Molecular analysis of Annexin expression in cancer.

BACKGROUND: Uptake of apoptotic cells induces a tolerogenic phenotype in phagocytes and promotes peripheral tolerance. The highly conserved Annexin core domain, present in all members of the Annexin family, becomes exposed on the apoptotic cell-surface and triggers tolerogenic signalling in phagocytes via the Dectin-1 receptor. Consequently, Annexins exposed on tumour cells upon cell death are expected to induce tolerance towards tumour antigens, inhibiting tumour rejection. METHODS: Expression analysis for all Annexin family members was conducted in cancer cell lines of diverse origins. Presentation of Annexins on the cell surface during apoptosis of cancer cell lines was investigated using surface washes and immunoblotting. Expression data from the GEO database was analysed to compare Annexin levels between malignant and healthy tissue. RESULTS: Six Annexins at least were consistently detected on mRNA and protein level for each investigated cell line. AnxA1, AnxA2 and AnxA5 constituted the major part of total Annexin expression. All expressed Annexins translocated to the cell surface upon apoptosis induction in all cell lines. Human expression data indicate a correlation between immune infiltration and overall Annexin expression in malignant compared to healthy tissue. CONCLUSIONS: This study is the first comprehensive analysis of expression, distribution and presentation of Annexins in cancer.

Vaccines targeting self-antigens: mechanisms and efficacy-determining parameters.

We recently showed that it is possible to compromise tumor vessel function and, as a consequence, suppress growth of aggressive preclinical tumors by immunizing against the tumor vascular markers extra domain-A (ED-A) or -B (ED-B) of fibronectin, using a fusion protein consisting of the ED-A or ED-B peptide fused to bacterial thioredoxin. To address the mechanism behind fusion protein-induced immunization and the specific contribution of the different vaccine constituents to elicit an anti-self-antibody response, we immunized mice with modified or unmodified self-antigens, combined with different adjuvant components, and analyzed antibody responses by ELISA in sera. Several essential requirements to circumvent tolerance were identified: (1) a potent pattern recognition receptor agonist like an oligonucleotide containing unmethylated cytosine and guanine dinucleotides (CpG); (2) a depot adjuvant to keep the CpG at the site of injection; and (3) the presence of foreign sequences in the vaccine protein. Lack of either of these factors abolished the anti-self-response (P = 0.008). In mice genetically deficient for type I IFN signaling, there was a 60% reduction in the anti-self-response compared with wild-type (P = 0.011), demonstrating a key role of this pathway in CpG-induced circumvention of self-tolerance. Identification of these mechanistic requirements to generate a potent anti-self-immune response should significantly aid the design of efficient, specific, and safe therapeutic cancer vaccines.

Influence of physical activity on T1 and T2* relaxation times of healthy Achilles tendons at 3T.

PURPOSE: To evaluate longitudinal (T1) and transverse (T2*) relaxation times at different Achilles tendon locations (insertion, mid-portion, and musculotendinous area) in a cohort of subjects with variable tendon straining activity, but without any signs of tendinopathy. MATERIALS AND METHODS: T1 and T2* relaxation times of the Achilles tendon were measured using an ultrashort echo time (UTE) imaging sequence with flip angle and echo time variation on a 3T whole-body clinical magnetic resonance (MR) scanner. Twenty-four tendons of 12 healthy volunteers and 18 tendons of nine healthy recreational long-distance runners (average weekly running distance of 47.4 ± 5.3 km) were enrolled. RESULTS: Mean T1 values of the whole tendon showed no marked group differences, whereas T2* relaxation times of runners were significantly longer (mean percentage increase 15.7 ± 4.9%; P = 0.0181) compared to controls. Regarding group differences for the investigated insertional, mid-portion, and musculotendinous area of the Achilles tendon, only the mid-portion of the Achilles tendon in the running group presented significantly longer T2* values (mean percentage increase 29.1 ± 23.0%; P = 0.0420) in comparison to the control group. CONCLUSION: Prolonged T2* values especially in the mid-portion of the Achilles tendon of runners are likely to reflect an adaptation of the tendon microstructure to repetitive tendon straining activity.

Diagnostic value of T1 and T2 * relaxation times and off-resonance saturation effects in the evaluation of Achilles tendinopathy by MRI at 3T.

PURPOSE: To evaluate and compare the diagnostic value of T1 , T2 * relaxation times and off-resonance saturation ratios (OSR) in healthy controls and patients with different clinical and morphological stages of Achilles tendinopathy. MATERIAL AND METHODS: Forty-two healthy Achilles tendons and 34 tendons of 17 patients with symptomatic and asymptomatic tendinopathy were investigated clinically with conventional magnetic resonance imaging (MRI) sequences on a 3T whole-body MR scanner and a dynamic ultrasound examination. In addition, T1 and T2 * relaxation times were assessed using an ultrashort echo time (UTE) imaging sequence with flip angle and echo time variation. For the calculation of OSR values a Gaussian off-resonance saturation pulse (frequency offset: 750-5000 Hz) was used. The diagnostic value of the derived MR values was assessed and compared using receiver operating characteristic (ROC) curves. RESULTS: ROC curves demonstrate the highest overall test performance for OSR values at 2000 Hz off-resonance in differentiating slightly (OSR-2000 [AUC: 0.930] > T2 * [AUC: 0.884] > T1 [AUC: 0.737]) and more severe pathologically altered tendon areas (OSR-2000 [AUC: 0.964] > T2 * [AUC: 0.917] > T1 [AUC: 0.819]) from healthy ones. CONCLUSION: OSR values at a frequency offset of 2000 Hz demonstrated a better sensitivity and specificity for detecting mild and severe stages of tendinopathy compared to T2 * and particularly when compared to T1 relaxation times.

Evaluation of a six-week whole-body vibration intervention on neuromuscular performance in older adults.

Research in the field of whole-body vibration (WBV) for the enhancement of neuromuscular performance is becoming increasingly popular. However, additional understanding of optimal WBV training protocols is still necessary to develop optimal and effective training and prevention concepts, especially for elderly people. The intention of this study was to evaluate a 6-week WBV intervention program based on optimal vibration loads adapted from the literature on lower-limb strength parameters and performance, as well as on perceived exertion according to a subjective rating. A total of 21 older adults were allocated randomly into either a WBV training or control group (CO). Before and after the intervention period, jump height was measured during a countermovement jump. In addition, isolated isokinetic maximal knee extension and flexion strength, mean power, and work were recorded using a motor-driven dynamometer. Borg's scale for rating of perceived exertion was used to evaluate the intensity of WBV exercises within each training session. After the intervention period, jump height increased by 18.55% (p < 0.001) in the WBV group, whereas values of the CO remained unchanged. There were no statistically significant differences in isokinetic maximal strength, mean power, or work values in knee extension or flexion (all p > 0.05). Finally, the subjective perceived exertion of the WBV exercises and respective training parameters ranged between moderate rating levels of 7 and 13 of Borg's scale. Our data show that WBV is a feasible and safe training program for elderly people to increase multijoint strength performance of the lower limbs during a countermovement jump. This could help to determine the potential of WBV programs in training of the elderly to prevent age-related reduction of neuromuscular performance.

Acute effects of whole-body vibration on trunk and neck muscle activity in consideration of different vibration loads.

The intention of this study was to systematically analyze the impact of biomechanical parameters in terms of different peak-to-peak displacements and knee angles on trunk and neck muscle activity during whole-body vibration (WBV). 28 healthy men and women (age 23 ± 3 years) performed four static squat positions (2 peak-to-peak displacements x 2 knee angles) on a side alternating vibration platform with and without vibration stimulus. Surface electromyography (EMG) was used to record the neuromuscular activity of the erector spinae muscle, the rectus abdominis muscle, and of the splenius muscle. EMG levels normalized to maximal voluntary contractions ranged between 3.2 - 27.2 % MVC during WBV. The increase in muscle activity caused by WBV was significant, particularly for the back muscles, which was up to 19.0 % MVC. The impact of the factor 'condition' (F-values ranged from 13.4 to 132.0, p ≤ 0.001) and of the factor 'peak-to-peak displacement' (F-values ranged from 6.4 to 69.0 and p-values from < 0.001 to 0.01) were statistically significant for each muscle tested. However, the factor 'knee angle' only affected the back muscles (F-value 10.3 and 7.3, p ≤ 0.01). The results of this study should give more information for developing effective and safe training protocols for WBV treatment of the upper body. Key pointsThe maximum levels of muscle activity were significantly reached at high amplitudes at a vibration frequency of 30 Hz.WBV leads to a higher muscle activation of the lower back muscles than of the abdominal muscles.Both knee angles of 30° and 45° have similar effects on the vibration load and represent safe positions to prevent any actual harm.Certain combinations of the biomechanical variables have similar effects on the level of muscle activity.

Short-term exercise-induced changes in hydration state of healthy Achilles tendons can be visualized by effects of off-resonant radiofrequency saturation in a three-dimensional ultrashort echo time MRI sequence applied at 3 Tesla.

PURPOSE: Off-resonant RF saturation influences signal intensity dependent on free and bound water fractions as well as the macromolecular content. The extent of interaction between these compartments can be evaluated by using the off-resonance saturation ratio (OSR). Combined with UTE sequences quantification of OSR even in tendinous tissues with extremely fast signal decay is possible. The aim of this prospective study was to investigate short-term exercise-induced effects of hydration state of the Achilles tendon by means of OSR and tendon volume. MATERIALS AND METHODS: Measurements of OSR and tendon volume before and after ankle-straining activity were performed in seven healthy male volunteers (median age 29 years) using a 3D UTE sequence with implemented off-resonance saturation pulse at 3T (off-resonance frequency 2/3 kHz) and by an automated contour detection in isotropic T2-weighted MR images with sub-millimeter resolution, respectively. Different tendon regions were evaluated. Reproducibility of OSR was measured in subsequent imaging sets. Root-mean-square-deviation (RMSD) and coefficient of variations (CV) were determined. RESULTS: RMSD of OSR in resting position were between 0.006 and 0.01 for different tendon regions and off-resonance frequencies (CV 2 to 3%). A significant increase (P < 0.05) of OSR after exercise was seen in all tendon regions except at the insertion (off-resonance frequency 3 kHz). Tendon volume was decreased significantly after ankle-straining activity (P = 0.003). CONCLUSION: The observed decreased tendon volume and increased OSR directly after exercise indicates a short-term change in tendinous proton compartments, most likely a loss of free water molecules within the tendon.

A 12-week exercise program for patients with hip osteoarthritis has no influence on gait parameters: A secondary analysis of a randomized controlled trial.

BACKGROUND: Hip osteoarthritis is a major musculoskeletal disorder in the elderly. Evidence is given for the efficacy of exercise interventions in terms of self-reported physical functioning and pain. However, it has not yet been fully examined whether exercises influence gait. RESEARCH QUESTION: The purpose of this RCT therefore was to evaluate effects of a 12-week exercise program on kinematic gait variables in subjects with hip osteoarthritis. METHODS: 210 participants were randomly assigned to exercise, non-treated control, or placebo ultrasound groups. The 12-week exercise intervention combined a weekly group session with home-based exercises (2/week), which entailed exercises for motor learning, flexibility, strengthening, and balance. Placebo ultrasound was given once a week. A 6-camera motion capture system was used for data collection. Data were derived from shod walking at self-selected speeds. Spatio-temporal and hip and knee joint angles of the stance phase were calculated. Data were averaged across five trials. Measurements were taken prior to and immediately after the intervention period. ANOVA/Kruskall-Wallis-Tests were used to analyze between-group effects for differences between test days. Pairwise comparisons were subsequently conducted in case of significant model effects. Data were analyzed per protocol (n = 185). RESULTS: No statistically significant differences were detected for any of the outcome measures. SIGNIFICANCE: Although hip muscle strength and gait quality are related, strength training of the hip-surrounding musculature without specific gait training elements cannot improve spatio-temporal gait characteristics or hip and knee joint angles in subjects with mild to moderate hip osteoarthritis. If gait should explicitly be improved through exercise, interventions must incorporate a relevant portion of gait-related tasks.

Associations between biomechanical and clinical/anthropometrical factors and running-related injuries among recreational runners: a 52-week prospective cohort study.

BACKGROUND: The purpose of this exploratory study was to investigate whether runners with certain biomechanical or clinical/anthropometrical characteristics sustain more running-related injuries than runners with other biomechanical or clinical/anthropometrical characteristics. METHODS: The study was designed as a prospective cohort with 52-weeks follow-up. A total of 224 injury-free, recreational runners were recruited from the Gothenburg Half Marathon and tested at baseline. The primary exposure variables were biomechanical and clinical/anthropometrical measures, including strength, lower extremity kinematics, joint range of motion, muscle flexibility, and trigger points. The primary outcome measure was any running-related injury diagnosed by a medical practitioner. Cumulative risk difference was used as measure of association. A shared frailty approach was used with legs as the unit of interest. A total of 448 legs were included in the analyses. RESULTS: The cumulative injury incidence proportion for legs was 29.0% (95%CI = 24.0%; 34.8%). A few biomechanical and clinical/anthropometrical factors influence the number of running-related injuries sustained in recreational runners. Runners with a late timing of maximal eversion sustained 20.7% (95%CI = 1.3; 40.0) more injuries, and runners with weak abductors in relation to adductors sustained 17.3% (95%CI = 0.8; 33.7) more injuries, compared with the corresponding reference group. CONCLUSIONS: More injuries are likely to occur in runners with late timing of maximal eversion or weak hip abductors in relation to hip adductors.

Dectin-1 Binding to Annexins on Apoptotic Cells Induces Peripheral Immune Tolerance via NADPH Oxidase-2.

Uptake of apoptotic cells (ACs) by dendritic cells (DCs) and induction of a tolerogenic DC phenotype is an important mechanism for establishing peripheral tolerance to self-antigens. The receptors involved and underlying signaling pathways are not fully understood. Here, we identify Dectin-1 as a crucial tolerogenic receptor binding with nanomolar affinity to the core domain of several annexins (annexin A1, A5, and A13) exposed on ACs. Annexins bind to Dectin-1 on a site distinct from the interaction site of pathogen-derived ß-glucans. Subsequent tolerogenic signaling induces selective phosphorylation of spleen tyrosine kinase (SYK), causing activation of NADPH oxidase-2 and moderate production of reactive oxygen species. Thus, mice deficient for Dectin-1 develop autoimmune pathologies (autoantibodies and splenomegaly) and generate stronger immune responses (cytotoxic T cells) against ACs. Our data describe an important immunological checkpoint system and provide a link between immunosuppressive signals of ACs and maintenance of peripheral immune tolerance.

Validation of an inertial measurement unit for the quantification of rearfoot kinematics during running.

BACKGROUND: The popular protocol used to study running motion suffers from problems that lead to a limited ability to generalize the obtained results. Inertial measurement units (IMU) appear to be promising in increasing ecological validity of the collected data. However, quantifying running kinematics utilizing IMU signals is complex and potentially affected by several well-established and less well-known errors. RESEARCH QUESTION: The purpose of this study was to examine the validity of kinematic variables obtained from a single, shoe-mounted IMU using an opto-electronic motion analysis reference system. METHODS: 51 recreational runners were analyzed, performing a single continuous run at three different speeds (10, 12, 15 km/h) on a treadmill. Descriptive statistics (Bland & Altman analysis, box plots, scatter plots) were employed to analyze the agreement between the two instruments. RESULTS: The findings of this study revealed considerable systematic and large random disagreement, which, in turn, is characterized by substantial inter-individual differences in the error distribution. These discrepancies may partly be explained by differences in foot strike behavior, resulting in varying degrees of vibration impact acting on the IMU. SIGNIFICANCE: Advances in IMU technology, as well as exploring new application approaches and signal processing strategies, might enhance the usability of IMUs in analyzing running kinematics.

Using the variability of continuous relative phase as a measure to discriminate between healthy and injured runners.

Several studies have used variability of continuous relative phase (CRP) to investigate overuse injuries, since low variability is thought to be related to running injuries. This study investigates whether the analysis of CRP variability leads to additional information about possible differences or similarities between healthy and injured runners. Further, a decision about future applications of CRP variability should be based on the ability to implement and interpret data. 18 healthy female runners (CO) and 18 female runners who suffered from iliotibial band syndrome (ITBS) were evaluated by calculating CRP variability for 4 coupling pairs. Besides analyzing continuous variability of CRP, we also averaged it for the whole stance phase and for four predefined stance phase intervals. Confidence intervals were displayed and independent t-tests for comparing the two groups were conducted. During initial and terminal stance phase as well as after heel-off an increase in CRP variability was detected for both groups of runners. In contrast, the foot flat period was characterized by stable joint coordination and a decrease in variability. This paper presents possible interpretations of CRP variability but no statistically significant differences in CRP variability were found between the two groups of runners. Despite the missing statistical significance, a relationship between high CRP variability and injury seems to be conceivable, since the injured runners demonstrated an increased variability for all couplings in the first half of the stance phase. Further application of CRP variability in biomechanical research is essential to determine whether a relationship exists between injury and coordination variability.

Ultrahigh-quality photonic crystal cavity in GaAs.

Membrane-based photonic crystal GaAs cavities with a double-heterostructure design were investigated. They show the highest quality factors for GaAs photonic crystal cavities to date, exhibiting quality factors higher than 10(5). The resonances of these cavities were fine-tuned in steps smaller than 2 nm by digital etching.