#METABOLOMICS WORKBENCH barbaraggcarvalho_20240620_035113 DATATRACK_ID:4943 STUDY_ID:ST003276 ANALYSIS_ID:AN005364 PROJECT_ID:PR002028 VERSION 1 CREATED_ON June 20, 2024, 6:45 am #PROJECT PR:PROJECT_TITLE Metabolomics analysis of human spermatozoa reveals impaired metabolic pathways PR:PROJECT_TITLE in asthenozoospermia PR:PROJECT_SUMMARY Background: Infertility is a major health issue, affecting 15% of PR:PROJECT_SUMMARY reproductive-age couples with male factors contributing to 50% of cases. PR:PROJECT_SUMMARY Asthenozoospermia, or low sperm motility, is a common cause of male infertility PR:PROJECT_SUMMARY with complex etiology, involving genetic and metabolic alterations, PR:PROJECT_SUMMARY inflammation, and oxidative stress. However, the molecular mechanisms behind low PR:PROJECT_SUMMARY motility are unclear. In this study, we used a metabolomics approach to identify PR:PROJECT_SUMMARY metabolic biomarkers and pathways involved in sperm motility. Methods: We PR:PROJECT_SUMMARY compared the metabolome and lipidome of spermatozoa of men with normozoospermia PR:PROJECT_SUMMARY (n = 44) and asthenozoospermia (n = 22) using untargeted LC-MS and the PR:PROJECT_SUMMARY metabolome of seminal fluid using 1H-NMR. Additionally, we evaluated the seminal PR:PROJECT_SUMMARY fluid redox status to assess the oxidative stress in the ejaculate. Results: We PR:PROJECT_SUMMARY identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in PR:PROJECT_SUMMARY the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of PR:PROJECT_SUMMARY the spermatozoa’s metabolomics and lipidomics data showed a clear separation PR:PROJECT_SUMMARY between groups. Spermatozoa of asthenozoospermic men presented lower levels of PR:PROJECT_SUMMARY several amino acids, and increased levels of energetic substrates and PR:PROJECT_SUMMARY lysophospholipids. However, the metabolome and redox status of the seminal fluid PR:PROJECT_SUMMARY was not altered in asthenozoospermia. Conclusions: Our results indicate impaired PR:PROJECT_SUMMARY metabolic pathways associated with redox homeostasis and amino acid, energy, and PR:PROJECT_SUMMARY lipid metabolism in asthenozoospermia. Taken together, these findings suggest PR:PROJECT_SUMMARY that the metabolome and lipidome of human spermatozoa are key factors PR:PROJECT_SUMMARY influencing their motility and that oxidative stress exposure during PR:PROJECT_SUMMARY spermatogenesis or sperm maturation may be in the etiology of decreased motility PR:PROJECT_SUMMARY in asthenozoospermia. PR:INSTITUTE University of Aveiro PR:DEPARTMENT Department of Chemistry PR:LAST_NAME Guerra-Carvalho PR:FIRST_NAME Bárbara PR:ADDRESS Campus Universitário de Santiago, 3810-193 Aveiro, Portugal PR:EMAIL barbaraggcarvalho@gmail.com PR:PHONE 234 370 360 PR:PROJECT_COMMENTS Study part 2 of 2 #STUDY ST:STUDY_TITLE Metabolomics analysis of human spermatozoa reveals impaired metabolic pathways ST:STUDY_TITLE in asthenozoospermia ST:STUDY_SUMMARY Background: Infertility is a major health issue, affecting 15% of ST:STUDY_SUMMARY reproductive-age couples with male factors contributing to 50% of cases. ST:STUDY_SUMMARY Asthenozoospermia, or low sperm motility, is a common cause of male infertility ST:STUDY_SUMMARY with complex etiology, involving genetic and metabolic alterations, ST:STUDY_SUMMARY inflammation, and oxidative stress. However, the molecular mechanisms behind low ST:STUDY_SUMMARY motility are unclear. In this study, we used a metabolomics approach to identify ST:STUDY_SUMMARY metabolic biomarkers and pathways involved in sperm motility. Methods: We ST:STUDY_SUMMARY compared the metabolome and lipidome of spermatozoa of men with normozoospermia ST:STUDY_SUMMARY (n = 44) and asthenozoospermia (n = 22) using untargeted LC-MS and the ST:STUDY_SUMMARY metabolome of seminal fluid using 1H-NMR. Additionally, we evaluated the seminal ST:STUDY_SUMMARY fluid redox status to assess the oxidative stress in the ejaculate. Results: We ST:STUDY_SUMMARY identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in ST:STUDY_SUMMARY the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of ST:STUDY_SUMMARY the spermatozoa’s metabolomics and lipidomics data showed a clear separation ST:STUDY_SUMMARY between groups. Spermatozoa of asthenozoospermic men presented lower levels of ST:STUDY_SUMMARY several amino acids, and increased levels of energetic substrates and ST:STUDY_SUMMARY lysophospholipids. However, the metabolome and redox status of the seminal fluid ST:STUDY_SUMMARY was not altered in asthenozoospermia. Conclusions: Our results indicate impaired ST:STUDY_SUMMARY metabolic pathways associated with redox homeostasis and amino acid, energy, and ST:STUDY_SUMMARY lipid metabolism in asthenozoospermia. Taken together, these findings suggest ST:STUDY_SUMMARY that the metabolome and lipidome of human spermatozoa are key factors ST:STUDY_SUMMARY influencing their motility and that oxidative stress exposure during ST:STUDY_SUMMARY spermatogenesis or sperm maturation may be in the etiology of decreased motility ST:STUDY_SUMMARY in asthenozoospermia. ST:INSTITUTE University of Aveiro ST:DEPARTMENT Department of Chemistry ST:LAST_NAME Guerra-Carvalho ST:FIRST_NAME Bárbara ST:ADDRESS Campus Universitário de Santiago, 3810-193 Aveiro, Portugal ST:EMAIL barbaraggcarvalho@gmail.com ST:PHONE 234 370 360 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 57 ST:NUM_MALES 57 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE 20 to 49 years old SU:GENDER Male SU:HUMAN_INCLUSION_CRITERIA Participants enrolled in this study were randomly recruited among men who SU:HUMAN_INCLUSION_CRITERIA attended fertility consultations at the Centre for Reproductive Genetics SU:HUMAN_INCLUSION_CRITERIA Professor Alberto Barros (Porto, Portugal) and the Centro Hospitalar SU:HUMAN_INCLUSION_CRITERIA Universitário de Santo António (Porto, Portugal) from March to June 2021. SU:HUMAN_INCLUSION_CRITERIA Adult men (age ≥ 18 years) diagnosed with asthenozoospermia (AS, sperm total SU:HUMAN_INCLUSION_CRITERIA motility < 42%) were recruited for AS group and men with normal sperm parameters SU:HUMAN_INCLUSION_CRITERIA (sperm concentration ≥ 15 million cells/mL, sperm count ≥ 39 million cells SU:HUMAN_INCLUSION_CRITERIA and sperm total motility ≥ 42%) were recruited for the control group SU:HUMAN_INCLUSION_CRITERIA (normozoospermia (NZ) group). SU:HUMAN_EXCLUSION_CRITERIA Men with a history of abusing drugs or alcoholism, under treatment or exposed to SU:HUMAN_EXCLUSION_CRITERIA drugs known to interfere with fertility, diagnosed with acute genital SU:HUMAN_EXCLUSION_CRITERIA inflammatory disease or with systemic disease known to affect fertility before SU:HUMAN_EXCLUSION_CRITERIA sperm collection, survivors of cancer treatment, or men diagnosed with SU:HUMAN_EXCLUSION_CRITERIA leukocytospermia were excluded from this study. Smoking was not included as an SU:HUMAN_EXCLUSION_CRITERIA exclusion criterion for this study. #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - N3 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=3 SUBJECT_SAMPLE_FACTORS - N4 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=4 SUBJECT_SAMPLE_FACTORS - N14 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=14 SUBJECT_SAMPLE_FACTORS - N15 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=15 SUBJECT_SAMPLE_FACTORS - N22 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=22 SUBJECT_SAMPLE_FACTORS - N24 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=24 SUBJECT_SAMPLE_FACTORS - N26 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=26 SUBJECT_SAMPLE_FACTORS - N27 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=27 SUBJECT_SAMPLE_FACTORS - N28 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=28 SUBJECT_SAMPLE_FACTORS - N29 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=29 SUBJECT_SAMPLE_FACTORS - N33 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=33 SUBJECT_SAMPLE_FACTORS - N34 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=34 SUBJECT_SAMPLE_FACTORS - N39 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=39 SUBJECT_SAMPLE_FACTORS - N43 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=43 SUBJECT_SAMPLE_FACTORS - N44 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=44 SUBJECT_SAMPLE_FACTORS - N49 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=49 SUBJECT_SAMPLE_FACTORS - N53 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=53 SUBJECT_SAMPLE_FACTORS - N56 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=56 SUBJECT_SAMPLE_FACTORS - N58 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=58 SUBJECT_SAMPLE_FACTORS - N64 Group:Asthenozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=64 SUBJECT_SAMPLE_FACTORS - N2 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=2 SUBJECT_SAMPLE_FACTORS - N5 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=5 SUBJECT_SAMPLE_FACTORS - N6 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=6 SUBJECT_SAMPLE_FACTORS - N7 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=7 SUBJECT_SAMPLE_FACTORS - N8 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=8 SUBJECT_SAMPLE_FACTORS - N10 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=10 SUBJECT_SAMPLE_FACTORS - N11 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=11 SUBJECT_SAMPLE_FACTORS - N12 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=12 SUBJECT_SAMPLE_FACTORS - N13 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=13 SUBJECT_SAMPLE_FACTORS - N16 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=16 SUBJECT_SAMPLE_FACTORS - N18 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=18 SUBJECT_SAMPLE_FACTORS - N19 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=19 SUBJECT_SAMPLE_FACTORS - N20 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=20 SUBJECT_SAMPLE_FACTORS - N21 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=21 SUBJECT_SAMPLE_FACTORS - N23 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=23 SUBJECT_SAMPLE_FACTORS - N25 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=25 SUBJECT_SAMPLE_FACTORS - N30 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=30 SUBJECT_SAMPLE_FACTORS - N31 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=31 SUBJECT_SAMPLE_FACTORS - N35 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=35 SUBJECT_SAMPLE_FACTORS - N36 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=36 SUBJECT_SAMPLE_FACTORS - N38 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=38 SUBJECT_SAMPLE_FACTORS - N40 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=40 SUBJECT_SAMPLE_FACTORS - N42 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=42 SUBJECT_SAMPLE_FACTORS - N45 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=45 SUBJECT_SAMPLE_FACTORS - N46 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=46 SUBJECT_SAMPLE_FACTORS - N47 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=47 SUBJECT_SAMPLE_FACTORS - N48 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=48 SUBJECT_SAMPLE_FACTORS - N50 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=50 SUBJECT_SAMPLE_FACTORS - N51 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=51 SUBJECT_SAMPLE_FACTORS - N52 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=52 SUBJECT_SAMPLE_FACTORS - N55 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=55 SUBJECT_SAMPLE_FACTORS - N57 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=57 SUBJECT_SAMPLE_FACTORS - N59 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=59 SUBJECT_SAMPLE_FACTORS - N61 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=61 SUBJECT_SAMPLE_FACTORS - N63 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=63 SUBJECT_SAMPLE_FACTORS - N68 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=68 SUBJECT_SAMPLE_FACTORS - N70 Group:Normozoospermia | Sample source:Human seminal fluid RAW_FILE_NAME(Raw file name)=70 #COLLECTION CO:COLLECTION_SUMMARY Human seminal samples were obtained through masturbation into a specific sterile CO:COLLECTION_SUMMARY container after 2 to 4 days of sexual abstinence. After collection, samples were CO:COLLECTION_SUMMARY left at 37°C until complete liquefaction and sperm parameters (semen pH, CO:COLLECTION_SUMMARY ejaculate volume, sperm concentration, sperm count and sperm motility) were CO:COLLECTION_SUMMARY analyzed according to the WHO guidelines 5. After routine semen analysis, CO:COLLECTION_SUMMARY spermatozoa were separated from the seminal fluid through centrifugation at CO:COLLECTION_SUMMARY 500.g for 5 min. Seminal fluid was collected into new centrifuge tubes and CO:COLLECTION_SUMMARY stored at -80°C until further use. CO:SAMPLE_TYPE Seminal plasma CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Participants enrolled in this study were randomly recruited among men who TR:TREATMENT_SUMMARY attended fertility consultations at the Centre for Reproductive Genetics TR:TREATMENT_SUMMARY Professor Alberto Barros (Porto, Portugal) and the Centro Hospitalar TR:TREATMENT_SUMMARY Universitário de Santo António (Porto, Portugal) from March to June 2021. TR:TREATMENT_SUMMARY Adult men (age ≥ 18 years) diagnosed with asthenozoospermia (AS, sperm total TR:TREATMENT_SUMMARY motility < 42%) were recruited for AS group and men with normal sperm parameters TR:TREATMENT_SUMMARY (sperm concentration ≥ 15 million cells/mL, sperm count ≥ 39 million cells TR:TREATMENT_SUMMARY and sperm total motility ≥ 42%) were recruited for the control group TR:TREATMENT_SUMMARY (normozoospermia (NZ) group). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Seminal fluid samples were allowed to thaw at room temperature and homogenized SP:SAMPLEPREP_SUMMARY using a vortex mixer. Then, samples were centrifuged at 20000.g for 20 min at SP:SAMPLEPREP_SUMMARY 4°C to remove any insoluble particles. After centrifugation, 45 µL of seminal SP:SAMPLEPREP_SUMMARY fluid was diluted in 135 µL of deuterated water and 45 µL of a 10 mM sodium SP:SAMPLEPREP_SUMMARY fumarate in 0.2 M phosphate buffer (pH 7.0) deuterated water (99.9%) solution. #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME 500 MHz Bruker Avance III HD NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:NMR_COMMENTS The samples were transferred to 3-mm NMR tubes (Norell, Landisville, NJ, USA) NM:NMR_COMMENTS and analyzed using the NOR5X3INSB optimizer inserts (Norell, Landisville, NJ, NM:NMR_COMMENTS USA) on a 500 MHz Bruker Avance III HD spectrometer equipped with a 5 mm TXI NM:NMR_COMMENTS probe (Bruker Corporation, Billerica, MA, USA), working at 298K. NM:NMR_COMMENTS Solvent-suppressed 1D-1H-NMR spectra were acquired using a zgpr pulse sequence, NM:NMR_COMMENTS with a sweep width of 7 kHz, relaxation delay of 7 s, pulse angle of 30°, NM:NMR_COMMENTS acquisition time of 2.3 s and 64 scans. Spectra were processed by applying a NM:NMR_COMMENTS line broadening of 0.2 Hz before Fourier transformation, and manually phased and NM:NMR_COMMENTS baseline corrected. Sodium fumarate (6.50 ppm) was used as an internal reference NM:NMR_COMMENTS for chemical shifts and metabolite quantification. The NUTS-Pro NMR software NM:NMR_COMMENTS (Acorn NMR, Inc, Fremont, CA, USA) was used for the processing of the spectra NM:NMR_COMMENTS and the quantification of the metabolites. Peaks were assigned by comparing the NM:NMR_COMMENTS acquired spectra with reference spectra in Chenomx (Chenomx Inc. Edmonton, NM:NMR_COMMENTS Canada) and the Human Metabolome Database (HMDB) V 5.0. 1H-NMR metabolomics data NM:NMR_COMMENTS was analyzed using Metaboanalyst 5.0 online (https://www.metaboanalyst.ca/). NM:NMR_COMMENTS Missing data was replaced by feature mean and data was log-transformed. NM:SPECTROMETER_FREQUENCY 500 MHz NM:NMR_PROBE 5 mm TXI probe NM:NMR_SOLVENT D2O NM:NMR_TUBE_SIZE 3mm NM:PULSE_SEQUENCE zgpr NM:WATER_SUPPRESSION Yes NM:RECEIVER_GAIN 203 NM:OFFSET_FREQUENCY 4.696 ppm NM:PRESATURATION_POWER_LEVEL 50 dB NM:TEMPERATURE 25 NM:NUMBER_OF_SCANS 64 NM:DUMMY_SCANS 2 NM:ACQUISITION_TIME 2.3 sec NM:RELAXATION_DELAY 7 sec NM:SPECTRAL_WIDTH 14 ppm NM:LINE_BROADENING 0.2 Hz NM:CHEMICAL_SHIFT_REF_STD Fumarate (doublet, 6.50 ppm) #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS normalized areas NMR_METABOLITE_DATA_START Samples N3 N4 N14 N15 N22 N24 N26 N27 N28 N29 N33 N34 N39 N43 N44 N49 N53 N56 N58 N64 N2 N5 N6 N7 N8 N10 N11 N12 N13 N16 N18 N19 N20 N21 N23 N25 N30 N31 N35 N36 N38 N40 N42 N45 N46 N47 N48 N50 N51 N52 N55 N57 N59 N61 N63 N68 N70 Factors Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Asthenozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Group:Normozoospermia | Sample source:Human seminal fluid Valine 1.557298228 1.498209103 1.151759494 1.814212234 1.492183491 1.71638431 1.367857361 1.054400692 1.028445999 1.720465445 1.472109282 1.353239129 1.907637726 1.452500573 1.466188021 1.329226167 1.122430648 1.118904657 1.415759565 0.770333106 1.20723935 1.325574409 1.588194867 1.688778658 1.360905235 1.313232954 1.559450156 1.55684766 1.138211027 0.765084649 1.536192992 1.468890344 1.299091034 1.278025441 1.062273693 0.909989239 1.5381087 1.449208143 1.734928339 1.426686516 0.958685433 1.423416884 1.160615349 1.185872009 1.476285625 1.65519315 0.781489711 1.698225438 0.795134632 0.481167402 0.740659843 0.723691048 1.259318 1.51833323 1.521660614 1.555407041 1.86125389 Isoleucine 1.477884346 1.362994312 1.069599583 1.725144454 1.463199039 1.63959744 1.303077191 1.119993825 0.925903119 1.658092075 1.374291498 1.219941387 1.832911509 1.320652985 1.388165342 1.241678817 1.137615719 0.919005477 1.239467564 0.712980432 1.119562088 1.302898764 1.510016043 1.593478557 1.31907301 1.217349743 1.441867001 1.444146049 1.035616485 0.731986159 1.384059228 1.374949867 1.239796199 1.187279392 1.002308551 0.784903212 1.554512065 1.374563022 1.679934055 1.320665154 0.883892839 1.36875044 1.122622473 1.057429848 1.461106195 1.617173693 0.719567616 1.460020744 0.78773875 0.43601527 0.638622323 0.602947661 1.059102206 1.29157472 1.333666413 1.449332522 1.726618532 Alanine 1.38729956 1.318737922 0.952893395 1.581091883 1.324424151 1.393948081 1.235063009 0.790029634 0.778867221 1.588589463 1.280133618 1.247056511 1.767159945 1.205444455 1.191637446 1.086439701 0.864291792 0.838497913 1.237102244 0.758361712 1.072752968 1.091451299 1.433361992 1.530761106 1.236468523 1.084353203 1.37052609 1.436483405 0.999291861 0.645074886 1.281147518 1.257934675 1.109043581 1.050175 0.924374269 0.78283833 1.261160001 1.276593257 1.555776678 1.284800272 0.790810907 1.244796612 1.01350255 0.93409963 1.253404646 1.41510237 0.576179602 1.482274136 0.606945839 0.077664998 0.34265357 0.460806652 0.9999278 1.267695034 1.339944947 1.40451405 1.622271735 Glutamate 1.893542472 1.935233225 1.601172965 2.186637983 1.947931224 2.226865651 1.856711983 1.446456804 1.345023697 2.121274861 2.012838183 1.962583451 2.376728178 1.870074068 1.825478309 1.75409185 1.566105449 1.528801989 1.959103786 1.528011489 1.570417895 1.789048225 1.949457914 2.18384891 1.954001387 1.741975166 2.019544029 2.043172668 1.754856841 1.406181625 2.07533636 1.943780642 1.738333104 1.666749452 1.525626436 1.441743241 2.002495676 1.976694186 2.234378561 1.9690946 1.356960855 1.877319297 1.619831174 1.598120721 1.96812775 2.161150772 1.254878534 2.183546618 1.301935137 1.143773313 1.200378622 1.28964181 1.559705004 2.036483714 2.042051726 2.081956842 2.315680101 Glutamine 1.759687378 1.511368299 1.087283473 1.839411087 1.567156824 1.867952468 1.472190768 1.165115628 1.096479303 1.861097637 1.540070156 1.34968402 1.913811523 1.431613627 1.511768162 1.407540608 1.229089691 1.122973896 1.462342303 0.809438096 1.339240664 1.431858407 1.655874645 1.749665401 1.404937374 1.375881374 1.668817721 1.720718524 1.201070783 -1.812346047 1.697455522 1.621022198 1.286831434 1.352389469 1.119728118 1.536502253 1.620954499 1.387728864 1.762376684 1.448132484 1.536502253 1.418677587 1.260941625 1.264788758 1.627147883 1.719051048 0.806787944 1.694342583 0.935179568 0.576198544 0.728848164 1.536502253 1.322165603 1.782812006 1.657471776 1.590783185 2.055546595 Citrate 2.320707884 2.382701242 2.124412724 2.243993888 2.193741363 1.907366901 2.120415901 1.588668091 1.726191939 2.318203989 2.076064615 2.648764638 2.493824555 2.054436027 2.101970374 2.066825167 1.557282887 1.612666284 2.215822247 1.879587028 1.964283299 1.869804545 2.361836584 2.325188589 2.399671448 1.824869836 1.941763488 1.908712721 2.417851792 2.1933517 2.076060567 1.999174514 1.7516802 1.907163464 1.892884519 1.775320597 1.735187889 2.063439486 2.327507544 2.248963 1.619745243 2.058520376 1.791319713 1.616024511 2.216684356 2.407611541 1.809929841 2.070809812 1.722919867 1.117334598 1.078064518 1.428804958 1.858399216 2.04432544 2.203743669 2.387336726 2.364975541 Aspartate 1.4539629 1.323864612 0.903044397 1.652127613 1.066442023 1.555282326 1.277674909 0.786951916 0.7909858 1.647670767 1.198145279 1.412729405 1.857982398 1.155857918 1.228958067 1.062602838 0.787817269 0.738660403 1.319146453 0.317741211 1.089580232 1.000083922 1.452229519 1.566736531 1.348233543 0.951041448 1.291591053 1.199549499 1.118056355 0.869479229 1.171810908 1.234731474 1.041164863 1.016886095 1.02233611 0.809029991 1.470785965 1.376745485 1.655475725 1.270891996 0.673374282 1.38289417 0.948463852 1.14451356 1.182632972 1.468251168 0.616732908 1.439792102 0.56917987 -1.812346047 0.704461638 0.483244445 1.058004281 1.386350258 1.386048676 1.481452478 1.780951081 Asparagine 1.363603095 1.366058802 1.134902333 1.7683728 1.405466968 1.781156278 1.477162703 0.75558872 0.93790789 1.828995007 1.272561757 1.359442093 1.90211119 1.462104083 1.472265732 1.231123263 1.010893307 1.033582908 1.488444302 0.724143083 1.319923726 1.130082179 1.652143599 1.708124613 1.335748566 1.374812106 1.605365004 1.537731672 1.037244725 0.782171234 1.432155273 1.462381397 1.359443746 1.301233051 0.966005095 0.828463594 1.553948407 1.366494261 1.752746089 1.315250015 0.87482649 1.393730881 1.211342224 1.143153927 1.353094168 1.545020645 0.73236234 1.736411173 0.813076845 0.338194814 0.513018303 0.522306903 1.164499574 1.612323952 1.534553445 1.663462781 1.952617901 Creatine 0.914738144 0.958567395 0.61238345 1.119310294 0.885085552 1.383193871 0.882932023 0.729650788 0.720937589 1.330280726 1.061644411 0.686034529 1.083469026 1.066351765 0.89836452 0.807392318 0.832474285 0.609634531 0.773453445 0.417703146 0.679608558 0.951841489 0.961203312 1.226198257 0.708354173 0.806416519 1.166050935 1.11610859 0.447925198 0.380162496 1.100692535 1.027478407 0.617771628 0.74551091 0.446354858 0.426340837 1.079694103 1.018319145 1.166683956 1.131569298 0.589963413 0.883178676 0.771684307 0.722927078 1.077480145 1.226641063 0.330361157 1.174147114 0.527021479 0.430616026 0.279604945 0.390219376 0.446844672 1.263400174 1.275407311 1.022012159 1.51323301 Choline 2.097681563 1.974212294 1.684610582 2.439069716 2.160051932 2.306934898 1.828828263 1.673864255 1.598538893 2.222000222 2.177595482 1.804244209 2.22966533 1.940278476 1.867911731 1.964526464 1.729675375 1.58522876 2.026536956 1.321197682 1.634529337 1.867238334 1.989091918 2.297607933 1.83578776 1.909938091 2.213414086 2.135771266 1.702924997 1.210616929 2.178967426 2.129061634 1.801115589 1.776226878 1.577954247 1.474374523 2.138067002 2.001629685 2.272065101 2.057191695 1.584262844 1.6983524 1.861441215 1.760701883 2.086219039 2.222354053 1.373785414 2.341125281 1.418396278 1.293542049 1.493123614 1.446178171 1.606837964 2.176812987 2.008483669 1.916314935 2.330324737 Glycine 1.695520815 1.723193505 1.348825159 1.933925841 1.615011918 1.855556373 1.587317146 1.185112607 1.226665765 1.919638293 1.692798628 1.585974526 2.115050376 1.599497909 1.617635538 1.471668945 1.18306644 1.238706978 1.578762608 1.043400451 1.427407005 1.54417362 1.802060462 1.861237109 1.58547282 1.383853628 1.675176689 1.779432666 1.328428261 1.064149719 1.665286297 1.627216865 1.419348072 1.395223522 1.28997129 1.178379314 1.569064021 1.632908999 1.927749187 1.609550115 1.15169385 1.645009538 1.351292258 1.248735415 1.684661532 1.796961805 1.014467181 1.777354142 0.972079957 0.771194285 0.838032597 0.914355231 1.387699671 1.62553606 1.707017819 1.703370615 2.00245579 Serine 1.985377454 1.934283783 1.578416701 2.260946609 1.977830728 2.160303382 1.830848997 1.471761394 1.510528189 2.196423221 1.923017811 1.806631693 2.321494648 1.829650773 1.871569096 1.754622697 1.579009045 1.475571223 1.894956601 1.290129726 1.65239937 1.79351634 2.08691738 2.175950087 1.792631259 1.798633704 1.960288621 2.023600248 1.597808339 1.22688757 1.999379895 1.915032431 1.817212275 1.691948896 1.449943164 1.345743514 1.931803962 1.854029592 2.216663314 1.843335316 1.428356408 1.887927379 1.624390784 1.564979536 1.932323687 2.13618641 1.216547223 2.005041876 1.330434482 0.969117213 1.087332533 1.101754943 1.589711061 1.898167871 1.959585173 2.032744106 2.315225882 Glucose 1.005306205 0.829805786 0.383753507 1.106619447 0.942176248 1.194752469 0.857740663 0.590680111 0.293298352 0.938755641 0.731277189 1.020417125 1.020310831 0.731546741 0.782311716 0.814554924 0.678664435 0.606436046 0.830669582 0.239776552 0.621556033 1.017939257 0.821615249 1.138842341 0.686111544 0.808380038 1.014057679 1.063409287 0.578493682 0.112934514 0.864044878 0.96885628 0.628685568 0.412595712 0.350098736 0.418955091 1.111048675 0.818417208 0.881085411 0.863110654 0.43659072 0.648093623 0.8019378 0.386418198 0.930937378 1.068648333 0.514242182 1.091716392 0.227576937 0.405060793 0.295145787 -0.115049527 0.354865282 0.949544591 0.949461528 0.840879044 1.236604146 Uridine 1.289890296 0.862318069 0.943807316 1.224848694 1.195853757 1.376773617 1.001917653 0.97218304 1.071149127 1.411450118 1.041747014 1.225697235 1.438479868 0.97527291 1.179439625 1.195093984 1.08084742 0.751040505 1.239930238 1.219317158 1.059682925 1.431997562 1.125763951 1.590335595 1.087586069 1.294856998 1.443415797 1.583786542 0.665160861 0.702772416 1.42072109 1.3918683 1.076947204 0.328974921 0.548960396 1.013833753 1.50018766 1.166384623 0.988985522 1.452242804 0.309045023 1.335706593 0.993442279 1.0290312 1.253246646 1.30056077 0.854163871 1.493071766 1.012810112 1.062408029 0.162740125 0.371525944 1.015820158 0.791051758 1.448850797 1.059838422 1.567018348 Tyrosine 1.091987522 1.147287166 0.864317041 1.281817841 1.106942364 1.26988888 0.963521679 0.863540007 0.751569386 1.225113692 1.237767306 1.120707609 1.424127564 0.954157369 1.04218351 1.079737732 0.923615888 0.833717935 1.193767964 0.984019217 0.818713962 1.169554853 1.095870335 1.354789507 1.008257946 1.012270068 1.208812763 1.28304105 0.874092749 0.828973395 1.320660258 1.223917415 0.944608109 0.778560578 0.906432002 0.844123049 1.228109272 1.12047821 1.227437967 1.23713936 0.644208511 1.005828044 0.938562707 0.897140091 1.153340712 1.286536864 0.624207749 1.295156735 0.747880972 0.801880382 0.653883191 0.528041022 0.800435695 1.067125096 1.259649405 1.023054756 1.369712006 Phenylalanine 0.847397066 0.889274621 0.670520664 0.84542696 0.791514363 0.943795649 0.760323041 0.439178215 0.396947252 1.048829563 0.940354992 0.657953751 1.237905646 0.588974246 0.848327047 0.648155758 0.358932391 0.387550599 0.726843195 0.341802075 0.684728763 0.701613469 1.032446422 0.998660836 0.785700906 0.6588445 0.874935079 1.043889425 0.464530239 0.466544408 0.861531533 0.807333689 0.687236715 0.644432865 0.506046411 0.328502872 0.886352681 0.775477042 1.05044821 0.794291424 0.329517693 0.841002819 0.49771173 0.425915851 0.784896173 0.969640128 0.12030554 0.75921943 0.191866913 -0.078237612 0.001963872 -0.152878507 0.477655469 0.656626806 0.906161282 0.826893431 1.062651508 Pyroglutamate 0.993914745 1.11898588 0.866980929 0.695023022 1.137862855 1.063590893 0.994591423 1.043865832 0.96780332 1.6743816 1.588727891 0.411169485 1.35510101 0.996351333 1.248954228 0.802743909 1.258983718 0.819389746 0.967722036 0.94845862 1.102186136 1.273900774 1.517548119 1.518362047 0.60697314 0.760879929 0.860191125 0.080640312 -0.024406003 0.4700246 1.063590893 1.121838037 0.83884499 1.140448747 -0.481673841 -0.151625167 1.063590893 0.711583182 1.298427907 0.652943228 0.857732863 1.249162855 1.063590893 0.486581647 0.569366727 0.753211514 0.38683831 1.063590893 0.375613489 1.063590893 -1.812346047 0.460368447 0.939673801 1.063590893 0.900461248 1.333928024 -0.012969048 Fructose 0.951395918 1.495273011 0.937787756 1.129250729 1.057133808 1.526655423 1.116517887 0.989659277 0.961938498 1.65345236 1.525521632 0.89461992 1.209617831 1.106534278 1.055719637 0.743717179 1.282423359 0.878275523 1.274104286 0.547001071 0.718221909 0.92764927 1.195174547 1.663462703 0.987745957 0.943608694 1.515979766 1.572321301 -0.510234365 0.770273567 1.505961603 1.658201865 0.745000963 0.909803177 -0.375153051 -0.328793412 1.478101164 1.23960686 1.233082024 1.374919884 0.69970508 0.749471751 1.023379795 0.887064293 1.046646394 1.44190221 0.645887037 1.660482473 0.443706773 0.715304518 0.645979698 0.681782494 0.249029674 1.707001018 1.349854957 1.234322825 1.557348537 Lactate 1.497516648 1.394212318 1.033541348 1.604445727 1.654825615 2.254966385 0.997118823 1.187482371 0.929556879 1.50059428 1.682866094 0.93899528 1.721346031 1.298572867 1.466411867 1.118194071 0.930715615 1.052742862 1.439554884 1.651893859 1.034135404 1.663156225 1.411579634 1.716001091 1.653162871 1.353213971 1.597186948 1.616930227 1.278199842 0.807706437 1.51291714 1.395159351 1.302328299 1.08274537 1.732157309 1.519306361 1.690178788 1.761671651 1.764904618 1.891579665 0.742075271 1.616439149 1.433588292 1.61872771 1.70617013 1.865348266 0.898205167 1.870046841 0.905428578 0.78449252 1.021239294 1.017701644 1.103662833 1.803368928 1.810467069 1.460577151 1.904147626 sn-3-GPC 1.439654931 1.090094838 1.210261559 1.483300326 1.229161677 1.610864118 1.039226797 0.782459632 0.752560422 1.28527943 1.503347224 0.563680619 1.705568516 1.651348483 1.257852802 1.419132819 0.894577572 0.857781135 1.302695364 0.479030135 0.821507366 1.260337945 1.359650502 1.68952824 1.233866067 1.201195603 1.401831176 1.408349687 1.084626189 0.99281953 1.35048496 1.260323575 1.031853946 0.833961606 1.194806692 1.025298901 1.351552778 1.436623423 1.640571081 1.391318232 0.888520023 1.008970925 1.013689603 1.25803907 1.220908418 1.712680319 0.50221644 1.477642036 0.514431438 0.231165031 0.742564077 0.835038762 1.227755028 1.469337326 1.232508253 1.169852869 1.619573705 Histidine 1.303908974 1.337457476 0.98607867 1.110101324 1.237505493 1.582488428 1.103447762 0.964940488 0.874064696 1.469446481 1.350206008 0.905185408 1.584863031 1.166417437 1.285738076 1.062451031 1.128376614 0.907876732 1.15304316 0.72922797 1.013319893 1.275998618 1.441615578 1.523654345 1.10550444 1.143082296 1.449805608 1.501972752 0.854559412 0.958778829 1.469011093 1.350223754 1.144687947 1.135074987 0.80025194 0.631017498 1.463327044 1.236740484 1.464789958 1.250040527 0.716718854 1.220519872 0.994632112 1.021202433 1.260119921 1.421821623 0.538664139 1.496059944 0.755321596 0.525752157 0.667260369 0.599304995 0.897544623 1.381064051 1.289666231 1.172461518 1.509111702 Threonine 1.268268079 1.483867089 0.754815039 1.14779308 1.255511862 1.641946309 1.338793937 0.664422966 0.788699926 1.795628297 1.409569771 1.302677964 1.828410616 1.32718279 1.281213128 1.083212847 0.842668125 0.924471804 1.236873483 0.630284969 1.040127016 0.905356758 1.473511655 1.665232386 1.239684447 1.024866482 1.435461441 1.375103373 1.056035637 1.067086243 1.40232515 1.382947321 1.188702459 1.134653492 0.954340428 0.914296576 1.435243349 1.358514688 1.720925052 1.262904425 0.727206281 1.288184048 1.12981848 1.056404289 1.37049671 1.605523172 0.533203726 1.641931446 0.560770367 0.785041128 0.392026588 0.59518566 1.144943817 1.477091864 1.406542852 1.527575957 1.852260303 Cadaverine 1.627516839 1.533888742 1.23736792 1.50010091 1.440026479 1.789282804 1.484067641 1.075211524 1.083216384 1.826894053 1.546953362 1.392007261 1.960581963 1.478925326 1.521678165 1.351680085 1.139069392 1.129726061 1.475499821 0.829836767 1.327197728 1.379639258 1.676279718 1.653806737 1.461493825 1.386466342 1.621264319 1.674659476 1.144525985 1.221440353 1.624886414 1.509354437 1.378502432 1.326670306 1.113929325 0.958156898 1.553703495 1.528100067 1.835138714 1.484310198 1.039577349 1.478116895 1.157491121 1.111505433 1.531035153 1.723361581 0.856749988 1.732655711 0.932680246 0.658168714 0.806335024 0.546463977 1.26092242 1.595903705 1.524607403 1.632418314 1.911023736 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Formula HMDB code Valine C5H11NO2 HMDB0000883 Isoleucine C6H13NO2 HMDB0000172 Alanine C3H7NO2 HMDB0000161 Glutamate C5H9NO4 HMDB0000148 Glutamine C5H10N2O3 HMDB0000641 Citrate C6H8O7 HMDB0000094 Aspartate C4H7NO4 HMDB0000191 Asparagine C4H8N2O3 HMDB0000168 Creatine C4H9N3O2 HMDB0000064 Choline C5H14NO HMDB0000097 Glycine C2H5NO2 HMDB0000123 Serine C3H7NO3 HMDB0000187 Glucose C6H12O6 HMDB0000122 Uridine C9H12N2O6 HMDB0000296 Tyrosine C9H11NO3 HMDB0000158 Phenylalanine C9H11NO2 HMDB0000159 Pyroglutamate C5H7NO3 HMDB0000267 Fructose C6H12O6 HMDB0000660 Lactate C3H6O3 HMDB0001311 sn-3-GPC C8H21NO6P HMDB0000086 Histidine C6H9N3O2 HMDB0000177 Threonine C4H9NO3 HMDB0000167 Cadaverine C5H14N2 HMDB0002322 METABOLITES_END #END