Stability boots for the treatment of Achilles tendon injuries: Gait analysis of healthy participants.

BACKGROUND: Achilles tendon injuries are commonly treated with stability boots that secure the ankle at a specific position and aim to reduce loading of the tendon. These boots allow full weight bearing by limiting the range of movement. It is, however, unknown, to what extent these boots can reduce tendon loading and if the biomechanics are altered during walking. RESEARCH QUESTION: How do orthopedic boots influence lower extremity biomechanics during walking? METHODS: For this cross-sectional study, ten healthy participants walked with three orthopedic boots (Oped Vacoped, Kuenzli Ortho Rehab Absolut, Orthotech Variostabil) commonly used to treat Achilles tendon injuries. Kinematics and kinetics of the lower extremity of the booted leg and spatiotemporal parameters of both sides were collected using motion-capturing system and dynamometry. Each boot was tested in the maximally plantarflexed position. Group differences between boot conditions were analyzed by means of repeated-measures ANOVA and post-hoc paired t-test. RESULTS: The boot dorsiflexion range of motion differed significantly between boots with Vacoped (1.8° (0.3)) showing the smallest range, followed by Kuenzli (5.0° (1.3)) and Orthotech (7.9° (1.7)). Orthotech displayed a higher peak plantarflexion moment (1.36 Nm/kg (0.09)) than both Kuenzli (1.06 Nm/kg (0.12)) and Vacoped (1.04 Nm/kg (0.14)). Concerning loading over time, significant differences in the plantarflexion impulse were found, with the highest impulse in Vacoped (0.42 Nms/kg (0.06)), followed by Orthotech (0.29 Nms/kg (0.03)) and Kuenzli (0.25 Nms/kg (0.05)). In addition, asymmetries were seen in stance and step length for the booted and contralateral sides. SIGNIFICANCE: The lower extremity biomechanics were affected by the boots, with Kuenzli showing the lowest joint loading, Vacoped the smallest joint motion and Orthotech the most natural gait pattern. Future research is needed to determine the most relevant variable expressing the risk of re-rupture of the Achilles tendon in order to conclude which boot may be most favorable to use in clinical practice.

Substrate screening identifies a novel target sequence for the proteasomal activity regulated by ionizing radiation.

The screening for treatment-induced enzyme activities offers the opportunity to discover important regulatory mechanisms and the identification of potential targets for anti-cancer therapies. A novel screening technique was applied to screen substrate peptide sequences for proteolytic activities up- or down-regulated by ionizing radiation in tumor cells. One specific substrate sequence was cleaved in control cell extracts but to a smaller extent in irradiated cell extracts and investigated in detail. Based on protease-class-specific inhibitory studies and cleavage site analysis a potent warhead-inhibitor was synthesized and used to identify the proteasome as the protease of interest. The investigated sequence shows high homology to a regulatory site of nucleoporin 50, an element of the nuclear pore complex, and site specific cleavage of nucleoporin 50 was determined in vitro suggesting a novel link between the ionizing radiation-regulated proteasome and nuclear protein shuttling.

Temperature sensitivity of phospho-Ser(473)-PKB/AKT.

The phospho-PKB/Akt status is often used as surrogate marker to measure activation of the PI3K/Akt/mTOR signal transduction pathway. Though, inconsistencies of the p-Ser(473)-PKB/Akt status have raised doubts in the validity of p-Ser(473)-PKB/Akt phosphorylation as endpoint. Here, we determined that p-Ser(473)-PKB/Akt but not p-Thr(308)-PKB/Akt phosphorylation is highly temperature sensitive. p-Ser(473)-PKB/Akt phosphorylation was rapidly reduced to levels below 50% on exposure to 20-25 degrees C in murine and human cell lines including cells expressing constitutively active PI3K or lacking PTEN. Down-regulation of p-Ser(473)-PKB/Akt was reversible and re-exposure to physiological temperature resulted in increased p-Ser(473)-PKB/Akt phosphorylation levels. Phosphatase activity at low temperature was sustained at 75% baseline level and phosphatase inhibition prevented p-Ser(473)-PKB/Akt dephosphorylation induced by the low temperature shift. Interestingly temperature-dependent deregulation of the p-Ser(473)-PKB/Akt status was also observed in response to irradiation. Thus our data demonstrate that minimal additional stress factors deregulate the PI3K/Akt-survival pathway and the p-Ser(473)-PKB/Akt status as experimental endpoint.

A Traceless Selection: Counter-selection System That Allows Efficient Generation of Transposon and CRISPR-modified T-cell Products.

Recent years have seen major breakthroughs in genome-engineering systems, such as transposon-mediated gene delivery systems and CRISPR-Cas9-mediated genome-editing tools. In these systems, transient expression of auxiliary genes is responsible for permanent genomic modification. For both systems, it would be valuable to select for cells that are likely to undergo stable genome modification. Importantly, in particular for clinical applications of genome-engineered cell products, it will also be of importance to remove those cells that, due to random vector integration, display an unwanted stable expression of the auxiliary gene. Here, we develop a traceless selection system that on the one hand allows efficient enrichment of modified cells, and on the other hand can be used to select against cells that retain expression of the auxiliary gene. The value of this system to produce highly enriched-auxiliary gene-free cell products is demonstrated.

Current concepts for the combined treatment modality of ionizing radiation with anticancer agents.

In current applied radiobiology, there exists a tremendous effort in basic and translational research to identify novel treatment modalities combining ionizing radiation with anticancer agents. This is mainly due to the highly improved molecular understanding of intrinsic radioresistance and the profiling of cellular stress responses to irradiation during recent years. Ionizing radiation not only damages DNA but also affects multiple cellular components that induce a multi-layered stress response. The treatment responses can be restricted to the individual cell level but might also be part of an intercellular stress communication network. Both DNA damage-induced signaling (which results in cell cycle arrest and induction of the DNA-repair machinery) and also ionizing radiation-induced signal transduction cascades, which are generated at cellular sites distant from and independent of DNA-damage, represent interesting targets for anticancer treatment modalities to sensitize for ionizing radiation. Due to the lack of molecular knowledge classic radiobiology assembled the cellular and tissue responses into four groups (4 R's of radiotherapy) which describe biological factors influencing the treatment response to fractionated radiotherapy. These classic 4 R's are Repair, Reassortment, Repopulation and Reoxygenation. With the tremendous progress in molecular oncology we now begin to understand theses factors on the molecular level. At the same time this classification may guide modern molecular radiobiologists to identify novel pharmaceuticals and antisignaling agents which can modulate the treatment response to irradiation. In this review we describe current approaches to sensitize tumor cells with novel anticancer agents along the lines of these 4 R's.