Diagnostic method for aesthetic deterioration of the skin

Abstract

The present invention relates to a method for diagnosing aesthetic deterioration of a subject's skin, in particular aesthetic deterioration associated with pollution, comprising the step (a) of determining the level of at least one marker in a skin sample of the subject, the marker being selected from the group consisting of: (i) bacteria of the species Propionibacterium acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brevibacterium, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Parococcus, bacteria of the family Rhodobacteraceae and bacteria of the genus Fusobacterium, and (ii) metabolites of these bacteria selected from the group consisting of 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid / 2-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, α-ketoglutaric acid, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, citraconic acid / pentylene Diacids, fructose, fumaric acid, γ-glutamylalanine, γ-glutamylglutamine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamylserine, γ-glutamylthreonine, γ-glutamyltryptophan, γ-glutamylvaline, glutaric acid, glyceric acid, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenine, lactic acid, linoleoylethanolamide, malic acid, maleic acid, malonic acid, maltose, methionine sulfoxide, methylsuccinic acid, N-acetylalanine, N-acetyarginine, N-acetylaspartic acid, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylthreonine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelic acid, proline, salicylic acid, sebacic acid, serine, suberic acid, succinic acid, undecanedioic acid, and S-amino-ω caprolactam.

Description

Technical Field

[0001] The present invention relates to the diagnosis of aesthetic degradation of the skin, in particular pollution-related aesthetic degradation of the skin. Background Art

[0002] Many factors can lead to imperfections in the skin and its complexion. These imperfections in the complexion of the skin are the cause of increasingly frequent consultations in aesthetic care centers or at the dermatologist's office.

[0003] Among the external factors that may influence the skin tone, there may be mentioned exposure to the sun, exposure to changes in temperature and / or humidity, exposure to pollutants or cigarette smoke. Among the internal factors that may influence the skin tone, there may be mentioned stress, fatigue, hormonal changes, dehydration of the epidermis, changes in the skin barrier function, aging or excess sebum secretion.

[0004] These various factors can blur the complexion, making it uneven, dull, waxy or yellowish, and can lead to, or even favour, the presence of pigment disorders.

[0005] However, in order to prevent and / or treat these aesthetic deteriorations of the skin, it is useful to understand their causes, in particular from the perspective of the factors mentioned above.

[0006] Pollution peaks often occur in some urban environments. In daily life, especially in urban areas, individuals may be exposed to multiple insults from different air pollutants to keratin materials, especially the skin.

[0007] Among the known pollutants, mention may firstly be made of heavy metals, exhaust gases which have become a major challenge for large cities, and also fine particles or polycyclic aromatic hydrocarbons, such as benzopyrene or benzanthracene.

[0008] These pollutants will significantly lead to the deposition of particles on the epidermal surface on the skin and can cause, among other consequences, a dull complexion and / or pigmentation disorders.

[0009] Identifying exposure to pollution as a factor that contributes to aesthetic deteriorations of the skin may allow for more effective prevention and / or treatment of such deteriorations.

[0010] Therefore, there is a great need for diagnostic methods for aesthetic deterioration of the skin, especially aesthetic deterioration related to pollution. Summary of the invention

[0011] The present invention satisfies this need.

[0012] The present invention arises from the inventors' unexpected discovery that skin samples of individuals exposed to chronic pollution (based on high levels of pollutants detected in their hair samples) show significantly higher levels of certain bacteria and certain metabolites of these bacteria relative to individuals not exposed to such pollution.

[0013] The present invention therefore relates to a method for diagnosing aesthetic deterioration of the skin of a subject, in particular aesthetic deterioration associated with pollution, comprising: step (a) determining the level of at least one marker in a skin sample of the subject, said at least one marker being selected from the group consisting of: (i) bacteria comprising a nucleic acid encoding a 16S rRNA sequence that is at least 90% identical to the sequence SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9, and (ii) metabolites of these bacteria,Selected from 3-hydroxy-3-methylglutarate, 3-methylglutarate / 2-methylglutarate, 4-guanidinobutanoate, 4-imidazoleacetate, 5-oxoproline, aconitrate, adipic acid, alanine, α-ketoglutaric acid alpha-cetoglutarate), arabinonic acid / xylonic acid, azelaic acid (azelate), beta-citrylglutamate, choline, cis-urocanate, citraconate / glutaconate, fructose, fumarate, gamma-glutamylalanine, gamma-glutamylglutamine, gamma-glutamylglycine, gamma-glutamylisoleucine, gamma-glutamylleucine, gamma-glutamylfilament amino acid, γ-glutamylthreonine, γ-glutamyltryptophan, γ-glutamylvaline, glutarate, glycerate, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenate, lactate, linoleylethanolamide, malate, maleate, malonate, maltose, methionine sulfoxide, methylsuccinate ccinate), N-acetylarginine, N-acetylalanine, N-acetylaspartic acid, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylthreonine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelate, proline, salicylate, sebacate, serine, suberate, succinate, undecanedioate, and S-amino-ω caprolactam.

[0014] In a specific embodiment, the at least one marker is selected from the group consisting of: (i) bacteria of the species Propionibacterim acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brachybacterium, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Parococcus, bacteria of the family Rhodobacteraceae and bacteria of the genus Fusobacterium, and (ii) metabolites of these bacteria selected from the group consisting of 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid / 2-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, α-ketoglutaric acid, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, Citraconic acid / glutaconic acid, fructose, fumaric acid, γ-glutamylalanine, γ-glutamylglutamine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamylserine, γ-glutamylthreonine, γ-glutamyltryptophan, γ-glutamylvaline, glutaric acid, glyceric acid, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenine, lactic acid, linoleoylethanolamide, malic acid, malic acid Acid, malonic acid, maltose, methionine sulfoxide, methylsuccinic acid, N-acetylarginine, N-acetylalanine, N-acetylaspartic acid, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylthreonine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelic acid, proline, salicylic acid, sebacic acid, serine, suberic acid, succinic acid, undecanedioic acid and S-amino-ω caprolactam.

[0015] In another specific embodiment, the at least one marker is selected from the group consisting of: (i) bacteria of the species Micrococcus luteus and bacteria of the species Paracoccus sp.; and (ii) metabolites of these bacteria selected from the group consisting of kynurenine, 4-imidazoleacetic acid, maleic acid, ornithine, 4-guanidinobutyric acid, cis-urocanic acid, malonate, γ-glutamylleucine, N-acetylarginine and glycerol-3-phosphate.

[0016] Another object of the present invention relates to a method for assessing the exposure of a subject's skin to pollution, the method comprising: step (a), determining the level of at least one marker in a skin sample of the subject, the at least one marker being selected from the group consisting of: (i) bacteria, the bacteria comprising a 16S nucleotide sequence that is at least 90% identical to the sequence SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9. rRNA sequences, and (ii) metabolites of these bacteria selected from the group consisting of 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid / 2-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, α-ketoglutaric acid, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, citraconic acid / glutaconic acid, fructose, fumaric acid, γ-glutamylalanine, γ-glutamylglutamine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamylserine, γ-glutamylthreonine, γ- Glutamyltryptophan, gamma-glutamylvaline, glutaric acid, glyceric acid, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenine, lactic acid, linoleoylethanolamide, malic acid, maleic acid, malonic acid, maltose, methionine sulfoxide, methylsuccinic acid, N-acetylglycine, N-acetylalanine, N-acetylaspartic acid, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylthreonine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelic acid, proline, salicylic acid, sebacic acid, serine, suberic acid, succinic acid, undecanedioic acid, and S-amino-ω caprolactam.

[0017] Detailed description of the invention

[0018] Marker microorganisms

[0019] The marker used in the context of the present invention can be selected from the group consisting of bacteria comprising a nucleic acid encoding a 16S rRNA sequence that is at least 90% identical to the sequence SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9.

[0020] "16S rRNA" here means the ribosomal RNA that forms the small subunit of the ribosome of prokaryotes. The gene encoding 16S rRNA is called "16S rDNA". Their sequence is highly used due to its structure and is very conserved in all bacteria. In fact, it is composed of a series of conserved domains, complementary regions of universal primers used to sequence this gene, and a group of other sequence parts that are unique to bacteria, which are called signature sequences (species, genus, family, order).

[0021] Therefore, the marker used in the context of the present invention can be selected from the group consisting of bacteria comprising a 16S rRNA sequence that is at least 90% identical to the sequence SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 or SEQ ID NO:9.

[0022] In the context of the present invention, "consistency percentage" is calculated using a full sequence comparison (that is, two sequences are compared on their entire sequence). Methods for comparing the consistency of two or more sequences are well known to those skilled in the art. For example, the "needle" program can be used, wherein the "needle" program uses the Needleman-Wunsch full sequence alignment algorithm (Needleman and Wunsch (1970) J.Mol.Biol.48:443-453) to find the best comparison (including room) between two sequences by considering their entire sequence. The Needle program can be derived from, for example, the Internet site ebi.ac.uk. The consistency percentage according to the present invention is preferably calculated by the needle program (full sequence) EMBOSS under the "Gap Open" parameter equal to 10.0, the "Gap Extend" parameter equal to 0.5 and the DNAFULL matrix.

[0023] In a specific embodiment, the marker used in the context of the present application can be selected from the group consisting of bacteria, wherein the bacteria comprises a nucleic acid encoding a 16S rRNA sequence that is at least 91% identical to the sequence SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9, in particular at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 98%, at least 99%, at least 99.5%, at least 99.9% or at least 100% identical.

[0024] Sequence SEQ ID NO: 1 is a representative sequence of 16S rDNA of bacteria of the species Propionibacterim acnes.

[0025] Sequence SEQ ID NO: 2 is a representative sequence of 16S rDNA of bacteria of the Micrococcaceae family.

[0026] Sequence SEQ ID NO: 3 is a representative sequence of 16S rDNA of bacteria of the genus Brachybacterium.

[0027] Sequence SEQ ID NO: 4 is a representative sequence of 16S rDNA of bacteria of the genus Brevibacterium.

[0028] Sequence SEQ ID NO: 5 is a representative sequence of 16S rDNA of bacteria of the order Burkholderiales.

[0029] Sequence SEQ ID NO: 6 is a representative sequence of 16S rDNA of bacteria of the genus Parococcus.

[0030] Sequence SEQ ID NO: 7 is a representative sequence of 16S rDNA of bacteria of the family Rhodobacteraceae.

[0031] Sequence SEQ ID NO: 8 is a representative sequence of 16S rDNA of bacteria of the genus Fusobacterium.

[0032] Sequence SEQ ID NO: 9 is a representative sequence of 16S rDNA of bacteria of the species Micrococcus luteus.

[0033] Thus, in a particular embodiment, the marker used in the context of the present invention is chosen from the group consisting of bacteria of the species Propionibacterim acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brachybacterium, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Parococcus, bacteria of the family Rhodobacteraceae and bacteria of the genus Fusobacterium.

[0034] "Bacteria of the species Propionibacterim acnes or Cutibacterium acnes" herein refers to anaerobic and saprophytic, acne-associated, rod-shaped, Gram-positive bacteria belonging to the order Actinomycetales, the family Propionibacteriaceae and the genus Propionibacterium.

[0035] "Bacteria of the Micrococcaceae family" herein refers to spherical Gram-positive cocci and clustered, catalase+. The Micrococcaceae family includes the following genera: Acaricomes, Arthrobacter, Auridibacter, Citricoccus, Enteroactinococcus, Kocuria, Micrococcus, Nesterenkonia, Renibacterium, Rothia, Sinomonas, Tersicoccus, Yaniella, and Zhihengliuella.

[0036] In a specific embodiment, the bacterium of the family Micrococcaceae is a bacterium of the genus Micrococcus.

[0037] "Bacteria of the genus Micrococcus" herein refers to Gram-positive bacteria with a diameter of 0.5 to 2 micrometers, usually aggregated in tetrads or irregular clusters, aerobic, with oxidative metabolism, possessing catalase and chemoorganotrophy.

[0038] The genus Micrococcus includes the following species: Micrococcus luteus, Micrococcus lylae, Micrococcus varians, Micrococcus roseus, Micrococcus agilis, Micrococcus kristinae, Micrococcus nishinomiyaensis, Micrococcus sedentarius, Micrococcus halobius, Micrococcus antarticus, Micrococcus cohnii, Micrococcus endophyticus, Micrococcus flavus, Micrococcus lactis, Micrococcus terreus, and Micrococcus yunnanensis.

[0039] In a specific embodiment, the bacterium of the family Micrococcaceae is a bacterium of the species Micrococcus luteus.

[0040] "Bacteria of the genus Brachybacterium" herein refers to non-motile Gram-positive bacteria of the family Dermabacteraceae, the typical species being Brachybacterium faecium.

[0041] "Bacteria of the genus Brevibacterium" herein refers to Gram-positive bacteria of the order Actinomycetales and the family Brevibacteriaceae.

[0042] "Bacteria of the order Burkholderiales" herein refers to Gram-negative Proteobacteria. The order Burkholderiales includes the families Alcaligenaceae, Burkholderiaceae, Comamonadadeae, Oxalobacteraceae, and Sutterellaceae.

[0043] "Bacteria of the family Rhodobacteraceae" herein refers to Gram-negative Proteobacteria of the order Rhodobacterales.

[0044] The family Erythrobacteria includes the following genera: Actibacterium, Acuticoccus, Aestuariibius, Aestuariicoccus, Aestuariivita, Agaricicola, Ahrensia, Albidovulum, Albimonas, Albirhodobacter, Aliiroseovarius, Aliisedimentitalea, Alkalimicrobium, Amaricoccus, Amylibacter, Antarctobacter, Aquicoccus, Aquimixticola, Ascidiaceihabitans, Boseongicola, Brevirhabdus, Catellibacterium, Celeribacter, Cereibacter, Citreicella, Citreimonas, Cognatishimia, Cognatiyoonia, Confluentimicrobium, Cribrihabitans, Defluviimonas, Dinoroseobacter, Donghicola, Epiba cterium, Falsirhodobacter, Flavimaricola, Frigidibacter, Gaetbulicola, Gemmobacter, Haematobacter, Halocynthiibacter, Halodurantibacterium, Halovulum, Hasllibacter, Hirschia, Huaishuia, Hwanghaeicola, Hyphomonas, Jannaschia, Jhaorihella, Kandeliimicrobium, Ketogulonicigenium, Labrenzia, Lacimonas, Leisingera, Lentibacter, Limibaculum, Limimaricola, Litoreibacter, Litorimicrobium, Litorisediminicola, Litorisediminivivens, Litorivivens, Loktanella, Lutimaribacter, Maliponia, Mameliella, Mangrovicoccus,Maribius, Maricaulis, Marimonas, Marinibacterium, Marinovum, Maritimibacter, Marivita, Marivivens, Methylarcula, Monaibacterium, Nautella, Neptunicoccus, Nereida, Nesiotobacter, Nioella, Nitropelagi, Oceanibulbus, Oceanicaulis, Oceanicella, Oceanicola, Oceaniglobus, Oceaniovalibus, Octadecabacter, Pacificibacter, Paenirhodobacter, Palleronia, Pannonibacter, Paracoccus, Paradonghicola, Paraphaeobacter, Pararhodobacter, Pelagibaca, Pelagicola, Pelagimonas, Phaeobacter, Planktomarina, Planktotalea, Plastorhodobacter, Pleomorphobacterium, Polymorphum, Pontibaca, Ponticoccus, Pontivivens, Primorskyibacter, Profundibacterium, Pseudaestuariivita, Pseudodonghicola, Pseudohalocynthiibacter, Pseudomaribius, Pseudooceanicola, Pseudooctadecabacter, Pseudopelagicola, Pseudophaeobacter, Pseudorhodobacter, Pseudoroseicyclus, Pseudoroseovarius, Pseudoruegeria, Pseudoseohaeicola, Pseudovibrio, Psychromarinibacter, Puniceibacterium, Rhodobaca, Rhodobaculum, Rhodosalinus, Rhodovulum, Roseibaca, RoseibacteriumRoseibacula, Roseibium, Roseicitreum, Roseicyclus, Roseinatronobacter, Roseisalinus, Roseivivax, Roseobacter, Roseovarius, Rubellimicrobium, Rubribacterium um, Rubricella, Rubrimonas, Ruegeria, Sagittula, Salinihabitans, Sali novum、Salipiger、Sedimentitalea、Sediminimonas、Seohaeicola、Shimia、Silicibacte r、Silicimonas、Sinorhodobacter、Staleya、Stappia、Sulfitobacter、Tabrizi cola、Tateyamaria、Thalassobacter、Thalassobius、Thalassococcus、Thiobacimonas、T hyoclava, Thiosphaera, Tranquillimonas, Tritonibacter, Tropicibacter, Tropicimonas, Vadicella, Wenxinia, Xinfangfangia, Xuhuaishua, Yangia, Yoonia, Young.

[0045] "Bacteria of the genus Paracoccus" refers to Gram-negative bacteria of the family Erythrobacterium, and a typical species is Paracoccus dendrificans. The genus Paracoccus includes the following species: Paracoccus acridae, Paracoccus aerius, Paracoccus aestuarii, Paracoccus aestuariivivens, Paracoccus alcaliphilus, Paracoccus alimentarius, Paracoccus alkenifer, Paracoccus aminophilus, Paracoccus aminovorans, Paracoccus angustae, Paracoccus bengalensis, Paracoccus caeni, Paracoccus carotinifaciens, Paracoccus cavernae, Paracoccus chinensis, Paracoccus communis, Paracoccus contaminans, Paracoccus denitrificans, Paracoccus fistulariae, Paracoccus fontiphilus, Paracoccus haeundaensis, Paracoccus halodenitrificans, Paracoccus halophilus, Paracoccus hibisci, Paracoccus hibiscisoli, Paracoccus homiensis, Paracoccus huijuniae, Paracoccus isoporae, Paracoccus kocurii, Paracoccus kondratievae, Paracoccus koreensis, Paracoccus laeviglucosivorans, Paracoccus limosus, Paracoccus litorisideminis, Paracoccus lutimaris, Paracoccus mangrovi, Paracoccus marcusii, Paracoccus marinus, Paracoccus methylutens, Paracoccus niistensis, Paracoccus pacificus, Paracoccuspanacisoli, Paracoccus pantotrophus, Paracoccusrhizosphaerae, Paracoccus saliphilus, Paracoccus sanguinis, Paracoccussediminis, Paracoccus seriniphilus, Paracoccus solventivorans, Paracoccussordidisoli, Paracoccus sphaerophysae, Paracoccus stylophorae, Paracoccus sulfuroxidans, Paracoccus thiocyanatus, Paracoccus tibetensis, Paracoccusversutus, Paracoccus yeei, Paracoccus zeaxanthinifaciens and Paracoccus sp. strains.

[0046] In a specific embodiment, the bacterium of the genus Paroccocus is a strain of Paracoccus sp.

[0047] "Bacteria of the genus Fusobacterium" herein refers to anaerobic filamentous Gram-negative bacteria of the family Fusobacteriaceae. The genus Fusobacterium includes the following species: Fusobacterium necrophorum, Fusobacterium nucleatum, Fusobacterium polymorphum, and Fusobacterium novum.

[0048] In a specific embodiment, the marker used in the context of the present invention is selected from the group consisting of the following bacterial species: Micrococcus luteus and Paracoccus sp.

[0049] Marker metabolites

[0050] The marker used in the context of the present invention may be selected from the group consisting of metabolites, in particular metabolites produced by the bacteria defined above in the section "marker microorganisms", selected from the group consisting of 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid / 2-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, α-ketoglutaric acid, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, citraconic acid / glutaconic acid, fructose, fumaric acid, γ-glutamylalanine, γ-glutamylglutamine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamyl acylserine, γ-glutamylthreonine, γ-glutamyltryptophan, γ-glutamylvaline, glutaric acid, glyceric acid, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenine, lactic acid, linoleoylethanolamide, malic acid, maleic acid, malonic acid, maltose, methionine sulfoxide, methylsuccinic acid, N-acetylglycine, N-acetylalanine, N-acetylaspartic acid, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylthreonine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelic acid, proline, salicylic acid, sebacic acid, serine, suberic acid, succinic acid, undecanedioic acid, and S-amino-ω caprolactam.

[0051] In a specific embodiment, the metabolite is selected from kynurenine, 4-imidazoleacetic acid, maleic acid, ornithine, 4-guanidinobutyric acid, cis-urocanic acid, malonate, gamma-glutamylleucine, N-acetylarginine, and glycerol-3-phosphate.

[0052] As is well known to those skilled in the art, one or more of these metabolites may be produced by one or more of the bacteria defined above in the "Marker Microbiome" section.

[0053] Indeed, bacteria of the species P. acnes produce proline, alanine, glycine, serine, choline, lactic acid, malic acid, maleic acid, α-ketoglutarate and succinic acid as metabolites.

[0054] Bacteria of the family Micrococcaceae produce alanine, serine, fructose, and lactic acid as metabolites.

[0055] Bacteria of the genus Brevibacterium produce proline, glycine, serine, fructose, and maltose as metabolites.

[0056] Bacteria of the genus Brevibacterium produce proline, alanine, serine, ornithine, choline, fructose, maltose, fumaric acid, lactic acid, malic acid, glutaric acid, α-ketoglutaric acid, succinic acid, malonic acid, aconitic acid, adipic acid, salicylic acid, and sebacic acid as metabolites.

[0057] Bacteria of the order Burkholderiales produce proline, alanine, glycine, serine, lactate, malate, and succinate as metabolites.

[0058] Bacteria of the genus Paracoccus produce proline, alanine, glycine, serine, ornithine, choline, fructose, maltose, fumaric acid, lactic acid, malic acid, α-ketoglutaric acid, succinic acid, maleic acid, malonic acid, aconitic acid, and N-acetylphenylalanine as metabolites.

[0059] Bacteria of the family Rhodobacteraceae produce alanine, glycine, glutamine, ornithine, and succinate as metabolites.

[0060] Bacteria of the genus Fusobacterium produce proline, alanine, glycine, serine, ornithine, choline, fructose, maltose, fumaric acid, lactic acid, malic acid, glutaric acid, α-ketoglutaric acid, and succinic acid as metabolites.

[0061] The metabolites produced by these different bacteria are typically described in Palazzi et al. (2018) Environ. Int. 121: 1341-1354, Thierry et al. (2011) Int. J. Food Microbiol. 149: 19-27, Krulwich et al. (1976) J. Bacteriol. 127: 179-183, Mori et al. (1987) Agricultural Biol. Chem. 51: 129-138, Levy et al. (2017) Current Op. in Microbiol. 25: 8-15, Jin et al. (2014) Mol. Pharmacol. 85: 777-788, Hug et al. (1999) J. Photochem. Photobiol. B 50: 66-73 and Wolfe et al. (2014) Mol. Pharmacol. 85: 777-788. al. (2016) Curr. Genet. 62:335-341.

[0062] Thus, in a specific embodiment, aconitic acid is produced by a bacterium of the genus Parococcus and / or by a bacterium of the genus Brevibacterium.

[0063] In a specific embodiment, adipic acid is produced by bacteria belonging to the genus Brevibacterium.

[0064] In a specific embodiment, alanine is produced by bacteria of the species Propionibacterium acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Paracoccus, bacteria of the family Rhodobacteraceae, and / or bacteria of the genus Fusobacterium.

[0065] In a specific embodiment, alpha-ketoglutarate is produced by bacteria of the species Propionibacterium acnes, bacteria of the genus Brevibacterium, bacteria of the genus Paracoccus and / or bacteria of the genus Fusobacterium.

[0066] In specific embodiments, choline is produced by bacteria of the species Propionibacterium acnes, Brevibacterium, Paracoccus, and / or Fusobacterium.

[0067] In a specific embodiment, fructose is produced by bacteria of the species Propionibacterium acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brevibacterium, bacteria of the genus Brevibacterium, bacteria of the genus Paracoccus and / or bacteria of the genus Fusobacterium.

[0068] In a specific embodiment, the fumaric acid is produced by a bacterium of the genus Brevibacterium, a bacterium of the genus Paracoccus, and / or a bacterium of the genus Fusobacterium.

[0069] In a specific embodiment, the gamma-glutamylglutamine is produced by a bacterium of the family Rhodobacteraceae.

[0070] In a specific embodiment, glutaric acid is produced by bacteria of the genus Brevibacterium and / or bacteria of the genus Fusobacterium.

[0071] In a specific embodiment, glycine is produced by bacteria of the species Propionibacterium acnes, bacteria of the genus Brevibacterium, bacteria of the orders Burkholderiales, bacteria of the genus Paracoccus, bacteria of the family Rhodobacteraceae, and / or bacteria of the genus Fusobacterium.

[0072] In a specific embodiment, the lactic acid is produced by bacteria of the species Propionibacterium acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brevibacterium, bacteria of the orders Burkholderiales, bacteria of the genus Paracoccus and / or bacteria of the genus Fusobacterium.

[0073] In a specific embodiment, the malic acid is produced by bacteria of the species Propionibacterium acnes, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Paracoccus, and / or bacteria of the genus Fusobacterium.

[0074] In a specific embodiment, the maleic acid is produced by bacteria of the species Propionibacterium acnes and / or bacteria of the genus Paracoccus.

[0075] In a specific embodiment, malonate is produced by bacteria of the genus Brevibacterium and / or bacteria of the genus Paracoccus.

[0076] In a specific embodiment, maltose is produced by bacteria of the genus Brevibacterium, bacteria of the genus Brevibacterium, bacteria of the genus Paracoccus and / or bacteria of the genus Fusobacterium.

[0077] In a specific embodiment, N-acetylphenylalanine is produced by a bacterium of the genus Paracoccus.

[0078] In specific embodiments, ornithine is produced by a bacterium of the genus Brevibacterium, a bacterium of the genus Paracoccus, a bacterium of the family Rhodobacteraceae, and / or a bacterium of the genus Fusobacterium.

[0079] In a specific embodiment, proline is produced by bacteria of the species Propionibacterium acnes, bacteria of the genus Brevibacterium, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Paracoccus, and / or bacteria of the genus Fusobacterium.

[0080] In a specific embodiment, the salicylic acid is produced by Brevibacterium spp.

[0081] In a specific embodiment, sebacic acid is produced by bacteria of the genus Brevibacterium.

[0082] In a specific embodiment, serine is produced by bacteria of the species Propionibacterium acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brevibacterium, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Paracoccus, and / or bacteria of the genus Fusobacterium.

[0083] In specific embodiments, succinic acid is produced by bacteria of the species Propionibacterium acnes, bacteria of the genus Brevibacterium, bacteria of the orders Burkholderiales, bacteria of the genus Paracoccus, bacteria of the family Rhodobacteraceae, and / or bacteria of the genus Fusobacterium.

[0084] Diagnostic Methods

[0085] The diagnostic method according to the invention is a method for diagnosing aesthetic deterioration of the skin, in particular aesthetic deterioration associated with pollution.

[0086] Aesthetic deterioration of the skin is in particular selected from aesthetic pigmentation disorders, lack of radiance and uneven skin tone.

[0087] "Aesthetic pigmentary disorders" are defined herein as aesthetic imperfections of the skin caused by insufficient or absent pigment or by excessive or uneven distribution of pigment. Aesthetic pigmentary disorders include, in particular, uneven skin color, solar lentigo, melasma and pigmented spots.

[0088] In a specific embodiment, the aesthetic deterioration of the skin is selected from pigmented spots and uneven skin color.

[0089] Skin refers in particular to the skin of the face (especially the skin of the cheeks and / or forehead), the skin of the décolleté, the skin of the neck, the skin of the arms and forearms. More preferably, skin refers to the skin of the face (especially the cheeks) and / or the skin of the neck.

[0090] In specific embodiments, the aesthetic degradation of the skin is associated with, in particular due to, pollution.

[0091] "Pollution" herein refers to exposure to particles of matter, particularly exposure to polycyclic aromatic hydrocarbons (PAHs).

[0092] In a specific embodiment, pollution refers to exposure to particulate matter, particularly exposure to PAH, resulting in the following levels of PAH and PAH metabolites in the subject's hair:

[0093] The level of 2-OH-phenanthrene was at least 1.8 times the control level;

[0094] The level of 3-OH-fluorene was at least 1.6 times that of the control level;

[0095] The level of 3-OH-phenanthrene was at least 1.7 times the control level;

[0096] The level of Bb-fluoranthene was at least 1.8 times the control level;

[0097] The levels of benzo-ghi-perylene were at least 1.7 times the control levels;

[0098] The level of fluoranthene is at least 2 times the control level, and / or

[0099] The level of pyrene was at least 1.6 times the control level;

[0100] The control level generally refers to the level of the compound in hair samples of subjects living in less polluted cities, particularly cities where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year.

[0101] The diagnostic method according to the present invention comprises: step (a), determining the level of at least one marker in a skin sample of a subject, wherein the at least one marker is selected from the group consisting of: (i) bacteria as defined in the "Marker Microorganisms" section above, and (ii) metabolites as defined in the "Marker Metabolites" section above.

[0102] The level of the at least one marker may be determined by any suitable technique.

[0103] In a specific embodiment, the marker is a bacterium as defined above in the "Marker Microorganisms" section and the level of the at least one marker is determined by measuring the level of the corresponding 16S rDNA gene.

[0104] Preferably, the marker is a bacterium as defined above in the section "Marker Microorganisms", and the level of the at least one marker is determined by PCR amplification and sequencing of the V1-3 region of 16S rDNA.

[0105] Typically, genomic bacterial DNA present in the skin sample is extracted and then amplified by PCR using primers targeting the V1-3 region of the bacterial 16S rDNA. The 16S rDNA amplicons obtained are sequenced, allowing the corresponding bacteria to be identified. The levels of each bacterial 16S rDNA identified are usually measured in parallel using quantitative PCR.

[0106] In another specific embodiment, said marker is a metabolite as defined above in the "Marker Metabolites" section, and the level of said at least one marker is determined by liquid chromatography-mass spectrophotometer (LC / MS / MS).

[0107] Typically, skin samples are extracted and subjected to liquid chromatography-mass spectrometry (LC / MS / MS) analysis. The various metabolites present are typically identified based on retention index, mass correspondence with data in a library, and "forward" and reverse MS / MS scores relative to standards, and the levels of metabolites are typically measured by quantifying the peaks obtained by LC / MS / MS using the area under the primary MS ion curve.

[0108] In a specific embodiment, the diagnostic method according to the present invention further comprises the steps consisting of:

[0109] (b) comparing the level of the at least one marker determined in step (a) with a control, and

[0110] (c) determining, based on the comparison in step (b), whether the subject's skin exhibits aesthetic degradation, in particular, exhibits aesthetic degradation associated with contamination.

[0111] In a specific embodiment, the control is a reference value.

[0112] In a specific embodiment, the reference value is determined by the mean value of the level of said marker in a defined population (eg, an age-defined and / or skin-type-defined population).

[0113] In a specific embodiment, the reference value is the mean value of the marker level in a population of subjects, in particular subjects as defined below, living in a less polluted city, in particular a city with an air quality index below 100 for less than 100 days, in particular less than 85 days per year.

[0114] In a specific embodiment, the subject's skin is diagnosed as exhibiting aesthetic degradation, particularly pollution-related aesthetic degradation, when:

[0115] - Levels of Propionibacterium acnes species bacteria determined in the subject's skin sample that are at least 0.9 times the control level;

[0116] - Levels of Micrococcaceae bacteria identified in the subject's skin specimen that are at least 2.2 times higher than the control level;

[0117] - The level of Brevibacterium bacteria determined in the subject's skin sample is at least 1.7 times higher than the control level;

[0118] - Levels of Brevibacterium bacteria identified in the subject's skin specimen that are at least 2.4 times higher than the control level;

[0119] - The level of Burkholderiales bacteria determined in the subject's skin specimen is at least 1.8 times the control level;

[0120] - The level of Parococcus bacteria identified in the subject's skin specimen is at least 1.4 times the control level;

[0121] - the level of Rhodobacteraceae bacteria determined in the subject's skin sample is at least 1.4 times the control level; and / or

[0122] - the level of Fusobacterium bacteria determined in the subject's skin sample is at least 0.7 times the control level;

[0123] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0124] The level of a marker is typically determined by measuring the level of the corresponding 16S rDNA gene, particularly by PCR amplification combined with sequencing the V1-3 region of the 16S rDNA, generally as described above.

[0125] In a specific embodiment, the subject's skin is diagnosed as exhibiting aesthetic degradation, particularly pollution-related aesthetic degradation, when:

[0126] - The level of Micrococcus luteus bacteria determined in the subject's skin sample is at least 1.01 times the control level, and / or

[0127] - Levels of Paracoccus bacteria identified in the subject's skin specimen that are at least 1.4 times the control level;

[0128] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0129] The level of a marker is typically determined by measuring the level of the corresponding 16S rDNA gene, particularly by PCR amplification combined with sequencing the V1-3 region of the 16S rDNA, generally as described above.

[0130] In a specific embodiment, the subject's skin is diagnosed as exhibiting aesthetic degradation, particularly pollution-related aesthetic degradation, when:

[0131] - 3-hydroxy-3-methylglutaric acid levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0132] - 3-methylglutaric acid / 2-methylglutaric acid levels determined in the subject's skin sample are at least 1.3 times the control level;

[0133] - 4-guanidinobutyric acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0134] - 4-imidazoleacetic acid levels determined in the subject's skin specimens that are at least 1.4 times the control level;

[0135] - 5-oxoproline levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0136] - Aconitic acid level determined in the subject's skin sample is at least 1.4 times the control level;

[0137] - Adipic acid level determined in the subject's skin sample is at least 1.2 times the control level;

[0138] - Alanine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0139] - alpha-ketoglutarate levels determined in the subject's skin sample that are at least 1.1 times the control level;

[0140] - Arabinonic acid / xylonic acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0141] - A level of azelaic acid determined in the subject's skin sample is at least 1.2 times the control level;

[0142] - β-citrylglutamate levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0143] - Choline levels determined in the subject's skin sample are at least 1.2 times the control level;

[0144] - CSA levels determined in the subject's skin sample are at least 1.5 times the control level;

[0145] - Citraconic acid / glutaconic acid levels determined in the subject's skin sample are at least 1.3 times the control level;

[0146] - fructose levels determined in the subject's skin sample that are at least 1.1 times the control level;

[0147] - fumaric acid levels determined in the subject's skin sample that are at least 1.2 times the control level;

[0148] - Gamma-glutamylalanine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0149] - Gamma-glutamylglutamine levels determined in the subject's skin sample are at least 1.2 times the control level;

[0150] - Gamma-glutamylglycine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0151] - Gamma-glutamylisoleucine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0152] - Gamma-glutamylleucine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0153] - Gamma-glutamylserine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0154] - γ-glutamylthreonine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0155] - Gamma-glutamyltryptophan levels determined in the subject's skin sample are at least 1.4 times the control level;

[0156] - Gamma-glutamylvaline levels determined in the subject's skin sample are at least 1.4 times the control level;

[0157] - glutaric acid levels determined in the subject's skin sample that are at least 1.1 times the control level;

[0158] - Glyceric acid levels determined in the subject's skin sample are at least 1.1 times the control level;

[0159] - Glycerol-3-phosphate levels determined in the subject's skin sample are at least 1.6 times the control level;

[0160] - Glycine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0161] - Isovalerylglycine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0162] - Kynurenine levels determined in the subject's skin sample are at least 2.3 times the control level;

[0163] - Lactic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0164] - Linoleoylethanolamide levels determined in the subject's skin sample are at least 1.3 times the control level;

[0165] - Malic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0166] - Maleic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0167] - Malonic acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0168] - Maltose levels determined in the subject's skin sample are at least 1.6 times the control level;

[0169] - Methionine sulfoxide levels determined in the subject's skin specimens are at least 1.2 times the control level;

[0170] - Levels of methylsuccinate determined in the subject's skin sample that are at least 1.3 times the control level;

[0171] - N-acetylarginine levels determined in the subject's skin sample that are at least 1.6 times the control level;

[0172] - N-acetylalanine levels determined in the subject's skin sample are at least 1.2 times the control level;

[0173] - N-acetylaspartate levels determined in the subject's skin sample that are at least 1.4 times the control level;

[0174] - N-acetylglycine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0175] - N-acetylhistidine levels determined in the subject's skin sample are at least 1.2 times the control level;

[0176] - N-acetylphenylalanine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0177] - N-acetylthreonine determined in the subject's skin sample is at least 1.1 times the control level;

[0178] - N-acetylvaline levels determined in the subject's skin sample that are at least 1.2 times the control level;

[0179] - Oleamide levels determined in the subject's skin sample are at least 1.3 times the control level;

[0180] - ornithine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0181] - Palmitamide levels determined in the subject's skin sample are at least 1.4 times the control level;

[0182] - pimelic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0183] - Proline levels determined in the subject's skin sample are at least 1.3 times the control level;

[0184] - Salicylic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0185] - sebacic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0186] - serine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0187] - Suberic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0188] - Succinate levels determined in the subject's skin sample are at least 1.2 times the control level;

[0189] - the level of undecanedioic acid determined in the subject's skin sample is at least 1.3 times the control level, and / or

[0190] - S-amino-ω caprolactam levels determined in the subject's skin specimens that are at least 2.3 times the control level;

[0191] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0192] The levels of markers are typically determined by liquid chromatography-mass spectrometry (LC / MS / MS), generally as described above.

[0193] In a specific embodiment, the subject's skin is diagnosed as exhibiting aesthetic degradation, particularly pollution-related aesthetic degradation, when:

[0194] - Kynurenine levels determined in the subject's skin sample are at least 2.3 times the control level;

[0195] - 4-imidazoleacetic acid levels determined in the subject's skin specimens that are at least 1.4 times the control level;

[0196] - Maleic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0197] - ornithine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0198] - 4-guanidinobutyric acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0199] - Citracenic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0200] - Malonic acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0201] - Gamma-glutamylleucine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0202] - N-acetylarginine levels determined in the subject's skin sample are at least 1.6 times the control level, and / or

[0203] - Glycerol-3-phosphate levels determined in the subject's skin sample are at least 1.6 times the control level;

[0204] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0205] The levels of markers are typically determined by liquid chromatography-mass spectrometry (LC / MS / MS), generally as described above.

[0206] According to a preferred embodiment, the skin sample of the subject used in the diagnostic method according to the present invention is collected from the subject's skin, preferably from the subject's face, in particular from the subject's cheek and / or forehead, preferably in a non-invasive manner. Preferably, the skin sample is from the stratum corneum.

[0207] The stratum corneum is the outermost layer of the epidermis and includes the surface of the skin. It is mainly composed of dead cells.

[0208] According to an embodiment, the diagnostic method according to the invention comprises the step of collecting a skin sample from a subject. This step is preferably performed in a non-invasive manner, and in particular in a manner that does not require local anesthesia. According to a preferred embodiment, the skin sample is collected by rubbing the skin surface or using a method such as The sample collection step is performed using the sticky surface of the plate.

[0209] In a specific embodiment, the skin sample is collected using The disk is collected.

[0210] "Subject" herein refers to a human being preferably aged between 25 and 45. Preferably, the subject is a female. Preferably, the subject is of Asian type.

[0211] The present invention also relates to a method for the cosmetic treatment of the skin of a subject showing aesthetic deterioration, in particular related to pollution, comprising the steps of:

[0212] A) Diagnosing a subject showing aesthetic deterioration of the skin, in particular aesthetic deterioration associated with pollution, by carrying out the diagnostic method according to the invention.

[0213] B) If the subject is diagnosed as showing aesthetic degradation of the skin, in particular aesthetic degradation related to pollution, treating the skin of said subject with a cosmetic composition capable of reducing and / or slowing down the aesthetic degradation of the skin.

[0214] Methods for assessing skin exposure to pollution

[0215] The present invention also relates to a method for assessing a subject's skin exposure to pollution, comprising: step (a), determining the level of at least one marker in a skin sample of the subject, the at least one marker being selected from the group consisting of: (i) bacteria as defined in the "Marker Microorganisms" section above and (ii) metabolites as defined in the "Marker Metabolites" section above.

[0216] The contamination is as defined above in the section "Diagnostic methods".

[0217] The determination of step (a) is preferably performed as described above for the diagnostic method.

[0218] The skin sample is as defined above in the "Diagnostic Methods" section.

[0219] The subject is as defined above in the "Diagnostic Methods" section.

[0220] In a specific embodiment, the evaluation method according to the present invention further comprises the steps consisting of:

[0221] (b) comparing the level of the at least one marker determined in step (a) with a control, and

[0222] (c) determining whether the subject has experienced skin exposure to the contaminant based on the comparison in step (b).

[0223] In certain embodiments, the control is a reference value.

[0224] In a specific embodiment, the reference value is determined by the mean value of the level of said marker in a defined population (eg, an age-defined and / or skin-type-defined population).

[0225] In a specific embodiment, the reference value is the mean value of the marker level in a population of subjects, in particular subjects as defined below, living in a less polluted city, in particular a city with less than 100, in particular less than 85 days per year with an air quality index below 100.

[0226] In a specific embodiment, a subject is determined to have experienced skin exposure to pollution if:

[0227] - Levels of Propionibacterium acnes species bacteria determined in the subject's skin sample that are at least 0.9 times the control level;

[0228] - Levels of Micrococcaceae bacteria identified in the subject's skin specimen that are at least 2.2 times higher than the control level;

[0229] - The level of Brevibacterium bacteria determined in the subject's skin specimen is at least 1.7 times the control level;

[0230] - Levels of Brevibacterium bacteria identified in the subject's skin specimen that are at least 2.4 times higher than the control level;

[0231] - The level of Burkholderiales bacteria determined in the subject's skin specimen is at least 1.8 times the control level;

[0232] - The level of Parococcus bacteria identified in the subject's skin sample is at least 1.4 times the control level;

[0233] - Levels of Rhodobacteraceae identified in the subject's skin specimen that are at least 1.4 times higher than the control level;

[0234] - the level of Fusobacterium bacteria determined in the subject's skin sample is at least 0.7 times the control level;

[0235] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0236] The level of a marker is typically determined by measuring the level of the corresponding 16S rDNA gene, particularly by PCR amplification combined with sequencing the V1-3 region of the 16S rDNA, generally as described above.

[0237] In a specific embodiment, a subject is determined to have experienced skin exposure to pollution if:

[0238] - The level of Micrococcus luteus bacteria determined in the subject's skin sample is at least 1.01 times the control level, and / or

[0239] - Levels of Paracoccus bacteria identified in the subject's skin specimen that are at least 1.4 times the control level;

[0240] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0241] The level of a marker is typically determined by measuring the level of the corresponding 16S rDNA gene, particularly by PCR amplification combined with sequencing the V1-3 region of the 16S rDNA, generally as described above.

[0242] In a specific embodiment, a subject is determined to have experienced skin exposure to pollution if:

[0243] - 3-hydroxy-3-methylglutaric acid levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0244] - 3-methylglutaric acid / 2-methylglutaric acid levels determined in the subject's skin sample are at least 1.3 times the control level;

[0245] - 4-guanidinobutyric acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0246] - 4-imidazoleacetic acid levels determined in the subject's skin specimens that are at least 1.4 times the control level;

[0247] - 5-oxoproline levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0248] - Aconitic acid level determined in the subject's skin sample is at least 1.4 times the control level;

[0249] - Adipic acid level determined in the subject's skin sample is at least 1.2 times the control level;

[0250] - Alanine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0251] - alpha-ketoglutarate levels determined in the subject's skin sample that are at least 1.1 times the control level;

[0252] - Arabinonic acid / xylonic acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0253] - A level of azelaic acid determined in the subject's skin sample is at least 1.2 times the control level;

[0254] - β-citrylglutamate levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0255] - Choline levels determined in the subject's skin sample are at least 1.2 times the control level;

[0256] - CSA levels determined in the subject's skin sample are at least 1.5 times the control level;

[0257] - Citraconic acid / glutaconic acid levels determined in the subject's skin sample are at least 1.3 times the control level;

[0258] - fructose levels determined in the subject's skin sample that are at least 1.1 times the control level;

[0259] - fumaric acid levels determined in the subject's skin sample that are at least 1.2 times the control level;

[0260] - Gamma-glutamylalanine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0261] - Gamma-glutamylglutamine levels determined in the subject's skin sample are at least 1.2 times the control level;

[0262] - Gamma-glutamylglycine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0263] - Gamma-glutamylisoleucine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0264] - Gamma-glutamylleucine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0265] - Gamma-glutamylserine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0266] - γ-glutamylthreonine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0267] - Gamma-glutamyltryptophan levels determined in the subject's skin sample are at least 1.4 times the control level;

[0268] - Gamma-glutamylvaline levels determined in the subject's skin sample are at least 1.4 times the control level;

[0269] - glutaric acid levels determined in the subject's skin sample that are at least 1.1 times the control level;

[0270] - Glyceric acid levels determined in the subject's skin sample are at least 1.1 times the control level;

[0271] - Glycerol-3-phosphate levels determined in the subject's skin sample are at least 1.6 times the control level;

[0272] - Glycine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0273] - Isovalerylglycine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0274] - Kynurenine levels determined in the subject's skin sample are at least 2.3 times the control level;

[0275] - Lactic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0276] - Linoleoylethanolamide levels determined in the subject's skin sample are at least 1.3 times the control level;

[0277] - Malic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0278] - Maleic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0279] - Malonic acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0280] - Maltose levels determined in the subject's skin sample are at least 1.6 times the control level;

[0281] - Methionine sulfoxide levels determined in the subject's skin specimens are at least 1.2 times the control level;

[0282] - Levels of methylsuccinate determined in the subject's skin sample that are at least 1.3 times the control level;

[0283] - N-acetylarginine levels determined in the subject's skin sample that are at least 1.6 times the control level;

[0284] - N-acetylalanine levels determined in the subject's skin sample are at least 1.2 times the control level;

[0285] - N-acetylaspartate levels determined in the subject's skin sample that are at least 1.4 times the control level;

[0286] - N-acetylglycine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0287] - N-acetylhistidine levels determined in the subject's skin sample are at least 1.2 times the control level;

[0288] - N-acetylphenylalanine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0289] - N-acetylthreonine determined in the subject's skin sample is at least 1.1 times the control level;

[0290] - N-acetylvaline levels determined in the subject's skin sample that are at least 1.2 times the control level;

[0291] - Oleamide levels determined in the subject's skin sample are at least 1.3 times the control level;

[0292] - ornithine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0293] - Palmitamide levels determined in the subject's skin sample are at least 1.4 times the control level;

[0294] - pimelic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0295] - Proline levels determined in the subject's skin sample are at least 1.3 times the control level;

[0296] - Salicylic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0297] - sebacic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0298] - serine levels determined in the subject's skin sample are at least 1.3 times the control level;

[0299] - Suberic acid levels determined in the subject's skin sample are at least 1.2 times the control level;

[0300] - Succinate levels determined in the subject's skin sample are at least 1.2 times the control level;

[0301] - the level of undecanedioic acid determined in the subject's skin sample is at least 1.3 times the control level, and / or

[0302] - S-amino-ω caprolactam levels determined in the subject's skin specimens that are at least 2.3 times the control level;

[0303] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0304] The levels of markers are typically determined by liquid chromatography-mass spectrometry (LC / MS / MS), generally as described above.

[0305] In a specific embodiment, a subject is determined to have experienced skin exposure to pollution if:

[0306] - Kynurenine levels determined in the subject's skin sample are at least 2.3 times the control level;

[0307] - 4-imidazoleacetic acid levels determined in the subject's skin specimens that are at least 1.4 times the control level;

[0308] - Maleic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0309] - ornithine levels determined in the subject's skin sample that are at least 1.3 times the control level;

[0310] - 4-guanidinobutyric acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0311] - Citracenic acid levels determined in the subject's skin sample are at least 1.5 times the control level;

[0312] - Malonic acid levels determined in the subject's skin sample are at least 1.4 times the control level;

[0313] - Gamma-glutamylleucine levels determined in the subject's skin sample are at least 1.4 times the control level;

[0314] - N-acetylarginine levels determined in the subject's skin sample are at least 1.6 times the control level, and / or

[0315] - Glycerol-3-phosphate levels determined in the subject's skin sample are at least 1.6 times the control level;

[0316] The control level generally refers to the level of the marker in a skin sample of a subject living in a less polluted city, particularly a city where the air quality index is below 100 for less than 100 days (particularly less than 85 days) per year, and

[0317] The levels of markers are typically determined by liquid chromatography-mass spectrometry (LC / MS / MS), generally as described above.

[0318] The present invention is described in more detail by means of the following figures and examples.

[0319] Sequence Description

[0320]

[0321]

[0322] BRIEF DESCRIPTION OF THE DRAWINGS

[0323] Figure 1 : Appearance and severity (mean score) of spreading yellow spots on the cheeks and forehead in women in the GP1 and GP2 groups as defined in the Examples.

[0324] Figure 2 : The severity of the spreading macules in each group, with a clinical score of 1 to 4 (4 being the most severe). DETAILED DESCRIPTION

[0325] Example

[0326] The following example shows the identification of a signature consisting of 60 metabolites and 8 microorganisms that were significantly modulated in skin samples of individuals exposed to chronic pollution (based on high levels of pollutants detected in their hair samples).

[0327] Methods and Results

[0328] Identification of “contaminated” and “uncontaminated” groups

[0329] Two groups of individuals, referred to hereafter as Group 1 (GP1) and Group 2 (GP2), were derived from a comprehensive unsupervised multivariate analysis of PAH (polycyclic aromatic hydrocarbons) data blocks, metabolites, and bacterial microbiome.

[0330] Skin samples from 42 women (GP1) exposed to high levels of pollution were collected The women were compared with 45 women (GP2) who were exposed to relatively lower levels of pollution. Both groups of women were in good health and aged 25 to 45 years. In group 1 (GP1), 30 / 42 women had lived in polluted cities in China for at least 15 years. In group 2 (GP2), 42 / 45 women had lived in less polluted cities for at least 15 years.

[0331] Polluted and unpolluted cities were selected based on significantly different air quality indices (AQI) over a period of one year.

[0332] Here, pollution was defined as exposure to particulate matter and was determined by analyzing PAHs and PAH metabolites in hair samples from these women (Palazzi et al. (2018) Environment International 121:1341-1354).

[0333] PAH and PAH metabolites were quantified in pg / ml and compared between the two groups. V-test analysis was performed and fold change (FC) was calculated. Statistical analysis is described in a separate section.

[0334] surface :FCPAH and PAH metabolites, GP1 and GP2

[0335]

[0336] Women in GP1 had a higher incidence of the clinical symptom “spreading yellow patches or uneven skin tone” on the cheeks and forehead compared with women in GP2 (see Figure 1 On a scale of 1 to 4, the severity of this clinical symptom is higher in GP1 than in GP2 (see Figure 2 ).

[0337] Analysis of metabolites

[0338] For GP1 and GP2 All samples were analyzed using a non-targeted approach.

[0339] In summary, samples were extracted and divided into aliquots for analysis by liquid chromatography-mass spectrometry (LC / MS / MS) and polar liquid chromatography platforms. Appropriate software was used to map ions to a standard custom library that was used to identify metabolites and quantify metabolites by peak area integration.

[0340] Therefore, biochemical identification is based on 3 criteria: retention index within the narrow retention window of the proposed identification, mass that corresponds exactly to the library ±10ppm, and "positive" and negative MS / MS scores between the experimental data and the authentic standards. The MS / MS scores are based on a comparison of the ions present in the experimental spectrum and the ions present in the library. Although there may be similarities between molecules based on just one of these factors, using three data points allows for differentiation and distinction of biochemical molecules.

[0341] More than 3300 commercially available purified standard compounds are obtained and recorded in LIMS to determine their analytical properties for analysis on all platforms. Additional mass spectrometry entries are created for structurally unnamed biochemical molecules, which are identified by their recurring properties (chromatograms and mass spectra). These compounds may be identified by obtaining corresponding purification standards in the future or by conventional structural analysis. Peak quantification is performed using the area under the primary MS ion curve.

[0342] Conventional T-tests were used to determine statistical significance.

[0343] Multiple processing procedures are performed to ensure high-quality data sets for statistical analysis and data interpretation. Quality control and processing procedures are designed to ensure accurate and consistent identification of true chemical entities and elimination of those representing systematic artifacts, false assignments, and background noise.

[0344] surface :FC metabolites, GP1 and GP2

[0345]

[0346]

[0347] Evaluation of bacterial microorganisms

[0348] Participants had not received any antibiotics or systemic antifungal drugs in the month before sampling, did not have any serious skin diseases, and had not used any skin or systemic treatments for depigmentation / whitening in the 3 months before sampling, or any exfoliating products in the month before sampling.

[0349] They were asked to wash their face with the provided neutral soap without antimicrobial compounds for 3 days (once a day) before sampling. The last shampoo and soap application was 48 hours and 24 hours before sampling, respectively. No other products were allowed to be used on the scalp, hair, and face until sampling was completed.

[0350] Sampling of the microbiome was performed in a controlled environment at 22°C and 60% humidity. Samples for microbiome analysis were collected using sterile dry cotton swabs heated to 150°C and pre-moistened with ST solution (0.15M NaCl with 0.1% Tween 20). For cheek samples, the swabs were soaked in collection buffer and rubbed vigorously on the cheek for 60 seconds to cover a 1 cm × 2 cm surface. After sampling, each swab was placed in a microtube and immediately frozen in liquid nitrogen and stored at -80°C before extracting genomic DNA (DNAg).

[0351] Profiling of bacterial 16S rDNA was performed as follows:

[0352] -Preparation of amplicon samples for 16S rRNA gene sequencing

[0353] PowerSoil was used according to the manufacturer's instructions and with modifications as disclosed in Leung et al. (2014) Appl. Environ. Microbiol. 80:6760-6770. DNAg was extracted using a separation kit (MO BIO Laboratories, Carlsbad, CA, USA). In addition, after eluting C6, the eluate was passed through the same filter column again to increase the yield. A negative control of water without DNA was extracted in parallel. PCR was performed in triplicate for each DNAg sample using primers targeting the V1-3 region of the bacterial 16S rRNA, thereby making the image of the skin bacterial community obtained more accurate (Meisel et al. (2016) J. Invest. Dermatol. 136: 947-956). For analysis of 16S rRNA, amplicon PCR and index PCR were performed on a PCR 7500 fast real-time PCR system (Applied Biosystems, Foster City, CA, USA), and the amplicons were purified using DNA / RNA purification beads (SeqMatic, Fremont, CA, USA). Sequencing was performed on an Illumina Library preparation and paired-end sequencing of 300 bp bacterial nucleic acids on the platform.

[0354] - Processing of rRNA 16S gene sequences and bioinformatics analysis

[0355] Paired bacterial and fungal reads were merged in .fastq format using the “-fastq_mergeairs” command in USEARCH, respectively. The merged reads were filtered for quality control using the “-fastq_filter” command in USEARCH, with a maximum expected error rate of 0.01. The merged reads were trimmed at 45 bp, eliminating shorter reads. The filtered reads were OTU grouped at 97% sequence identity using the UPARSE algorithm (Edgar (2013) Nature Methods 10:996-998), and the “assign_taxonomy.py” command in QIIME (version 1.9) was used to provide taxonomic information for sequences representing bacterial OTUs against the SILVA database (128 outputs). OTUs present in more than 5% of the taxonomic lines of the negative control were considered potential contaminants (Leung et al. (2018) Microbiome 6:26) and eliminated from the data set. In addition, chimeras, chloroplasts, and mitochondrial OTUs were also eliminated. After quality control and elimination of bad reads, a total of 9,656,916 bacterial reads remained.

[0356] surface : FC bacterial microbiome, GP1 and GP2

[0357]

[0358] Statistical analysis

[0359] In their original scale, the measured values ​​of metabolites were normalized according to the raw surface counts, and then each metabolite was scaled to obtain a median equal to 1, and missing values ​​were imputed as the minimum value. Statistical analyses were performed on the transformed values.

[0360] Multi-block statistical analysis was performed to correlate metabolites, cheek-sampled microbiome data, and PAH data in a unified model. This analysis enabled the inventors to identify relevant groups of individuals and to use χ2 correlations based on the type of clinical score. 2 The groups were characterized by means of clinical variables.

[0361] More precisely, sparse generalized canonical correlation analysis (Witten et al. (2008) Biostatistics 10: 515-534) was performed to select descriptors that were significantly associated with the covariance / correlation structure between blocks. Then, based on the selected descriptors, a regularized PCA consensus called MAXVAR-A (Tenenhaus et al. (2017) Psychometrika 82: 737-777) was implemented to construct a consensus space. Finally, hierarchical grouping on the consensus space enabled the discovery of two feature groups GP1 and GP2.

[0362] Comparisons between groups were performed using Student's T test based on block descriptors. Sequence Listing <110> L'Oreal <120> Diagnostic method for aesthetic deterioration of the skin <130> BET 20P1982 <150> FR19 06831 <151> 2019-06-24 <160> 9 <170> PatentIn Version 3.5 <210> 1 <211> 450 <212> DNA <213> Propionibacterium acnes <400> 1 agagtttgat cctggctcag gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 ggaaaggccc tgcttttgtg gggtgctcga gtggcgaacg ggtgagtaac acgtgagtaa 120 cctgcccttg actttggggat aacttcagga aactggggct aataccggat aggagctcct 180 gctgcatggt gggggttgga aagtttcggc ggttggggat ggactcgcgg cttatcagct 240 tgttggtggg gtagtggctt accaaggctt tgacgggtag ccggcctgag agggtgaccg 300 gccacattgg gactgagata cggcccagac tcctacggga ggcagcagtg gggaatattg 360 cacaatgggc ggaagcctga tgcagcaacg ccgcgtgcgg gatgacggcc ttcgggttgt 420 aaaccgcttt cgcctgtgac gaagcgtgag 450 <210> 2 <211> 450 <212> DNA <213> Artificial Sequence <220> <223> Micrococcaceae 16S rDNA <400> 2 agagtttgat cctggctcag gatgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 gatgaagccc agcttgctgg gtggattagt ggcgaacggg tgagtaacac gtgagtaacc 120 tgcccttgac tctgggataa gcctgggaaa ctgggtctaa tactggatat gacttctcac 180 cgcatggtgg ggggtggaaa gatttattgg tcttggatgg actcgcggcc tatcagcttg 240 ttggtgaggt aatggctcac caaggcgacg acgggtagcc ggcctgagag ggtgaccggc 300 cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg gaatattgca 360 caatgggcgc aagcctgatg cagcgacgcc gcgtgaggga tgacggcctt cgggttgtaa 420 acctctttca gtagggaaga agccttcggg 450 <210> 3 <211> 450 <212> DNA <213> Artificial sequence <220> <223> Brevibacterium 16S rDNA <400> 3 agagtttgat cctggctcag gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 gatgacgaca gggcttgcct tgtctgatta gtggcgaacg ggtgagtaac acgtgagtaa 120 cctgcccccc tcttcgggat aacctcggga aatcggggct aataccggat atgacttcgc 180 accgcatggt gtgttgtgga aagtttttcg gtgggggatg gactcgcggc ctatcagttt 240 gttggtgagg taatggctca ccaaggcgat gacgggtagc cggcctgaga gggcgaccgg 300 ccacactggg actgagacac ggcccagact cctacgggag gcagcagtgg ggaatattgc 360 acaatgggcg aaagcctgat gcagcgacgc cgcgtggggg atgacggcct tcgggttgta 420 aacctctttc agtagggaag aagcgaaagt 450 <210> 4 <211> 450 <212> DNA <213> Artificial sequence <220> <223> Brevibacterium 16S rDNA <400> 4 agagtttgat cctggctcag gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 gctgaagcct gcagcttgct gtgggtggat gagtggcgaa cgggtgagta acacgtgagt 120 aacctgcccc tgactctggg ataagcccgg gaaactgggt ctaataccgg atatgaccat 180 tctccgcatg gagtggtggt ggaaagtttt tcggttgggg atgggctcgc ggcctatcag 240 cttgttggtg gggtgatggc ctaccaaggc gacgacgggt agccggcctg agagggcgac 300 cggccacact gggactgaga cacggcccag actcctacgg gaggcagcag tggggaatat 360 tgcacaatgg gggaaaccct gatgcagcga cgccgcgtgg gggatgacgg ccttcgggtt 420 gtaaacctct ttcagtaggg aagaagcctt 450 <210> 5 <211> 450 <212> DNA <213> Artificial Sequence <220> <223> Burkholderiales 16S rDNA <400> 5 agagtttgat cctggctcag attgaacgct ggcggcatgc cttacacatg caagtcgaac 60 ggcagcatga tctagcttgc tagattgatg gcgagtggcg aacgggtgag taatacatcg 120 gaacgtgccc tgtagtgggg gataactagt cgaaagatta gctaataccg catacgacct 180 gagggtgaaa gtgggggacc gcaaggcctc atgctatagg agcggccgat gtctgattag 240 ctagttggtg gggtaaaggc ccaccaaggc gacgatcagt agctggtctg agaggacgat 300 cagccacact gggactgaga cacggcccag actcctacgg gaggcagcag tggggaattt 360 tggacaatgg gggcaaccct gatccagcaa tgccgcgtgt gtgaagaagg ccttcgggtt 420 gtaaagcact tttgtccgga aagaaatcgc 450 <210> 6 <211> 450 <212> DNA <213> Artificial Sequence <220> <223> Paracoccus 16S rDNA <400> 6 agagtttgat cctggctcag aacgaacgct ggcggcaggc ctaacacatg caagtcgagc 60 gcgcccttcg gggtgagcgg cggacgggtg agtaacgcgt gggaacgtgc ccttctctgc 120 ggaatagcct cgggaaactg agagtaatac cgcatacgcc ctttggggga aagatttatc 180 ggagaaggat cggcccgcgt tggattaggt agttggtggg gtaatggccc accaagccga 240 cgatccatag ctggtttgag aggatgatca gccacactgg gactgagaca cggcccagac 300 tcctacggga ggcagcagtg gggaatctta gacaatgggg gcaaccctga tctagccatg 360 ccgcgtgagt gatgaaggcc ttagggttgt aaagctcttt cagctgggaa gataatgacg 420 gtaccagcag aagaagcccc ggctaactcc 450 <210> 7 <211> 450 <212> DNA <213> Artificial sequence <220> <223> Rhodobacter 16S rDNA <400> 7 agagtttgat cctggctcag aacgaacgct ggcggcaggc ttaacacatg caagtcgagc 60 gagatcttcg ggtctagcgg cggacgggtg agtaacgcgt gggaacgtgc cctttgctac 120 ggaatagtcc tgggaaactg ggggtaatac cgtatacgcc cttttgggga aagatttatc 180 ggcgaaggat cggcccgcgt tggattaggt agttggtggg gtaatggcct accaagccga 240 cgatccatag ctggtttgag aggatgatca gccacactgg gactgagaca cggcccagac 300 tcctacggga ggcagcagtg gggaatctta gacaatgggg gcaaccctga tctagccatg 360 ccgcgtgagt gatgaaggcc ttagggttgt aaagctcttt cagctgggaa gataatgacg 420 gtaccagcag aagaagcccc ggctaactcc 450 <210> 8 <211> 450 <212> DNA <213> Artificial sequence <220> <223> Fusobacterium 16S rDNA <400> 8 agagtttgat cctggctcag gatgaacgct gacagaatgc ttaacacatg caagtctact 60 tgaacttcgg tttgggtggc ggacgggtga gtaacgcgta aagaacttgc ctcacagtta 120 gggacaacat ttggaaacga atgctaatac ctgatattat gattttaggg catcctaaga 180 ttatgaaagc tatatgcgct gtgagagagc tttgcgtccc attagctagt tggagaggta 240 acggctcacc aaggcgatga tgggtagccg gcctgagagg gtgaacggcc acaaggggac 300 tgagacacgg cccttactcc tacgggaggc agcagtgggg aatattggac aatggaccaa 360 aagtctgatc cagcaattct gtgtgcacga tgacgttttt cggaatgtaa agtgctttca 420 gttgggaaga aaaaaatgac ggtaccaaca 450 <210> 9 <211> 450 <212> DNA <213> Micrococcus luteus <400> 9 agagtttgat cctggctcag gatgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 gatgaagccc agcttgctgg gtggattagt ggcgaacggg tgagtaacac gtgagtaacc 120 tgcccttaac tctgggataa gcctgggaaa ctgggtctaa taccggatag gagcgtccac 180 cgcatggtgg gtgttggaaa gatttatcgg ttttggatgg actcgcggcc tatcagcttg 240 ttggtgaggt aatggctcac caaggcgacg acgggtagcc ggcctgagag ggtgaccggc 300 cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg gaatattgca 360 caatgggcgc aagcctgatg cagcgacgcc gcgtgaggga tgacggcctt cgggttgtaa 420 acctctttca gtagggaaga agcgaaagtg 450

Claims

1. A method for confirming aesthetic deterioration of a subject's skin due to chronic exposure to polycyclic aromatic hydrocarbons (PAHs), wherein the method is used for purposes other than disease diagnosis, comprising: Step (a), determining the level of at least one marker in a skin sample of the subject, the marker being selected from the group consisting of: (i) a bacterium comprising a 16S nucleotide sequence that is at least 95% identical to the sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8; rRNA sequences, and (ii) metabolites of these bacteria selected from the group consisting of 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid / 2-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, citraconic acid / glutaconic acid, fumaric acid, γ-glutamylalanine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamylserine, γ-glutamylthreonine, γ-glutamylthreonine, γ-glutamyl-3-hydroxy-4-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, citraconic acid / glutaconic acid, fumaric acid, γ-glutamylalanine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamylserine, γ-glutamylthreonine, γ-glutamyl-3-hydroxy-4-methylglutaric acid, 4- Glutamyltryptophan, gamma-glutamylvaline, glyceric acid, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenine, lactic acid, linoleoylethanolamide, malic acid, maleic acid, malonic acid, methionine sulfoxide, methylsuccinic acid, N-acetylalanine, N-acetylaspartic acid, N-acetyarginine, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelic acid, proline, salicylic acid, sebacic acid, serine, suberic acid, undecanedioic acid, and S-amino-ω caprolactam.

2. The confirmation method according to claim 1, wherein: The at least one marker is selected from the group consisting of: (i) bacteria of the species Propionibacterim acnes, bacteria of the family Micrococcaceae, bacteria of the genus Brachybacterium, bacteria of the genus Brevibacterium, bacteria of the order Burkholderiales, bacteria of the genus Paracoccus, bacteria of the family Rhodobacteraceae and bacteria of the genus Fusobacterium, and (ii) metabolites of these bacteria selected from the group consisting of 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid / 2-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, arabinonic acid / xylonic acid, azelaic acid, β-citric acid acylglutamate, choline, cis-urocanic acid, citraconic acid / glutaric acid, fumaric acid, gamma-glutamylalanine, gamma-glutamylglycine, gamma-glutamylisoleucine, gamma-glutamylleucine, gamma-glutamylserine, gamma-glutamylthreonine, gamma-glutamyltryptophan, gamma-glutamylvaline, glyceric acid, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenine, lactic acid, linoleoylethanolamide, malic acid, maleic acid, malonic acid, methionine sulfoxide, methylsuccinic acid, N-acetylalanine, N-acetylaspartic acid, N-acetyarginine, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelic acid, proline, salicylic acid, sebacic acid, serine, suberic acid, undecanedioic acid, and S-amino-ω caprolactam.

3. The method according to claim 1, wherein: The at least one marker is selected from the group consisting of bacteria of the Paracoccus sp. strain, and metabolites selected from the group consisting of kynurenine, 4-imidazoleacetic acid, maleic acid, ornithine, 4-guanidinobutyric acid, cis-urocanic acid, malonate, γ-glutamylleucine, N-acetylarginine and glycerol-3-phosphate.

4. The method according to claim 1, further comprising the steps of: (b) comparing the level of the at least one marker measured in step (a) with a control, and (c) determining whether the subject's skin exhibits aesthetic degradation based on the comparison of step (b).

5. The method according to any one of claims 1 to 4, wherein: The at least one marker is a bacterium as defined in any one of claims 1 to 3, and the level of the at least one marker is determined by measuring the level of the corresponding 16S rRNA gene.

6. The method according to any one of claims 1 to 4, wherein: The at least one marker is a metabolite as defined in any one of claims 1 to 3, and the level of the at least one marker is determined by liquid chromatography-mass spectrometry.

7. The method according to any one of claims 1 to 4, wherein: The skin samples were Plate to collect.

8. The method according to any one of claims 1 to 4, wherein: The aesthetic deterioration of the skin is selected from the group consisting of aesthetic pigmentation disorders, lack of radiance and uneven skin tone.

9. A method for confirming that a subject's skin has been exposed to pollution due to chronic exposure to polycyclic aromatic hydrocarbons (PAHs), the method being used for purposes other than disease diagnosis, comprising: Step (a), determining the level of at least one marker in a skin sample of the subject, wherein the at least one marker is selected from the group consisting of: (i) bacteria, wherein the bacteria comprises a 16S rRNA sequences, and (ii) metabolites of these bacteria selected from the group consisting of 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid / 2-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, citraconic acid / glutaconic acid, fumaric acid, γ-glutamylalanine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamylserine, γ-glutamylthreonine, γ-glutamylthreonine, γ-glutamyl-3-hydroxy-4-methylglutaric acid, 4-guanidinobutyric acid, 4-imidazoleacetic acid, 5-oxoproline, aconitic acid, adipic acid, alanine, arabinonic acid / xylonic acid, azelaic acid, β-citrolglutamate, choline, cis-urocanic acid, citraconic acid / glutaconic acid, fumaric acid, γ-glutamylalanine, γ-glutamylglycine, γ-glutamylisoleucine, γ-glutamylleucine, γ-glutamylserine, γ-glutamylthreonine, γ-glutamyl-3-hydroxy-4-methylglutaric acid, 4- Glutamyltryptophan, gamma-glutamylvaline, glyceric acid, glycerol-3-phosphate, glycine, isovalerylglycine, kynurenine, lactic acid, linoleoylethanolamide, malic acid, maleic acid, malonic acid, methionine sulfoxide, methylsuccinic acid, N-acetylalanine, N-acetylaspartic acid, N-acetyarginine, N-acetylglycine, N-acetylhistidine, N-acetylphenylalanine, N-acetylvaline, oleamide, ornithine, palmitamide, pimelic acid, proline, salicylic acid, sebacic acid, serine, suberic acid, undecanedioic acid, and S-amino-ω caprolactam.

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