263 research outputs found

    Assessment of different anther culture approaches to produce doubled haploids in cucumber (Cucumis sativus L.)

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    [EN] Cucumber is one of the most important vegetable crops worldwide, which makes it a good candidate to produce doubled haploid (DH) lines to accelerate plant breeding. Traditionally, these approaches involved induction of gynogenesis or parthenogenesis with irradiated pollen, which carries some disadvantages compared to androgenesis. Despite this, studies on anther/microspore cultures in cucumber are surprisingly scarce. Furthermore, most of them failed to unambiguously demonstrate the haploid origin of the individuals obtained. In this work we focused on anther cultures using two cucumber genotypes, different previously published protocols for anther culture, different in vitro culture variants to make it more efficient, and most importantly, a combination of flow cytometry and microsatellite molecular markers to evaluate the real androgenic potential and the impact of anther wall tissue proliferation. We developed a method to produce DH plants involving a bud pretreatment at 4 C, a 35 C treatment to anthers, culture with BAP and 2,4-D, and induction of callus morphogenesis by an additional 35 C treatment and sequential culture first in liquid medium in darkness and second in solid medium with light. We also found that factors such as genotype, proliferation of anther wall tissues, orientation of anthers in the culture medium and growth regulator composition of the initial anther culture medium have a remarkable impact. Our rate of chromosome doubling (81%) was high enough to exclude additional chromosome doubling steps. Together, our results present androgenesis as an improvable but yet more convenient alternative to traditional gynogenesis and parthenogenesis-based approaches.Thanks are due to all the whole staff of the Cell Biology Group for helping and training AA during his stay in the group. This work was supported by Grant AGL2017- 88135-R to JMSS from Spanish Ministerio de Economı´a y Competitividad (MINECO) jointly funded by FEDER.Asadi, A.; Zebarjadi, A.; Abdollahi, MR.; Seguí-Simarro, JM. (2018). Assessment of different anther culture approaches to produce doubled haploids in cucumber (Cucumis sativus L.). Euphytica. 214(216):1-17. https://doi.org/10.1007/s10681-018-2297-xS117214216Abdollahi MR, Najafi S, Sarikhani H, Moosavi SS (2016) Induction and development of anther-derived gametic embryos in cucumber (Cucumis sativus L.) by optimizing the macronutrient and agar concentrations in culture medium. Turk J Biol 40(3):571–579Ashok Kumar HG, Murthy HN (2004) Effect of sugars and amino acids on androgenesis of Cucumis sativus. Plant Cell, Tissue Organ Cult 78(3):201–208. https://doi.org/10.1023/b:ticu.0000025637.56693.68Bai B, Su YH, Yuan J, Zhang XS (2013) Induction of somatic embryos in arabidopsis requires local YUCCA expression mediated by the down-regulation of ethylene biosynthesis. Mol Plant 6(4):1247–1260. https://doi.org/10.1093/mp/sss154Claveria E, Garcia-Mas J, Dolcet-Sanjuan R (2005) Optimization of cucumber doubled haploid line production using in vitro rescue of in vivo induced parthenogenic embryos. J Am Soc Hortic Sci 130(4):555–560Corral-Martínez P, Nuez F, Seguí-Simarro JM (2011) Genetic, quantitative and microscopic evidence for fusion of haploid nuclei and growth of somatic calli in cultured ms1035 tomato anthers. Euphytica 178(2):215–228. https://doi.org/10.1007/s10681-010-0303-zDanin-Poleg Y, Reis N, Tzuri G, Katzir N (2001) Development and characterization of microsatellite markers in Cucumis. Theor Appl Genet 102(1):61–72. https://doi.org/10.1007/s001220051618Dong Y-Q, Zhao W-X, Li X-H, Liu X-C, Gao N-N, Huang J-H, Wang W-Y, Xu X-L, Tang Z-H (2016) Androgenesis, gynogenesis, and parthenogenesis haploids in cucurbit species. Plant Cell Rep. https://doi.org/10.1007/s00299-016-2018-7FAOSTAT (2018) http://faostat.fao.org. Accessed July 2018Ficcadenti N, Sestili S, Annibali S, Di Marco M, Schiavi M (1999) In vitro gynogenesis to induce haploid plants in melon Cucumis melo L. Genet Breed 53:255–257Gałązka J, Niemirowicz-Szczytt K (2013) Review of research on haploid production in cucumber and other cucurbits. Folia Hortic. https://doi.org/10.2478/fhort-2013-0008Hamidvand Y, Abdollahi MR, Chaichi M, Moosavi SS (2013) The effect of plant growth regulators on callogenesis and gametic embryogenesis from anther culture of cucumber (Cucumis sativus L.). Int J Agric Crop Sci 5(10):1089Kurtar ES, Balkaya A, Kandemir D (2016) Evaluation of haploidization efficiency in winter squash (Cucurbita maxima Duch.) and pumpkin (Cucurbita moschata Duch.) through anther culture. Plant Cell, Tissue Organ Cult 127(2):497–511. https://doi.org/10.1007/s11240-016-1074-6Lotfi M, Alan AR, Henning MJ, Jahn MM, Earle ED (2003) Production of haploid and doubled haploid plants of melon (Cucumis melo L.) for use in breeding for multiple virus resistance. Plant Cell Rep 21(11):1121–1128Metwally EI, Moustafa SA, El-Sawy BI, Shalaby TA (1998) Haploid plantlets derived by anther culture of Cucurbita pepo. Plant Cell, Tissue Organ Cult 52(3):171–176. https://doi.org/10.1023/a:1005908326663Mohamed M, Refaei E (2004) Enhanced haploids regeneration in anther culture of summer squash (Curcurbita pepo L.). Cucurbit Genet Coop Rep 27:57–60Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479Parra-Vega V, Renau-Morata B, Sifres A, Seguí-Simarro JM (2013) Stress treatments and in vitro culture conditions influence microspore embryogenesis and growth of callus from anther walls of sweet pepper (Capsicum annuum L.). Plant Cell, Tissue Organ Cult 112(3):353–360. https://doi.org/10.1007/s11240-012-0242-6Rakha M, Metwally E, Moustafa S, Etman A, Dewir Y (2012) Evaluation of regenerated strains from six Cucurbita interspecific hybrids obtained through anther and ovule in vitro cultures. Aust J Crop Sci 6(1):23–30Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81(24):8014–8018. https://doi.org/10.1073/pnas.81.24.8014Sauton A, Dumas de Vaulx R (1987) Obtention de plantes haploides chez melon (Cucumis melo L.) par gynogenese indute par du pollen irraidié. Agronomie 7:141–148Seguí-Simarro JM (2016) Androgenesis in solanaceae. In: Germanà MA, Lambardi M (eds) In vitro embryogenesis. Methods in molecular biology, vol 1359. Springer, New York, pp 209–244. https://doi.org/10.1007/978-1-4939-3061-6_9Seguí-Simarro JM, Nuez F (2006) Androgenesis induction from tomato anther cultures: callus characterization. Acta Hort 725:855–861Seguí-Simarro JM, Nuez F (2007) Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers. J Exp Bot 58(5):1119–1132Seguí-Simarro JM, Nuez F (2008) Pathways to doubled haploidy: chromosome doubling during androgenesis. Cytogenet Genome Res 120(3–4):358–369. https://doi.org/10.1159/000121085Shalaby TA (2006) Embryogenesis and plantlets regeneration from anther culture of squash plants (Cucurbita pepo L.) as affected by different genotypes. J Agric Res Tanta Univ 32(1):173–183Song H, Lou QF, Luo XD, Wolukau JN, Diao WP, Qian CT, Chen JF (2007) Regeneration of doubled haploid plants by androgenesis of cucumber (Cucumis sativus L.). Plant Cell, Tissue Organ Cult 90(3):245–254. https://doi.org/10.1007/s11240-007-9263-ySteward FC, Mapes MO, Mears K (1958) Growth and organized development of cultured cells. II. Organization in cultures grown from freely suspended cells. Am J Bot 45(10):705–708Su YH, Zhao XY, Liu YB, Zhang CL, O’Neill SD, Zhang XS (2009) Auxin-induced WUS expression is essential for embryonic stem cell renewal during somatic embryogenesis in arabidopsis. Plant J 59(3):448–460. https://doi.org/10.1111/j.1365-313X.2009.03880.xSuprunova T, Shmykova N (2008) In vitro induction of haploid plants in unpollinated ovules, anther and microspore culture of Cucumis sativus. In: Cucurbitaceae 2008: proceedings of the IXth Eucarpia meeting on genetics and breeding of cucurbitaceae, pp 371–374Xie M, Qin L-Y, Pan J-S, He H-L, Wu A-Z, Cai R (2005) Flower morphogenesis and microspore development versus anther culture of cucumber. Acta Bot Boreal-Occid Sin 25(6):1096Zhan Y, Chen J-F, Malik AA (2009) Embryoid induction and plant regeneration of cucumber (Cucumis sativus L.) through microspore culture. Acta Hort Sin 36(2):221–22

    Optimization of the conditions for production of synthetic seeds by encapsulation of axillary buds derived from minituber sprouts in potato (Solanum tuberosum)

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    [EN] Synthetic seed technology is a convenient alternative to conventional multiplication in potato. In this work, we studied and optimized the process of alginate encapsulation of axillary buds derived from potato minituber sprouts (PMSs). We assayed different concentrations of sodium alginate, CaCl2 and matrix culture media, different explant sizes, different concentrations of 24-epibrassinolide (EBr) applied at different stages of the encapsulation process, different planting substrates and different cold storage periods in order to determine the best conditions for encapsulation in cvs. Sante and Agria. The conditions that resulted in the greatest regrowth rates and speed in both cultivars involved the choice of 2 3 mmlong buds, a 2 days pre-culture of buds in culture medium supplemented with 10-6 M EBr, encapsulation in 3 % sodium alginate with 1 % CaCl2 and full-strength MS culture medium, regrowth in solid MS culture medium and then transfer to coco peat (coir fiber pith) for conversion into plantlets. We also found that buds encapsulated under these conditions maintained the initial viability rates for up to 120 days in Sante and 90 days in Agria , although regrowth speed decreased after 60 days in both cultivars. We demonstrate the possibility of producing synthetic seeds efficiently using axillary buds derived from PMSs.Ghambarali, S.; Abdollahi, MR.; Zolnorian, H.; Moosavi, SS.; Seguí-Simarro, JM. (2016). Optimization of the conditions for production of synthetic seeds by encapsulation of axillary buds derived from minituber sprouts in potato (Solanum tuberosum). Plant Cell Tissue and Organ Culture (PCTOC). 126(3):449-458. doi:10.1007/s11240-016-1013-6S4494581263Adriani M, Piccioni E, Standardi A (2000) Effect of different treatments on the conversion of ‘Hayward’ kiwifruit synthetic seeds to whole plants following encapsulation of in vitro-derived buds. N Z J Crop Hortic Sci 28:59–67Ahmad N, Anis M (2010) Direct plant regeneration from encapsulated nodal segments of Vitex negundo. Biol Plant 54:748–752Azpeitia A, Chan JL, Saenz L, Oropeza C (2003) Effect of 22(S),23(S)-homobrassinolide on somatic embryogenesis in plumule explants of Cocos nucifera (L.) cultured in vitro. J Hortic Sci Biotech 78:591–596Brosa C (1999) Biological effects of brassinosteroids. Crit Rev Biochem Mol Biol 34:339–358Bustam S, Sinniah UR, Kadir MA, Zaman FQ, Subramaniam S (2012) Selection of optimal stage for protocorm-like bodies and production of artificial seeds for direct regeneration on different media and short term storage of Dendrobium Shavin White. Plant Growth Regul 69:215–224Corral-Martínez P, Seguí-Simarro JM (2014) Refining the method for eggplant microspore culture: effect of abscisic acid, epibrassinolide, polyethylene glycol, naphthaleneacetic acid, 6-benzylaminopurine and arabinogalactan proteins. Euphytica 195:369–382Divi UK, Rahman T, Krishna P (2010) Brassinosteroid-mediated stress tolerance in Arabidopsis shows interactions with abscisic acid, ethylene and salicylic acid pathways. BMC Plant Biol 10:151Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42Estrada R, Tovar P, Dodds JH (1986) Induction of in vitro tubers in a broad range of potato genotypes. Plant Cell Tissue Organ Cult 7:3–10Faisal M, Anis M (2007) Regeneration of plants from alginate-encapsulated shoots of Tylophora indica (Burm. f.) Merrill, an endangered medicinal plant. J Hortic Sci Biotechnol 82:351–354Fujioka S, Yokota T (2003) Biosynthesis and metabolism of brassinosteroids. Ann Rev Plant Biol 54:137–164Gantait S, Kundu S, Ali N, Sahu NC (2015) Synthetic seed production of medicinal plants: a review on influence of explants, encapsulation agent and matrix. Acta Physiol Plant 37:1–12Grove MD, Spencer GF, Rohwedder WK, Mandava N, Worley JF, Warthen JD, Steffens GL, Flippenanderson JL, Cook JC (1979) Brassinolide, a plant growth-promoting steroid isolated from Brassica napus pollen. Nature 281:216–217Hung CD, Trueman SJ (2012a) Alginate encapsulation of shoot tips and nodal segments for short-term storage and distribution of the eucalypt Corymbia torelliana × C. citriodora. Acta Physiol Plant 34:117–128Hung CD, Trueman SJ (2012b) Preservation of encapsulated shoot tips and nodes of the tropical hardwoods Corymbia torelliana × C. citriodora and Khaya senegalensis. Plant Cell, Tissue Organ Cult 109:341–352Jones ED (1988) A current assessment of in vitro culture and other rapid multiplication methods in North America and Europe. 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    Production of doubled haploid plants from anther cultures of borage (Borago officinalis L.) by the application of chemical and physical stress

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    [EN] Anther culture can be used as a powerful tool to produce doubled haploid (DH) lines in medicinal plants, thus accelerating breeding programs. In the particular case of borage (Borago officinalis L.), a method to produce DH plants has not been yet published. In this work we evaluated the effect of different culture media and of different chemical (colchicine and n-butanol) and physical stresses (centrifugation and electroporation) on androgenesis induction and plant regeneration in borage anther cultures. We found that the highest response can be obtained with culture medium containing B5 salts and NLN vitamins, the addition of 200 mg/l colchicine during 4 days, a pretreatment of anthers with 0.2% n-butanol for 5 hours, or the application to anthers of single physical stresses (either centrifugation at 300 g or a 100 v electrical shock, but not combined). This is the first report on the production of DH plants in borage. Together, the results presented hereby can be used as a basic framework for large-scale generation of DH plants in this important medicinal species.Hoveida, Z.; Abdollahi, MR.; Mirzaie-Asl, A.; Moosavi, SS.; Seguí-Simarro, JM. (2017). Production of doubled haploid plants from anther cultures of borage (Borago officinalis L.) by the application of chemical and physical stress. Plant Cell Tissue and Organ Culture (PCTOC). 130(2):369-378. doi:10.1007/s11240-017-1233-4S3693781302Alemanno L, Guiderdoni E (1994) Increased doubled haploid plant regeneration from rice (Oryza sativa L.) anthers cultured on colchicine-supplemented media. Plant Cell Rep 13:432–436Bayliss K, Wroth J, Cowling W (2004) Pro-embryos of Lupinus spp. produced from isolated microspore culture. Crop Pasture Sci 55:589–593Castillo AM, Cistué L, Vallés MP, Soriano M (2009) Chromosome doubling in monocots. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. 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    Solvent-Free Preparation of 1,8-Dioxo-Octahydroxanthenes Employing Iron Oxide Nanomaterials

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    In this study, 1,8-dioxo-octahydroxanthenes were prepared employing a simple, effective and environmentally sound approach utilizing an iron oxide nanocatalyst under solventless conditions. The proposed iron oxide nanomaterial exhibited high product yields, short reaction times and a facile work-up procedure. The synthesized catalyst was also found to be highly stable and reusable under the investigated conditions (up to twelve consecutive cycles) without any significant loss in its catalytic activity

    Induction of androgenesis and production of haploid embryos in anther cultures of borage (Borago officinalis L.)

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    [EN] Borage (Borago officinalis L.) is an important medicinal plant with different culinary, pharmaceutical and industrial properties. Unfortunately, there are no published reports on the establishment of protocols to produce DHs in this species up to now. In this work, we show for the first time the induction of borage microspores to become embryogenic calli, from which haploid embryos are produced. In addition, we evaluated the effect of using different flower bud sizes, carbon sources, concentrations of 2,4-D and BAP, cold (4 A degrees C) pretreatments and heat shock treatments. Production of total calli, embryogenic calli and callus-derived embryos was differently affected by the different parameters studied. Our results showed that the use of 5-7 mm-long flower buds, a cold (4 A degrees C) pretreatment during 4 days, a 32 A degrees C heat shock for 3 days, and the addition of 3 % maltose and 2 mgl(-1) 2,4-D and 1 mgl(-1) BAP to the culture medium, was beneficial for embryo production. Overall, this work demonstrates that DH technology is possible in borage, and opens the door for future improvements needed to finally obtain borage DH plants.Eshaghi, ZC.; Abdollahi, MR.; Moosavi, SS.; Deljou, A.; Seguí-Simarro, JM. (2015). Induction of androgenesis and production of haploid embryos in anther cultures of borage (Borago officinalis L.). Plant Cell, Tissue and Organ Culture. 122:321-329. doi:10.1007/s11240-015-0768-5S321329122Abdollahi MR, Moieni A, Javaran MJ (2004) Interactive effects of shock and culture density on embryo induction in isolated microspore culture of Brassica napus L. cv. 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    Global, regional, and national burden of disorders affecting the nervous system, 1990–2021:a systematic analysis for the Global Burden of Disease Study 2021

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    BackgroundDisorders affecting the nervous system are diverse and include neurodevelopmental disorders, late-life neurodegeneration, and newly emergent conditions, such as cognitive impairment following COVID-19. Previous publications from the Global Burden of Disease, Injuries, and Risk Factor Study estimated the burden of 15 neurological conditions in 2015 and 2016, but these analyses did not include neurodevelopmental disorders, as defined by the International Classification of Diseases (ICD)-11, or a subset of cases of congenital, neonatal, and infectious conditions that cause neurological damage. Here, we estimate nervous system health loss caused by 37 unique conditions and their associated risk factors globally, regionally, and nationally from 1990 to 2021.MethodsWe estimated mortality, prevalence, years lived with disability (YLDs), years of life lost (YLLs), and disability-adjusted life-years (DALYs), with corresponding 95% uncertainty intervals (UIs), by age and sex in 204 countries and territories, from 1990 to 2021. We included morbidity and deaths due to neurological conditions, for which health loss is directly due to damage to the CNS or peripheral nervous system. We also isolated neurological health loss from conditions for which nervous system morbidity is a consequence, but not the primary feature, including a subset of congenital conditions (ie, chromosomal anomalies and congenital birth defects), neonatal conditions (ie, jaundice, preterm birth, and sepsis), infectious diseases (ie, COVID-19, cystic echinococcosis, malaria, syphilis, and Zika virus disease), and diabetic neuropathy. By conducting a sequela-level analysis of the health outcomes for these conditions, only cases where nervous system damage occurred were included, and YLDs were recalculated to isolate the non-fatal burden directly attributable to nervous system health loss. A comorbidity correction was used to calculate total prevalence of all conditions that affect the nervous system combined.FindingsGlobally, the 37 conditions affecting the nervous system were collectively ranked as the leading group cause of DALYs in 2021 (443 million, 95% UI 378–521), affecting 3·40 billion (3·20–3·62) individuals (43·1%, 40·5–45·9 of the global population); global DALY counts attributed to these conditions increased by 18·2% (8·7–26·7) between 1990 and 2021. Age-standardised rates of deaths per 100 000 people attributed to these conditions decreased from 1990 to 2021 by 33·6% (27·6–38·8), and age-standardised rates of DALYs attributed to these conditions decreased by 27·0% (21·5–32·4). Age-standardised prevalence was almost stable, with a change of 1·5% (0·7–2·4). The ten conditions with the highest age-standardised DALYs in 2021 were stroke, neonatal encephalopathy, migraine, Alzheimer's disease and other dementias, diabetic neuropathy, meningitis, epilepsy, neurological complications due to preterm birth, autism spectrum disorder, and nervous system cancer.InterpretationAs the leading cause of overall disease burden in the world, with increasing global DALY counts, effective prevention, treatment, and rehabilitation strategies for disorders affecting the nervous system are needed.FundingBill &amp; Melinda Gates Foundation.<br/

    Advice from the Scientific Advisory Board of the Organisation for the Prohibition of Chemical Weapons on riot control agents in connection to the Chemical Weapons Convention

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    Compounds that cause powerful sensory irritation to humans were reviewed by the Scientific Advisory Board (SAB) of the Organisation for the Prohibition of Chemical Weapons (OPCW) in response to requests in 2014 and 2017 by the OPCW Director-General to advise which riot control agents (RCAs) might be subject to declaration under the Chemical Weapons Convention (the Convention). The chemical and toxicological properties of 60 chemicals identified from a survey by the OPCW of RCAs that had been researched or were available for purchase, and additional chemicals recognised by the SAB as having potential RCA applications, were considered. Only 17 of the 60 chemicals met the definition of a RCA under the Convention. These findings were provided to the States Parties of the Convention to inform the implementation of obligations pertaining to RCAs under this international chemical disarmament and non-proliferation treaty.Peer reviewe

    Global, regional, and national burden of disorders affecting the nervous system, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

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    Background: Disorders affecting the nervous system are diverse and include neurodevelopmental disorders, late-life neurodegeneration, and newly emergent conditions, such as cognitive impairment following COVID-19. Previous publications from the Global Burden of Disease, Injuries, and Risk Factor Study estimated the burden of 15 neurological conditions in 2015 and 2016, but these analyses did not include neurodevelopmental disorders, as defined by the International Classification of Diseases (ICD)-11, or a subset of cases of congenital, neonatal, and infectious conditions that cause neurological damage. Here, we estimate nervous system health loss caused by 37 unique conditions and their associated risk factors globally, regionally, and nationally from 1990 to 2021. Methods: We estimated mortality, prevalence, years lived with disability (YLDs), years of life lost (YLLs), and disability-adjusted life-years (DALYs), with corresponding 95% uncertainty intervals (UIs), by age and sex in 204 countries and territories, from 1990 to 2021. We included morbidity and deaths due to neurological conditions, for which health loss is directly due to damage to the CNS or peripheral nervous system. We also isolated neurological health loss from conditions for which nervous system morbidity is a consequence, but not the primary feature, including a subset of congenital conditions (ie, chromosomal anomalies and congenital birth defects), neonatal conditions (ie, jaundice, preterm birth, and sepsis), infectious diseases (ie, COVID-19, cystic echinococcosis, malaria, syphilis, and Zika virus disease), and diabetic neuropathy. By conducting a sequela-level analysis of the health outcomes for these conditions, only cases where nervous system damage occurred were included, and YLDs were recalculated to isolate the non-fatal burden directly attributable to nervous system health loss. A comorbidity correction was used to calculate total prevalence of all conditions that affect the nervous system combined. Findings: Globally, the 37 conditions affecting the nervous system were collectively ranked as the leading group cause of DALYs in 2021 (443 million, 95% UI 378–521), affecting 3·40 billion (3·20–3·62) individuals (43·1%, 40·5–45·9 of the global population); global DALY counts attributed to these conditions increased by 18·2% (8·7–26·7) between 1990 and 2021. Age-standardised rates of deaths per 100 000 people attributed to these conditions decreased from 1990 to 2021 by 33·6% (27·6–38·8), and age-standardised rates of DALYs attributed to these conditions decreased by 27·0% (21·5–32·4). Age-standardised prevalence was almost stable, with a change of 1·5% (0·7–2·4). The ten conditions with the highest age-standardised DALYs in 2021 were stroke, neonatal encephalopathy, migraine, Alzheimer's disease and other dementias, diabetic neuropathy, meningitis, epilepsy, neurological complications due to preterm birth, autism spectrum disorder, and nervous system cancer. Interpretation: As the leading cause of overall disease burden in the world, with increasing global DALY counts, effective prevention, treatment, and rehabilitation strategies for disorders affecting the nervous system are needed. Funding: Bill & Melinda Gates Foundation.info:eu-repo/semantics/publishedVersio
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