Angiogenin Released from ABCB5+ Stromal Precursors Improves Healing of Diabetic Wounds by Promoting Angiogenesis.

Severe angiopathy is a major driver for diabetes-associated secondary complications. Knowledge on the underlying mechanisms essential for advanced therapies to attenuate these pathologies is limited. Injection of ABCB5+ stromal precursors at the edge of nonhealing diabetic wounds in a murine db/db model, closely mirroring human type 2 diabetes, profoundly accelerates wound closure. Strikingly, enhanced angiogenesis was substantially enforced by the release of the ribonuclease angiogenin from ABCB5+ stromal precursors. This compensates for the profoundly reduced angiogenin expression in nontreated murine chronic diabetic wounds. Silencing of angiogenin in ABCB5+ stromal precursors before injection significantly reduced angiogenesis and delayed wound closure in diabetic db/db mice, implying an unprecedented key role for angiogenin in tissue regeneration in diabetes. These data hold significant promise for further refining stromal precursors-based therapies of nonhealing diabetic foot ulcers and other pathologies with impaired angiogenesis.

TLR4-dependent shaping of the wound site by MSCs accelerates wound healing.

We here address the question whether the unique capacity of mesenchymal stem cells to re-establish tissue homeostasis depends on their potential to sense pathogen-associated molecular pattern and, in consequence, mount an adaptive response in the interest of tissue repair. After injection of MSCs primed with the bacterial wall component LPS into murine wounds, an unexpected acceleration of healing occurs, clearly exceeding that of non-primed MSCs. This correlates with a fundamental reprogramming of the transcriptome in LPS-treated MSCs as deduced from RNAseq analysis and its validation. A network of genes mediating the adaptive response through the Toll-like receptor 4 (TLR4) pathway responsible for neutrophil and macrophage recruitment and their activation profoundly contributes to enhanced wound healing. In fact, injection of LPS-primed MSCs silenced for TLR4 fails to accelerate wound healing. These unprecedented findings hold substantial promise to refine current MSC-based therapies for difficult-to-treat wounds.

The kinase domain of CK1δ can be phosphorylated by Chk1.

Members of the casein kinase 1 (CK1) family are key regulators in numerous cellular signal transduction pathways and in order to prevent the development of certain diseases, CK1 kinase activity needs to be tightly regulated. Modulation of kinase activity by site-specific phosphorylation within the C-terminal regulatory domain of CK1δ has already been shown for several cellular kinases. By using biochemical methods, we now identified residues T161, T174, T176, and S181 within the kinase domain of CK1δ as target sites for checkpoint kinase 1 (Chk1). At least residues T176 and S181 show full conservation among CK1δ orthologues from different eukaryotic species. Enzyme kinetic analysis furthermore led to the hypothesis that site-specific phosphorylation within the kinase domain finally contributes to fine-tuning of CK1δ kinase activity. These data provide a basis for the extension of our knowledge about the role of site-specific phosphorylation for regulation of CK1δ and associated signal transduction pathways.

Biomechanical and histological analyses of the fracture healing process after direct or prolonged reduction.

BACKGROUND: Reduction of femoral shaft fractures remains a challenging problem in orthopaedic surgery. Robot-assisted reduction might ease reduction and fracture treatment. However, the influence of different reduction pathways on patients' physiology is not fully known yet. Therefore, the aim of this study was to examine the biomechanics and histology of fracture healing after direct and prolonged robot-assisted reduction in an in vivo rat model. METHODS: 144 male CD® rats were randomly assigned to 12 groups. Each animal received an external fixator and an osteotomy on the left femoral shaft. On the fourth postoperative day, the 1× reduction groups received a single reduction maneuver, whereas the 10× reduction groups received the same reduction pathway with ten repetitions. The control groups did not undergo any reduction maneuvers. Animals were killed after 1, 2, 3 and 4 weeks, respectively, and the composition of the fracture gap was analyzed by µCT and non-decalcified histology. Biomechanical properties were investigated by a three-point bending test, and the bone turnover markers PINP, bCTx, OPG, sRANKL, TRACP-5b, BALP, and OT/BGP were measured. RESULTS: One week after the reduction maneuver, µCT analysis showed a higher cortical bone volume in the 1× reduction group compared to the 10× reduction group. Biomechanically, the control group showed higher maximum force values measured by three-point bending test compared to both reduction groups. Furthermore, less collagen I formation was examined in the 10× reduction group compared to the control group after 1 week of fracture healing. PINP concentration was decreased in 10× reduction group after 1 week compared to control group. The same trend was seen after 3 weeks. CONCLUSION: A single reduction maneuver has a beneficial effect in the early phase of the fracture healing process compared to repeated reduction maneuvers. In the later phase of fracture healing, no differences were found between the groups.

Repetitive reduction lead to significant elevated IL-6 and decreased IL-10 levels in femoral osteotomies: A quantitative analysis of a robot-assisted reduction process in a rat model.

INTRODUCTION: The field of robot-assisted fracture reduction has been developed by several research groups over more than one decade by now, with the main goals of increasing the fracture reduction accuracy. However, the influence of different reduction paths to patients' physiology is not fully known yet. The aim of our study was to compare the impacts of a robot-assisted direct reduction path versus an artificially prolonged reduction path by measuring the cytokine responses in an in vivo rat model. MATERIALS AND METHODS: Thirty-six male CD(©) rats were assigned into three groups with an external fixator and osteotomy on the left femur. Seven days later, the robot was attached and one group was reduced in a single attempt, while the other group underwent 10 attempts by the robot. The third group was the control group without reduction. Before, and as well as 6, 24 and 48h after the reduction process blood samples were collected. IL-1, IL-6, IL-10, IL-17, and MCP-1 concentrations where analysed via ELISA or cytometric bead assay. Muscle biopsies in the osteotomy area were collected 48h after the reduction process for histological analyses. Statistical significance was set at p≤0.05. RESULTS: Analysis of the cytokines showed that the pro-inflammatory cytokine IL-6 of the Ten-Attempts reduction group significantly increased 6h after reduction compared to the control group. IL-6 further showed markedly elevated levels 6h after surgery in the Ten-Attempts reduction group compared to the Single-Attempt reduction group. On the anti-inflammatory side, IL-10 showed a significant decrease in the Ten-Attempts reduction group 6h after reduction compared to the Single-Attempt reduction and control group. Muscle biopsies showed a significant increase of pathological changes in both reduction groups and an increase in the severity of bleedings of the Ten-Attempts reduction group compared to the Single-Attempt reduction and control group. CONCLUSION: A direct and gentle reduction procedure as feasible by the aid of a robot is preferable over a prolonged reduction in terms of cytokine response and tissue changes.

Simultaneous Bilateral Quadriceps Tendon Rupture following Long-Term Low-Dose Nasal Corticosteroid Application.

Simultaneous bilateral quadriceps tendon rupture is a very rare injury, which was previously only described in slightly more than 100 cases in the English literature. Occurrence after minor trauma is predominantly associated with certain medical conditions including chronic diseases and long-term use of certain drugs. We report the case of a 61-year-old healthy patient who sustained a simultaneous bilateral quadriceps tendon rupture following minor trauma. Medical history was completely clear except of a long-term nasal corticosteroid medication due to allergic rhinitis.

Prolonged postsurgical recovery period and adverse effects of a leptin application in endotoxemic obese rodents.

AIMS: Increasing evidence suggests that the adipokine leptin plays a role in modulating immune responses and mediating the link between metabolism and immune system. Obese patients are more susceptible to infections than normal weight individuals. To define the pathophysiological role of leptin during endotoxemia, we examined the effects of leptin on energy metabolism, hemodynamics and quality of life in normal weight and diet-induced obese rats by means of radio-telemetry. MAIN METHODS: Telemetric-transmitter and a central venous catheter were implanted in male Lewis rats. All animals performed two experiments. First, an intravenous injection of 500µlkg(-1) leptin or vehicle (isotonic saline) was performed. After an infusion time of 30min an i.v. bolus of 0.2ml saline over 1min was injected. In the second phase, infusion of placebo or 500µlkg(-1) leptin and an i.v. bolus injection of 100µlkg(-1)Escherichia coli endotoxin were performed. Mean arterial blood pressure (MAP), locomotor activity and electromyogram were recorded via radio-telemetry. Food and water consumption were assessed daily. Quality of life tests were performed at specific times. KEY FINDINGS: Obese animals displayed a prolonged postsurgical recovery period. No benefit could be observed by exogenous leptin in endotoxemic lean or obese animals regarding nutrition balance and locomotor activity. However, leptin treatment even destabilized MAP in obese endotoxemic animals. SIGNIFICANCE: These data demonstrate the necessity to differentiate between normal weight and obese individuals when targeting novel therapeutic strategies for endotoxemia and point out the body weight dependent postsurgical recovery period.

Hybrid and electric low-noise cars cause an increase in traffic accidents involving vulnerable road users in urban areas.

Due to resource scarcity, the number of low-noise and electric cars is expected to increase rapidly. The frequent use of these cars will lead to a significant reduction of traffic related noise and pollution. On the other hand, due to the adaption and conditioning of vulnerable road users the number of traffic accidents involving pedestrians and bicyclists is postulated to increase as well. Children, older people with reduced eyesight and the blind are especially reliant on a combination of acoustic and visual warning signals with approaching or accelerating vehicles. This is even more evident in urban areas where the engine sound is the dominating sound up to 30 kph (kilometres per hour). Above this, tyre-road interaction is the main cause of traffic noise. With the missing typical engine sound a new sound design is necessary to prevent traffic accidents in urban areas. Drivers should not be able to switch the sound generator off.

Injury patterns of seniors in traffic accidents: A technical and medical analysis.

AIM: To investigate the actual injury situation of seniors in traffic accidents and to evaluate the different injury patterns. METHODS: Injury data, environmental circumstances and crash circumstances of accidents were collected shortly after the accident event at the scene. With these data, a technical and medical analysis was performed, including Injury Severity Score, Abbreviated Injury Scale and Maximum Abbreviated Injury Scale. The method of data collection is named the German In-Depth Accident Study and can be seen as representative. RESULTS: A total of 4430 injured seniors in traffic accidents were evaluated. The incidence of sustaining severe injuries to extremities, head and maxillofacial region was significantly higher in the group of elderly people compared to a younger age (P < 0.05). The number of accident-related injuries was higher in the group of seniors compared to other groups. CONCLUSION: Seniors are more likely to be involved in traffic injuries and to sustain serious to severe injuries compared to other groups.

Distinct effects of NPY13-36, a specific NPY Y2 agonist, in a model of rodent endotoxemia on leukocyte subsets and cytokine levels.

Even now, sepsis remains a major problem in modern clinical medicine, leading to systemic inflammatory response including altered leukocyte subset distribution and increased cytokine release. As immune cells are known to express NPY receptors, we investigated the effects of a specific NPY Y(2) receptor agonist (NPY(13-36)) and/or the corresponding Y(2) receptor antagonist BIIE0246 treatment on blood (by FACS analyses) and tissue (by immunohistochemistry) leukocyte subsets as well as on levels of IL-4, IL-6, IL-10, TNF-α, INF-γ (by Cytometric Bead Array) in healthy and acutely endotoxemic rats. Results show a significant decrease in blood monocytes after LPS challenge in endotoxemic control animals (by 93%), in endotoxemic NPY(13-36) treated animals (by 83%) and in endotoxemic BIIE0246 treated animals (by 88%) as compared to the corresponding healthy controls. Endotoxemic control animals showed a significant increase of TNF-α (by 98%) as compared to the healthy control group. A treatment with NPY(13-36) significantly stabilized TNF-α level in endotoxemic animals. This study indicates distinct subset- and cytokine-specific in vivo effects induced by an NPY Y(2) receptor specific treatment after a short-term LPS challenge.

Peripheral but not central leptin treatment increases numbers of circulating NK cells, granulocytes and specific monocyte subpopulations in non-endotoxaemic lean and obese LEW-rats.

Leptin, a hormone mainly generated by adipocytes, acts centrally in the hypothalamus to regulate body weight and energy expenditure. However, there is strong evidence that leptin is also involved in cell-mediated immunity and cytokine crosstalk. In the present study the effects of diet-induced obesity and central and peripheral leptin treatment on leukocyte subsets and cytokine production was investigated. Leptin was injected either intravenously (i.v.) or intracerebroventricularly (i.c.v.) in male endotoxaemic or vehicle-treated healthy LEW-rats. Numbers of blood leukocyte subsets were analysed by FACS and cytokines (TNF-alpha and IL-6) by ELISA. Results showed that peripheral rather than central leptin treatment was able to significantly increase numbers of granulocytes, NK cells and monocytes. Three-colour staining revealed that the increase of ED9(+) monocytes was most likely due to the mobilization of two distinct monocyte subsets, predominantly ED9(+)CD4(-)NKR-P1A(+) and ED9(+)CD4(+)NKR-P1A(+). ELISA analysis revealed significantly elevated TNF-alpha levels in obese animals compared to their lean littermates, while IL-6 failed to show notable changes. In conclusion, the data of the present study revealed that leptin application induces a nutrition- and application-site dependent increase of circulating NK cells, granulocytes and specific monocyte subsets.