RelA/NF-kappaB recruitment on the bax gene promoter antagonizes p73-dependent apoptosis in costimulated T cells

The balance between antiapoptotic and proapoptotic proteins of the Bcl-2 family is critical in determining the fate of T cells in response to death stimuli. Proapoptotic genes, such as bax, are generally regulated by the p53 family of transcription factors, whereas NF-kappaB subunits can activate the transcription of antiapoptotic Bcl-2 members. Here, we show that CD28 activation protects memory T cells from irradiation-induced apoptosis by both upregulating bcl-xL and inhibiting bax gene expression. We found that p73, but not p53, binds to and trans-activates the bax gene promoter in irradiated T cells. The activation of RelA/NF-kappaB subunit in CD28 costimulated T cells and its binding onto the bax gene promoter results in suppression of bax transcription and decrease in both p73 and RNA polymerase II recruitment in vivo. RelA recruitment on the bax gene promoter is also accompanied by the lost of p300 binding and the parallel appearance of histone deacetylase-1-containing complexes. These findings identify RelA/NF-kappaB as a critical regulator of T-cell survival by affecting the balance of Bcl-2 family members

Tumor sensitizing function and mechanism of action of a RasGAP derived peptide

Cancer is the second cause of death after cardio-vascular diseases in economically developed countries. Two of the most commonly used anti-cancer therapies are chemo and radiotherapy. Despite the remarkable advances made in term of delivery and specificity of these two anti-tumor regimens, their toxicity towards healthy tissue remains a limitation. A promising approach to overcome this obstacle would be the utilization of therapeutic peptides that specifically augment the sensitivity of tumoral cells to treatments. Lower therapeutical doses would then be required to kill malignant cells, limiting toxic effects on healthy tissues. It was previously shown in our laboratory that the caspase-3 generated fragment N2 of RasGAP is able to potentiate the genotoxin-induced apoptosis selectively in cancer cells. In this work we show that fragment N2 strictly requires a cytoplasmic localization to deliver its pro-apoptotic effect in genotoxin-treated cancer cells. The tumor sensitizing capacity of fragment N2 was found to reside within the 10 amino acid sequence 317-326. Our laboratory earlier demonstrated that a peptide corresponding to amino acids 317 to 326 of RasGAP fused to the TAT cell permeable moiety, called TAT-RasGAP317.326, is able to sensitize cancer cells, but not normal cells, to genotoxin-induced apoptosis. In the present study we describe the capacity of TAT-RasGAP 317.326 to sensitize tumors to both chemo and radiotherapy in an in vivo mouse model. The molecular mechanism underlying the TAT-RasGAP 317.326-mediated sensitization starts now to be elucidated. We demonstrate that G3BP1, an endoribonuclease binding to amino acids 317-326 of RasGAP, is not involved in the sensitization mechanism. We also provide evidence showing that TAT-RasGAP3 17-326 potentiates the genotoxin-mediated activation of Bax in a tBid-dependent manner. Altogether our results show that TAT-RasGAP 317.326 could be potentially used in cancer therapy as sensitizer, in order to improve the efficacy of chemo and radiotherapy and prolong the life expectancy of cancer patients. Moreover, the understanding of the TAT-RasGAP317.326 mode of action might help to unravel the mechanisms by which cancer cells resist to chemo and radiotherapy and therefore to design more targeted and efficient anti-tumoral strategies

Revisiting G3BP1 as a RasGAP binding protein: sensitization of tumor cells to chemotherapy by the RasGAP 317-326 sequence does not involve G3BP1.

RasGAP is a multifunctional protein that controls Ras activity and that is found in chromosomal passenger complexes. It also negatively or positively regulates apoptosis depending on the extent of its cleavage by caspase-3. RasGAP has been reported to bind to G3BP1 (RasGAP SH3-domain-binding protein 1), a protein regulating mRNA stability and stress granule formation. The region of RasGAP (amino acids 317-326) thought to bind to G3BP1 corresponds exactly to the sequence within fragment N2, a caspase-3-generated fragment of RasGAP, that mediates sensitization of tumor cells to genotoxins. While assessing the contribution of G3BP1 in the anti-cancer function of a cell-permeable peptide containing the 317-326 sequence of RasGAP (TAT-RasGAP₃₁₇₋₃₂₆), we found that, in conditions where G3BP1 and RasGAP bind to known partners, no interaction between G3BP1 and RasGAP could be detected. TAT-RasGAP₃₁₇₋₃₂₆ did not modulate binding of G3BP1 to USP10, stress granule formation or c-myc mRNA levels. Finally, TAT-RasGAP₃₁₇₋₃₂₆ was able to sensitize G3BP1 knock-out cells to cisplatin-induced apoptosis. Collectively these results indicate that G3BP1 and its putative RasGAP binding region have no functional influence on each other. Importantly, our data provide arguments against G3BP1 being a genuine RasGAP-binding partner. Hence, G3BP1-mediated signaling may not involve RasGAP

Checkpoints in TNF-Induced Cell Death: Implications in Inflammation and Cancer.

Tumor necrosis factor (TNF) is a proinflammatory cytokine that coordinates tissue homeostasis by regulating cytokine production, cell survival, and cell death. However, how life and death decisions are made in response to TNF is poorly understood. Many inflammatory pathologies are now recognized to be driven by aberrant TNF-induced cell death, which, in most circumstances, depends on the kinase Receptor-interacting serine/threonine-protein kinase 1 (RIPK1). Recent advances have identified ubiquitin (Ub)-mediated phosphorylation of RIPK1 as belonging to crucial checkpoints for cell fate in inflammation and infection. A better understanding of these checkpoints might lead to new approaches for the treatment of chronic inflammatory diseases fueled by aberrant RIPK1-induced cell death, and/or reveal novel strategies for anticancer immunotherapies, harnessing the ability of RIPK1 to trigger immunogenic cell death

Acute primary repair of the anterior cruciate ligament with anterolateral ligament augmentation

Acute injuries of the anterior cruciate ligament are often associated with concurrent injuries to the structures of the anterolateral complex, specifically the anterolateral ligament. Some injury patterns of the anterior cruciate ligament involve tearing of the majority of the ligament from the femoral origin, leaving a large, viable ligament remnant. In these patients, a repair of the anterior cruciate ligament back to the femoral origin can be undertaken. Subsequently, percutaneous repair of the anterolateral ligament can be performed through anatomical, percutaneous suture tape augmentation. The combined technique of anterior cruciate ligament repair with anterolateral ligament reinforcement is presented

Combined acl and segond repair in combined acute proximal acl tears and segond fracture

A renewed interest in anterior cruciate ligament preservation has been noted using arthroscopic primary repair in patients with proximal tears, but the main concern remained the control of the rotational instability. Segond fracture occurs in less than 10% of cases of acute anterolateral instability, but it can result in continued rotation instability. The aim of this study is to describe the surgical technique to acutely repair both the anterior cruciate ligament and Segond fracture in the acute setting

Non-invasive computer navigation can quantify the pivot shift maneuver with good to excellent reliability in healthy volunteers

Purpose: The aim of this study was to determine the inter- and intra-observer reliability of knee laxity assessment using a non-invasive navigation system in a population of healthy young athletes. It was hypothesized that knee laxity parameters recorded using non-invasive computer navigation would demonstrate good inter- and intra- observer reliability. Methods: Healthy volunteers aged between 18 to 30 years were recruited to the study. Static and dynamic knee laxity parameters including anterior tibial translation and tibial rotation during the pivot shift test were recorded on awake patients using non-invasive computer navigation by two independent observers: at the first visit each athlete was evaluated by the consultant and resident surgeons independently; 6 weeks after the first visit all the participants were re-tested only by the resident surgeon. Inter- and intra-observer reliability was calculated and then interpreted according to Cicchetti’s criteria. Results: One hundred healthy volunteers were recruited to the study, of these 38 were women (38%), and the average age was 25.5 ± 2.4 years. According to Cicchetti’s criteria the intra- and inter-observer reliability for static measurements were fair for anterior tibial translation (0.572 and 0.529, respectively) and excellent for total passive tibial rotation (0.859 and 0.883, respectively). For the dynamic measurements of translation and rotation during the pivot shift maneuver both measurements demonstrated good to excellent reliability with intra and inter observer reliability ranging from 0.684 to 0.936. Conclusion: Non-invasive navigation for the assessment of knee laxity is associated with fair to excellent inter- and intra-observer reliability in a population of healthy volunteers

Quick recovery and no arthrofibrosis in acute anterior cruciate ligament reconstruction. A prospective trial of early versus delayed reconstruction

Background. Anterior cruciate ligaments tears is one of the most frequent orthopae- dics and sports medicine injuries in the athletically active population and timing of reconstruction represents a debated topic. The aim of the study is to compare range of motion (ROM) recovery and clinical outcomes between patients operated for acute reconstruction (maximum 2 weeks injury-surgery interval) and delayed reconstruction (minimum 3 weeks injury-surgery interval). Methods. A total of 52 patients were prospectively involved in the study. 26 patients underwent acute reconstruction and 26 delayed reconstruction. A standard physical examination with Lachman and Pivot shift test and a passive ROM measurement with a goniometer were performed at each follow-up (2, 4, 8, 12 and 24 weeks postoper- atively). Clinical outcomes were measured at final follow-up using Knee Injury and Osteoarthritis outcome score (KOOS), Tegner Lysholm Score and International Knee Documentation Committe (IKDC 2000) and KT-1000 evaluation. Single-leg hop test and thigh circumference measurement were performed at final follow-up. Results. Both groups showed no statistically significant differences regarding the ROM. Full ROM was achieved 12 weeks after surgery in both groups. The mean IKDC was 98.7 and 95.2; the mean Tegner Lysholm was 100 and 93.8 and the mean KOOS was 99 and 95.5 in the acute group and delayed ACLR group respectively. Conclusions. There were no differences between acute and delayed anterior cruci- ate ligament reconstruction regarding the risk of arthrofibrosis and clinical outcomes. Acute reconstruction can be performed safely with no increased risk of arthrofibrosis

In-office needle arthroscopic assessment after primary ACL repair. Short-term results in 15 patients

Purpose: In-office needle arthroscopy has been reported as a diagnostic tool for different knee pathologies. In addi- tion, ACL repair has seen a resurgence with the advent of innovative orthopedic devices. The aim of this study was to assess clinical, radiological, and in-office needle arthroscopic findings in 15 adult patients who underwent acute (within 14 days from injury) anterior cruciate ligament (ACL) repair. Methods: Fifteen patients voluntarily participated in the study. A second-look arthroscopy was performed with an in-office needle arthroscopy at an average of 7.2 months after the primary repair. The parameters included in the investigation were the continuity of the anatomical footprint of the repaired ACL, subjective assessment of the ACL tension with the probe, and synovial coverage of the ACL. All patients had a Magnetic Resonance Imaging (MRI) at 6 months after repair and an arthrometric evaluation with the KT-1000. Clinical evaluation with the scores, Tegner Lysholm Knee Scoring Scale (TLKSS), the Knee Injury and Osteoarthritis Outcome Score (KOOS), and International Knee Documentation Committee (IKDC) was performed at the final follow-up of 2 years. Moreover, a correlation between the characteristics of ACL appearance at the time of the second look in-office needle arthroscopy, MRI and KT-1000 was performed. Results: The mean TLKSS was 97.86, the mean KOOS was 98.08 and the mean subjective IKDC was 96.71. The objec- tive IKDC was A in 10 patients and B in 5 patients. ACL healing was graded as A in 11 patients and B in 4 patients. Synovial coverage was graded as good in 10 patients and fair in 5 while MRI assessment showed a type I ACL in 10 patients, type II in 4 patients and type III in 1 patient. Conclusion: In-office needle arthroscopy is a reliable tool to assess the condition of a repaired ACL. In addition, ACL repair performed in acute proximal tears demonstrated excellent clinical results