner inclusion complex was prepared by freeze-drying, as previously described by Azzi et al. [START_REF] Azzi | First evaluation of drug-in-cyclodextrin-in-liposomes as an encapsulating system for nerolidol[END_REF]. Briefly, an aqueous solution of HP-β-CD (25 mM) containing an excess of nerolidol was kept under magnetic stirring at 300 rpm for 24 h at room temperature. The suspension was then filtered (0.45 µm, cellulose acetate membrane) to remove the excess of nerolidol. The filtrate was frozen at -80 °C, and lyophilized. HP-β-CD/Ner complex in powder form was stored at 4 °C until usage. The amount of nerolidol encapsulated in the cyclodextrin cavity was determined by HPLC as described previously by Azzi et al. [START_REF] Azzi | First evaluation of drug-in-cyclodextrin-in-liposomes as an encapsulating system for nerolidol[END_REF], and the result was expressed as mass of nerolidol (µg) per mg of powder (μg Ner /mg powder ).

Study of the antibacterial activity of nerolidol and HP-β-CD/Ner complex 2.5.1. Determination of MIC and MBC values

Bacterial cultures were prepared as previously described in section 2.3.1, in presence of nerolidol dissolved in DMSO, and added at a final concentration ranging from 0.1 to 3500 µM. The MIC was determined as the minimal concentration at which no bacterial growth was observed in MRS broth, whereas the MBC was determined as the minimal concentration at which no bacterial growth was observed on agar. Each experiment was performed in triplicate and under sterile conditions. On the other hand, HP-β-CD was investigated for its capacity to replace DMSO for nerolidol solubilization in aqueous solution. The inclusion complex was tested at a final concentrations of nerolidol ranging between 50 and 4000 μM. Bacterial cultures (5 ml) exempt of any agent, or containing DMSO (25 µl) or blank HP-β-CD added in similar amounts to that of the inclusion complex, served as controls.

Time-kill analysis

Time-kill assay was performed in triplicate on nerolidol and HP-β-CD/Ner complex. Cultures were prepared as described in section 2.3.1. Nerolidol solubilized in DMSO and the complex were added to cultures at their respective MBC values. Starting from an initial bacterial concentration of 10 5 CFU/ml (5 log CFU/ml), bacterial growth was followed during 22 h in cultures maintained at 37 °C under anaerobic conditions. The viable plate count was determined at different time intervals using the spread plate method [START_REF] Sanders | Aseptic Laboratory Techniques : Plating Methods 2. Streak Plate Procedure : Isolation of bacterial colonies using the quadrant method[END_REF]. Therefore, 100 μl of samples with appropriate dilutions was spread on MRS agar. The plates were incubated under L. fermentum optimal growth conditions for 22 h. The colonies were then counted and the bacterial concentration was determined.

Statistical Analysis

Statistical analysis was performed using the Student T test. P values equal or less than 0.05 were considered statistically significant.

Results

Antibacterial activity of terpenic and phenolic compounds

Terpenic compounds exhibited a significantly higher antibacterial activity against L. fermentum compared to phenolic compounds, with α-terpinene being the only exception. In fact at 3500µM, 16 among the 17 tested terpenes were bactericidal, whereas α-terpinene strongly inhibited the proliferation of L. fermentum (89.1%) (Table 1). Eugenol and its ester derivative eugenyl acetate were the only phenolic compounds exhibiting a strong bactericidal activity against L. fermentum at 3500 μM. Indeed, eugenol and eugenyl acetate exhibited a bactericidal activity of 100% and 95%, respectively, within 22 h of incubation at 37 °C under anaerobic conditions (Table 2).

Among the phenolic compounds, trans-anethole, estragole, and isoeugenol showed a significant anti-proliferative activity against L. fermentum at the highest concentration (3500 μM), where bacterial growth inhibition was higher than 88% (Table 2). At this concentration, p-coumaric acid, trans-ferulic acid, and anisole showed a weak bacterial growth inhibition of 21.9, 17.3, and 16.1%, respectively (Table 2). Whereas, quercetin, o-cresol, and resveratrol possessed no antibacterial activity against L. fermentum (Table 2). In fact, quercetin and resveratrol were insoluble in MRS at this concentration, which was marked by their strong precipitation.

The stronger antibacterial property of terpenes compared to phenolic compounds was highlighted by the strong bactericidal and anti-proliferative effect of different studied terpenes at 500 μM.

Indeed, at this concentration, the two phenolic compounds, eugenol and eugenyl acetate, showed no antibacterial activity against L. fermentum (Table 2). On the other hand, β-pinene, bornyl acetate, linalyl acetate, and nerolidol exhibited a total bactericidal activity at 500 μM (Table 1).

Camphene, p-cymene, limonene, α-phellandrene, α-pinene, and γ-tepinene showed a total inhibition of L. fermentum proliferation but were not bactericidal (Table 1). Furthermore, βcaryophyllene, geraniol, and menthol showed a bacterial inhibition percentage higher than 75%, compared to control, whereas borneol demonstrated a weak anti-proliferative activity (28.5%) (Table 1). However, linalool and camphor, which exhibited a bactericidal activity against L.

fermentum at 3500 μM, showed no antibacterial activity at 500 μM (Table 1).

When β-pinene, bornyl acetate, linalyl acetate, and nerolidol were tested at 250 μM, nerolidol was the only compound exhibiting a bactericidal activity (100%) (Table 1). Whereas, β-pinene, bornyl acetate, and linalyl acetate showed a strong anti-proliferative activity against L.

fermentum at 250 µM, as no bacterial growth was observed in MRS broth within 22 h of incubation (Table 1). Nerolidol maintained a strong bactericidal activity (100%) against L.

fermentum at 100 µM, whereas, β-pinene strongly inhibited bacterial proliferation (~91%), and bornyl acetate and linalyl acetate showed no antibacterial activity (Table 1). Thus, among the 28 tested compounds, nerolidol was retained for further studies as it was the most effective studied compound against L. fermentum.

Antibacterial activity of nerolidol

Nerolidol was the only molecule that exhibits a bactericidal activity against L. fermentum at a low concentration (100 µM) (Tables 1 and2). Therefore, the study was taken further to determine the MIC and MBC values of nerolidol. Nerolidol exhibited a strong antibacterial activity against L. fermentum with low MIC (25 µM; 5.56 mg/l) and MBC (50 µM; 11.12 mg/l) values. Moreover, L. fermentum survival was approximately 1% at a nerolidol concentration of concentration.

The antibacterial activity profile (bactericidal or bacteriostatic) of nerolidol was evaluated using the MBC to MIC ratio. The in vitro antimicrobial activity of nerolidol can be described as bactericidal as the MBC to MIC ratio (MBC/MIC= 2) is lower than 4 [START_REF] Nkanwen | Antibacterial agents from the leaves of Crinum purpurascens herb (Amaryllidaceae)[END_REF].

Antibacterial activity of HP-β-CD/Ner complex

The amount of nerolidol in the freeze-dried inclusion complex, determined by HPLC, was 40 μg Ner /mg powder . The antibacterial activity of HP-β-CD/Ner complex was investigated for the first time against L. fermentum in culture medium under the optimal conditions for bacterial growth.

HP-β-CD without nerolidol had no effect on the bacterial growth (data not shown). The MIC and MBC values of HP-β-CD/Ner against L. fermentum were 100 and 200 μM, respectively. At 50 μM, a 4 log increment was observed in the bacterial concentration within 22 h of incubation (data not shown).

Nerolidol and HP-β-CD/Ner complex time-kill analysis

A time-kill analysis was conducted to determine the time required to achieve a total bacterial kill in presence of nerolidol solubilized in DMSO at 50 μM and HP-β-CD/Nero complex at 200 μM (MBC). Free nerolidol exhibited a bactericidal activity against L. fermentum within the first few hours of incubation (Fig. 3). Indeed, a 1.44 log reduction of L. fermentum concentration was observed within 4 h of incubation. The bacterial concentration continued to decrease over time, as a 2.82 log reduction was obtained after 10 h. At 16 h of incubation, no viable cells were observed.

In presence of HP-β-CD/Ner complex (200 μM), the bacterial concentration decreased by 1.22 log and 2.44 log after 4 and 10 h, respectively, and a total bacterial death was obtained within 12 h (Fig. 3).

Discussion

Antibacterial efficiency of nerolidol

Plants produce a wide range of antimicrobial agents highly desired by consumers due to their natural origin. However, many of these antimicrobials are only effective at high concentrations (for example at millimolar range), thus exhibiting a weak activity compared to common antibiotics [START_REF] Lewis | Prospects for plant-derived antibacterials[END_REF]. In this study, nerolidol exhibited the highest antibacterial activity among the 28 tested terpenic and phenolic compounds. The outstanding antibacterial potency of nerolildol against L. fermentum was marked by the low MIC (25 μM; 5.56 mg/l) and MBC (50 μM; 11.12 mg/l) values, as well as the rapid bactericidal activity (Fig. 3). This compound acts by disrupting and by damaging the bacterial cell membrane, and by interfering with genes responsible for the microbe pathogenicity [START_REF] Chan | Nerolidol: A sesquiterpene alcohol with multi-faceted pharmacological and biological activities[END_REF]. Besides, Brehm-Stecher and Johnson [START_REF] Brehm-Stecher | Sensitization of Staphylococcus aureus and Escherichia coli to antibiotics by the sesquiterpenoids nerolidol, farnesol, bisabolol, and apritone[END_REF] demonstrated the permeabilizing effect of nerolidol on L. fermentum membrane. The permeabilization of the bacterial membrane leads to the leakage of the cytoplasmic molecules, thus causing cell lysis [START_REF] Togashi | Antibacterial activities against Staphylococcus aureus of terpene alcohols with aliphatic carbon chains[END_REF]. Also, the disruption of the bacterial membrane would allow the permeation of exogenous molecules into the bacterial cytoplasm [START_REF] Brehm-Stecher | Sensitization of Staphylococcus aureus and Escherichia coli to antibiotics by the sesquiterpenoids nerolidol, farnesol, bisabolol, and apritone[END_REF]. Moreover, Brehm-Stecher and Johnson [START_REF] Brehm-Stecher | Sensitization of Staphylococcus aureus and Escherichia coli to antibiotics by the sesquiterpenoids nerolidol, farnesol, bisabolol, and apritone[END_REF] suggested that the permeabilizing activity of nerolidol may be due to its structural resemblance to the lipids of the bacterial membrane. This was previously highlighted by Cornwell and Barry [START_REF] Cornwell | Sesquiterpene components of volatile oils as skin penetration enhancers for the hydrophilic permeant 5-fluorouracil[END_REF] which attributed the enhancement of skin penetration by nerolidol to its long hydrocarbon tail which promotes the interaction of the molecule with the interior of the cell bilayer.

Nerolidol is a sesquiterpene alcohol widely used in the food industry as a flavoring agent. It demonstrated a potent antimicrobial activity against some fungi, in addition to different Grampositive and Gram-negative bacterial strains, such as Staphylococcus aureus (MIC: 3.9 mg/l; 50 mg/l) [START_REF] Kubo | Antimicrobial activity of green tea flavor components and their combination effects[END_REF][START_REF] Hada | Inhibitory effects of terpenes on the growth of Staphylococcus aureus[END_REF][START_REF] Tao | Antibacterial/antifungal activity and synergistic interactions between polyprenols and other lipids isolated from Ginkgo Biloba L. leaves[END_REF], Streptococcus mutans, Propionibacterium acnes (MIC: 25 mg/l) [START_REF] Kubo | Antimicrobial activity of green tea flavor components and their combination effects[END_REF],

Salmonella enterica (MIC: 15.6 mg/l) [START_REF] Tao | Antibacterial/antifungal activity and synergistic interactions between polyprenols and other lipids isolated from Ginkgo Biloba L. leaves[END_REF], Trichophyton mentagrophytes (MIC: 12.5 mg/l) [START_REF] Kubo | Antimicrobial activity of green tea flavor components and their combination effects[END_REF],

and Aspergillus niger (15.6 mg/l) [START_REF] Tao | Antibacterial/antifungal activity and synergistic interactions between polyprenols and other lipids isolated from Ginkgo Biloba L. leaves[END_REF]. On the other hand, nerolidol was able to enhance the susceptibility of S. aureus and Escherichia coli to antibiotics, including ciprofloxacin, erythromycin, gentamicin, vancomycin [START_REF] Simoes | Enhancement of Escherichia coli and Staphylococcus aureus antibiotic susceptibility using sesquiterpenoids[END_REF], and amoxicilline/clavulanic acid [START_REF] Gonçalves | Evaluation of the mutagenicity of sesquiterpenic compounds and their influence on the susceptibility towards antibiotics of two clinically relevant bacterial strains[END_REF]. In this study, nerolidol antibacterial potency against L. fermentum (MIC: 25 μM; 5.56 mg/l) was weaker than that of gentamicin [START_REF] Egervärn | Antibiotic susceptibility profiles of Lactobacillus reuteri and Lactobacillus fermentum[END_REF] and chloramphenicol [START_REF] Klare | Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use[END_REF], and close to that of novobiocin [START_REF] Gfeller | Sequence and genetic organization of the 19.3kb erythromycin-and dalfopristin-resistance plasmid pLME300 from Lactobacillus fermentum ROT1[END_REF] and Nalkyldimethylbenzyl ammonium chloride [START_REF] Oliva-Neto | Susceptibility of Saccharomyces cerevisiae and lactic acid bacteria from the alcohol industry to several antimicrobial compounds[END_REF] (Table 3). On the other hand, L. fermentum strains were less sensitive to well-known antibiotics including vancomycin [START_REF] Klare | Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use[END_REF][START_REF] Florez | Antimicrobial susceptibility profiles of 32 type strains of Lactobacillus, Bifidobacterium, Lactococcus and Streptococcus spp[END_REF], teicoplanin [START_REF] Klare | Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use[END_REF],

streptomycin [START_REF] Egervärn | Antibiotic susceptibility profiles of Lactobacillus reuteri and Lactobacillus fermentum[END_REF][START_REF] Klare | Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use[END_REF][START_REF] Florez | Antimicrobial susceptibility profiles of 32 type strains of Lactobacillus, Bifidobacterium, Lactococcus and Streptococcus spp[END_REF], erythromycin, tobramycin, clindamycin, kanamycin, polymixin B [START_REF] Egervärn | Antibiotic susceptibility profiles of Lactobacillus reuteri and Lactobacillus fermentum[END_REF][START_REF] Gfeller | Sequence and genetic organization of the 19.3kb erythromycin-and dalfopristin-resistance plasmid pLME300 from Lactobacillus fermentum ROT1[END_REF][START_REF] Florez | Antimicrobial susceptibility profiles of 32 type strains of Lactobacillus, Bifidobacterium, Lactococcus and Streptococcus spp[END_REF], metronidazole, and nitrofurantoin [START_REF] Neto | Chemical inhibition of the contaminant Lactobacillus fermentum from distilleries producing fuel bioethanol[END_REF] (Table 3).

Antibacterial assay of HP-β-CD/Ner complex

Cyclodextrin inclusion complexes have been widely studied for their capacity to enhance the stability and solubility of antimicrobials [START_REF] Zhao | Identification of cyclodextrin inclusion complex of chlorogenic acid and its antimicrobial activity[END_REF][START_REF] Li | Identification of a cyclodextrin inclusion complex of antimicrobial peptide CM4 and its antimicrobial activity[END_REF][START_REF] Dima | Encapsulation of Coriander essential oil in beta-cyclodextrin: Antioxidant and antimicrobial properties evaluation[END_REF].

Some studies reported similar antibacterial activities of free and complexed antimicrobials (e.g. clarithromycin, chlorogenic acid, peptide CM4) [START_REF] Zhao | Identification of cyclodextrin inclusion complex of chlorogenic acid and its antimicrobial activity[END_REF][START_REF] Li | Identification of a cyclodextrin inclusion complex of antimicrobial peptide CM4 and its antimicrobial activity[END_REF][START_REF] Salem | Efficacies of cyclodextrin-complexed and liposome-encapsulated clarithromycin against Mycobacterium avium complex infection in human macrophages[END_REF], while others proved modifications of the potency of the antimicrobial agent following complexation with cyclodextrins. Compared to the free form, the inclusion complex of Hyptis martiusii essential oil [START_REF] Andrade | Physico-chemical characterization and antibacterial activity of inclusion complexes of Hyptis martiusii Benth essential oil in β-cyclodextrin[END_REF] and coriander essential oil [START_REF] Dima | Encapsulation of Coriander essential oil in beta-cyclodextrin: Antioxidant and antimicrobial properties evaluation[END_REF] showed a lower antimicrobial activity, whereas that of carvacrol showed a higher antimicrobial activity [START_REF] Santos | Characterization of carvacrol betacyclodextrin inclusion complexes as delivery systems for antibacterial and antioxidant applications[END_REF].

The incorporation of nerolidol into HP-β-CD inclusion complex increased the MIC and MBC

values by 4 fold. Azzi et al. [START_REF] Azzi | First evaluation of drug-in-cyclodextrin-in-liposomes as an encapsulating system for nerolidol[END_REF] studied the release of nerolidol from the inclusion complex in water and at room temperature by dialysis. Around 45% of nerolidol were released from HP-β-CD/Ner complex within 8 h, followed by a slow release rate over 7 days [START_REF] Azzi | First evaluation of drug-in-cyclodextrin-in-liposomes as an encapsulating system for nerolidol[END_REF]. In fact, when HPβ-CD/Ner complex is added at 50 μM, the actual concentration of nerolidol interacting with L.

fermentum in MRS is significantly lower than 50 μM during the whole experiment, thus explaining the ability of the inoculated bacteria to proliferate (data not shown). However, at 200 μM the concentration of nerolidol in the culture medium should be approximately 100 μM after 8 to 10 h of incubation, which could explain the rapid drop in the bacterial concentration (Fig. 3).

The slow release and the photo-protection of nerolidol provided by the encapsulation systems [START_REF] Azzi | First evaluation of drug-in-cyclodextrin-in-liposomes as an encapsulating system for nerolidol[END_REF] would prevent and limit the proliferation of spoilage bacteria in food products.

Relationship between the antibacterial activity of terpenic and phenolic compounds and their structural and physicochemical parameters

Various parameters may modulate the activity of antibacterial agents, including the drug hydrophobicity, the presence of functional groups, the bacterial cell envelope characteristics, and the incubation conditions (for instance, aeration).

The influence of hydrophobicity on the antibacterial activity of molecules

It has been reported that molecules with high hydrophobicity exhibit a high affinity to the lipophilic structures in the target microorganisms, such as the bacterial membrane [START_REF] Sikkema | Mechanisms of membrane toxicity of hydrocarbons[END_REF].

Therefore, they may disrupt the membrane integrity, thus affecting membrane permeability and enzymes activity [START_REF] Sikkema | Mechanisms of membrane toxicity of hydrocarbons[END_REF]. However, many exceptions could be revealed from this work, which suggests that the hydrophobicity is not always the key parameter governing the antibacterial activity; and some structural features could be taken into consideration when analyzing the structure-antibacterial activity relationship.

Nerolidol, which has shown the strongest antibacterial activity against L. fermentum, has a LogP value (5.33-5.36) considerably higher than that of the other tested molecules, except βcaryophyllene (6.87) (Table 1). The latter, showed a modest antibacterial activity against L.

fermentum as it lost its bactericidal potential at 500 μM (Table 1). In fact, β-caryophyllene have been found to exhibit a weak antibacterial activity against different Gram-positive and Gramnegative bacteria [START_REF] Selestino Neta | Effects of β-caryophyllene and Murraya paniculata essential oil in the murine hepatoma cells and in the bacteria and fungi 24-h time-kill curve studies[END_REF].

On the other hand, the antibacterial potency of the compounds exhibiting an antibacterial activity at a concentration lower than 500 μM was influenced by hydrophobicity (Table 1). Indeed, βpinene showed a higher antibacterial activity than the less hydrophobic molecules, bornyl acetate and linalyl acetate (Table 1). The acetylation of linalool and borneol increased their hydrophobicity and their antibacterial activity (Table 1). Indeed, bornyl acetate and linalyl acetate retained their total anti-proliferative activity at 250 μM, whereas borneol and linalool lost partially and totally the activity at 500 μM, respectively (Table 1). Similarly, Knobloch et al.

[57] reported a higher inhibition of H + -translocation by linalyl acetate, compared to linalool.

Also, Dorman and Deans [START_REF] Dorman | Antimicrobial agents from plants: antibacterial activity of plant volatile oils[END_REF] observed a higher antibacterial activity of bornyl acetate compared to borneol against a wide range of bacterial strains, among which Lactobacillus plantarum.

Moreover, phenolic compounds with LogP values close to or lower than 2, such as anisole (2.11) and o-cresol (1.95-1.98), showed no or a weak antibacterial activity against L. fermentum (Table 2).

Acyclic monoterpenoids showed an increased antibacterial activity with the increment of their LogP value. Indeed, linalyl acetate showed a higher antibacterial activity than geraniol, which was more potent than linalool (Table 1). This was also observed for bicyclic monoterpenes, as borneol and camphor had the lowest LogP values, and showed the weakest antibacterial activities (Table 1). However, despite β-pinene not being the most hydrophobic compound in the bicyclic monoterpenes chemical class, it exhibited the strongest antibacterial activity (Table 1).

For monocyclic monoterpenes, limonene, α-phelladrene, and γ-terpinene, having higher LogP values than menthol, showed a stronger antibacterial activity (Table 1). α-Terpinene was found to be an exception as it showed the weakest antibacterial potency, despite a high LogP value (Table 1).

Structure-activity analysis of terpenic compounds

Different studies have demonstrated the role of terpenoid functional groups in the antimicrobial activity. Carvacrol showed a better antimicrobial activity compared to its derivatives, carvacrol methyl ether and p-cymene, which lack the hydroxyl group [START_REF] Dorman | Antimicrobial agents from plants: antibacterial activity of plant volatile oils[END_REF]. On the other hand, Kotan et al.

[59] reported a better antibacterial activity for alcohol derivatives of oxygenated monoterpenes, when compared to ketone and acetate derivatives.

The weak antibacterial activity of β-caryophyllene, despite its high hydrophobicity, may be due to the absence of a hydrophilic functional group in the chemical structure of the molecule. The combination of a lipophilic character of the skeleton and the presence of a hydrophilic functional group was found to be important for the antimicrobial activity of essential oils components [START_REF] Kalemba | Antibacterial and antifungal properties of essential oils[END_REF].

The structural features of nerolidol are in line with the previous findings. Indeed, the antibacterial activity of aliphatic terpene alcohols was demonstrated to be dependent on the hydrophobic chain length starting from the carbon connected to the hydroxyl group [START_REF] Togashi | Antibacterial activities against Staphylococcus aureus of terpene alcohols with aliphatic carbon chains[END_REF][START_REF] Kubo | Antibacterial activity of long-chain alcohols: The role of hydrophobic alkyl groups[END_REF]. In fact, farnesol (C 12 ) exhibited a stronger antibacterial activity than nerolidol (C 10 ) followed by geraniol (C 8 ), whereas linalool (C 6 ) showed no antibacterial activity against Staphylococcus aureus [START_REF] Kubo | Antibacterial activity of long-chain alcohols: The role of hydrophobic alkyl groups[END_REF]. Similarly, Togashi et al. [START_REF] Togashi | Antibacterial activities against Staphylococcus aureus of terpene alcohols with aliphatic carbon chains[END_REF] reported a very weak antibacterial activity of geraniol and linalool against S. aureus. Additionally, farnesol (C 12 ), nerolidol (C 10 ), and plaunotol (C 11 ), showed a strong antibacterial activity against S. aureus, in that order [START_REF] Togashi | Antibacterial activities against Staphylococcus aureus of terpene alcohols with aliphatic carbon chains[END_REF][START_REF] Inoue | The antibacterial effects of terpene alcohols on Staphylococcus aureus and their mode of action[END_REF]. Also, no or weak antibacterial activity against S. aureus was reported for alcohols with chains containing more than 12 carbon atoms like farnesylacetol (C 14 ) [START_REF] Kubo | Antibacterial activity of long-chain alcohols: The role of hydrophobic alkyl groups[END_REF], geranylgeraniol, and phytol (C 16 ) [START_REF] Togashi | Antibacterial activities against Staphylococcus aureus of terpene alcohols with aliphatic carbon chains[END_REF].

Therefore, to exhibit a potent antibacterial effect, the authors suggested that the chain, starting from the hydroxyl group, should contain from 10 to 12 carbon atoms [START_REF] Togashi | Antibacterial activities against Staphylococcus aureus of terpene alcohols with aliphatic carbon chains[END_REF], or less than 12 carbon atoms but as close to 12 as possible [START_REF] Kubo | Antibacterial activity of long-chain alcohols: The role of hydrophobic alkyl groups[END_REF]. Although the previous studies were conducted on S.

aureus, our study supports the pattern of the antibacterial activity of terpene alcohols against L.

fermentum. Indeed, nerolidol (C 10 ; sesquiterpene) exhibited a stronger antibacterial activity than geraniol (C 8 ; monoterpene), the latter being more potent than linalool (C 6 ; monoterpene) (Table 1).

Structure-activity analysis of phenolic compounds

The importance of the propenyl side chain was noted among the tested phenolic compounds, as eugenol, trans-anethole, isoeugenol, and estragole, showed a significantly higher antibacterial activity against L. fermentum compared to anisole (Table 2). The propenyl side chain is absent in anisole compared to eugenol, trans-anethole, isoeugenol, and estragole (Fig. 2), which could be the reason for the weaker antibacterial activity of anisole.

On the other hand, eugenol showed the highest antibacterial activity among the tested phenolic compounds against the Gram-positive bacterium L. fermentum. However, Gharib et al. [START_REF] Gharib | Interaction of selected phenylpropenes with dipalmitoylphosphatidylcholine membrane and their relevance to antibacterial activity[END_REF] reported a higher antibacterial activity for anethole and estragole compared to eugenol, against the Gram-negative bacterium Escherichi coli. Indeed, the hydrophobicity of the compound seemed to play a role in the potency of the antibacterial activity against E. coli, as anethole and estragole have a higher hydrophobicity than eugenol [START_REF] Gharib | Interaction of selected phenylpropenes with dipalmitoylphosphatidylcholine membrane and their relevance to antibacterial activity[END_REF] (Table 2). Whereas, the hydrophobicity of phenylpropenes did not seem to influence their antibacterial activity against the Gram-positive bacterium, L. fermentum. Therefore, our results strongly suggest the presence of other factors that modulate the antibacterial activity of phenylpropenes against L. fermentum. The higher antibacterial activity of eugenol compared to its ester, eugenyl acetate (Table 2), highlights the importance of the hydroxyl group in the phenolic structure (Fig. 2). Additionally, the absence of the hydroxyl group in trans-anethole and estragole structure (Fig. 2) could explain the lower antibacterial activity against L. fermentum, compared to eugenol (Table 2). Indeed, the hydroxyl group of eugenol and isoeugenol has been found to reinforce the lipid membrane-fluidizing effect, compared to anethole and estragole [START_REF] Gharib | Interaction of selected phenylpropenes with dipalmitoylphosphatidylcholine membrane and their relevance to antibacterial activity[END_REF]. Also, the bacterial membrane characteristics play a crucial role in the differential bacterial susceptibility to a given antibacterial agent [START_REF] Shrivastava | Characterization of enhanced antibacterial effects of novel silver nanoparticles[END_REF][START_REF] Tamboli | Mechanistic antimicrobial approach of extracellularly synthesized silver nanoparticles against gram positive and gram negative bacteria[END_REF][START_REF] Nazzaro | Effect of essential oils on pathogenic bacteria[END_REF]. In fact, eugenol has previously shown lower MIC values against the Gram-positive bacteria Staphylococcus aureus (MIC= 2.5 mg/l) and Bacillus subtilis (MIC= 1.25 mg/l), compared to E. coli (MIC= 5 mg/l) [START_REF] Liang | Effects of cyclodextrins on the antimicrobial activity of plant-derived essential oil compounds[END_REF].

Both ferulic acid and p-coumaric acid exhibited a weak anti-proliferative activity against L.

fermentum in MRS broth (pH ~6.2) (Table 2). The antibacterial activity of hydroxycinnamic acids depends on pH, which controls the concentration of their undissociated form. The undissociated form can easily penetrate the cytoplasmic membrane of the bacterium [START_REF] Russell | Another explanation for the toxicity of fermentation acids at low pH: anion accumulation versus uncoupling[END_REF]. The pKa values of ferulic acid and p-coumaric acid are between 4 and 5, thus a greater proportion of their dissociated forms is found at the pH of the culture medium (~6.2). In fact, the antibacterial activity of ferulic acid and p-coumaric acid was found to increase in culture media at pH values below 6 [START_REF] Sánchez-Maldonado | Structure-function relationships of the antibacterial activity of phenolic acids and their metabolism by lactic acid bacteria[END_REF]. Moreover, L. fermentum is able to metabolize ferulic acid and coumaric acid into less potent metabolites, such as phloretic acid, p-vinyl phenol, or dihydroferulic acid [START_REF] Sánchez-Maldonado | Structure-function relationships of the antibacterial activity of phenolic acids and their metabolism by lactic acid bacteria[END_REF]. This also may explain the weak antibacterial activity of both acids against L. fermentum.

Conclusion

In this study, 28 terpenic and phenolic compounds were screened against L. fermentum, an ubiquitously present bacterium which could cause spoilage of different food products. Among the tested compounds, nerolidol exhibited the strongest antibacterial activity marked by the low MIC and MBC values. Moreover, a total bacterial kill was obtained within 16 h in presence of nerolidol (50 μM). The antibacterial activity of nerolidol was dependent on different factors, including the hydrophobicity of the compound, as well as the position of the hydroxyl group.

Compared to nerolidol solubilized in DMSO, HP-β-CD/Ner inclusion complex exhibited 4 fold higher MIC and MBC values and a more rapid bactericidal activity. Indeed, HP-β-CD/Ner inclusion complex was proven effective against L. fermentum in culture medium. The high demand for the replacement of synthetic food additives by natural molecules encourages the application of nerolidol in food. Moreover, due to the physico-chemical limitations of the application of natural bioactive components in food, the use of encapsulation systems has recently became a widely investigated novel approach for the delivery of bioactive compounds in food products. Therefore, further studies could be realized to investigate the antibacterial activity of free and encapsulated nerolidol in different food products including fruit juices, alcoholic beverages, and milk products. This evaluation would lead to a better understanding of the antibacterial potency of natural molecules such as nerolidol, under their free and complexed form, in various types of food matrices and under various conditions. 2.9 [START_REF] Sensch | Effects of inhibition of calcium and potassium currents in guinea-pig cardiac contraction: comparison of β-caryophyllene oxide, eugenol, and nifedipine[END_REF] 1.51 [START_REF] Hansch | Exploring QSAR: Hydrophobic, electronic, and steric constants[END_REF] 3.04 [START_REF] Griffin | Determination of octanol-water partition coefficient for terpenoids using reversed-phase high-performance liquid chromatography[END_REF] 1.82 [START_REF] Rothwell | Experimental determination of octanol-water partition coefficients of quercetin and related flavonoids[END_REF] 3.10 [85] 2.45 [START_REF] Sensch | Effects of inhibition of calcium and potassium currents in guinea-pig cardiac contraction: comparison of β-caryophyllene oxide, eugenol, and nifedipine[END_REF] 3.0961 [START_REF] Kfoury | Effect of cyclodextrin complexation on phenylpropanoids' solubility and antioxidant activity[END_REF] 1.975 [START_REF] Ritter | Partition coefficients of some environmentally important organic compounds between 1-octanol and water from reversed-phase highperformance liquid chromatography[END_REF] 2.818 [START_REF] Kfoury | Effect of cyclodextrin complexation on phenylpropanoids' solubility and antioxidant activity[END_REF] 2.29 [START_REF] Reiner | Lipophilicity of some GABAergic phenols and related compounds determined by HPLC and partition coefficients in different systems[END_REF] 1.249 [START_REF] Kfoury | Effect of cyclodextrin complexation on phenylpropanoids' solubility and antioxidant activity[END_REF] 3.06 [START_REF] Wesołowska | Interaction of the chemopreventive agent resveratrol and its metabolite, piceatannol, with model membranes[END_REF] 2.73 [START_REF] Ben Arfa | Antimicrobial activity of carvacrol related to its chemical structure[END_REF] 2.99 [START_REF] Griffin | Determination of octanol-water partition coefficient for terpenoids using reversed-phase high-performance liquid chromatography[END_REF]

2. 3 . 3 .

 33 Determination of the bactericidal activity of the natural molecules Cultures showing a total inhibition of bacterial proliferation were analyzed in duplicate by spreading an aliquot of 100 µl on MRS agar. The bacterial concentration in the control was determined by enumeration. The bactericidal capacity was then evaluated according to the decrease in the initial bacterial concentration and calculated as follows: Bacterial kill (%) = , where [bac] m and [bac] c are the bacterial concentrations in the tube containing the molecule and the control tube, respectively.

Fig. 1 .

 1 Fig.1. Chemical structure of terpenic compounds.

Fig. 2 .

 2 Fig. 2. Chemical structure of phenolic compounds.

Fig. 3 .

 3 Fig. 3. Nerolidol (50 µM) and HP-β-CD/Ner complex (200 µM Ner) time-kill analysis against L.

Fig. 1

 1 Fig. 1

Fig. 3

 3 Fig. 3

Table 2 : Bactericidal and anti-proliferative activity of phenolic compounds against L. fermentum.

 2

	trans-Anethole	Anisole	p-Coumaric	acid	o-Cresol	Estragole	Eugenol	Eugenyl acetate	trans-Ferulic	acid	Isoeugenol	Quercetin	Resveratrol
	3.31[80]				1.95[73]	3.13[80]	2.40[83]
	LogP	2.11[81] 1.43[82]

Acknowledgments

The authors thank the Research Funding Program at the Lebanese for supporting this project.

Conflict of Interest

The authors declare no conflict of interest.


Reframing the creative city debate from locally-based artistic activities. The case of independent films scenes in Paris suburb

Keywords: independent cinema, artistic scene, Paris, creative city, creative industries

The purpose of this article is to open a discussion on utilitarian approaches to the territorial embeddedness of creative activities, using the concept of "scene. This concept highlights the desire to create as a defining feature of artistic creation, compared to other human activities. The article presents the results of a survey conducted with stakeholders in the independent film scenes in north-eastern Paris. It stresses the importance of self-training and selfproduction in the emergence of these independent cinema scenes, and highlights the role played by cultural policies in their structuring. Yet these independent scenes seem disconnected from urban and economic development policies, despite their support for creative industries. The concept of scene offers an alternative theoretical framework to analyse and understand the local anchoring of an artistic community led by the share of aesthetic or moral values: for the ones, a certain vision of filmmaking based and traditional techniques, for the others, a political will to voice as postcolonial subjects.

Reframing the creative city debate from locally-based artistic activities.

The case of independent films scenes in Paris suburb

The film industry's territorial embeddedness and the structural changes that this industry has undergone have been the subject of several publications, some of which have contributed to forging the analytical framework for the structuring of clusters of creative industries. These clusters are reinforced by endogenous territorial resources, be they geographical (landscapes, local history), economic (an entrepreneurial and financial ecosystem), socio-technical (technological industries, skilled labour) or political (tax incentives, provision of space) [START_REF] Frédéric | Mutations economiques, transformations urbaines, identité territoriale : San Francisco et l'industrie du cinéma[END_REF]. The studies reported in the literature shed light on the ways in which technological and artistic environments are intertwined and inform one another. They also show how relations of interdependence in the industry are reinforcing the trend towards the clustering of audiovisual firms [START_REF] Storper | The Transition to Flexible Specialisation in the US Film Industry : External Economies, the Division of Labour, and the Crossing of Industrial Divides[END_REF][START_REF] Storper | Flexible Specialisation and Regional Industrial Agglomeration: the Case of the US Motion Picture Industry[END_REF][START_REF] Leriche | Hollywood, un siècle d'industrie cinématographique[END_REF], De Propis, Hypponen 2008). While they have emerged in distinct theoretical contexts [START_REF] Pilati | Cité Créative et District Culturel; Une Analyse Des Thèses En Présence[END_REF][START_REF] Tommaso | Cultural clusters and districts: the state of the art[END_REF], the terms "cluster", "ecosystem", and "district" are often used interchangeably to describe forms and relationships between stakeholders in an economic sector within a territory. They are also used to analyse how policy makers and economic and urban developers try, with differing degrees of success, to create fiscal, real estate and infrastructural conditions that encourage their development. These theories often serve to design or defend policies aimed at supporting these sectors when they seem to bear the promise of economic development [START_REF] Greffe | Le développement de l'Ile de France par la création de districts culturels Paris: Puca, Ministère de l'équipement[END_REF][START_REF] Molho | Du Cluster à la Scène: Glissement sémantique dans le cas nantais[END_REF][START_REF] Cooke | Creative Cities, Cultural Clusters and Local Economic Development[END_REF][START_REF] Lazzeretti | Reasons for clustering of creative industries in Italy and Spain[END_REF][START_REF] Santagata | Cultural districts, property rights and sustainable economic growth[END_REF], even though the policies are sometimes out of touch with the realities, needs and practices of the cultural industry stakeholders themselves [START_REF] Lefevre | Entrepreneurs musicaux et territoires. Les clusters culturels sous l'emprise des politiques publiques et des acteurs locaux[END_REF]Morteau, 2016;Currid 2009). Artists and professionals in creative circles are increasingly challenging these policies (d'Ovidio, Morato 2017) as their impact on the cost of living in cities (and rising rents in particular) is directly detrimental to them [START_REF] Vivant | Creatives in the City. Urban Contradictions of the Creative City[END_REF].

The primacy of utilitarian approach to creative activities' territorial embeddedness obscures that which makes art unique compared to other human activities: the desire to create.. A survey on film production networks and self-production showed that this singularity of the act of creation, as an expression of self, is crucial to a comprehensive understanding of their local involvement and of the gap between these practices and the local policies for economic development through culture. On the fringes of mainstream or commercial cinema, other forms of cinema are developing underground, which are rarely visible to regular distribution channels. They are characterized by economic structures that are fragile and usually selfproduced. The emergence of these independent film scenes is driven by strong aesthetic choices and made possible by public mechanisms designed to support artistic creation and education programs. This article presents two aesthetic scenes of this French independent cinema. Both are embedded in a particular territory, the north-eastern suburbs of Paris, which plays a crucial part in their aesthetic choices and structuring process. For some of these filmmakers, this territory is historically the birthplace of cinema and of its techniques, which they want to keep alive in a context of digitization. For others, it is a symbol of the banlieues1 as a place of spatial, social and symbolic relegation of discriminated populations. This article presents the results of an in-depth investigation carried out under a research project that investigated the relationships between the different artistic worlds located north-east of Paris. Some thirty semi-structured interviews were conducted with stakeholders on the independent film scenes (filmmakers, producers, support policy operators, festival and cinema programmers, observers) and managers of independent cultural venues. This study was a follow-up to previous research conducted by the authors on creative industry clusters, and was informed by the ethnographic observations of one of the authors during a research residency at a cultural institution in the area. This empirical work leads us to a discussion on the relevance of standard explanatory models for creative ecosystems and of operational materialization of the notion of a creative city. Andy Pratt has repeatedly highlighted the inherent contradictions of creative city policies, pointing to tensions between the social and artistic implications of cultural policy and economic development objectives (2010,2011).

We propose to move away from the term "creativity", which refers to modelling the phases prior to innovation processes [START_REF] Paris | Manager La Créativité. Innover En s'inspirant de Pixar[END_REF], in order to return to the intimate and aesthetic meaning of artistic creation. To this end, we use the concept of scene from the sociology of culture, to put aesthetics back at the centre of the relationship between an artistic practice and a territory. This concept also stresses the importance of aesthetic proximity in the emergence, structuring and embedding of networks of stakeholders and practices in independent filmmaking. The concept refers to people, practices and objects that revolve around a particular cultural domain (a style of music or a literary genre, for example) [START_REF] Straw | Systems of articulation, logics of change: communities and scenes in popular music studies[END_REF]. This perspective emphasizes the structuring role of aesthetic proximity in the creation of an artistic scene, that is, a network of stakeholders embedded in a territory where an artistic form emerges through its history, its aesthetics and its public policies. The concept of the scene was defined by sociologists of culture [START_REF] Guy | La notion de « scène », entre sociologie de la culture et sociologie urbaine: genèse, actualités et perspectives[END_REF]Guibert 2014, Woo et al 2015) and was also exported to urban studies [START_REF] Lloyd | Neo-Bohemia: Art and Neighborhood Redevelopment in Chicago[END_REF][START_REF] Daniel | Scenes, innovation and urban Development[END_REF][START_REF] Daniel | Scenes, innovation and urban Development[END_REF]. It offers a model for analysing the relationship between an artistic movement, its audience, the territory in which it is embedded, and its atmosphere (Silver et al. 2011, Emin and[START_REF] Diao | Complexité et auto organisation en entrepreneuriat collectif: analyse d'une scène musicale locale[END_REF]. The scene evokes, on the one hand, the territorialized organization of an artistic activity that spawns novelty (new styles, new genres) and, on the other, a particular urban atmosphere related to the way in which cultural activities are embedded in a territory, and to values shared by various stakeholders in this territory (Ambrosino and Sagot-Duvauroux 2018 ; [START_REF] Borelli | Création d'une ambiance. Le cas du Raval de Barcelone[END_REF][START_REF] Daniel | Scenes, innovation and urban Development[END_REF]. A scene is therefore an ecosystem comprising stakeholders, places, networks and values, as well as customs and protocols that are inherent to a domain which is both the product of a territory and a component of its identity. The notion of scene also presents the possibility of dissociating aesthetic issues from the economic issues raised by the notion of independent cinema. An activity's independence is often understood in relation to the dominant companies in the industry. The economic independence of so-called independent cinema is however very relative, due to the role played by the Centre National du Cinéma et de l'image animée (CNCnational motion picture institute) in financing French cinema, compounded by the oligopolistic organization of the industry (Augros, 2013). To speak of independent film scenes highlights the aesthetic and artistic implications of these forms of filmmaking which, at least at the time of the investigation, were not market driven. Faced with the vertical logics of the film industry (Creton, 2003)2 , individuals organize themselves to create independent films in any genres, which are produced outside media conglomerates and demand a certain autonomy with regard to commercial cinema's production methods, conventions and policies. Here, independent film refers not to a style or an aesthetic, but rather to a practice that claims to go against mainstream cinema (Hurault-Paupe, Murillo 2013).

Much like Emin and Guibert ( 2017), who use this understanding of scene to study the workings of stakeholders in relation to a musical subculture (punk) in a French rural area, we analyse the structuring of two scenes of independent film in northeast Paris. Their emergence stemmed from very different expectations for filmmaking: either to defend radical aesthetic and technical choices, or to use film as a medium for political discourse and to change the image of the banlieues and their inhabitants. We first describe national and local specificities regarding the role played by public policies in supporting cinematographic creation, and then explain how the independent filmmakers organize to carry out their film projects and broadcast them on the fringes of the mainstream. The concept of scene offers a framework to understand the dialectic relationship between local embeddedness and international distribution. These case studies question to the implicit expectations of public policies to support creative industries.

A TERRITORY OF CINEMAS

The French case is an ideal example to reconsider the place of aesthetics in local artistic dynamics. Support for artistic creation is at the heart of cultural policies, and its relationship with economic policies to support creative industries is at times ambiguous [START_REF] Alexandre | La règle de l'exception, L'écologie du cinéma français[END_REF].

The historical role of public authorities in supporting cinema is essential to the emergence of these independent scenes. The French State and local authorities have long implemented regulatory policies aimed at protecting film creation and promoting its plurality, by tempering the effects of the market while supporting the cinema economy (Vernier, 2004). This is reflected in the way the Centre National du Cinéma et de l'Image Animée (CNC) structures the industry, in indirect policies to support artistic employment through the unemployment insurance scheme for intermittent workers in the entertainment industry, and in a training system in public schools [START_REF] Laurent | Retour sur les origines du système d'aide à la production cinématographique en France[END_REF]. Additionally, as in other countries, many local authorities encourage filming on their territory (via tax incentives or administrative facilities) to support the employment of professionals in the filmmaking and visual effects industries. Thus, compared to many other countries, French cinema enjoys access to resources and production conditions that facilitate aesthetic risk taking a priori. Yet, for various reasons, many filmmakers develop their film projects outside the usual production and distribution channels.

In the northeast of Paris3 , incentivizing policies for the cinema and audiovisual production industries have contradictory objectives that are reflected in the organizational split between culture and youth policies on the one hand, and urban and economic development policies on the other. In recent years, the economic and urban development strategies of Plaine Commune have been structured with "culture and creation" as keywords (Préfet de région et al, 2012). This vision plans the creation of new workspaces for the creative industries, support for the development of these activities, and the inclusion of artistic projects in real estate developments. It can be seen as a differentiation strategy in a context of competition between territories within the Parisian metropolis itself, to attract real estate investments towards more or less ambitious urban regeneration projects [START_REF] Aubry | The promotion of creative industries as a tool for urban planning: The case of the Territoire de la Culture et de la Création in Paris Region[END_REF]. In calling Plaine Commune a "territory of culture and creation", local authorities emphasized the historical embeddedness of audiovisual industries in the territory (where the first studios were created at the beginning of the 20th century) and the presence of a dense artistic network (Plaine Commune, Notre Atelier Commun et Polau 2010; Thomas 2010; Verdalle, Rot, Sauguet 2008).

As warehouses were converted to television studios in Plaine Saint-Denis in the 1980s and filming or post-production equipment rental companies have settled there, this territory tends to have an important place in the audiovisual industry [START_REF] Camors | Les industries culturelles en Ile-de-France[END_REF][START_REF] Camors | Les industries créatives en Ile de France: Un nouveau regard sur la métropole[END_REF]). Yet these activities remain dependent on decisions taken in other areas (Paris and its western suburbs) where the dominant players in this industry (television channels, financiers, producers) are located [START_REF] Scott | French Cinema. Economy, Policy and Place in the Making of a Cultural-Products Industry[END_REF]. Despite its historical roots and the importance of this territory for the audiovisual industry, the economic weight of the industry (and of cultural industries in general) remains marginal, both locally [START_REF] Lebeau | Une "banlieue créative" dans le Grand Paris?[END_REF]) and with regard to the entire metropolis [START_REF] Camors | Les industries culturelles en Ile-de-France[END_REF][START_REF] Camors | Les industries créatives en Ile de France: Un nouveau regard sur la métropole[END_REF]. Public support policies for the project to set up film studios led by EuropaCorp (called La Cité du Cinéma) can thus be seen as a way to incentivize this industry and, in the framework of a territorial communication strategy, also as a promotional argument for neighbouring real estate developments.

In parallel with these actions carried out by the authorities in charge of economic and urban development, this territory is marked by a strong political will for cultural democratization, social inclusion and support for creation. This can be explained by the tradition of municipal communism. With regard to cinema, this is materialized by an image education program in schools and a network of more than twenty public cinema theatres which put on art-house (art-

DESIRES FOR CINEMA

Far from the objectives of economic policies aimed at creative industries clusters, cultural and educational policies contribute to the development of independent film scenes by training and professionalizing young filmmakers and technicians. They support a more diverse cinematographic representations of banlieues as much as the expression of a singular artistic perspective. They have allowed the emergence of independent film scenes and new cinematographic aesthetic forms marked by the desire to make films differently, unfettered by the economic contingencies (and temporalities) of the market and the industry, and thus to support a unique voice, gaze and relationship to art.

Using the camera to experiment

Since 2014, an association of filmmakers (L'Abominable) has been occupying a building in Plaine Commune, earmarked for demolition. This temporary use space, in a peripheral yet accessible area, is a spatial translation of the association's project: to be on the fringes, in uncertain spaces, to carry out artistic projects whose audiences can only be small, and which will only be screened in dedicated spaces and festivals. The pun in its name l'Abominable -which could read as "Labo minable" (pathetic lab) -refers to photographic development laboratories and experimental laboratories, as this association is part of the DIY movement that emerged in alternative circles in the 1990s, at a time of technological upheaval and digital transition [START_REF] Cervulle | Le Cinéma Expérimental en France : Economie, Structure et acteurs[END_REF]. It operates on a cooperative basis and provides filmmakers and visual artists with the tools to work on analog media, allowing them to process their negative or reversal films, apply visual effects or change formats, edit, work on sound, and make copies. New members are trained in the use of these machines, to share and keep these endangered techniques alive. The type of post-production equipment it provides is becoming rare and new generations of directors and technicians are unfamiliar with it. Yet learning to use it guarantees creative autonomy.

"The aim is to allow people to become autonomous. There is a lot of emphasis on this: we do not provide services. We are not there to...we are not providers. People come to do their work, to share tools, to help each other. In other words, you go and see people who are available here who will be able to give a hand if you don't know how to use a title bench. You're going to be trained and the aim is to become more and more autonomous. That's a very special mentality" Project Manager, L'Abominable.

These filmmakers, most of whom were trained in prestigious art schools, defend aesthetic and technical choices that run counter to prevailing trends. In the age of digitization, they choose to work the old-fashioned way, with conventional photographic equipment. By reappropriating the tools of production, they defend an independent craft and art, a practice that is linked to the materiality of photographic film and its. They produce filmic works of art with very limited budgets and outside the usual distribution channels. These artists claim to represent experimental cinema and stand against established writing and aesthetics. This type of cinema is an aesthetic practice and a statement, in times when the film industry is undergoing upheaval triggered by the advent of digital technology. The aim is therefore to appropriate and teach methods and know-how before they are lost, to revive the tools of photographic film in order to show a different kind of cinema [START_REF] Cervulle | Le Cinéma Expérimental en France : Economie, Structure et acteurs[END_REF].

L'Abominable's choice to settle in the Plaine Commune area is due partly to sheer opportunity: the availability of cheap premises close to Paris. It also allowed the association to recover equipment discarded by neighbouring studios and post-production laboratories (such as Studio Éclair). The location moreover offers proximity to other stakeholders in the independent artistic scenes located in the area (e.g. Mains d'oeuvre, Les Laboratoires d'Aubervilliers). This experimental film scene remains insular and propelled mainly by self-produced directors and specific distribution channels [START_REF] Cervulle | Le Cinéma Expérimental en France : Economie, Structure et acteurs[END_REF]. Films that are post-produced at L'Abominable are often distributed by experimental film cooperatives, by the artists themselves, or by art galleries. The films are shown in specific circuits on a local, national (Fidé in Marseille) and international (Doc Lisboa, Viennale) scale. These festivals' audiences belong to networks of insiders. Films are also distributed within the contemporary art world, from the most institutional to the most independent venues. In this multi-scalar dynamics, the visibility and survival of this type of cinema depends largely on the ability of its stakeholders to join international networks while benefiting from the resources of the Parisian metropolis.

Using the camera to fight prejudice

Using the camera to carry the voices of underprivileged communities and take a different look at these stigmatized areas is the motivation of another independent films scene that journalist Claire Diao calls Double Vague (Double Wave)(2017). The name Double Vague refers to the image of a wave rolling in from the fringes, that is, from the banlieues to the centre. It is headed by a new generation of French filmmakers of immigrant descent who proudly claim their dual postcolonial identity. The term Double Vague is also an explicit reference to the Nouvelle Vague5 , although these directors do not identify with it. Instead, they usually refer to American cinema where they do find the ethnic diversity that has long been lacking on French screens. Their desires to film is fuelled by the will to offer an alternative representation of minorities and to break with the caricatured images of the banlieues conveyed by the media, especially during the 2005 riots6 . "From 2005, after the crisis in the [hoods], there was this awareness: the image that the media conveys of us is not the right one. We have to move. As much on the side of directors as on the side of festivals. We need to create places where we can show something else." A Double Wave producer To this end, and with an urgent need to film and create, these self-trained directors organize themselves without seeking first to integrate mechanisms of public support for cinema production 7 . They thus breathe new life into methods of training, production and distribution. Having access neither to the training provided by public film schools, which are extremely selective, nor to that of expensive private schools, Double Vague filmmakers train on the fringes of the system. Most of them are self-trained. The desire for filming and the need for a means of expression are urgent: they pick up a camera and shoot; their friends improvised themselves actors; their neighbourhood is the natural setting for their films. They use social networks to post their videos and benefit from a broadcasting window that was previously inaccessible, with hopes of being spotted by a major production company. Some gain experience in practical filmmaking and training workshops given by associations subsidized by local authorities, described in the first part of the paper. As they do at l'Abominable, former members who attended these activities go on to train a new generation by volunteering in the structures that supported them or by creating their own schools. Others belong to hip-hop scene: they shoot rap music videos for friends, make hip-hop culture the subject of a film, draw inspiration from the types of organization they observe in hip-hop (peer recognition; creating associations, festivals, labels and production companies; distribution through non-commercial and online channels). In keeping with the spirit of hip-hop, these artists act spontaneously, readily taking up the microphone, the camera, the spray paint, the public space.

" Filmmakers have been inspired by hip-hop in that you don't wait for permission, you make the films you want to make. And they have tried to understand how it works to do the same thing: set up an association, a production company, a festival... What's more, many of them are former rappers. There's a total filiation side to it. In the approach there are really a lot of common points. In hip-hop, they created their own labels, their own producers, they started to have gold records, the mainstream started were shaken by riots. Our respondents were deeply unsettled by U.S. media's use of the term "no-go zones" or "muslim riots" to describe them. 7 These forms of autonomous organization of the means of production are reminiscent of the Black is Beautiful movement in the United States (Rolland-Diamond, C. (2016). Black America: Une histoire des luttes pour l'égalité et la justice (XIXe-XXIe siècle). Paris: La Découverte.).

to take an interest in them when they saw that they were economically important." A Double Wave producer.

Through these self-training and self-production practices, directors are inventing a new way of shooting and making films, adapting editing and production schedules to the availability of resources. Directors promote their first films themselves via social media and crowd funding platforms. They benefit from local distribution channels, in particular local festivals that allow films to reach their first audience. These filmmakers are thus potentially able to access a form of recognition and possibly a producer, a prize or press coverage that can act as a springboard.

Their professionalization is reflected in the growing number of production companies and in a change in the scale of their distribution channels (through national and international festivals). Some dedicated workspaces had been set up in the area, such as Commune Image (a business incubator, co-working space and maker space dedicated to cinema) or Médialab 93 (a media incubator created by the advertising agency BETC when it moved there). The emergence of the Additionally, along with the establishment of the Cité du Cinéma, the Ecole de la Cité was created (in 2012) by several private producers, to serve as an alternative curriculum for out-of-school youth. EuropaCorp has also created a dedicated branch to the production of Double Vague films (UrbanFilm). These changes did not come about on their own; they were provoked by the stakeholders of these independent scenes in search of recognition and funding. When they give speeches at awards ceremonies, when books by or portraits of Double Wave filmmakers are published, or when they produce works that serve as manifestos to denounce the criteria for allocating fundings, or put out calls "to decolonize the arts"9 , they are moving towards recognition as stakeholders in the art world, in a process of artifying a film production led by self-trained directors (Heinich, Shapiro 2012). Likewise, the Manifeste des Labos, written in 2002, aimed to blow the whistle on the need to support experimental films, which is tossed about between the funding channels of visual arts and of cinema. The CNC has since recognized the existence of this hybrid cinema and now funds part of the activities of l'Abominable and other labos.

DISCUSSION: THE SCENE AS A TOOL TO RETHINK THE LOCAL EMBEDDEDNESS OF CREATIVE ACTIVITIES

The north-eastern suburbs of Paris have become a place for independent film scenes. This is the result of a series of factors that we have outlined here by conceptualizing the relationship between artistic creation and territory in terms of scenes, in order to critically examine policies for creative industries support and the presuppositions underpinning them. Yes, this area offers affordable premises close to Paris, the place to be for a newcoming artist. But this is not the only reason why this appeals filmmakers on the fringes. experiences and culture. The concept of scenes offers an alternative theoretical framework to analyse and understand the local anchoring of an artistic community. Following [START_REF] Straw | Systems of articulation, logics of change: communities and scenes in popular music studies[END_REF], the scene helps to focus on the territory as a space of shared aesthetic or moral values: for the ones, a certain vision of filmmaking based and traditional techniques, for the others, a political will to voice as postcolonial subjects. This emphasis the creation itself, what motivates artists to invent new aesthetics and new organizational forms that make them possible. The territory is the soil for the emergence of new forms of cinema, within a group of peers who share aesthetic or political affinities. Making films differently and/or with the aim to change the representations of the territory drive the filmmakers to create and invent their own production structures. These scenes emerged outside of the dominant logic of film production, fuelled by a sense of urgency, either to save equipment and techniques (experimental cinema) or to voice a discourse to break up territorial prejudice (Double Vague). They have benefited from local cultural policies for artistic creation. The scene is also a stage: this implies a relationship between artists and their audience (Bellevance and Guibert 2014). Here, public support for art creation meet youth policies through the local programs for image education. For instance, directors who have benefited from its support are required to give talks or workshops in local schools to explain their work. Their recognition is also owing to the presence of an insider audience who keeps an eye open for novelties and, through various distribution channels, follows these artists and their films (specialized festivals, artistic squats, contemporary art venues, public space and Internet). Indeed, within the Paris metropolis, there are numerous show spaces forming an audience for alternative artistic proposition, as an amateur and as a fan. The scene reminds the importance of the local atmosphere as source of inspiration for artists that contribute to the making of local subculture or style [START_REF] Lloyd | Neo-Bohemia: Art and Neighborhood Redevelopment in Chicago[END_REF], Silver, Clarke 2016). Finally, the scene relates both to interactions at the micro level and to global cultural flows, emphasizing the need to adopt a multi-scale perspective on creative environments. The emergence and structuring of independent film scenes should thus be understood within a dialectic frame, between the territory in which they are embedded (north-eastern Paris) and an international space of production, distribution and recognition. Analysing the emergence of new forms of cinema in terms of scene rather than cluster underlines the predominant role in this movement of cultural policies (image education, support for creation and distribution) rather than economic development policies (tax incentives, ad hoc real estate developments). Indeed, the filmmakers of these independent scenes find resources, spaces, inspiration and support in the territory. However, their projects take shape outside of urban planning and economic development policies set up to encourage the formation of a cluster of creative activities benefitting wellestablished players in the cinema and audiovisual industry.

  et-essai) films and films by local directors (Augros 2013) . Local authorities support art creation and projects that do not meet economic criteria with programs such as author residencies, training and initiation workshops, support for creative short films, broadcasting through festivals, etc. Various operators -be they public (Office Municipal de la Jeunesse d'Aubervilliers), para-public (cinema 93) or non-profit organizations (Côté Court, Périphérie, Espace Khiasma, L'Abominable) -offer networking and training spaces for future professionals. Several filmmakers from independent scenes thus shot their first films thanks to training and support mechanisms financed by local authorities. Their films were screened at subsidized local festivals 4 and shown in public theatres.

  Double Vague and its critical acclaim 8 has prompted stakeholders in the French film industry to create new mechanisms, such as the Fond Images de la diversité (Images of Diversity Fund) set up by the CNC in 2007. La Résidence is a new training program proposed by elitist public cinema school La Fémis since 2015 aimed at young filmmakers who do not meet the usual requirements to apply. However, La Résidence remains a parallel and shorter curriculum which does not offer the same training or networking opportunities.

Translator's note: while "banlieue" translates directly to "suburb" in terms of geography, the term has taken on a connotation in France that is the very opposite of the American "white picket fence" ideal of middle-class suburbs. When the French term appears in this text, it is to be understood in a sense which, to an American reader, would probably best translate to "the projects".

Before it can be launched, a film must go through the various stages of the film-making industry, from its conception to its release: production (gathering and combining all the resources necessary to produce a film), distribution (commercially promoting the product by choosing its release date, the number of copies and the level of promotional investment), and release in cinemas. At each of these stages, many players are involved: studios, producers, distributors, broadcasters, etc.

That is to say mainly Plaine Commune, a group of nine towns located in the north of Paris and the département (county) of Seine-Saint-Denis.

Génération Court, Côté Court, Cinébanlieue, Les Pépites du Cinéma, Regards Jeunes sur la Cité, Urban Film Festival

The Nouvelle Vague is the avant-garde movement in French cinema in the 1960s in which Agnes Varda, François Truffaut, Jean-Luc Godard and others participated.

In the fall of 2005, after two teenagers died in an encounter with police, many working-class neighbourhoods

Laurence Lascary's film L'Ascension was commercially very successful, and sold over a million tickets. Others, such as Djinn Carrenard (Donoma) or Pascal Tessaud (Rue des cités) have garnered attention from critics, and some, like Houda Benyanima (Divines), Alice Diop (Vers la tendresse) or Ladj Ly (Les Misérables) have won prestigious prizes.

This is the title of a book published by the similarly named collective: Décoloniser les arts.