Immuno- and constitutive proteasome crystal structures reveal differences in substrate and inhibitor specificity.

Constitutive proteasomes and immunoproteasomes shape the peptide repertoire presented by major histocompatibility complex class I (MHC-I) molecules by harboring different sets of catalytically active subunits. Here, we present the crystal structures of constitutive proteasomes and immunoproteasomes from mouse in the presence and absence of the epoxyketone inhibitor PR-957 (ONX 0914) at 2.9 Å resolution. Based on our X-ray data, we propose a unique catalytic feature for the immunoproteasome subunit ß5i/LMP7. Comparison of ligand-free and ligand-bound proteasomes reveals conformational changes in the S1 pocket of ß5c/X but not ß5i, thereby explaining the selectivity of PR-957 for ß5i. Time-resolved structures of yeast proteasome:PR-957 complexes indicate that ligand docking to the active site occurs only via the reactive head group and the P1 side chain. Together, our results support structure-guided design of inhibitory lead structures selective for immunoproteasomes that are linked to cytokine production and diseases like cancer and autoimmune disorders.

Global signal peptide profiling reveals principles of selective Sec61 inhibition.

Cotransins target the Sec61 translocon and inhibit the biogenesis of an undefined subset of secretory and membrane proteins. Remarkably, cotransin inhibition depends on the unique signal peptide (SP) of each Sec61 client, which is required for cotranslational translocation into the endoplasmic reticulum. It remains unknown how an SP's amino acid sequence and biophysical properties confer sensitivity to structurally distinct cotransins. Here we describe a fluorescence-based, pooled-cell screening platform to interrogate nearly all human SPs in parallel. We profiled two cotransins with distinct effects on cancer cells and discovered a small subset of SPs, including the oncoprotein human epidermal growth factor receptor 3 (HER3), with increased sensitivity to the more selective cotransin, KZR-9873. By comparing divergent mouse and human orthologs, we unveiled a position-dependent effect of arginine on SP sensitivity. Our multiplexed profiling platform reveals how cotransins can exploit subtle sequence differences to achieve SP discrimination.

Proteasome composition in immune cells implies special immune­cell­specific immunoproteasome function.

Immunoproteasomes are a special class of proteasomes, which can be induced with IFN-γ in an inflammatory environment. In recent years, it became evident that certain immune cell types constitutively express high levels of immunoproteasomes. However, information regarding the basal expression of proteolytically active immunoproteasome subunits in different types of immune cells is still rare. Hence, we quantified standard proteasome subunits (ß1c, ß2c, ß5c) and immunoproteasome subunits (LMP2, MECL-1, LMP7) in the major murine (CD4+ T cells, CD8+ T cells, CD19+ B cells, CD11c+ dendritic cells, CD49d+ natural killer cells, Ly-6G+ neutrophils) and human immune cell (CD4+ T cells, CD8+ T cells, CD19+ B cells, CD1c+CD141+ myeloid dendritic cells, CD56+ natural killer cells, granulocytes) subsets. The different human immune cell types were isolated from peripheral blood and the murine immune cell subsets from spleen. We found that proteasomes of most immune cell subsets mainly consist of immunoproteasome subunits. Our data will serve as a reference and guideline for immunoproteasome expression and imply a special role of immunoproteasomes in immune cells.

Signal peptide mimicry primes Sec61 for client-selective inhibition.

Preventing the biogenesis of disease-relevant proteins is an attractive therapeutic strategy, but attempts to target essential protein biogenesis factors have been hampered by excessive toxicity. Here we describe KZR-8445, a cyclic depsipeptide that targets the Sec61 translocon and selectively disrupts secretory and membrane protein biogenesis in a signal peptide-dependent manner. KZR-8445 potently inhibits the secretion of pro-inflammatory cytokines in primary immune cells and is highly efficacious in a mouse model of rheumatoid arthritis. A cryogenic electron microscopy structure reveals that KZR-8445 occupies the fully opened Se61 lateral gate and blocks access to the lumenal plug domain. KZR-8445 binding stabilizes the lateral gate helices in a manner that traps select signal peptides in the Sec61 channel and prevents their movement into the lipid bilayer. Our results establish a framework for the structure-guided discovery of novel therapeutics that selectively modulate Sec61-mediated protein biogenesis.

The International Olympic Committee framework on fairness, inclusion and nondiscrimination on the basis of gender identity and sex variations does not protect fairness for female athletes.

The International Olympic Committee (IOC) recently published a framework on fairness, inclusion, and nondiscrimination on the basis of gender identity and sex variations. Although we appreciate the IOC's recognition of the role of sports science and medicine in policy development, we disagree with the assertion that the IOC framework is consistent with existing scientific and medical evidence and question its recommendations for implementation. Testosterone exposure during male development results in physical differences between male and female bodies; this process underpins male athletic advantage in muscle mass, strength and power, and endurance and aerobic capacity. The IOC's "no presumption of advantage" principle disregards this reality. Studies show that transgender women (male-born individuals who identify as women) with suppressed testosterone retain muscle mass, strength, and other physical advantages compared to females; male performance advantage cannot be eliminated with testosterone suppression. The IOC's concept of "meaningful competition" is flawed because fairness of category does not hinge on closely matched performances. The female category ensures fair competition for female athletes by excluding male advantages. Case-by-case testing for transgender women may lead to stigmatization and cannot be robustly managed in practice. We argue that eligibility criteria for female competition must consider male development rather than relying on current testosterone levels. Female athletes should be recognized as the key stakeholders in the consultation and decision-making processes. We urge the IOC to reevaluate the recommendations of their Framework to include a comprehensive understanding of the biological advantages of male development to ensure fairness and safety in female sports.

Strength and Conditioning (S&C) Practices of Judo Athletes and S&C Coaches: A Survey-Based Investigation.

ABSTRACT: Weldon, A, Cloak, R, Kirk, C, Ruddock, A, Langan-Evans, C, Detanico, D, Loturco, I, and Kons, R. Strength and conditioning (S&C) practices of judo athletes and S&C coaches: A survey-based investigation. J Strength Cond Res 38(4): e160-e173, 2024-The benefits of strength and conditioning (S&C) for improving judo performance and reducing injuries have been widely studied. However, the S&C practices used and perspectives held by those delivering S&C have yet to be elucidated. Therefore, this study investigated the S&C practices and perspectives of judokas and S&C coaches working within judo. Forty-two judokas and 9 S&C coaches completed an online survey comprising 6 sections: (a) written informed consent; (b) background information; (c) education, qualifications, and prescription; (d) views on S&C; (e) exercise selection; and (f) issues and improvements. Frequency analysis was used to report responses to fixed-response questions and thematic analysis for open-ended questions. Results indicated that S&C coaches were primarily responsible for delivering S&C programs (60%), and S&C information was predominantly sourced from S&C coaches (43%). Strength and conditioning was deemed very important for randori (78-88%), overall judo performance (67-79%), and judo fitness (62-78%). Similarly, S&C was considered very important for the development of speed and power (76-89%), strength (71-89%), and injury reduction (69-78%). Novel findings were also observed, such as integrating judo-specific training within S&C practice, which may be partly explained by more S&C coaches holding judo belts (67%) than S&C qualifications (11%). This study supports practitioners delivering S&C in judo by offering a base of information to critique or align with their existing S&C practices and perspectives. Furthermore, our results may help identify potential gaps between methods used, proposed guidelines, and actual practice, facilitating the development of research and education resources tailored to the current climate.

Effective therapy of polymyositis in mice via selective inhibition of the immunoproteasome.

Polymyositis (PM) is a chronic autoimmune inflammatory myopathy resulting in muscle weakness. The limited approved therapies and their poor efficacy contribute to its comorbidity. We investigated the therapeutic use of ONX 0914 and KZR-616, selective inhibitors of the immunoproteasome, in C protein-induced myositis (CIM), a mouse model of PM that closely resembles the human disease. Diseased mice (day 13 postimmunization) were treated with 10 mg/kg ONX 0914, KZR-616, or vehicle on alternate days until day 28. Endpoints included muscle strength assessed by a grip strength meter, serum creatine kinase activity, histology, and immunohistochemistry analysis. Treatment with ONX 0914 or KZR-616 prevented the loss of grip strength in mice after CIM induction, while vehicle-treated animals displayed progressive muscle weakness. Immunoproteasome inhibition lowered PM-associated leukocyte infiltration of the muscle and prevented increased serum creatine kinase levels. LMP7-deficient mice were resistant to CIM induction, as they showed no alterations in grip strength or creatine kinase (CK) levels or muscular alterations. In conclusion, selective inhibition of the immunoproteasome displays therapeutic efficacy in a preclinical mouse model of PM with suppression of muscle inflammation and preservation of muscle strength. Positive results from this study support the rationale for using KZR-616 in clinical studies.

Intra-unit reliability and movement variability of submission grappling external load as measured by torso mounted accelerometery.

Submission grappling consists of skills and movements used in combat sports to physically control opponents whilst trying to apply choke holds and joint locks. There is currently no accepted method of monitoring external load in grappling-based sports due to the absence of key variables such as distance, velocity or time. The primary aim of this study was to determine whether PlayerLoad is a reliable variable for measuring external load of submission grappling movements, with a secondary aim of determining the between repetition variance of submission grappling movements. 7 experienced submission grapplers were recruited. Each wore a torso mounted Catapult® Optimeye S5 microelectromechanical systems (MEMS) device and completed 5 repetitions of each of the following: 4 submission techniques; 5 transition techniques; 2 guard pass techniques; 2 takedown techniques. Accumulated PlayerLoad (PLdACC) was recorded as a marker of absolute load, with accumulated PlayerLoad per minute (PLdACC∙min-1) representing relative load. Reliability of each was assessed using intraclass correlation coefficient (ICC(3,1)) (≥ 0.70). Between repetition movement variation was assessed via coefficient of variation with 95% confidence intervals (CV, 95%CI) (acceptable ≤ 15%, good ≤ 10%). PLdACC ICC(3,1) range = 0.78-0.98, with CV range = 9-22%. PLdACC∙min-1 ICC(3,1) range = 0.83-0.98, with CV range = 11-19%. Though several variables displayed CV > 15%, all had 95%CI lower boundaries ≤ 15%. Whilst PlayerLoad was found to be a reliable measure for submission grappling, relatively high CVs across most techniques examined suggest PlayerLoad may not be appropriate for measuring changes in external load for individual movements in submission grappling. However, it may prove a useful tool for monitoring the external load of full, grappling-based, training sessions within an individual.

Inhibition of the Sec61 translocon overcomes cytokine-induced glucocorticoid resistance in T-cell acute lymphoblastic leukaemia.

Glucocorticoid (GC) resistance is a poor prognostic factor in T-cell acute lymphoblastic leukaemia (T-ALL). Interleukin-7 (IL-7) mediates GC resistance via GC-induced upregulation of IL-7 receptor (IL-7R) expression, leading to increased pro-survival signalling. IL-7R reaches the cell surface via the secretory pathway, so we hypothesized that inhibiting the translocation of IL-7R into the secretory pathway would overcome GC resistance. Sec61 is an endoplasmic reticulum (ER) channel that is required for insertion of polypeptides into the ER. Here, we demonstrate that KZR-445, a novel inhibitor of Sec61, potently attenuates the dexamethasone (DEX)-induced increase in cell surface IL-7R and overcomes IL-7-induced DEX resistance.

Role of Epoxide Hydrolases and Cytochrome P450s on Metabolism of KZR-616, a First-in-Class Selective Inhibitor of the Immunoproteasome.

KZR-616 is an irreversible tripeptide epoxyketone-based selective inhibitor of the human immunoproteasome. Inhibition of the immunoproteasome results in anti-inflammatory activity in vitro and based on promising therapeutic activity in animal models of rheumatoid arthritis and systemic lupus erythematosus KZR-616 is being developed for potential treatment of multiple autoimmune and inflammatory diseases. The presence of a ketoepoxide pharmacophore presents unique challenges in the study of drug metabolism during lead optimization and clinical candidate profiling. This study presents a thorough and systematic in vitro and cell-based enzymatic metabolism and kinetic investigation to identify the major enzymes involved in the metabolism and elimination of KZR-616. Upon exposure to liver microsomes in the absence of NADPH, KZR-616 and its analogs were converted to their inactive diol derivatives with varying degrees of stability. Diol formation was also shown to be the major metabolite in pharmacokinetic studies in monkeys and correlated with in vitro stability results for individual compounds. Further study in intact hepatocytes revealed that KZR-616 metabolism was sensitive to an inhibitor of microsomal epoxide hydrolase (mEH) but not inhibitors of cytochrome P450 (P450) or soluble epoxide hydrolase (sEH). Primary human hepatocytes were determined to be the most robust source of mEH activity for study in vitro. These findings also suggest that the exposure of KZR-616 in vivo is unlikely to be affected by coadministration of inhibitors or inducers of P450 and sEH. SIGNIFICANCE STATEMENT: This work presents a thorough and systematic investigation of metabolism and kinetics of KZR-616 and related analogs in in vitro and cell-based enzymatic systems. Information gained could be useful in assessing novel covalent proteasome inhibitors during lead compound optimization. These studies also demonstrate a robust source in vitro test system that correlated with in vivo pharmacokinetics for KZR-616 and two additional tripeptide epoxyketones.

Pre-competition body mass loss characteristics of Brazilian jiu-jitsu competitors in the United Kingdom.

BACKGROUND: Brazilian jiu-jitsu (BJJ) is a grappling-based combat sport in which competitors engage in pre-competition acute 'weight' loss (AWL) and rapid 'weight' loss (RWL) to achieve the body mass (BM) required for their desired division. AWL/RWL practices of UK BJJ competitors have not previously been reported. AIM: Our aim in this study was to determine the prevalence, magnitude and stakeholder influences of AWL and RWL amongst BJJ participants in the United Kingdom (UK). A secondary aim was to explore whether there is any influence of time spent in the sport or competition frequency on AWL/RWL practices. METHODS: In this study we used the rapid weight loss questionnaire (RWLQ) adapted for BJJ to determine the prevalence and magnitude of AWL/RWL in UK BJJ, the prevalence of methods used and the key stakeholder influences on these practices. As a secondary investigation we aimed to determine whether there was any effect of age starting BJJ on AWL/RWL. RESULTS: Of 115 completed responses, 59% stated they performed AWL/RWL before competition. Mean BM loss for this competition was 1.9 ± 3.8 kg (2.3 ± 4.6%), with 34% of participants starting BM loss 3-7 days prior and 16% starting 0-2 days prior. Methods used tend to be achieving calorie deficit via exercise and diet rather than hypohydration, with little advice from formally qualified personnel. Participants who perform AWL/RWL started training (BF10 = 199, d = .72) and competing (BF10 = 107, d = .68) in BJJ younger than those who do not perform AWL/RWL. CONCLUSIONS: AWL/RWL is prevalent in UK BJJ, but not at the magnitude of other combat sports or countries. Though negative effects of extreme hypohydration are unlikely, there may be a higher chance of eating disorders in BJJ, particularly due to the young age of AWL/RWL commencement.

Proteasome Inhibitors and Their Pharmacokinetics, Pharmacodynamics, and Metabolism.

The proteasome is responsible for mediating intracellular protein degradation and regulating cellular function with impact on tumor and immune effector cell biology. The proteasome is found predominantly in two forms, the constitutive proteasome and the immunoproteasome. It has been validated as a therapeutic drug target through regulatory approval with 2 distinct chemical classes of small molecular inhibitors (boronic acid derivatives and peptide epoxyketones), including 3 compounds, bortezomib (VELCADE), carfilzomib (KYPROLIS), and ixazomib (NINLARO), for use in the treatment of the plasma cell neoplasm, multiple myeloma. Additionally, a selective inhibitor of immunoproteasome (KZR-616) is being developed for the treatment of autoimmune diseases. Here, we compare and contrast the pharmacokinetics (PK), pharmacodynamics (PD), and metabolism of these 2 classes of compounds in preclinical models and clinical studies. The distinct metabolism of peptide epoxyketones, which is primarily mediated by microsomal epoxide hydrolase, is highlighted and postulated as a favorable property for the development of this class of compound in chronic conditions.

Co-inhibition of immunoproteasome subunits LMP2 and LMP7 is required to block autoimmunity.

Cells of hematopoietic origin express high levels of the immunoproteasome, a cytokine-inducible proteasome variant comprising the proteolytic subunits LMP2 (ß1i), MECL-1 (ß2i), and LMP7 (ß5i). Targeting the immunoproteasome in pre-clinical models of autoimmune diseases with the epoxyketone inhibitor ONX 0914 has proven to be effective. ONX 0914 was previously described as a selective LMP7 inhibitor. Here, we show that PRN1126, developed as an exclusively LMP7-specific inhibitor, has limited effects on IL-6 secretion, experimental colitis, and experimental autoimmune encephalomyelitis (EAE). We demonstrate that prolonged exposure of cells with ONX 0914 leads to inhibition of both LMP7 and LMP2. Co-inhibition of LMP7 and LMP2 with PRN1126 and LMP2 inhibitors LU-001i or ML604440 impairs MHC class I cell surface expression, IL-6 secretion, and differentiation of naïve T helper cells to T helper 17 cells, and strongly ameliorates disease in experimental colitis and EAE. Hence, co-inhibition of LMP2 and LMP7 appears to be synergistic and advantageous for the treatment of autoimmune diseases.

The physical demands of mixed martial arts: A narrative review using the ARMSS model to provide a hierarchy of evidence.

The physical demands of mixed martial arts (MMA) training and competition is not yet well quantified. The Applied Research Model for the Sport Sciences (ARMSS) provides a framework through which to conduct sport science, determining pertinent questions to test research findings in real-world settings. The aim of this review was to evaluate MMA research within the context of ARMSS to critically analyse our understanding of the physical requirements of MMA training and competition. Research databases were searched, with 70 peer-reviewed articles being discussed in relation to the specific stage of the ARMSS in which their results best fit. MMA research was found to be mostly foundational and descriptive in nature and has generally not developed along systematic lines. The internal and external loads and responses to training and competition have not been adequately identified. Therefore, it is not currently possible to state which variables are key predictors of success, or how coaches can optimally manipulate these variables. We propose that MMA research be refocused to be conducted within ARMSS. Specifically, stage 2 studies describing the physical, physiological and technical demands of MMA training and competition, and stage 3 studies determining the physiological predictors of performance should be initially prioritised.

Worth the Weight? Post Weigh-In Rapid Weight Gain is Not Related to Winning or Losing in Professional Mixed Martial Arts.

Body mass (BM) manipulation via rapid weight loss (RWL) and rapid weight gain (RWG) is a common practice among mixed martial art (MMA) athletes to ensure qualification for the division in which the athlete wishes to compete. Professional MMA competitors in California are required to weigh in twice: 24 hr prior to competition and immediately prior to the bout after they have typically engaged in RWG. In analyzing data from five MMA events sanctioned by the Californian State Athletic Commission, the authors used Bayesian analyses to compare bout winners (n = 62) and losers (n = 62) in terms of in-competition BM (in kilograms) and the amount of BM regained between the two weigh-ins (in kilograms). These data do not support the hypothesis that differences in in-competition BM (Bayes factor [BF10] = 0.667, d = 0.23) or the amount of BM regained between the two weigh-ins (BF10 = 0.821, d = 0.23) determine winning or losing. In addition, there was no statistical difference between bouts ending via strikes, submission, or decision for either in-competition BM (BF10 = 0.686, ω2 < 0.01) or the amount of BM regained between the two weigh-ins (BF10 = 0.732, ω2 = 0.054). In conclusion, the authors report for the first time that the magnitude of RWG does not predict winning or losing in a professional cohort of MMA athletes. In addition, they also report that MMA athletes typically compete at a BM that is at least 1-2 divisions higher than the division in which they officially weighed-in. These analyses may provide impetus for governing bodies and coaches to enact changes at both professional and amateur levels to reduce negative health consequences associated with extreme RWL and RWG.

Effects of High-Intensity Interval Training in Combat Sports: A Systematic Review with Meta-Analysis.

Vasconcelos, BB, Protzen, GV, Galliano, LM, Kirk, C, and Del Vecchio, FB. Effects of high-intensity interval training in combat sports: A systematic review with meta-analysis. J Strength Cond Res 34(3): 888-900, 2020-Combat sports (CS) are intermittent by nature and high-intensity interval training (HIIT) has been used as a tool to maintain and improve physical fitness among CS athletes. The aim of this study was to perform a systematic review and meta-analysis about chronic effects of HIIT in CS athletes. An electronic search was performed in PubMed, Science Direct, and Google Scholar using the following Boolean criteria: ("CS" OR "martial arts" OR "judo" OR "taekwondo" OR "jiu jitsu" OR "boxing" OR "karate" OR "wrestling" OR "wushu" OR "kung fu") AND ("HIIT" OR "intermittent exercise" OR "sprint interval training" OR "repeated sprint training [RST]"). To be included, the studies needed to be original, involve CS athletes, present HIIT intervention protocol (HIIT, sprint interval training [SIT] or RST), and analyze chronic physiological outcomes. From 2,211 identified studies, after screening and eligibility evaluation, 12 studies were included in this review with meta-analysis. Aerobic (aerobic capacity, heart rate, and maximum oxygen uptake), anaerobic (peak and mean power in single and successive Wingate tests, and blood lactate concentration), and anthropometric outcomes (body mass and body fat percentage) were evaluated. Data of 255 subjects from 12 studies were assessed. Regarding methodological quality, 7 studies obtained 9-10/12 on the TESTEX scale. For the interventions, 5 studies used HIIT, 4 studies used RST protocols, one used SIT protocols, and one used an intermittent protocol that could not be classified. Relating to aerobic power, was found an increase in V[Combining Dot Above]O2max, with a mean difference (MD) of 2.83 ml·kg·min (CI 95% = 0.40-5.25; p < 0.001) for striking and 2.36 ml·kg·min (CI 95% = 1.05-3.66; p < 0.001) for grappling athletes. No differences on anaerobic peak power for striking (MD = 0.67 W; CI 95% = -0.43 to 1.77; p = 0.23) were found, and a statistical improvement for grappling athletes, (MD = 0.51 W; CI 95% = 0.03-0.98; p = 0.04) was found. Seven studies analyzed anthropometric variables, with differences for body mass in striking (MD = -0.93 kg; CI 95% = -1.68 to -0.19; p = 0.01) and no differences for grappling (MD = -0.09 kg; CI 95% = -2.80 to 2.62; p = 0.95). Differences in body fat percentage in striking (MD = 0.50%; CI 95% = 0.30-0.70; p < 0.001) and no differences in grappling (MD = -0.87%; CI 95% = -1.77 to 0.03; p = 0.06) were found. It was concluded that HIIT positively influences maximum oxygen uptake and anaerobic power in combat sport athletes, with a minor impact on body composition.

Immunoproteasome inhibition induces plasma cell apoptosis and preserves kidney allografts by activating the unfolded protein response and suppressing plasma cell survival factors.

Chronic antibody-mediated rejection is the leading cause of allograft dysfunction and loss after kidney transplantation, and current immunosuppressive regimens fail to target the plasma cells that produce alloantibodies. We previously showed that treatment with the immunoproteasome inhibitor ONX 0914 prevented the expansion of plasma cells and prevented chronic allograft nephropathy and organ failure after kidney transplantation in rats, but the mechanism has remained elusive. In the current study, we confirmed a long-term reduction in alloantibody production and improvements in allograft histology in rats treated with ONX 0914 or with the broad-spectrum proteasome inhibitor bortezomib. Plasma cells from allotransplanted rats expressed immunoproteasomes at high levels. Immunoproteasome inhibition with ONX 0914 led to ubiquitin-conjugate accumulation, activation of the unfolded protein response, and induction of apoptosis in plasma cells. In addition, ONX 0914 suppressed the expression of adhesion molecules (VLA-4 and LFA-1), plasma cell survival factors (APRIL and IL-6), and IFN-γ-inducible chemokines in bone marrow, while the APRIL receptor BCMA, the IL-6 receptor, and the chemokine receptors CXCR4 and CXCR3 were down-regulated on plasma cells. Taken together, immunoproteasome inhibition blocked alloantibody production by inducing apoptosis of plasma cells through activating the unfolded protein response and suppressing plasma cell survival factors in the bone marrow.

Diagnostic accuracy of intraoral photographic orthodontic records.

OBJECTIVE: The aims of this study were (1) to investigate how relevant intraoral photographs are to contemporary orthodontic diagnosis and (2) to assess orthodontists' ability to accurately diagnose angle classification and dental midlines using standardized intraoral photographs. METHODS: Study participants were orthodontists who completed a survey regarding photography protocols and their use of intraoral photographs for diagnosis. Each participant was randomized to complete 1 visual diagnostic task regarding either angle classification or midlines. Accuracy was compared across groups and camera angulations. RESULTS: In all, 80% of 192 respondents reported using photographs and clinic notes to plan orthodontic treatment; 50% also included dental casts. For the angle task, accuracy judging molar and canine classification was 79.9% and 51.3%, respectively with ideal standardized photographs. As camera angulation deviated, accuracy decreased significantly (P < 0.0001). For the midline task, accuracy judging the direction of deviation decreased with a small camera angulation change yet increased with a large change (P < 0.001). CONCLUSIONS: When using ideal intraoral photographs alone to diagnose angle classification and midline relationships, accuracy is not likely to be greater than 80%. As camera angulation becomes less ideal, by 15 degrees when judging angle classification or 4 degrees when judging midlines, accuracy is likely to significantly decrease. CLINICAL SIGNIFICANCE: For the clinician who wants to have the most accurate and complete records, our results suggest that intra-oral photos alone may not be adequate when it comes to judging occlusal relationships such as angle classification and esthetic parameters like midlines. When using ideal intraoral photographs to diagnose angle classification and midline relationships, accuracy is not likely to be greater than 80%. As camera angulation becomes less ideal, by as little as 15 degrees when judging angle classification or 4 degrees when judging midlines, accuracy is likely to decrease significantly. Understanding these limitations will allow clinicians to improve both their clinical photography technique and their diagnostic skills.

Immunoproteasome inhibition prevents chronic antibody-mediated allograft rejection in renal transplantation.

Chronic antibody-mediated rejection is the major cause of fading allograft function and loss after renal transplantation. Currently, pharmacological agents for the suppression of chronic antibody-mediated rejection are lacking. Non-selective proteasome inhibitors suppress antibody-mediated allograft rejection. However, extensive adverse side effects of these inhibitors severely limit their application. In contrast, immunoproteasome inhibition is effective in preclinical models of autoimmune diseases and was applied over weeks without obvious adverse side effects. ONX 0914, an immunoproteasome subunit LMP7 (ß5i)-selective inhibitor, impeded the chronic rejection of kidneys transplanted from Fischer to allogeneic Lewis rats. ONX 0914 inhibited immunoproteasome induction both in immune organs and renal allografts. Selective immunoproteasome inhibition reduced the numbers of B and plasma cells, and suppressed donor-specific alloantibody production. The infiltration of T cells, B cells and macrophages as well as interferon-γ, interleukin-17, IgG and complement deposition were reduced in renal allografts of ONX 0914-treated recipients. Chronic nephropathy was ameliorated and renal allograft function preserved, enabling long-term survival of recipients. Thus, our studies define a critical role of the immunoproteasome in chronic kidney allograft rejection and suggest immunoproteasome inhibition as a promising therapeutic approach to suppress chronic antibody-mediated rejection.

Inhibition and deficiency of the immunoproteasome subunit LMP7 attenuates LCMV-induced meningitis.

In addition to antigen processing, immunoproteasomes were recently shown to exert functions influencing cytokine production by monocytes and T cells, T-helper cell differentiation, and T-cell survival. Moreover, selective inhibition of the immunoproteasome subunit LMP7 ameliorated symptoms of autoimmune diseases including CD4(+) T-cell mediated EAE. In this study, we show that LMP7 also plays a crucial role in the pathogenesis of lymphocytic choriomeningitis virus (LCMV)-induced meningitis mediated by CTLs. Mice lacking functional LMP7 display delayed and reduced clinical signs of disease accompanied by a strongly decreased inflammatory infiltration into the brain. Interestingly, we found that selective inhibition and genetic deficiency of LMP7 affect the pathogenesis of LCMV-induced meningitis in a distinct manner. Our findings support the important role of LMP7 in inflammatory disorders and suggest immunoproteasome inhibition as a novel strategy against inflammation-induced neuropathology in the CNS.