Seminal plasma exosomes and male infertility: current progress and future directions

Male infertility remains a major global health concern. Conventional semen analysis offers limited explanatory power, with most cases remaining idiopathic, underscoring the urgent need for novel biomarkers and mechanisms. Exosomes, 30–150 nm extracellular vesicles that transfer proteins, RNAs, and bioactive molecules, have emerged as pivotal regulators of male reproductive function. Epididymosomes promote sperm maturation and capacitation by delivering proteins and antioxidants. Prostasomes act post-ejaculation, supporting capacitation, the acrosome reaction, and immune protection within the female reproductive tract. Testicular exosomes regulate early spermatogenesis, particularly by modulating spermatogonial stem-cell proliferation and differentiation. Seminal-vesicle exosomes influence motility and fertilization, while bulbourethral exosomes may modulate seminal pH, ion balance, and viscosity. Dysregulation of these exosomal pathways has been linked to oligozoospermia, asthenozoospermia, and teratozoospermia. This review integrates mechanistic and translational evidence across seminal plasma exosome subtypes, emphasizes their reproductive functions, and highlights their diagnostic and therapeutic potential, offering a foundation for exosome-based precision medicine in male infertility.

Seminal plasma exosomes; Epididymosomes; Prostasomes; Testicular exosomes; Male infertility; Sperm maturation; Sperm function; Intercellular communication; Biomarkers

[1] Agarwal A, Baskaran S, Parekh N, Cho CL, Henkel R, Vij S, et al. Male infertility. The Lancet. 2021; 397: 319–333.

[2] El-Hamd M, Aboeldahab S. Cell phone and male infertility: an update. Integrative Medicine in Nephrology and Andrology. 2018; 5: 1–5.

[3] Kumar S, Agrawal D, Sharma K, Swain T. Association of male infertility to metabolic syndrome and other related disorders. Integrative Medicine in Nephrology and Andrology. 2015; 2: 107–116.

[4] Boitrelle F, Shah R, Saleh R, Henkel R, Kandil H, Chung E, et al. The sixth edition of the WHO manual for human semen analysis: a critical review and SWOT analysis. Life. 2021; 11: 1368.

[5] van Niel G, D’Angelo G, Raposo G. Shedding light on the cell biology of extracellular vesicles. Nature Reviews Molecular Cell Biology. 2018; 19: 213–228.

[6] Gurung S, Perocheau D, Touramanidou L, Baruteau J. The exosome journey: from biogenesis to uptake and intracellular signalling. Cell Communication and Signaling. 2021; 19: 47.

[7] van Niel G, Carter DRF, Clayton A, Lambert DW, Raposo G, Vader P. Challenges and directions in studying cell–cell communication by extracellular vesicles. Nature Reviews Molecular Cell Biology. 2022; 23: 369–382.

[8] Höög JL, Lötvall J. Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy. Journal of Extracellular Vesicles. 2015; 4: 28680.

[9] Pan B, Johnstone RM. Fate of the transferrin receptor during maturation of sheep reticulocytes in vitro: selective externalization of the receptor. Cell. 1983; 33: 967–978.

[10] Raposo G, Nijman HW, Stoorvogel W, Liejendekker R, Harding CV, Melief CJ, et al. B lymphocytes secrete antigen-presenting vesicles. The Journal of Experimental Medicine. 1996; 183: 1161–1172.

[11] Valadi H, Ekström K, Bossios A, Sjöstrand M, Lee JJ, Lötvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nature Cell Biology. 2007; 9: 654–659.

[12] Krylova SV, Feng D. The machinery of exosomes: biogenesis, release, and uptake. International Journal of Molecular Sciences. 2023; 24: 1337.

[13] Kowalczyk A, Wrzecińska M, Czerniawska-Piątkowska E, Kupczyński R. Exosomes—spectacular role in reproduction. Biomedicine & Pharmacotherapy. 2022; 148: 112752.

[14] Wang H, Zhu Y, Tang C, Zhou Z, Wang Z, Li Z, et al. Reassessment of the proteomic composition and function of extracellular vesicles in the seminal plasma. Endocrinology. 2022; 163: bqab214.

[15] Aalberts M, Stout TAE, Stoorvogel W. Prostasomes: extracellular vesicles from the prostate. Reproduction. 2013; 147: R1–R14.

[16] Martin-DeLeon P. Epididymosomes: transfer of fertility-modulating proteins to the sperm surface. Asian Journal of Andrology. 2015; 17: 720–725.

[17] Khodamoradi K, Golan R, Dullea A, Ramasamy R. Exosomes as potential biomarkers for erectile dysfunction, varicocele, and testicular injury. Sexual Medicine Reviews. 2022; 10: 311–322.

[18] Simon C, Greening DW, Bolumar D, Balaguer N, Salamonsen LA, Vilella F. Extracellular vesicles in human reproduction in health and disease. Endocrine Reviews. 2018; 39: 292–332.

[19] Trams EG, Lauter CJ, Salem N III, Heine U. Exfoliation of membrane ecto-enzymes in the form of micro-vesicles. Biochimica et Biophysica Acta (BBA)—Biomembranes. 1981; 645: 63–70.

[20] Johnstone RM, Adam M, Hammond JR, Orr L, Turbide C. Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles (exosomes). Journal of Biological Chemistry. 1987; 262: 9412–9420.

[21] Poliakov A, Spilman M, Dokland T, Amling CL, Mobley JA. Structural heterogeneity and protein composition of exosome-like vesicles (prostasomes) in human semen. The Prostate. 2009; 69: 159–167.

[22] Samanta L, Parida R, Dias TR, Agarwal A. The enigmatic seminal plasma: a proteomics insight from ejaculation to fertilization. Reproductive Biology and Endocrinology. 2018; 16: 41.

[23] Sharma R, Agarwal A, Mohanty G, Du Plessis SS, Gopalan B, Willard B, et al. Proteomic analysis of seminal fluid from men exhibiting oxidative stress. Reproductive Biology and Endocrinology. 2013; 11: 85.

[24] Rolland AD, Lavigne R, Dauly C, Calvel P, Kervarrec C, Freour T, et al. Identification of genital tract markers in the human seminal plasma using an integrative genomics approach. Human Reproduction. 2013; 28: 199–209.

[25] Druart X, Rickard JP, Tsikis G, de Graaf SP. Seminal plasma proteins as markers of sperm fertility. Theriogenology. 2019; 137: 30–35.

[26] Théry C, Witwer KW, Aikawa E, Alcaraz MJ, Anderson JD, Andriantsitohaina R, et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. Journal of Extracellular Vesicles. 2018; 7: 1535750.

[27] Théry C, Regnault A, Garin J, Wolfers J, Zitvogel L, Ricciardi-Castagnoli P, et al. Molecular characterization of dendritic cell-derived exosomes. Journal of Cell Biology. 1999; 147: 599–610.

[28] Escola J, Kleijmeer MJ, Stoorvogel W, Griffith JM, Yoshie O, Geuze HJ. Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes. Journal of Biological Chemistry. 1998; 273: 20121–20127.

[29] Yu K, Xiao K, Sun QQ, Liu RF, Huang LF, Zhang PF, et al. Comparative proteomic analysis of seminal plasma exosomes in buffalo with high and low sperm motility. BMC Genomics. 2023; 24: 8.

[30] Baranyai T, Herczeg K, Onódi Z, Voszka I, Módos K, Marton N, et al. Isolation of exosomes from blood plasma: qualitative and quantitative comparison of ultracentrifugation and size exclusion chromatography methods. PLOS ONE. 2015; 10: e0145686.

[31] Yang C, Guo WB, Zhang WS, Bian J, Yang JK, Zhou QZ, et al. Comprehensive proteomics analysis of exosomes derived from human seminal plasma. Andrology. 2017; 5: 1007–1015.

[32] Colombo M, Moita C, van Niel G, Kowal J, Vigneron J, Benaroch P, et al. Analysis of ESCRT functions in exosome biogenesis, composition and secretion highlights the heterogeneity of extracellular vesicles. Journal of Cell Science. 2013; 126: 5553–5565.

[33] Baietti MF, Zhang Z, Mortier E, Melchior A, Degeest G, Geeraerts A, et al. Syndecan–syntenin–ALIX regulates the biogenesis of exosomes. Nature Cell Biology. 2012; 14: 677–685.

[34] Kowal J, Arras G, Colombo M, Jouve M, Morath JP, Primdal-Bengtson B, et al. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proceedings of the National Academy of Sciences. 2016; 113: E968–E977.

[35] Mathieu M, Névo N, Jouve M, Valenzuela JI, Maurin M, Verweij FJ, et al. Specificities of exosome versus small ectosome secretion revealed by live intracellular tracking of CD63 and CD9. Nature Communications. 2021; 12: 4389.

[36] Lozano-Ramos I, Bancu I, Oliveira-Tercero A, Armengol MP, Menezes-Neto A, Portillo HAD, et al. Size-exclusion chromatography-based enrichment of extracellular vesicles from urine samples. Journal of Extracellular Vesicles. 2015; 4: 27369.

[37] Lötvall J, Hill AF, Hochberg F, Buzás EI, Di Vizio D, Gardiner C, et al. Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. Journal of Extracellular Vesicles. 2014; 3: 26913.

[38] Murdica V, Giacomini E, Alteri A, Bartolacci A, Cermisoni GC, Zarovni N, et al. Seminal plasma of men with severe asthenozoospermia contain exosomes that affect spermatozoa motility and capacitation. Fertility and Sterility. 2019; 111: 897–908.e2.

[39] Gilany K, Minai-Tehrani A, Savadi-Shiraz E, Rezadoost H, Lakpour N. Exploring the human seminal plasma proteome: an unexplored gold mine of biomarker for male infertility and male reproduction disorder. Journal of Reproduction & Infertility. 2015; 16: 61–71.

[40] Candenas L, Chianese R. Exosome composition and seminal plasma proteome: a promising source of biomarkers of male infertility. International Journal of Molecular Sciences. 2020; 21: 7022.

[41] Tu C, Rudnick PA, Martinez MY, Cheek KL, Stein SE, Slebos RJC, et al. Depletion of abundant plasma proteins and limitations of plasma proteomics. Journal of Proteome Research. 2010; 9: 4982–4991.

[42] Goss DM, Vasilescu SA, Sacks G, Gardner DK, Warkiani ME. Microfluidics facilitating the use of small extracellular vesicles in innovative approaches to male infertility. Nature Reviews Urology. 2023; 20: 66–95.

[43] Eswaran N, Agaram Sundaram V, Rao KA, Thalaivarisai Balasundaram S. Simple isolation and characterization of seminal plasma extracellular vesicle and its total RNA in an academic lab. 3 Biotech. 2018; 8: 139.

[44] Sidhom K, Obi PO, Saleem A. A review of exosomal isolation methods: is size exclusion chromatography the best option? International Journal of Molecular Sciences. 2020; 21: 6466.

[45] An M, Wu J, Zhu J, Lubman DM. Comparison of an optimized ultracentrifugation method versus size-exclusion chromatography for isolation of exosomes from human serum. Journal of Proteome Research. 2018; 17: 3599–3605.

[46] Nordin JZ, Lee Y, Vader P, Mäger I, Johansson HJ, Heusermann W, et al. Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties. Nanomedicine. 2015; 11: 879–883.

[47] Wu Y, Wang Y, Lu Y, Luo X, Huang Y, Xie T, et al. Microfluidic technology for the isolation and analysis of exosomes. Micromachines. 2022; 13: 1571.

[48] Caradec J, Kharmate G, Hosseini-Beheshti E, Adomat H, Gleave M, Guns E. Reproducibility and efficiency of serum-derived exosome extraction methods. Clinical Biochemistry. 2014; 47: 1286–1292.

[49] Noboa-Velástegui J, León JC, Castro J, Fletes A, Madrigal P, Álvarez I, et al. Comparison of methods for isolating exosomes from plasma subjects with normal and high fat percentages. Life. 2025; 15: 410.

[50] Konoshenko MY, Lekchnov EA, Vlassov AV, Laktionov PP. Isolation of extracellular vesicles: general methodologies and latest trends. BioMed Research International. 2018; 2018: 8545347.

[51] Weng Y, Sui Z, Shan Y, Hu Y, Chen Y, Zhang L, et al. Effective isolation of exosomes with polyethylene glycol from cell culture supernatant for in-depth proteome profiling. The Analyst. 2016; 141: 4640–4646.

[52] Petrova T, Kalinina O, Aquino A, Grigoryev E, Dubashynskaya NV, Zubkova K, et al. Topographic distribution of miRNAs (miR-30a, miR-223, miR-let-7a, miR-let-7f, miR-451, and miR-486) in the plasma extracellular vesicles. Non-Coding RNA. 2024; 10: 15.

[53] Alameldin S, Costina V, Abdel-Baset HA, Nitschke K, Nuhn P, Neumaier M, et al. Coupling size exclusion chromatography to ultracentrifugation improves detection of exosomal proteins from human plasma by LC-MS. Practical Laboratory Medicine. 2021; 26: e00241.

[54] Djoseland O. Androgen metabolism by rat epididymis. 3. Effect of castration and anti-androgens. Steroids. 1976; 27: 47–64.

[55] Cao D, Min X, Su L, Luo C, Cheng H, Zhang S, et al. Yi-jing decoction ameliorates oligoasthenozoospermia by inhibiting the oxidative stress-p38 mitogen-activated protein kinase-mediated mitochondrial apoptosis pathway in leydig and sertoli cells. Integrative Medicine in Nephrology and Andrology. 2025; 12: e24-00058.

[56] O’Flaherty C. Orchestrating the antioxidant defenses in the epididymis. Andrology. 2019; 7: 662–668.

[57] Domeniconi RF, Souza ACF, Xu B, Washington AM, Hinton BT. Is the epididymis a series of organs placed side by side? Biology of Reproduction. 2016; 95: 10.

[58] Thimon V, Koukoui O, Calvo E, Sullivan R. Region-specific gene expression profiling along the human epididymis. Molecular Human Reproduction. 2007; 13: 691–704.

[59] Yanagimachi R, Kamiguchi Y, Mikamo K, Suzuki F, Yanagimachi H. Maturation of spermatozoa in the epididymis of the Chinese hamster. American Journal of Anatomy. 1985; 172: 317–330.

[60] Frenette G, Sullivan R. Prostasome-like particles are involved in the transfer of P25b from the bovine epididymal fluid to the sperm surface. Molecular Reproduction and Development. 2001; 59: 115–121.

[61] Sullivan R, Saez F. Epididymosomes, prostasomes, and liposomes: their roles in mammalian male reproductive physiology. Reproduction. 2013; 146: R21–R35.

[62] Baskaran S, Panner Selvam MK, Agarwal A. Exosomes of male reproduction. Advances in Clinical Chemistry. 2020; 73: 149–163.

[63] Sullivan R, Mieusset R. The human epididymis: its function in sperm maturation. Human Reproduction Update. 2016; 22: 574–587.

[64] James ER, Carrell DT, Aston KI, Jenkins TG, Yeste M, Salas-Huetos A. The role of the epididymis and the contribution of epididymosomes to mammalian reproduction. International Journal of Molecular Sciences. 2020; 21: 5377.

[65] Barrachina F, Battistone MA, Castillo J, Mallofré C, Jodar M, Breton S, et al. Sperm acquire epididymis-derived proteins through epididymosomes. Human Reproduction. 2022; 37: 651–668.

[66] Paul N, Talluri TR, Nag P, Kumaresan A. Epididymosomes: a potential male fertility influencer. Andrologia. 2021; 53: e14155.

[67] Thimon V, Frenette G, Saez F, Thabet M, Sullivan R. Protein composition of human epididymosomes collected during surgical vasectomy reversal: a proteomic and genomic approach. Human Reproduction. 2008; 23: 1698–1707.

[68] Nixon B, De Iuliis GN, Hart HM, Zhou W, Mathe A, Bernstein IR, et al. Proteomic profiling of mouse epididymosomes reveals their contributions to post-testicular sperm maturation. Molecular & Cellular Proteomics. 2019; 18: S91–S108.

[69] Boué F, Blais J, Sullivan R. Surface localization of P34H an epididymal protein, during maturation, capacitation, and acrosome reaction of human spermatozoa. Biology of Reproduction. 1996; 54: 1009–1017.

[70] Martin-DeLeon PA. Epididymal SPAM1 and its impact on sperm function. Molecular and Cellular Endocrinology. 2006; 250: 114–121.

[71] Taylor A, Robson A, Houghton BC, Jepson CA, Ford WCL, Frayne J. Epididymal specific, selenium-independent GPX5 protects cells from oxidative stress-induced lipid peroxidation and DNA mutation. Human Reproduction. 2013; 28: 2332–2342.

[72] Sullivan R. Epididymosomes: a heterogeneous population of microvesicles with multiple functions in sperm maturation and storage. Asian Journal of Andrology. 2015; 17: 726–729.

[73] D’Amours O, Frenette G, Caron P, Belleannée C, Guillemette C, Sullivan R. Evidences of biological functions of biliverdin reductase a in the bovine epididymis. Journal of Cellular Physiology. 2016; 231: 1077–1089.

[74] Oh JS, Han C, Cho C. ADAM7 is associated with epididymosomes and integrated into sperm plasma membrane. Molecules and Cells. 2009; 28: 441–446.

[75] Choi H, Han C, Jin S, Kwon JT, Kim J, Jeong J, et al. Reduced fertility and altered epididymal and sperm integrity in mice lacking ADAM7. Biology of Reproduction. 2015; 93: 70.

[76] Ronquist G. Prostasomes are mediators of intercellular communication: from basic research to clinical implications. Journal of Internal Medicine. 2012; 271: 400–413.

[77] Ronquist G, Hedström M. Restoration of detergent-inactivated adenosine triphosphatase activity of human prostatic fluid with concanavalin A. Biochimica et Biophysica Acta (BBA)—Enzymology. 1977; 483: 483–486.

[78] Stegmayr B, Ronquist G. Promotive effect on human sperm progressive motility by prostasomes. Urological Research. 1982; 10: 253–257.

[79] Ronquist G, Brody I. The prostasome: its secretion and function in man. Biochimica et Biophysica Acta (BBA)—Reviews on Biomembranes. 1985; 822: 203–218.

[80] Vojtech L, Woo S, Hughes S, Levy C, Ballweber L, Sauteraud RP, et al. Exosomes in human semen carry a distinctive repertoire of small non-coding RNAs with potential regulatory functions. Nucleic Acids Research. 2014; 42: 7290–7304.

[81] Arienti G, Carlini E, Nicolucci A, Cosmi EV, Santi F, Palmerini CA. The motility of human spermatozoa as influenced by prostasomes at various pH levels. Biology of the Cell. 1999; 91: 51–54.

[82] Ronquist G. Prostasomes: their characterisation: implications for human reproduction. Advances in Experimental Medicine and Biology. 2015; 86: 191–209.

[83] Fraser LR. The “switching on” of mammalian spermatozoa: molecular events involved in promotion and regulation of capacitation. Molecular Reproduction and Development. 2010; 77: 197–208.

[84] Park KH, Kim BJ, Kang J, Nam TS, Lim JM, Kim HT, et al. Ca2+ signaling tools acquired from prostasomes are required for progesterone-induced sperm motility. Science Signaling. 2011; 4: ra31.

[85] Tarazona R, Delgado E, Guarnizo MC, Roncero RG, Morgado S, Sánchez-Correa B, et al. Human prostasomes express CD48 and interfere with NK cell function. Immunobiology. 2011; 216: 41–46.

[86] Palmerini CA, Saccardi C, Carlini E, Fabiani R, Arienti G. Fusion of prostasomes to human spermatozoa stimulates the acrosome reaction. Fertility and Sterility. 2003; 80: 1181–1184.

[87] Ma Y, Ma Q, Sun Y, Chen X. The emerging role of extracellular vesicles in the testis. Human Reproduction. 2023; 38: 334–351.

[88] Lin Y, Fang Q, He Y, Gong X, Wang Y, Liang A, et al. Thy1-positive spermatogonia suppress the proliferation of spermatogonial stem cells by extracellular vesicles in vitro. Endocrinology. 2021; 162: bqab052.

[89] Mohammadi A, Shabani R, Bashiri Z, Rafiei S, Asgari H, Koruji M. Therapeutic potential of exosomes in spermatogenesis regulation and male infertility. Biology of the Cell. 2024; 116: e2300127.

[90] Tan F, Li X, Wang Z, Li J, Shahzad K, Zheng J. Clinical applications of stem cell-derived exosomes. Signal Transduction and Targeted Therapy. 2024; 9: 17.

[91] Izadi M, Dehghan Marvast L, Rezvani ME, Zohrabi M, Aliabadi A, Mousavi SA, et al. Mesenchymal stem-cell derived exosome therapy as a potential future approach for treatment of male infertility caused by chlamydia infection. Frontiers in Microbiology. 2021; 12: 785622.

[92] Li Q, Li H, Liang J, Mei J, Cao Z, Zhang L, et al. Sertoli cell-derived exosomal MicroRNA-486-5p regulates differentiation of spermatogonial stem cell through PTEN in mice. Journal of Cellular and Molecular Medicine. 2021; 25: 3950–3962.

[93] Liu Z, Cao K, Liao Z, Chen Y, Lei X, Wei Q, et al. Monophosphoryl lipid a alleviated radiation-induced testicular injury through TLR4-dependent exosomes. Journal of Cellular and Molecular Medicine. 2020; 24: 3917–3930.

[94] Choy KHK, Chan SY, Lam W, Jin J, Zheng T, Law TYS, et al. The repertoire of testicular extracellular vesicle cargoes and their involvement in inter-compartmental communication associated with spermatogenesis. BMC Biology. 2022; 20: 78.

[95] Fayezi S, Fayyazpour P, Norouzi Z, Mehdizadeh A. Strategies for mammalian mesenchymal stem cells differentiation into primordial germ cell-like cells: a review. Cell Journal. 2022; 24: 434–441.

[96] Mobarak H, Heidarpour M, Rahbarghazi R, Nouri M, Mahdipour M. Amniotic fluid-derived exosomes improved spermatogenesis in a rat model of azoospermia. Life Sciences. 2021; 274: 119336.

[97] Parra A, Padilla L, Lucas X, Rodriguez-Martinez H, Barranco I, Roca J. Seminal extracellular vesicles and their involvement in male (in)fertility: a systematic review. International Journal of Molecular Sciences. 2023; 24: 4818.

[98] Korbakis D, Schiza C, Brinc D, Soosaipillai A, Karakosta TD, Légaré C, et al. Preclinical evaluation of a TEX101 protein ELISA test for the differential diagnosis of male infertility. BMC Medicine. 2017; 15: 60.

[99] Salas-Huetos A, Blanco J, Vidal F, Grossmann M, Pons MC, Garrido N, et al. Spermatozoa from normozoospermic fertile and infertile individuals convey a distinct miRNA cargo. Andrology. 2016; 4: 1028–1036.

[100] Barceló M, Mata A, Bassas L, Larriba S. Exosomal microRNAs in seminal plasma are markers of the origin of azoospermia and can predict the presence of sperm in testicular tissue. Human Reproduction. 2018; 33: 1087–1098.

[101] Shen Y, You Y, Zhu K, Fang C, Chang D, Yu X. Exosomes in the f ield of reproduction: a scientometric study and visualization analysis. Frontiers in Pharmacology. 2022; 13: 1001652.

[102] Wang C, Swerdloff RS. Limitations of semen analysis as a test of male fertility and anticipated needs from newer tests. Fertility and Sterility. 2014; 102: 1502–1507.

[103] Agarwal A, Majzoub A, Esteves SC, Ko E, Ramasamy R, Zini A. Clinical utility of sperm DNA fragmentation testing: practice recommendations based on clinical scenarios. Translational Andrology and Urology. 2016; 5: 935–950.

[104] Liu S, Geng Q, Zhong C, Yu X, Hong Z, Yan B, et al. Linggui Yangyuan paste for patients with male infertility: a study protocol for a multicenter, double-blind, double-dummy, randomized controlled trial. Journal of Men’s Health. 2023; 19: 67–72.