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1.
Nature ; 2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38658755

RESUMO

RAD52 is important for the repair of DNA double-stranded breaks1,2, mitotic DNA synthesis3-5 and alternative telomere length maintenance6,7. Central to these functions, RAD52 promotes the annealing of complementary single-stranded DNA (ssDNA)8,9 and provides an alternative to BRCA2/RAD51-dependent homologous recombination repair10. Inactivation of RAD52 in homologous-recombination-deficient BRCA1- or BRCA2-defective cells is synthetically lethal11,12, and aberrant expression of RAD52 is associated with poor cancer prognosis13,14. As a consequence, RAD52 is an attractive therapeutic target against homologous-recombination-deficient breast, ovarian and prostate cancers15-17. Here we describe the structure of RAD52 and define the mechanism of annealing. As reported previously18-20, RAD52 forms undecameric (11-subunit) ring structures, but these rings do not represent the active form of the enzyme. Instead, cryo-electron microscopy and biochemical analyses revealed that ssDNA annealing is driven by RAD52 open rings in association with replication protein-A (RPA). Atomic models of the RAD52-ssDNA complex show that ssDNA sits in a positively charged channel around the ring. Annealing is driven by the RAD52 N-terminal domains, whereas the C-terminal regions modulate the open-ring conformation and RPA interaction. RPA associates with RAD52 at the site of ring opening with critical interactions occurring between the RPA-interacting domain of RAD52 and the winged helix domain of RPA2. Our studies provide structural snapshots throughout the annealing process and define the molecular mechanism of ssDNA annealing by the RAD52-RPA complex.

3.
Sports Med ; 2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38609697

RESUMO

OBJECTIVES: The aim of this study was to compare the incidence, severity, and burden of injury in starting and replacement players from professional men's teams of five rugby unions. METHODS: Match injuries of greater than 24 h time-loss (including data on the severity, match quarter, event, body region) and player minutes of match exposure data were collated for all starting and replacement players in the men's English Premiership, Welsh Pro14 (both 2016/17-2018/19 seasons), and Australian, New Zealand, and South African Super Rugby (all 2016-2018 seasons) teams. Injury incidences and mean injury burden (incidence × days missed) were calculated, and rate ratios (RRs) (95% confidence intervals [CIs]) were used to compare injury incidence and burden between starting (reference group) and replacement players. RESULTS: Overall injury incidence was not different between starters and replacements for all injuries (RR = 0.98, 95% CI 0.88-1.10), nor for concussions (RR = 0.85; 95% CI 0.66-1.11). Mean injury burden was higher for replacement players (RR = 1.31, 95% CI 1.17-1.46). Replacement injury incidence was lower than the starters in the third (RR = 0.68, 95% CI 0.51-0.92) and fourth (RR = 0.78, 95% CI 0.67-0.92) match quarters. Injury incidence was not different between starters and replacements for any match event or body region, but compared with starters, replacements' injury burden was higher in lower limbs (RR = 1.24, 95% CI 1.05-1.46) and in the tackled player (RR = 1.30, 95% CI 1.01-1.66). CONCLUSION: This study demonstrated a lower injury incidence in replacement players compared with starters in the second half of matches, with a higher injury burden for replacement players due to higher mean injury severity.

4.
J Sport Health Sci ; 13(3): 387-397, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38232900

RESUMO

BACKGROUND: Return to play (RTP) in elite rugby is managed using a 6-stage graduated RTP protocol, which can result in clearance to play within 1 week of injury. We aimed to explore how symptom, cognitive, and balance presentation and evolution during concussion screens 2 h (head injury assessment (HIA) 2) and 48 h (HIA3) after injury were associated with time to RTP) to identify whether a more conservative graduated RTP may be appropriate. METHODS: A retrospective cohort study was conducted in 380 concussed rugby players from elite men's rugby over 3 consecutive seasons. Players were classified as shorter or longer returns, depending on whether RTP occurred within 7 days (allowing them to be considered to play the match 1 week after injury) or longer than 8 days, respectively. Symptom, cognitive, and balance performance during screens was assessed relative to baseline (normal or abnormal) and to the preceding screen (improving or worsening). Associations between sub-test abnormalities and RTP time were explored using odds ratios (OR, longer vs. shorter). Median day absence was compared between players with abnormal or worsening results and those whose results were normal or improving. RESULTS: Abnormal symptom results during screens 2 h and 48 h after concussion were associated with longer return time (HIA2: OR = 2.21, 95% confidence interval (95%CI): 1.39-3.50; HIA3: OR = 3.30, 95%CI: 1.89-5.75). Worsening symptom number or severity from the time of injury to 2 h and 48 h post-injury was associated with longer return (HIA2: OR = 2.49, 95%CI: 1.36-4.58; HIA3: OR = 3.34, 95%CI: 1.10-10.15. Median days absence was greater in players with abnormal symptom results at both HIA2 and HIA3. Cognitive and balance performance were not associated with longer return and did not affect median days absence. CONCLUSION: Symptom presentation and evolution within 48 h of concussion were associated with longer RTP times. This may guide a more conservative approach to RTP, while still adhering to individualized concussion management principles.


Assuntos
Concussão Encefálica , Futebol Americano , Volta ao Esporte , Humanos , Concussão Encefálica/complicações , Masculino , Estudos Retrospectivos , Futebol Americano/lesões , Fatores de Tempo , Adulto Jovem , Equilíbrio Postural , Adulto , Traumatismos em Atletas , Cognição/fisiologia
5.
Clin J Sport Med ; 34(2): 121-126, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-37389460

RESUMO

OBJECTIVE: To describe shoulder-related injury rates (IRs), types, severity, mechanisms, and risk factors in youth ice hockey players during games and practices. DESIGN: Secondary analysis of data from a 5-year prospective cohort study, Safe-to-Play (2013-2018). SETTING: Canadian youth ice hockey. PARTICIPANTS: Overall, 6584 player-seasons (representing 4417 individual players) participated. During this period, 118 shoulder-related games and 12 practice injuries were reported. ASSESSMENT OF RISK FACTORS: An exploratory multivariable mixed-effects Poisson regression model examined the risk factors of body checking policy, weight, biological sex, history of injury in the past 12 months, and level of play. MAIN OUTCOME MEASURES: Injury surveillance data were collected from 2013 to 2018. Injury rates with 95% confidence interval (CI) were estimated using Poisson regression. RESULTS: The shoulder IR was 0.35 injuries/1000 game-hours (95% CI, 0.24-0.49). Two-thirds of game injuries (n = 80, 70%) resulted in >8 days of time-loss, and more than one-third (n = 44, 39%) resulted in >28 days of time-loss. An 83% lower rate of shoulder injury was associated with policy prohibiting body checking compared with leagues allowing body checking (incidence rate ratio [IRR], 0.17; 95% CI, 0.09-0.33). A higher shoulder IR was observed for those who reported any injury in the last 12-months compared with those with no history (IRR, 2.00; 95% CI, 1.33-3.01). CONCLUSIONS: Most shoulder injuries resulted in more than 1 week of time-loss. Risk factors for shoulder injury included participation in a body-checking league and recent history of injury. Further study of prevention strategies specific to the shoulder may merit further consideration in ice hockey.


Assuntos
Traumatismos em Atletas , Concussão Encefálica , Hóquei , Lesões do Ombro , Humanos , Adolescente , Canadá/epidemiologia , Traumatismos em Atletas/prevenção & controle , Concussão Encefálica/epidemiologia , Estudos Prospectivos , Ombro , Hóquei/lesões , Fatores de Risco , Incidência , Lesões do Ombro/epidemiologia
6.
Br J Sports Med ; 2023 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-37985003

RESUMO

OBJECTIVE: To examine differences in match and training musculoskeletal injury and concussion rates and describe mechanisms of concussion while considering previous playing experience in female and male Canadian high school Rugby Union ('rugby') players. METHODS: A 2-year prospective cohort study was completed in a high school league (n=361 females, 421 player-seasons; n=429 males, 481 player-seasons) in Calgary, Canada over the 2018 and 2019 rugby playing seasons. Baseline testing was completed at the start of each season and injury surveillance and individual player participation through session attendance was documented to quantify individual-level player exposure hours. Injury incidence rates (IRs) and incidence rate ratios (IRRs) were calculated using Poisson regression, offset by player exposure hours and clustered by team. RESULTS: Overall match IR for females was 62% higher than males (overall IRR=1.62, 95% CI: 1.20 to 2.18) and the overall training IR was twice as high for females (overall IRR=2.15, 95% CI: 1.40 to 3.32). The female match concussion IR was 70% higher than the males (concussion IRR=1.70, 95% CI: 1.08 to 2.69). Females had a 75% greater tackle-related IR compared with males (IRR=1.75, 95% CI: 1.20 to 2.56). Additionally, female tacklers had a twofold greater rate of injury compared with male tacklers (IRR=2.17, 95% CI: 1.14 to 4.14). Previous playing experience was not associated with tackle-related injury or concussion IRs. CONCLUSION: The rate of injury and concussion was significantly higher in females within this Canadian high school cohort. These results emphasise the need for development, implementation and evaluation of female-specific injury and concussion prevention strategies to reduce injury and concussion in female youth rugby.

7.
BMJ Open Sport Exerc Med ; 9(3): e001645, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37780130

RESUMO

Video analysis is a useful tool for injury surveillance in rugby union. There are few video analysis studies in the professional female game, with most studies published in the male elite/professional settings. Moreover, there is a sparsity of literature in youth rugby settings. The following narrative review outlines the strengths and limitations of the current video analysis literature for injury surveillance in youth rugby union, highlights the importance of video analysis for youth rugby player safety and welfare, and discusses recommendations for using video analysis to inform player safety in youth rugby.

8.
Nat Commun ; 14(1): 6809, 2023 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-37884503

RESUMO

Poly(ADP-ribose) polymerase (PARP) inhibitors are used in the clinic to treat BRCA-deficient breast, ovarian and prostate cancers. As their efficacy is potentiated by loss of the nucleotide salvage factor DNPH1 there is considerable interest in the development of highly specific small molecule DNPH1 inhibitors. Here, we present X-ray crystal structures of dimeric DNPH1 bound to its substrate hydroxymethyl deoxyuridine monophosphate (hmdUMP). Direct interaction with the hydroxymethyl group is important for substrate positioning, while conserved residues surrounding the base facilitate target discrimination. Glycosidic bond cleavage is driven by a conserved catalytic triad and proceeds via a two-step mechanism involving formation and subsequent disruption of a covalent glycosyl-enzyme intermediate. Mutation of a previously uncharacterised yet conserved glutamate traps the intermediate in the active site, demonstrating its role in the hydrolytic step. These observations define the enzyme's catalytic site and mechanism of hydrolysis, and provide important insights for inhibitor discovery.


Assuntos
Nucleotídeos , Humanos , Modelos Moleculares , Hidrólise , Domínio Catalítico , Catálise
9.
Anal Biochem ; 682: 115347, 2023 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-37821038

RESUMO

DNA molecules that contain single Holliday junctions have served as model substrates to investigate the pathway in which homologous recombination intermediates are processed. However, the preparation of DNA containing Holliday junctions in high yield remains a challenge. In this work, we used a nicking endonuclease to generate gapped DNA, from which α-structured DNA or figure-8 DNA were created via RecA-mediated reactions. The resulting DNA molecules were found to serve as good substrates for Holliday junction resolvases. The simplified method negates the requirement for radioactive labelling of DNA, making the generation of Holliday junction DNA more accessible to non-experts.


Assuntos
DNA Cruciforme , Proteínas de Escherichia coli , DNA Cruciforme/metabolismo , Proteínas de Escherichia coli/química , Endodesoxirribonucleases/química , Endodesoxirribonucleases/genética , Endodesoxirribonucleases/metabolismo , Escherichia coli/genética , DNA/química
10.
Mol Cell ; 83(16): 2925-2940.e8, 2023 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-37499663

RESUMO

Homologous recombination (HR) is essential for error-free repair of DNA double-strand breaks, perturbed replication forks (RFs), and post-replicative single-stranded DNA (ssDNA) gaps. To initiate HR, the recombination mediator and tumor suppressor protein BRCA2 facilitates nucleation of RAD51 on ssDNA prior to stimulation of RAD51 filament growth by RAD51 paralogs. Although ssDNA binding by BRCA2 has been implicated in RAD51 nucleation, the function of double-stranded DNA (dsDNA) binding by BRCA2 remains unclear. Here, we exploit single-molecule (SM) imaging to visualize BRCA2-mediated RAD51 nucleation in real time using purified proteins. We report that BRCA2 nucleates and stabilizes RAD51 on ssDNA either directly or through an unappreciated diffusion-assisted delivery mechanism involving binding to and sliding along dsDNA, which requires the cooperative action of multiple dsDNA-binding modules in BRCA2. Collectively, our work reveals two distinct mechanisms of BRCA2-dependent RAD51 loading onto ssDNA, which we propose are critical for its diverse functions in maintaining genome stability and cancer suppression.


Assuntos
Proteína BRCA2 , Rad51 Recombinase , Humanos , Proteína BRCA2/genética , Proteína BRCA2/metabolismo , Rad51 Recombinase/genética , Rad51 Recombinase/metabolismo , Proteínas de Ligação a DNA/metabolismo , DNA de Cadeia Simples/genética , DNA/metabolismo , Reparo do DNA , Ligação Proteica
11.
Front Sports Act Living ; 5: 1080356, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37334015

RESUMO

Objective: The tackle is the most injurious event in rugby league and carries the greatest risk of concussion. This study aims to replicate previous research conducted in professional men's rugby league by examining the association between selected tackle characteristics and head impact events (HIEs) in women's professional rugby league. Methods: We reviewed and coded 83 tackles resulting in an HIE and every tackle (6,318 tackles) that did not result in an HIE for three seasons (2018-2020) of the National Rugby League Women's (NRLW) competition. Tackle height, body position of the tackler and ball carrier, as well as the location of head contact with the other player's body were evaluated. Propensity of each situation that caused an HIE was calculated as HIEs per 1,000 tackles. Results: The propensity for tacklers to sustain an HIE was 6.60 per 1,000 tackles (95% CI: 4.87-8.92), similar to that of the ball carrier (6.13 per 1,000 tackles, 95% CI: 4.48-8.38). The greatest risk of an HIE to either the tackler or ball carrier occurred when head proximity was above the sternum (21.66 per 1,000 tackles, 95% CI: 16.55-28.35). HIEs were most common following impacts between two heads (287.23 HIEs per 1,000 tackles, 95% CI: 196.98-418.84). The lowest propensity for both tackler (2.65 per 1,000 tackles, 95% CI: 0.85-8.20) and ball carrier HIEs (1.77 per 1,000 tackles, 95% CI: 0.44-7.06) occurred when the head was in proximity to the opponent's shoulder and arm. No body position (upright, bent or unbalanced/off feet) was associated with an increased propensity of HIE to either tackler or ball carrier. Conclusions: In the NRLW competition, tacklers and ball carriers have a similar risk of sustaining an HIE during a tackle, differing from men's NRL players, where tacklers have a higher risk of HIEs. Further studies involving larger samples need to validate these findings. However, our results indicate that injury prevention initiatives in women's rugby league should focus on how the ball carrier engages in contact during the tackle as well as how the tackler executes the tackle.

12.
Br J Sports Med ; 57(12): 749-761, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37316182

RESUMO

OBJECTIVES: To evaluate prevention strategies, their unintended consequences and modifiable risk factors for sport-related concussion (SRC) and/or head impact risk. DESIGN: This systematic review and meta-analysis was registered on PROSPERO (CRD42019152982) and conducted according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. DATA SOURCES: Eight databases (MEDLINE, CINAHL, APA PsycINFO, Cochrane (Systematic Review and Controlled Trails Registry), SPORTDiscus, EMBASE, ERIC0 were searched in October 2019 and updated in March 2022, and references searched from any identified systematic review. ELIGIBILITY CRITERIA: Study inclusion criteria were as follows: (1) original data human research studies, (2) investigated SRC or head impacts, (3) evaluated an SRC prevention intervention, unintended consequence or modifiable risk factor, (4) participants competing in any sport, (5) analytic study design, (6) systematic reviews and meta-analyses were included to identify original data manuscripts in reference search and (7) peer-reviewed. Exclusion criteria were as follows: (1) review articles, pre-experimental, ecological, case series or case studies and (2) not written in English. RESULTS: In total, 220 studies were eligible for inclusion and 192 studies were included in the results based on methodological criteria as assessed through the Scottish Intercollegiate Guidelines Network high ('++') or acceptable ('+') quality. Evidence was available examining protective gear (eg, helmets, headgear, mouthguards) (n=39), policy and rule changes (n=38), training strategies (n=34), SRC management strategies (n=12), unintended consequences (n=5) and modifiable risk factors (n=64). Meta-analyses demonstrated a protective effect of mouthguards in collision sports (incidence rate ratio, IRR 0.74; 95% CI 0.64 to 0.89). Policy disallowing bodychecking in child and adolescent ice hockey was associated with a 58% lower concussion rate compared with bodychecking leagues (IRR 0.42; 95% CI 0.33 to 0.53), and evidence supports no unintended injury consequences of policy disallowing bodychecking. In American football, strategies limiting contact in practices were associated with a 64% lower practice-related concussion rate (IRR 0.36; 95% CI 0.16 to 0.80). Some evidence also supports up to 60% lower concussion rates with implementation of a neuromuscular training warm-up programme in rugby. More research examining potentially modifiable risk factors (eg, neck strength, optimal tackle technique) are needed to inform concussion prevention strategies. CONCLUSIONS: Policy and rule modifications, personal protective equipment, and neuromuscular training strategies may help to prevent SRC. PROSPERO REGISTRATION NUMBER: CRD42019152982.


Assuntos
Concussão Encefálica , Futebol Americano , Hóquei , Adolescente , Criança , Humanos , Concussão Encefálica/prevenção & controle , Rugby , Bases de Dados Factuais
13.
Nature ; 619(7970): 650-657, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37344587

RESUMO

Homologous recombination is a fundamental process of life. It is required for the protection and restart of broken replication forks, the repair of chromosome breaks and the exchange of genetic material during meiosis. Individuals with mutations in key recombination genes, such as BRCA2 (also known as FANCD1), or the RAD51 paralogues RAD51B, RAD51C (also known as FANCO), RAD51D, XRCC2 (also known as FANCU) and XRCC3, are predisposed to breast, ovarian and prostate cancers1-10 and the cancer-prone syndrome Fanconi anaemia11-13. The BRCA2 tumour suppressor protein-the product of BRCA2-is well characterized, but the cellular functions of the RAD51 paralogues remain unclear. Genetic knockouts display growth defects, reduced RAD51 focus formation, spontaneous chromosome abnormalities, sensitivity to PARP inhibitors and replication fork defects14,15, but the precise molecular roles of RAD51 paralogues in fork stability, DNA repair and cancer avoidance remain unknown. Here we used cryo-electron microscopy, AlphaFold2 modelling and structural proteomics to determine the structure of the RAD51B-RAD51C-RAD51D-XRCC2 complex (BCDX2), revealing that RAD51C-RAD51D-XRCC2 mimics three RAD51 protomers aligned within a nucleoprotein filament, whereas RAD51B is highly dynamic. Biochemical and single-molecule analyses showed that BCDX2 stimulates the nucleation and extension of RAD51 filaments-which are essential for recombinational DNA repair-in reactions that depend on the coupled ATPase activities of RAD51B and RAD51C. Our studies demonstrate that BCDX2 orchestrates RAD51 assembly on single stranded DNA for replication fork protection and double strand break repair, in reactions that are critical for tumour avoidance.


Assuntos
Microscopia Crioeletrônica , Proteínas de Ligação a DNA , Complexos Multiproteicos , Rad51 Recombinase , Proteínas Supressoras de Tumor , Humanos , Reparo do DNA , Replicação do DNA , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a DNA/ultraestrutura , Recombinação Homóloga , Rad51 Recombinase/química , Rad51 Recombinase/metabolismo , Rad51 Recombinase/ultraestrutura , Proteínas Supressoras de Tumor/química , Proteínas Supressoras de Tumor/metabolismo , Proteínas Supressoras de Tumor/ultraestrutura , Inibidores de Poli(ADP-Ribose) Polimerases , Neoplasias/genética , Neoplasias/prevenção & controle , Proteômica , Simulação por Computador , Complexos Multiproteicos/química , Complexos Multiproteicos/metabolismo , Complexos Multiproteicos/ultraestrutura , Quebras de DNA de Cadeia Dupla
14.
Sports Med ; 53(7): 1375-1393, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37191819

RESUMO

BACKGROUND: Rugby Union is a collision team sport played globally. Despite this, significant concerns have been raised regarding the sport's safety, particularly in youth players. Given this, a review of injury rates, risk factors and prevention strategies is required across different youth age groups as well as in males and females. OBJECTIVE: The objective of this systematic review (SR) and meta-analysis was to investigate injury and concussion rates, risk factors and primary prevention strategies in youth rugby. METHODS: To be included, studies were required to report either rates, risk factors or prevention strategies in youth rugby and to have a randomised controlled trial, quasi-experimental, cohort, case control, or ecological study design. Exclusion criteria included non-peer-reviewed grey literature, conference abstracts, case studies, previous systematic reviews and studies not written in English. Nine databases were searched. The full search strategy and list of sources are available and pre-registered on PROSPERO (Ref: CRD42020208343). Each study was assessed for risk of bias using the Downs and Black quality assessment tool. Meta-analyses were conducted using a DerSimonian Laird random effect model for each age group and sex. RESULTS: Sixty-nine studies were included in this SR. The match injury rates (using a 24-h time-loss definition) were 40.2/1000 match hours (95% CI 13.9-66.5) in males and 69.0/1000 match hours (95% CI 46.8-91.2) in females. Concussion rates were 6.2/1000 player-hours (95% CI 5.0-7.4) for males and 33.9/1000 player-hours (95% CI: 24.1-43.7) for females. The most common injury site was lower extremity (males) and the head/neck (females). The most common injury type was ligament sprain (males) and concussion (females). The tackle was the most common event associated with injury in matches (55% male, 71% females). Median time loss was 21 days for males and 17 days for females. Twenty-three risk factors were reported. The risk factors with the strongest evidence were higher levels of play and increasing age. Primary injury prevention strategies were the focus of only eight studies and included law changes (n = 2), equipment (n = 4), education (n = 1) and training (n = 1). The prevention strategy with the most promising evidence was neuromuscular training. The primary limitations included a broad range of injury definitions (n = 9) and rate denominators (n = 11) used, as well as a limited number of studies which could be included in the meta-analysis for females (n = 2). CONCLUSION: A focus on high-quality risk factor and primary prevention evaluation should be considered in future studies. Targeting primary prevention and stakeholder education remain key strategies in the prevention, recognition and management of injuries and concussions in youth rugby.


Assuntos
Traumatismos em Atletas , Concussão Encefálica , Futebol Americano , Feminino , Humanos , Masculino , Adolescente , Traumatismos em Atletas/epidemiologia , Traumatismos em Atletas/prevenção & controle , Traumatismos em Atletas/etiologia , Rugby , Futebol Americano/lesões , Concussão Encefálica/epidemiologia , Concussão Encefálica/prevenção & controle , Concussão Encefálica/complicações , Fatores de Risco , Incidência , Ensaios Clínicos Controlados Aleatórios como Assunto
16.
Cell Rep ; 42(2): 112062, 2023 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-36729836

RESUMO

Our genomes harbor conserved DNA sequences, known as common fragile sites (CFSs), that are difficult to replicate and correspond to regions of genome instability. Following replication stress, CFS loci give rise to breaks or gaps (termed CFS expression) where under-replicated DNA subsequently undergoes mitotic DNA synthesis (MiDAS). We show that loss of the structure-selective endonuclease GEN1 reduces CFS expression, leading to defects in MiDAS, ultrafine anaphase bridge formation, and DNA damage in the ensuing cell cycle due to aberrant chromosome segregation. GEN1 knockout cells also exhibit an elevated frequency of bichromatid constrictions consistent with the presence of unresolved regions of under-replicated DNA. Previously, the role of GEN1 was thought to be restricted to the nucleolytic resolution of recombination intermediates. However, its ability to cleave under-replicated DNA at CFS loci indicates that GEN1 plays a dual role resolving both DNA replication and recombination intermediates before chromosome segregation.


Assuntos
Replicação do DNA , DNA , Humanos , Sítios Frágeis do Cromossomo/genética , Replicação do DNA/genética , DNA/metabolismo , Proteínas de Ligação a DNA/metabolismo , Endonucleases/genética , Endonucleases/metabolismo , Instabilidade Genômica
17.
Adv Sci (Weinh) ; 10(12): e2204388, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36825683

RESUMO

Chromatin bridges connecting the two segregating daughter nuclei arise from chromosome fusion or unresolved interchromosomal linkage. Persistent chromatin bridges are trapped in the cleavage plane, triggering cytokinesis delay. The trapped bridges occasionally break during cytokinesis, inducing DNA damage and chromosomal rearrangements. Recently, Caenorhabditis elegans LEM-3 and human TREX1 nucleases have been shown to process chromatin bridges. Here, it is shown that ANKLE1 endonuclease, the human ortholog of LEM-3, accumulates at the bulge-like structure of the midbody via its N-terminal ankyrin repeats. Importantly, ANKLE1-/- knockout cells display an elevated level of G1-specific 53BP1 nuclear bodies, prolonged activation of the DNA damage response, and replication stress. Increased DNA damage observed in ANKLE1-/- cells is rescued by inhibiting actin polymerization or reducing actomyosin contractility. ANKLE1 does not act in conjunction with structure-selective endonucleases, GEN1 and MUS81 in resolving recombination intermediates. Instead, ANKLE1 acts on chromatin bridges by priming TREX1 nucleolytic activity and cleaving bridge DNA to prevent the formation of micronuclei and cytosolic dsDNA that activate the cGAS-STING pathway. It is therefore proposed that ANKLE1 prevents DNA damage and autoimmunity by cleaving chromatin bridges to avoid catastrophic breakage mediated by actomyosin contractile forces.


Assuntos
Cromatina , Endonucleases , Animais , Humanos , Endonucleases/química , Endonucleases/genética , Endonucleases/metabolismo , Actomiosina/genética , Actomiosina/metabolismo , Núcleo Celular/metabolismo , DNA/metabolismo , Dano ao DNA , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo
18.
Clin J Sport Med ; 33(3): 233-238, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-36730669

RESUMO

OBJECTIVE: To describe levels of sport specialization in Canadian high school students and investigate whether sport specialization and/or sport participation volume is associated with the history of musculoskeletal injury and/or concussion. DESIGN: Cross-sectional study. SETTING: High schools, Alberta, Canada. PARTICIPANTS: High school students (14-19 years) participating in various sports. INDEPENDENT VARIABLES: Level of sport specialization (high, moderate, low) and sport participation volume (hours per week and months per year). MAIN OUTCOME MEASURES: Twelve-month injury history (musculoskeletal and concussion). RESULTS: Of the 1504 students who completed the survey, 31% were categorized as highly specialized (7.5% before the age of 12 years). Using multivariable, negative, binomial regression (adjusted for sex, age, total yearly training hours, and clustering by school), highly specialized students had a significantly higher musculoskeletal injury rate [incidence rate ratio (IRR) = 1.36, 95% confidence interval (CI), 1.07-1.73] but not lower extremity injury or concussion rate, compared with low specialization students. Participating in one sport for more than 8 months of the year significantly increased the musculoskeletal injury rate (IRR = 1.27, 95% CI, 1.02-1.58). Increased training hours significantly increased the musculoskeletal injury rate (IRR = 1.18, 95% CI, 1.13-1.25), lower extremity injury rate (IRR = 1.16, 95% CI, 1.09-1.24), and concussion rate (IRR = 1.31, 95% CI, 1.24-1.39). CONCLUSIONS: Approximately one-third of Canadian high school students playing sports were categorized as highly specialized. The musculoskeletal injury rate was higher for high sport specialization students compared with low sport specialization students. Musculoskeletal injuries and concussion were also more common in students who train more and spend greater than 8 months per year in one sport.


Assuntos
Traumatismos em Atletas , Concussão Encefálica , Humanos , Criança , Traumatismos em Atletas/epidemiologia , Estudos Transversais , Fatores de Risco , Concussão Encefálica/epidemiologia , Estudantes , Alberta/epidemiologia , Atletas
19.
Clin J Sport Med ; 2023 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-36633403

RESUMO

OBJECTIVE: To examine injury and concussion rates, mechanisms, locations, and types of injury in Canadian high school male rugby. DESIGN: Prospective cohort study. SETTING: High school male rugby. PARTICIPANTS: A total of 429 high school players (2018: n = 225, 2019: n = 256) were recruited from 12 teams in 7 schools in Calgary, Canada. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Injury surveillance included baseline questionnaires, weekly exposure, and injury reports. Injuries included those requiring medical attention, resulted in time loss and/or inability to complete a session. Concussion was defined as per the fifth Consensus on Concussion in Sport, and all players with a suspected concussion were referred to a study sport medicine physician. RESULTS: A total of 134 injuries were captured, leading to an injury incidence rate (IR) of 57.9/1000 hours [95% confidence intervals (CIs): 45.4-73.8]. Median time loss was 6 days (range: 0-90). Injuries to the head were the most common (40%), followed by shoulder (12%) and ankle (10%). The concussion IR was 22.0/1000 hours (95% CIs: 15.9-30.4), which was the most common injury type (38%), followed by sprain (20%) and strain (15%). Sixty-five percent of injuries occurred in the tackle (ball carrier 35%, tackler 30%) and 76% of concussions (ball carrier 41%, tackler 35%). CONCLUSIONS: The rate of injury and concussion in Canadian youth high school male rugby is high, with tackle-related injuries and concussions the most common. Given this, there is a critical need for implementation of prevention strategies, in particular targeting concussion and the tackle event (eg, neuromuscular, tackle training, and law changes).

20.
J Sci Med Sport ; 26(1): 25-30, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36371396

RESUMO

OBJECTIVES: To investigate the influence of previous season match exposure on injury incidence and burden in elite men's rugby union. DESIGN: A three-season (2016-17 to 2018-19) retrospective cohort design was used to collect and analyse injury and exposure data across English Premiership rugby union teams. METHODS: Generalised linear mixed-effects models were used to model the influence of match exposure (all match involvements, match involvements of ≥20 mins, and full-game equivalents) upon match and training injury incidence and burden in the following season. RESULTS: Involvement in ≥31 matches within a season was associated with substantially increased match and training injury burden in the following season. Match exposure was not clearly associated with injury incidence in the following season. The increased match injury burden associated with higher match involvements appeared to be driven by an increased risk for older (>26 y) Forwards, whilst the increased training injury burden associated with higher match involvements appeared to be driven by an increased risk for older (>26 y) Backs. CONCLUSIONS: The present study demonstrates that all match involvements, regardless of duration, should be considered when exploring associations between match exposure and injury risk. High match involvements (≥ 31 matches) are associated with elevated injury burden, in both matches and training, in the following season. The physical and psychological load of players with high previous-season match exposure should be carefully managed.


Assuntos
Traumatismos em Atletas , Futebol Americano , Masculino , Humanos , Traumatismos em Atletas/epidemiologia , Traumatismos em Atletas/etiologia , Estudos Retrospectivos , Rugby , Futebol Americano/lesões , Incidência
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