The molecular basis of central element assembly within the synaptonemal complex during meiosis

PhD ThesisIn the first meiotic division, homologous chromosomes must pair, synapsis, exchange genetic material through crossing over, and then segregate to allow haploid germ cell production. This challenging process is triggered by double-strand break (DSB) induction, which activates repair machinery to generate recombination intermediates between homologues. These physical tethers guide synaptonemal complex (SC) assembly; a supramolecular lattice producing continuous synapsis between homologues. The SC provides the necessary three-dimensional structure for recombination intermediate resolution, including crossover formation, therefore is essential for meiotic division and thereby fertility. The underlying mammalian SC lattice is provided through self-assembly of SYCP1. However, a further five central element (CE) proteins are essential for full SC structure and function in vivo. A direct biochemical interaction was identified between human SYCP1 and CE protein SYCE3, revealing the sole physical link between SYCP1 and the CE. Light and Xray scattering experiments elucidated conformational re-modelling upon interacting, with SYCE3 modifying the SYCP1 lattice to achieve its regular incorporation. SYCE3 further interacts with, and thereby recruits, other CE proteins thus acting as a molecular glue for SC maturation. The CE SYCE2-TEX12 complex is recruited via SYCE3 binding, and undergoes fibrous assembly to achieve SC elongation. Through identification of two mutants that separately block SYCE2-TEX12 4:4 complex formation and its fibrous assembly, coupled with detailed biophysical analysis and two X-ray crystal structures, a structural model is proposed for the SYCE2-TEX12 complex and the mechanism of fibrous assembly. Analysis of mammalian SC assembly in vivo is severely hampered by the lack of a genetically tractable system. Whilst SC structure is conserved in yeast, protein constituents show no overt sequence similarity; thus, structural studies of the yeast SC were initiated to understand the underlying conservation. Yeast CE proteins, Ecm11-Gmc2, form a constitutive 2:2 complex with a structural model presented based upon solution scattering studies

Editorial commentary: The acrid bioscrew in anterior cruciate ligament reconstruction of the knee

Bioresorbable screws have been widely adopted for graft fixation in knee anterior cruciate ligament (ACL) reconstruction on the promise of screw resorbtion and replacement by bone. When considering the value of bioresorbable screws, it is imperative to understand that the “A” in PGA and PLA, the base ingredient of all bioscrews, is for Acid. All resorb by a process of hydrolysis; the speed and extent of resorbtion is determined by both the acidity of the screw and the environment in which it is placed. Regrettably, the promise of reliable, predictable screw resorbtion and replacement by bone remains elusive, despite the addition of “osteoconductive” materials. For the most part, bioresorbables are associated with good clinical outcomes for ACL reconstruction; they are not “dangerous,” just disappointing with respect to bony replacement. We propose that nonresorbing inert plastics such as PEEK (polyether ether ketone) may be better suited for the purpose of graft fixation devices for ACL reconstruction

Structural basis of meiotic chromosome synaptic elongation through hierarchical fibrous assembly of SYCE2-TEX12

The synaptonemal complex (SC) is a supramolecular protein assembly that mediates synapsis between homologous chromosomes during meiosis. SC elongation along the chromosome length (up to 24 μm) depends on its midline α-fibrous component SYCE2-TEX12. Here, we report X-ray crystal structures of human SYCE2-TEX12 as an individual building-block and upon assembly within a fibrous lattice. We combine these structures with mutagenesis, biophysics and electron microscopy to reveal the hierarchical mechanism of SYCE2-TEX12 fibre assembly. SYCE2-TEX12’s building-blocks are 2:2 coiled-coils which dimerise into 4:4 hetero-oligomers and interact end-to-end and laterally to form 10-nm fibres, which intertwine within 40-nm bundled micrometre-long fibres that define the SC’s midline structure. This assembly mechanism bears striking resemblance with intermediate filament proteins vimentin, lamin and keratin. Thus, SYCE2-TEX12 exhibits behaviour typical of cytoskeletal proteins to provide an α-fibrous SC backbone that structurally underpins synaptic elongation along meiotic chromosomes

5-Year survival of pediatric anterior cruciate ligament reconstruction with living donor hamstring tendon grafts

Background: It is well accepted that there is a higher incidence of repeat anterior cruciate ligament (ACL) injuries in the pediatric population after ACL reconstruction (ACLR) with autograft tissue compared with adults. Hamstring autograft harvest may contribute to the risk for repeat ACL injuries in this high functional demand group. A novel method is the use of a living donor hamstring tendon (LDHT) graft from a parent; however, there is currently limited research on the outcomes of this technique, particularly beyond the short term. Purpose/Hypothesis: The purpose was to determine the medium-term survival of the ACL graft and the contralateral ACL (CACL) after primary ACLR with the use of an LDHT graft from a parent in those aged less than 18 years and to identify factors associated with subsequent ACL injuries. It was hypothesized that ACLR with the use of an LDHT provides acceptable midterm outcomes in pediatric patients. Study Design: Case series; Level of evidence, 4. Methods: Between 2005 and 2014, 247 (of 265 eligible) consecutive patients in a prospective database, having undergone primary ACLR with the use of an LDHT graft and aged less than 18 years, were included. Outcomes were assessed at a minimum of 2 years after surgery including data on ACL reinjuries, International Knee Documentation Committee (IKDC) scores, and current symptoms, as well as factors associated with the ACL reinjury risk were investigated. Results: Patients were reviewed at a mean of 4.5 years (range, 24-127 months [10.6 years]) after ACLR with an LDHT graft. Fifty-one patients (20.6%) sustained an ACL graft rupture, 28 patients (11.3%) sustained a CACL rupture, and 2 patients sustained both an ACL graft rupture and a CACL rupture (0.8%). Survival of the ACL graft was 89%, 82%, and 76% at 1, 2, and 5 years, respectively. Survival of the CACL was 99%, 94%, and 86% at 1, 2, and 5 years, respectively. Survival of the ACL graft was favorable in patients with Tanner stage 1-2 at the time of surgery versus those with Tanner stage 3-5 at 5 years (87% vs 69%, respectively; hazard ratio, 3.7; P = .01). The mean IKDC score was 91.7. A return to preinjury levels of activity was reported by 59.1%. Conclusion: After ACLR with an LDHT graft from a parent in those aged less than 18 years, a second ACL injury (ACL graft or CACL injury) occurred in 1 in 3 patients. The 5-year survival rate of the ACL graft was 76%, and the 5-year survival rate of the CACL was 86%. High IKDC scores and continued participation in sports were maintained over the medium term. Importantly, there was favorable survival of the ACL graft in patients with Tanner stage 1-2 compared with patients with Tanner stage 3-5 over 5 years. Patients with Tanner stage 1-2 also had a significantly lower incidence of second ACL injuries over 5 years compared with those with Tanner stage 3-5, occurring in 1 in 5 patients. Thus, an LDHT graft from a parent is an appropriate graft for physically immature children

A randomized controlled trial of PEEK versus titanium interference screws for anterior cruciate ligament reconstruction with 2-year follow-up

Purpose: To compare the clinical performance of ACL reconstruction with PEEK and titanium interference screws at 2 years and to evaluate a novel method of measuring tunnel volume. Study Design: Randomized controlled trial; Level of evidence, 1. Methods: A total of 133 patients underwent arthroscopic ACL reconstruction with 4-strand hamstring autografts and were randomized to have titanium or PEEK interference screws for femoral and tibial tunnel fixation. At 2 years, subjective Lysholm and International Knee Documentation Committee scores were assessed and clinical examination performed. At 12 months, MRI was performed to assess graft incorporation and cyst formation, and a novel technique was employed to measure tunnel volumes. Results: There were no significant differences in graft rerupture rate, contralateral ACL rupture rate, subjective outcomes, or objective outcomes. In the titanium and PEEK groups, MRI demonstrated high overall rates of graft integration (96%-100% and 90%-93%, respectively) and ligamentization (89% and 84%) and low rates of synovitis (22% and 10%) and cyst formation (0%-18% and 13%-15%). There was a higher proportion of patients with incomplete graft integration within the femoral tunnel in the PEEK group as compared with the titanium group (10% vs 0%, P = .03); however, the authors suggest that metal artifact precluded proper assessment of the graft in the titanium group by MRI. Tunnel volumes also appeared to be equivalent in the 2 groups and were measured with a novel technique that was highly reproducible in the PEEK group secondary to the absence of flare. Conclusion: Two-year clinical analysis of PEEK interference screws for femoral and tibial fixation of ACL reconstructions showed equivalent clinical performance to titanium interference screws. Given the excellent mechanical characteristics, biological compatibility, and absence of metal artifact on MRI, PEEK has become our material of choice for interference screw fixation in ACL reconstruction

Fifteen-year survival of endoscopic anterior cruciate ligament reconstruction in patients aged 18 years and younger

Background: The current body of literature surrounding anterior cruciate ligament (ACL) survival and the variables contributing to further ACL injuries after primary ACL reconstruction in children and adolescents is limited, with no long-term evidence examining the incidence and contributing factors of further ACL injuries in this younger patient population. Purpose: To determine the long-term survival of the ACL graft and the contralateral ACL (CACL) after primary reconstruction in patients aged 18 years and to identify the factors that increase the odds of subsequent ACL injuries. Study Design: Case series; Level of evidence, 4. Methods: Patients having undergone primary ACL reconstruction at age 18 years between 1993 and 1998 who were included in a prospective database by a single surgeon were considered for this study. Single-incision endoscopic ACL reconstruction was performed with either an autologous bone–patellar tendon–bone graft or a hamstring tendon graft. At a minimum of 15 years after ACL reconstruction, patients completed a subjective survey involving the International Knee Documentation Committee (IKDC) questionnaire in addition to questions regarding current symptoms, further ACL injuries, family history of ACL injury, and current level of activity. Results: A total of 288 adolescents (age range, 13-18 years) met the inclusion criteria, of whom 242 (84%) were reviewed at a mean of 16 years and 6 months after ACL reconstruction. Of these patients, 75 (31%) sustained a further ACL injury: 27 (11.2%) suffered an ACL graft rupture, 33 suffered a CACL injury (13.6%), and 15 sustained both an ACL graft rupture and a CACL injury (6.2%) over 15 years. Survival of the ACL graft was 95%, 92%, 88%, 85%, and 83% at 1, 2, 5, 10, and 15 years, respectively, and survival of the CACL was 99%, 98%, 90%, 83%, and 81%, respectively. Survival of the ACL graft was less favorable in those with a family history of ACL injury than in those without a family history (69% vs 90%, respectively; hazard ratio [HR], 3.6; P = .001). Survival of the CACL was less favorable in male patients than in female patients (75% vs 88%, respectively; HR, 2.1; P = .03) and in those who returned to competitive team ball sports than in those who did not (78% vs 89%, respectively; HR, 2.3; P = .05). Conclusion: After ACL reconstruction in patients aged 18 years, a further ACL injury occurred in 1 in 3 patients over 15 years. The 15-year survival rate of the ACL graft was 83%, and the 15-year survival rate of the CACL was 81%. The ACL graft and CACL were most vulnerable within the first 5 years after index surgery. A family history of ACL rupture significantly increased the risk for ACL graft ruptures, and a CACL injury was more common in male patients and those who returned to team ball sports. High IKDC scores and continued participation in sports were maintained over the long term after ACL reconstruction in the adolescent population

Twenty-year outcomes of a longitudinal prospective evaluation of isolated endoscopic anterior cruciate ligament reconstruction with patellar tendon autografts

Background: Long term prospective follow up studies of single-incision endoscopic anterior cruciate ligament reconstruction are limited and may include confounding factors. Objectives: This longitudinal prospective study reports the outcome of isolated anterior cruciate ligament (ACL) reconstruction using middle-third patellar tendon autograft in 90 patients over 20 years. Study design: Case series; Level of evidence, 4. Methods: Between January 1993 and April 1994, 90 patients met study inclusion criteria, evaluation 1,2,3,4,5,7,10,15 and 20 years post-surgery. Exclusion criteria: associated ligamentous injury requiring surgery, previous meniscectomy; meniscal injury meniscectomy more than 1/3; chondral injury; and an abnormal contralateral knee. Results: At 20 years, 32(36%) patients had sustained another ACL injury, 8(9%) to the index limb and 27(29%) to the contralateral limb (3 injuring both knees). Mean IKDC score was 86, 50% participated in strenuous/very strenuous activities, kneeling pain was present in 63%. Radiographic degenerative change was found in 61%, 20% IKDC Grade C, 0% Grade D. IKDC clinical examination revealed 95% had a normal/nearly normal knee. Significant gender differences existed: females were less likely to re-injure the reconstructed ACL (18%v2%, p=0.01), reported poorer IKDC subjective score (90v83, p=0.03), had more activity related pain (57%v20%, p=0.02), and less likely to participate in strenuous activity (35v66, p=0.01). ACL graft survival was not related to age. Patients \u3c18years old had an increased odd ratio (3.2) for rupturing the contralateral ACL. Coronal graft angles \u3c17 degrees had increased risk of failure compared to those over 17 degrees (96% v 77%), by a factor of 8.5. Conclusion: Injury commonly occurred in the contralateral ACL than the reconstructed ACL graft, the most significant predictor of contralateral ACL injury is age under 18yrs. The most significant predictor of ACL graft rupture is a coronal graft angle of less than 17 degrees. Females had lower re- rupture rates, poorer subjective scores, decreased participation in strenuous activity, putting the graft at less risk of failure. Kneeling pain remained persistent over 20 years. Radiographic osteoarthritis was evident in 61% of subjects but symptomatic osteoarthritic symptoms were rarely reported

Feedback from activity trackers improves daily step count after knee and hip arthroplasty: A randomized controlled trial

Background: Commercial wrist-worn activity monitors have the potential to accurately assess activity levels and are being increasingly adopted in the general population. The aim of this study was to determine if feedback from a commercial activity monitor improves activity levels over the first 6 weeks after total hip arthroplasty (THA) or total knee arthroplasty (TKA). Methods: One hundred sixty-three consecutive subjects undergoing primary TKA or THAwere randomized into 2 groups. Subjects received an activity tracker with the step display obscured 2 weeks before surgery and completed patient-reported outcome measures (PROMs). On day 1 after surgery, participants were randomized to either the “feedback (FB) group” or the “no feedback (NFB) group.” The FB group was able to view their daily step count and was given a daily step goal. Participants in the NFB group wore the device with the display obscured for 2 weeks after surgery, after which time they were also able to see their daily step count but did not receive a formal step goal. The mean daily steps at 1, 2, 6 weeks, and 6 months were monitored. At 6 months after surgery, subjects repeated PROMs and daily step count collection. Results: Of the 163 subjects, 95 underwent THA and 68 underwent TKA. FB subjects had a significantly higher (P \u3c .03) mean daily step count by 43% in week 1, 33% in week 2, 21% in week 6, and 17% at 6 months, compared with NFB. The FB subjects were 1.7 times more likely to achieve a mean 7000 steps per day than the NFB subjects at 6 weeks after surgery (P ¼ .02). There was no significant difference between the groups in PROMs at 6 months. Ninety percent of FB and 83% of NFB participants reported that they were satisfied with the results of the surgery (P ¼ .08). At 6 months after surgery, 70% of subjects had a greater mean daily step count compared with their preoperative level. Conclusion: Subjects who received feedback from a commercial activity tracker with a daily step goal had significantly higher activity levels after hip and knee arthroplasty over 6 weeks and 6 months, compared with subjects who did not receive feedback in a randomized controlled trial. Commercial activity trackers may be a useful and effective adjunct after arthroplasty

Midterm outcomes of arthroscopic reduction and internal fixation of anterior cruciate ligament tibial eminence avulsion fractures with K-Wire fixation

Purpose: To determine the clinical and radiological outcomes of patients who underwent arthroscopic reduction and internal fixation of a tibial eminence avulsion fracture with Kirshner wires (K-wires) at a mean of 8 years following surgery. Methods: This was a retrospective study with prospectively collected data. Inclusion criteria consisted of patients who underwent arthroscopic reduction and internal fixation of tibial eminence fracture with K-wires between 1989 and 2015 at a minimum of 18 months follow-up. Assessment included the International Knee Documentation Committee Ligament Evaluation, Lysholm Knee Score, and clinical outcomes. Magnetic resonance imaging (MRI) was performed to evaluate the anterior cruciate ligament (ACL) and evidence of osteoarthritis. Results: A total of 48 participants met the inclusion criteria, and 32 were reviewed at a mean of 8 years (range, 18-260 months) after surgery. The mean age at the time of surgery was 24.5 years (10-55 years). Subsequent ACL injury occurred in 5 participants (10.4%) on the index knee and in 1 participant also on the contralateral knee; 86% had a normal examination, and no patients had \u3e5-mm side-to-side difference on instrumented testing. The mean International Knee Documentation Committee subjective score at 8 years was 86 (range, 40-100). On MRI scan assessment for osteoarthritic changes at final follow-up, 82% of participants had no evidence of chondral wear on the medial compartment and 73% had no changes in the lateral compartment according to Magnetic Resonance Image Osteoarthritis Knee Score classification. On MRI scan qualitative assessment of ACL and tibial eminence, 7 participants (32%) were found to have high signal at the fracture site. The mean medial tibial eminence height was 9.2 mm (range, 6.3 mm to 1.31 cm) and the lateral tibial eminence height was an average of 6.7 mm (range, 0.38-0.97 mm). Significant kneeling pain was reported by 8 participants (25%). Conclusions: This study indicates that internal fixation with K-wires is an acceptable approach to reduce tibial eminence avulsion fractures, providing excellent clinical and radiological outcomes at a minimum of 18 months of follow-up. Level of Evidence: Level IV, therapeutic case series

Transphyseal anterior cruciate ligament reconstruction using living parental donor hamstring graft: Excellent clinical results at 2 years in a cohort of 100 patients

Methods: One hundred consecutive juveniles undergoing ACL reconstruction with a living parental hamstring allograft were recruited prospectively and reviewed 2 years after ACL reconstruction with IKDC Knee Ligament Evaluation, and KT1000 instrumented laxity testing. Skeletally immature participants obtained annual radiographs until skeletal maturity, and long leg alignment radiographs at 2 years. Radiographic Posterior tibial slope was recorded. Results: Of 100 juveniles, the median age was 14 years (range 8-16) and 68% male. At surgery, 30 juveniles were graded Tanner 1 or 2, 21 were Tanner 3 and 49 were Tanner 4 or 5. There were no cases of iatrogenic physeal injury or leg length discrepancy on long leg radiographs at 2 years, despite a median increase in height of 8 cm. Twelve patients had an ACL graft rupture and 9 had a contralateral ACL injury. Of those without further ACL injury, 82% returned to competitive sports, IKDC ligament evaluation was normal in 52% and nearly normal in 48%. The median side to side difference on manual maximum testing with the KT1000 was 2 mm (range - 1 to 5). A radiographic PTS of 12° or more was observed in 49%. Conclusions: ACL reconstruction in the juvenile with living parental hamstring tendon allograft is a viable procedure associated with excellent clinical stability, patient-reported outcomes and return to sport over 2 years. Further ACL injury to the reconstructed and the contralateral knee remains a significant risk, with identical prevalence observed between the reconstructed and contralateral ACL between 12 and 24 months after surgery