#METABOLOMICS WORKBENCH bahgatfayed_20241123_224140 DATATRACK_ID:5388 STUDY_ID:ST003612 ANALYSIS_ID:AN005937 PROJECT_ID:PR002232 VERSION 1 CREATED_ON 12-09-2024 #PROJECT PR:PROJECT_TITLE Candida auris planktonic and dispersed cells Chromatographic analysis using PR:PROJECT_TITLE GC-MS PR:PROJECT_SUMMARY The emerging Candida auris (C. auris) has caused several outbreaks globally. PR:PROJECT_SUMMARY While several studies explored the resistant biofilm formed by C. auris, little PR:PROJECT_SUMMARY is known regarding the cells dispersed following biofilm formation. Herein, I PR:PROJECT_SUMMARY investigated and characterized the cells dispersed from C. auris biofilms. Cells PR:PROJECT_SUMMARY dispersed from biofilm developed in 96 well plate were isolated and counted. PR:PROJECT_SUMMARY GC-MS analysis indicated that dispersed cells exhibit altered metabolic profile PR:PROJECT_SUMMARY that enhance cells survivability under stress and nutrient limit condition. The PR:PROJECT_SUMMARY presented study is the first to explore C. auris dispersed cells and indicated PR:PROJECT_SUMMARY that they are not able to revert to the planktonic mode once released from the PR:PROJECT_SUMMARY biofilm. PR:INSTITUTE National Research Centre, Dokki, Egypt PR:LAST_NAME Fayed PR:FIRST_NAME Bahgat PR:ADDRESS Dokki, Giza, Cairo, 11433, Egypt PR:EMAIL bm.ezzat@nrc.sci.eg PR:PHONE +201098292083 PR:DOI http://dx.doi.org/10.21228/M8424B #STUDY ST:STUDY_TITLE Candida auris planktonic and dispersed cells Chromatographic analysis using ST:STUDY_TITLE GC-MS ST:STUDY_SUMMARY The emerging Candida auris (C. auris) has caused several outbreaks globally. ST:STUDY_SUMMARY While several studies explored the resistant biofilm formed by C. auris, little ST:STUDY_SUMMARY is known regarding the cells dispersed following biofilm formation. Herein, I ST:STUDY_SUMMARY investigated and characterized the cells dispersed from C. auris biofilms. Cells ST:STUDY_SUMMARY dispersed from biofilm developed in 96 well plate were isolated and counted. ST:STUDY_SUMMARY GC-MS analysis indicated that dispersed cells exhibit altered metabolic profile ST:STUDY_SUMMARY that enhance cells survivability under stress and nutrient limit condition. The ST:STUDY_SUMMARY presented study is the first to explore C. auris dispersed cells and indicated ST:STUDY_SUMMARY that they are not able to revert to the planktonic mode once released from the ST:STUDY_SUMMARY biofilm. ST:INSTITUTE National Research Centre, Dokki, Egypt ST:LAST_NAME Fayed ST:FIRST_NAME Bahgat ST:ADDRESS Dokki ST:EMAIL bm.ezzat@nrc.sci.eg ST:PHONE +201098292083 ST:SUBMIT_DATE 2024-11-23 #SUBJECT SU:SUBJECT_TYPE Yeast SU:SUBJECT_SPECIES Candida auris #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - sample tested Sample source:Candida auris dispersed cells | Geno type:Wild RAW_FILE_NAME(Raw_file_name)=RESULTS-S raw SUBJECT_SAMPLE_FACTORS - Control Sample source:Candida auris planktonic cells | Geno type:Wild RAW_FILE_NAME(Raw_file_name)=RESULTS-C raw #COLLECTION CO:COLLECTION_SUMMARY C. auris dispersed cell were allowed to grow in 250-mL Erlenmeyer flasks CO:COLLECTION_SUMMARY containing 50 mL of SD broth media for 3 days at 30°C. Following incubation, CO:COLLECTION_SUMMARY the supernatant was separated by centrifugation for 20 min at 4000 rpm. The CO:COLLECTION_SUMMARY resulting supernatant was extracted with 100 mL ethyl acetate. CO:SAMPLE_TYPE Candida auris #TREATMENT TR:TREATMENT_SUMMARY Isolation of cells dispersed from C. auris biofilm C. auris biofilms were TR:TREATMENT_SUMMARY developed by loading 1Ã106 cells/ml in RPMI-1640 supplemented with 2% glucose TR:TREATMENT_SUMMARY and then incubating for 24 h at 37°C in 96-well flat-bottomed microplates [17]. TR:TREATMENT_SUMMARY Following incubation, the supernatant containing floating cells was removed, the TR:TREATMENT_SUMMARY formed biofilm was washed 3 times with PBS, fresh media was added, and the TR:TREATMENT_SUMMARY biofilm was incubated for an additional 24 h under the same conditions. The next TR:TREATMENT_SUMMARY day, the cells dispersed from the biofilm in the upper supernatant were TR:TREATMENT_SUMMARY collected, counted by a hemocytometer and maintained in SD media or stored in TR:TREATMENT_SUMMARY 30% glycerol for further use. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY C. auris dispersed cell were allowed to grow in 250-mL Erlenmeyer flasks SP:SAMPLEPREP_SUMMARY containing 50 mL of SD broth media for 3 days at 30°C. Following incubation, SP:SAMPLEPREP_SUMMARY the supernatant was separated by centrifugation for 20 min at 4000 rpm. The SP:SAMPLEPREP_SUMMARY resulting supernatant was extracted with 100 mL ethyl acetate. After the SP:SAMPLEPREP_SUMMARY extraction step, the organic phases were dried on anhydrous MgSO4, filtered, and SP:SAMPLEPREP_SUMMARY concentrated under reduced pressure. The parent C. auris planktonic cells served SP:SAMPLEPREP_SUMMARY as control. #CHROMATOGRAPHY CH:INSTRUMENT_NAME Agilent 7890B CH:COLUMN_NAME Agilent DB5-MS (15m x 0.25mm, 0.25um) CH:COLUMN_TEMPERATURE 300°C CH:FLOW_GRADIENT nothing CH:FLOW_RATE nothing CH:SOLVENT_A nothing CH:SOLVENT_B nothing CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 7890B MS:INSTRUMENT_TYPE GC-MS MS:MS_TYPE EI MS:MS_COMMENTS The sample analysis began with the column held at 60°C for 5 minutes MS:MS_COMMENTS post-injection. The temperature was subsequently increased to 300°C at a rate MS:MS_COMMENTS of 20°C per minute, followed by a 5-minute hold. The injection was performed in MS:MS_COMMENTS split mode with a split ratio of 5:1 at 300°C. The MS scan range was 50â550 MS:MS_COMMENTS atomic mass units (AMU) under electron impact ionization (70 eV), with a solvent MS:MS_COMMENTS delay of 8.0 minutes. The constituents were identified by mass fragmentations MS:MS_COMMENTS using the NIST mass spectral search program for the NIST/EPA/NIH mass spectral MS:MS_COMMENTS library Version 2.2 (Jun 2014). MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Area Pct MS_METABOLITE_DATA_START Samples sample tested Control Factors Sample source:Candida auris dispersed cells | Geno type:Wild Sample source:Candida auris planktonic cells | Geno type:Wild 1_2_3_4-Butanetetrol_ 1-[2-(2-fluorophenyl)-2H-1_2_3-triazol-4-yl]-_ [1R-(1R*_2S*_3R*)]- 2.2251 (+)-1_2_3_4-Tetrahydroisoquinoline_ 1-benzyl-6_7-dimethoxy- 1.2848 13-Octadecenoic acid 0.3852 1_4_6-Trimethyl-1_2_3_3a_4_7_8_8a-octahydro-4_7-ethanoazulene 3.9742 1-[4-(Acridin-9-ylamino)-phenyl]-ethanone 2.4725 1-Ethoxy-2-propanol 2.2272 2.0329 1H-1_2_3-Triazolo[4_5-c]quinoline-1-hexanoic acid 0.5572 1-Octadecene 0.3986 1-Phenyl-2-(2-trimethylsilylcyclopropen-1-yl)ethanol 0.1934 [2-(2-Ethoxyethoxy)ethoxy]acetic acid 0.2965 2_4-Heptanedione_ 6-[[2-(2_3_8_8a-tetrahydro-5_8a-dimethyl-7-oxoimidazo[1_2-a]pyridin-1(7H)-yl)ethyl]imino]- 0.4353 2_4-Hexadienedioic acid 0.2943 2_6_10_14-Tetramethyl-7-(3-methylpent-4-enylidene) pentadecane 0.7039 2-Amino-4-nitrophenol 0.6552 0.4366 2-Butenedioic acid 2.5567 3.3092 2-Cyclohexen-1-one_ 3-[3-(4_4-dimethyl-4_5-dihydro-1_3-oxazol-2-yl)phenyl] 1.4553 2^-Deoxyribolactone 1.0069 2-(Ethylamino)ethanol_ N-pentafluoropropionyl_ O-trimethylsilyl 1.4055 2-Furanglycolic acid 3.0224 5.6076 2-Hydroxy-3-methylbutyric acid 0.4132 (2-Hydroxyethyl)trimethylsilyl methyl sulfide 0.4769 3_10-Dioxa-2_11-disiladodeca-5_7-diene_ 2_2_11_11-tetramethyl- 0.4649 3_5-Cyclocholestan-19-al_ 6-hydroxy-_ (6.beta.)- 1.4153 3_5-Dimethyl-2-phenylpyridine 1.1860 3-Hydroxy-3-phenylpropanoic acid 0.9058 3-Methylbenzyl alcohol 1.7887 3-Phenyllactic acid 1.3658 1.4679 4-Hexylphenol 1.8193 4-Hydroxybenzoic acid 0.3358 0.1455 4-Hydroxybutanoic acid 0.3026 4-Hydroxyphenyllactic acid 0.9302 1.4228 4-Phenyl-3-buten-2-ol 0.1518 5_14_23-Octadecatrien-14_15-diol 1.4181 5_6-Epoxyretinoic acid 1.8058 5-Hydroxy-2-methylpyridine 0.8189 5-Hydroxymethyl-2-furoic acid 0.6883 9-Octadecenoic acid 0.4473 0.8917 Acetamide 0.9232 Acetic acid 2.8062 Arachidonic acid 0.4275 1.0648 benzenamine_ 2_3_5_6-tetrafluoro-N_N-dimethyl- 1.8908 Benzonitrile_ 2-benzylthio-4-methoxy- 1.6721 2.3720 Bis(2-ethylhexyl) phthalate 5.8313 9.4822 Bis(pentamethylcyclopentadienyl)iron 3.6266 Bis(trimethylsilyl) 3_6_9_12-tetraoxatetradecane-1_14-dioate 2.5982 1.5865 Butanedioic acid 6.2011 6.6823 Carbitol 0.1844 Cholest-4-en-26-al_ 3-oxo-_ cyclic 26-(ethylene acetal) 0.3088 0.3720 cis-4-Trimethylsilyloxy-cyclohexyl(trimethylsilyl)carboxylate 1.0798 3.0974 Citric acid 2.4850 Cyclodocosane_ ethyl- 0.5696 Cyclohexene_ 3-butyl-3-methyl-1-trimethylsilyloxy- 0.6972 D-Arabino-Hexonic acid_ 3-deoxy-2_5_6-tris-O-(trimethylsilyl)-_ .gamma.-lactone 0.6249 0.4485 D-(+)-Arabitol 0.3812 Dihydroxyacetone 1.4153 Dodecane_ 1-(methoxymethoxy)- 0.2769 D-(+)-Ribono-1_4-lactone 0.5507 0.7383 Glyceric acid 1.5688 1.4406 Glycerol 0.1551 1.3409 Gulonic acid_ .gamma.-lactone 0.1513 0.8962 Heptacosane_ 1-chloro- 1.0062 Heptadecanoic acid 0.2167 Hexacosane 0.9089 Hexadeca-2_6_10_14-tetraen-1-ol_ 3_7_11_16-tetramethyl- 1.3311 Hydracrylic acid 1.3861 0.6538 Kojic acid 0.7209 Malic acid 2.8299 1.2667 Myristic acid 0.2689 Nonacosane 1.8403 Octacosane 1.8773 Palmitic Acid 1.7674 3.4313 Pentanedioic acid 0.4923 0.5274 Phenylephrine 0.3245 Phenylethyl Alcohol 4.8358 Propionic acid_ 3-iodo-_ heptadecyl ester 1.2140 Ribitol 0.2862 Stearic acid 0.3520 0.7313 Tetrapentacontane_ 1_54-dibromo- 0.4162 Triacontane 1.2337 Tricosane 0.1912 Trimethylsilyl O-(trimethylsilyl)isohomovanillate 1.1204 Tyrosol 3.1125 3.5699 Vanillic Acid 0.6273 0.5630 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant 1,2,3,4-Butanetetrol, 1-[2-(2-fluorophenyl)-2H-1,2,3-triazol-4-yl]-, [1R-(1R*,2S*,3R*)]- (+)-1,2,3,4-Tetrahydroisoquinoline, 1-benzyl-6,7-dimethoxy- 13-Octadecenoic acid 1,4,6-Trimethyl-1,2,3,3a,4,7,8,8a-octahydro-4,7-ethanoazulene 1-[4-(Acridin-9-ylamino)-phenyl]-ethanone 1-Ethoxy-2-propanol 1H-1,2,3-Triazolo[4,5-c]quinoline-1-hexanoic acid 1-Octadecene 1-Phenyl-2-(2-trimethylsilylcyclopropen-1-yl)ethanol [2-(2-Ethoxyethoxy)ethoxy]acetic acid 2,4-Heptanedione, 6-[[2-(2,3,8,8a-tetrahydro-5,8a-dimethyl-7-oxoimidazo[1,2-a]pyridin-1(7H)-yl)ethyl]imino]- 2,4-Hexadienedioic acid 2,6,10,14-Tetramethyl-7-(3-methylpent-4-enylidene) pentadecane 2-Amino-4-nitrophenol 2-Butenedioic acid 2-Cyclohexen-1-one, 3-[3-(4,4-dimethyl-4,5-dihydro-1,3-oxazol-2-yl)phenyl] 2'-Deoxyribolactone 2-(Ethylamino)ethanol, N-pentafluoropropionyl, O-trimethylsilyl 2-Furanglycolic acid 2-Hydroxy-3-methylbutyric acid (2-Hydroxyethyl)trimethylsilyl methyl sulfide 3,10-Dioxa-2,11-disiladodeca-5,7-diene, 2,2,11,11-tetramethyl- 3,5-Cyclocholestan-19-al, 6-hydroxy-, (6.beta.)- 3,5-Dimethyl-2-phenylpyridine 3-Hydroxy-3-phenylpropanoic acid 3-Methylbenzyl alcohol 3-Phenyllactic acid 4-Hexylphenol 4-Hydroxybenzoic acid 4-Hydroxybutanoic acid 4-Hydroxyphenyllactic acid 4-Phenyl-3-buten-2-ol 5,14,23-Octadecatrien-14,15-diol 5,6-Epoxyretinoic acid 5-Hydroxy-2-methylpyridine 5-Hydroxymethyl-2-furoic acid 9-Octadecenoic acid Acetamide Acetic acid Arachidonic acid benzenamine, 2,3,5,6-tetrafluoro-N,N-dimethyl- Benzonitrile, 2-benzylthio-4-methoxy- Bis(2-ethylhexyl) phthalate Bis(pentamethylcyclopentadienyl)iron Bis(trimethylsilyl) 3,6,9,12-tetraoxatetradecane-1,14-dioate Butanedioic acid Carbitol Cholest-4-en-26-al, 3-oxo-, cyclic 26-(ethylene acetal) cis-4-Trimethylsilyloxy-cyclohexyl(trimethylsilyl)carboxylate Citric acid Cyclodocosane, ethyl- Cyclohexene, 3-butyl-3-methyl-1-trimethylsilyloxy- D-Arabino-Hexonic acid, 3-deoxy-2,5,6-tris-O-(trimethylsilyl)-, .gamma.-lactone D-(+)-Arabitol Dihydroxyacetone Dodecane, 1-(methoxymethoxy)- D-(+)-Ribono-1,4-lactone Glyceric acid Glycerol Gulonic acid, .gamma.-lactone Heptacosane, 1-chloro- Heptadecanoic acid Hexacosane Hexadeca-2,6,10,14-tetraen-1-ol, 3,7,11,16-tetramethyl- Hydracrylic acid Kojic acid Malic acid Myristic acid Nonacosane Octacosane Palmitic Acid Pentanedioic acid Phenylephrine Phenylethyl Alcohol Propionic acid, 3-iodo-, heptadecyl ester Ribitol Stearic acid Tetrapentacontane, 1,54-dibromo- Triacontane Tricosane Trimethylsilyl O-(trimethylsilyl)isohomovanillate Tyrosol Vanillic Acid METABOLITES_END #END