Dark complexes of the Calvin-Benson cycle in a physiological perspective

: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) are two enzymes of the Calvin Benson cycle that stand out for some peculiar properties they have in common: (i) they both use the products of light reactions for catalysis (NADPH for GAPDH, ATP for PRK), (ii) they are both light-regulated through thioredoxins and (iii) they are both involved in the formation of regulatory supramolecular complexes in the dark or low photosynthetic conditions, with or without the regulatory protein CP12. In the complexes, enzymes are transiently inactivated but ready to recover full activity after complex dissociation. Fully active GAPDH and PRK are in large excess for the functioning of the Calvin-Benson cycle, but they can limit the cycle upon complex formation. Complex dissociation contributes to photosynthetic induction. CP12 also controls PRK concentration in model photosynthetic organisms like Arabidopsis thaliana and Chlamydomonas reinhardtii. The review combines in vivo and in vitro data into an integrated physiological view of the role of GAPDH and PRK dark complexes in the regulation of photosynthesis

Custom-made implants for massive acetabular bone loss: accuracy with CT assessment

Background: Custom-made implants are a valid option in revision total hip arthroplasty to address massive acetabular bone loss. The aim of this study was to assess the accuracy of custom-made acetabular implants between preoperative planning and postoperative positioning using CT scans. Methods: In a retrospective analysis, three patients who underwent an acetabular custom-made prosthesis were identified. The custom-made designs were planned through 3D CT analysis considering surgical points of attention. The accuracy of intended implants positioning was assessed by comparing pre- and postoperative CT analyzing the center of rotation (CoR), anteversion, inclination, screws, and implant surface in contact with the bone. Results: The three cases presented satisfactory accuracy in positioning. A malpositioning in the third case was observed due to the posterization of the CoR of the implant of more than 10 mm. The other CoR vectors considered in the third patient and all vectors in the other two cases fall within 10 mm. All the cases were positioned with a difference of less than 10° of anteversion and inclination with respect to the planning. Conclusions: The current case series revealed promising accuracy in the positioning of custom-made acetabular prosthesis comparing the planned implant in preoperative CT with postoperative CT

Validation of an optical, computer-assisted technique for intraoperative tracking of 3-dimensional canine stifle joint motion

Background: Cranial cruciate ligament (CCL) rupture is the most common orthopedic pathology in dog and in men. In human, optical computer-assisted technique is considered as a repeatable and reliable method for the biomechanical assessment of joint kinematics and laxity in case of CCL surgery. Aim: To evaluate the repeatability and reliability afforded by clinical tests in terms of laxity measured by means of a computer-assisted tracking system in two canine CCL conditions: CCL-Intact, CCL-Deficient. Methods: Fourteen fresh frozen canine stifles were passively subjected to Internal/External (IE) rotation at 120\ub0 of flexion and Cranial drawer test (CC). To quantify the repeatability and the reliability, intra-class correlation coefficient (ICC) and the mean percent error were evaluated (\u394 r %). Results: The study showed a very good intra-class correlation, before and after CCL resection for kinematics tests. It was found a minimum ICC = 0.73 during the IE rotation in CCL-Intact and a maximum value of ICC = 0.97 for the CC displacement in CC-Deficient. IE rotation with CCL-Intact is the condition with the greatest \u394 r % = 14%, while the lowest \u394 r % = 6% was obtained for CC displacement in CCL-Deficient. Conclusion: The presented work underlined the possibility of using a computer-assisted method also for biomechanical studies concerning stifle kinematics and laxity

Two-Dimensional and Three-Dimensional Biomechanical Factors During 90° Change of Direction are Associated to Non-Contact ACL injury in Female Soccer Players

Background The two-dimensional (2D) video-analysis of the change of direction (COD) technique has never been used to attempt to predict the risk of ACL injury in female football players. Hypothesis/Purpose The purpose of the present pilot study was to prospectively investigate the biomechanical predictors of ACL injury during a COD task in female football players using both gold standard 3D motion capture and a qualitative scoring system based on 2D video-analysis. Study Design Prospective cohort study Methods Sixteen competitive female football (soccer) players (age 21.4 ± 4.3) performed a series of pre-planned 90° COD tasks. 3D motion data was recorded through 10 stereophotogrammetric cameras and a force platform. 2D frontal and transverse plane joint kinematics were computed through video-analysis from three high-speed cameras. A scoring system based on five criteria was adopted: limb stability, pelvis stability, trunk stability, shock absorption, and movement strategy. The players were prospectively followed for the next two consecutive football seasons and the occurrence of severe knee injuries was registered. Results Four players (25%) experienced an ACL injury. In 3D analysis, ACL-injured players showed greater knee valgus, knee internal rotation, and lower knee flexion (p= 0.017 – 0.029). Lower hip flexion coupled with greater external rotation (p= 0.003 – 0.042), ankle eversion, and contralateral pelvic drop (p<0.001) were also noted. In 2D analysis, ACL-injured players showed greater internal foot rotation, contralateral pelvic drop, lower knee flexion, and contralateral trunk tilt (moderate-to-large effect size). Pelvis stability and trunk stability showed the highest predictive value towards ACL injury. Total score was significantly lower in ACL-injured players with a moderate effect size (d=0.45). Conclusions Both 3D and 2D methodologies depicted biomechanical risk factors and offered predictive insights towards the ACL injury risk. Awareness should rise in women’s football regarding the high risk of ACL injury and the strategies to assess and mitigate it

Systematic Video Analysis of Anterior Cruciate Ligament Injuries in Professional Male Rugby Players: Pattern, Injury Mechanism, and Biomechanics in 57 Consecutive Cases

Background: Anterior cruciate ligament (ACL) injuries represent a significant burden to rugby players. Improving our understanding of the patterns and biomechanics that result in ACL injury may aid in the design of effective prevention programs. Purpose: To describe, using video analysis, the mechanisms, situational patterns, and biomechanics of ACL injuries in professional rugby matches. Further aims were to document injuries according to pitch location and timing within the match. Study Design: Case series; Level of evidence, 4. Methods: A total of 62 ACL injuries were identified in players of the 4 most important rugby leagues across 4 consecutive seasons. We analyzed 57 (92%) injury videos for injury mechanism and situational patterns; biomechanical analysis was performed on indirect and noncontact ACL injuries only (38 cases available). Three reviewers independently evaluated each video. Results: More injuries occurred while attacking than defending (41 [72%] vs 16 [28%]; P <.01). Regarding mechanism, 18 (32%) injuries were direct contact; 15 (26%), indirect contact; and 24 (42%), noncontact. Most direct contact injuries involved being tackled directly to the knee (n = 10). Three situational patterns were identified for players who had a noncontact or indirect contact injury: offensive change of direction (COD) (n = 18), being tackled (n = 10), and pressing/tackling (n = 8). Injuries generally involved a knee-loading strategy in the sagittal plane, which was accompanied by knee valgus loading in most cases (94%). Overall, 73% of injuries occurred during the first 40 minutes of effective playing time. Conclusion: Most ACL injuries in professional male rugby players happened through a noncontact or indirect contact mechanism (68%). Three situational patterns were described, including offensive change of direction, being tackled, and pressing/tackling. Biomechanical analysis confirmed a multiplanar mechanism, with a knee-loading pattern in the sagittal plane accompanied by dynamic valgus. As most injuries occurred in the first 40 minutes, accumulated fatigue appears not to be a major risk factor for ACL injury

Good subjective outcomes, stable knee and high return to sport after tibial eminence avulsion fracture in children

Avulsion fracture of the tibial spine (TSA) is uncommon in children, although its incidence is increasing with the earlier practice of competitive sport activities. This study aims to report mid to long term outcomes in children who sustained a TSA, with a special focus on a return to sport activities. Skeletally immature patients with a TSA, treated in two orthopedic hospitals, were evaluated for range of motion and knee laxity using KT1000, KiRA and Rolimeter. The pediatric International Knee Documentation Committee score (Pedi-IKDC) and the Hospital for Special Surgery pediatric Functional Activity Brief Scale (Pedi-FABS) questionnaires were recorded during the latest visit. Forty-two children were included. Twenty-six were treated nonoperatively and 16 underwent surgery. At a mean follow-up of 6.9 ± 3.6 years, 36 patients completed the questionnaires and 23 patients were tested with arthrometers. Among them, 96% had normal knee laxity. The Pedi-IKDC score averaged 96.4 ± 5.7 points, while the mean Pedi-FABS was 22.2 ± 5.9 points, without statistically significant differences between groups. Twenty-eight patients (78%) returned to their previous level of sport activity (eight amateur, 13 competitive, seven elite athletes). Eight patients (22%) quit sport, mostly because of re-injury fear. If properly treated, pediatric TSAs achieve a high rate of successful healing, with complete restoration of knee stability and an early return to sport activities

Biomechanical analysis of two types of osseointegrated transfemoral prosthesis

In the last two decades, osseointegrated prostheses have been shown to be a good alternative for lower limb amputees experiencing complications in using a traditional socket-type prosthesis; however, restraining biomechanical issues, such as peri-prosthetic bone fractures or loosening, are present. To better understand and overcome these limiting issues, and thus reduce the number of implant failures, many studies have investigated the stress distribution on bone and implant during normal daily activities. The aim of this study was a biomechanical analysis of two different osseointegrated implants, a screw-type (OPRA) and a press fit system (OPL, Osseointegrated Prosthetic Limb), to evaluate the stresses generated in bone and prosthesis during a fall. In particular, four scenarios have been experimentally reproduced to determine the loads on the limb during different kinds of fall. For this purpose, a motion capture system and a force plate have been used. Numerical FEM (Finite Element Method) simulations have been performed to compare the behaviour of the OPRA and OPL systems in different fall scenarios. The obtained results showed that a fall backwards due to balance loss is the most stressful scenario among the ones analysed. As regards the comparison between OPRA and OPL devices, it emerged they have similar behaviours in terms of peak values of the stress, but the OPL implant generates larger high-stress areas in the distal femur as compared with the OPRA system

Increased Rotatory Laxity after Anterolateral Ligament Lesion in Anterior Cruciate Ligament- (ACL-) Deficient Knees: A Cadaveric Study with Noninvasive Inertial Sensors

The anterolateral ligament (ALL) has been suggested as an important secondary knee restrain on the dynamic laxity in anterior cruciate ligament- (ACL-) deficient knees. Nevertheless, its kinematical contribution to the pivot-shift (PS) phenomenon has not been clearly and objectively defined, and noninvasive sensor technology could give a crucial contribution in this direction. The aim of the present study was to quantify in vitro the PS phenomenon in order to investigate the differences between an ACL-deficient knee and an ACL+ALL-deficient knee. Ten fresh-frozen paired human cadaveric knees (n=20) were included in this controlled laboratory study. Intact, ACL-deficient, and ACL+ALL-deficient knees were subjected to a manual PS test quantified by a noninvasive triaxial accelerometer (KiRA, OrthoKey). Kinematic data (i.e., posterior acceleration of the tibial lateral compartment) were recorded and compared among the three statuses. Pairwise Student's t-test was used to compare the single groups (p<0.05). Intact knees, ACL-deficient knees, and ACL+ALL-deficient knees showed an acceleration of 5.3±2.1 m/s2, 6.3±2.3 m/s2, and 7.8±2.1 m/s2, respectively. Combined sectioning of ACL and ALL resulted in a statistically significant acceleration increase compared to both the intact state (p<0.01) and the ACL-deficient state (p<0.01). The acceleration increase determined by isolated ACL resection compared to the intact state was not statistically significant (p>0.05). The ALL sectioning increased the rotatory laxity during the PS after ACL sectioning as measured through a user-friendly, noninvasive triaxial accelerometer

The Thioredoxin-Regulated α-Amylase 3 of Arabidopsis thaliana Is a Target of S-Glutathionylation

Reactive oxygen species (ROS) are produced in cells as normal cellular metabolic by-products. ROS concentration is normally low, but it increases under stress conditions. To stand ROS exposure, organisms evolved series of responsive mechanisms. One such mechanism is protein S-glutathionylation. S-glutathionylation is a post-translational modification typically occurring in response to oxidative stress, in which a glutathione reacts with cysteinyl residues, protecting them from overoxidation. α-Amylases are glucan hydrolases that cleave α-1,4-glucosidic bonds in starch. The Arabidopsis genome contains three genes encoding α-amylases. The sole chloroplastic member, AtAMY3, is involved in osmotic stress response and stomatal opening and is redox-regulated by thioredoxins. Here we show that AtAMY3 activity was sensitive to ROS, such as H2O2. Treatments with H2O2 inhibited enzyme activity and part of the inhibition was irreversible. However, in the presence of glutathione this irreversible inhibition was prevented through S-glutathionylation. The activity of oxidized AtAMY3 was completely restored by simultaneous reduction by both glutaredoxin (specific for the removal of glutathione-mixed disulfide) and thioredoxin (specific for the reduction of protein disulfide), supporting a possible liaison between both redox modifications. By comparing free cysteine residues between reduced and GSSG-treated AtAMY3 and performing oxidation experiments of Cys-to-Ser variants of AtAMY3 using biotin-conjugated GSSG, we could demonstrate that at least three distinct cysteinyl residues can be oxidized/glutathionylated, among those the two previously identified catalytic cysteines, Cys499 and Cys587. Measuring the pKa values of the catalytic cysteines by alkylation at different pHs and enzyme activity measurement (pKa1 = 5.70 ± 0.28; pKa2 = 7.83 ± 0.12) showed the tendency of one of the two catalytic cysteines to deprotonation, even at physiological pHs, supporting its propensity to undergo redox post-translational modifications. Taking into account previous and present findings, a functional model for redox regulation of AtAMY3 is proposed