Complement C5a-Induced Changes in Neutrophil Morphology During Inflammation.

The complement and neutrophil defence systems, as major components of innate immunity, are activated during inflammation and infection. For neutrophil migration to the inflamed region, we hypothesized that the complement activation product C5a induces significant changes in cellular morphology before chemotaxis. Exposure of human neutrophils to C5a dose- and time-dependently resulted in a rapid C5a receptor-1 (C5aR1)-dependent shape change, indicated by enhanced flow cytometric forward-scatter area values. Similar changes were observed after incubation with zymosan-activated serum and in blood neutrophils during murine sepsis, but not in mice lacking the C5aR1. In human neutrophils, Amnis high-resolution digital imaging revealed a C5a-induced decrease in circularity and increase in the cellular length/width ratio. Biomechanically, microfluidic optical stretching experiments indicated significantly increased neutrophil deformability early after C5a stimulation. The C5a-induced shape changes were inhibited by pharmacological blockade of either the Cl-/HCO3--exchanger or the Cl- -channel. Furthermore, actin polymerization assays revealed that C5a exposure resulted in a significant polarization of the neutrophils. The functional polarization process triggered by ATP-P2X/Y-purinoceptor interaction was also involved in the C5a-induced shape changes, because pretreatment with suramin blocked not only the shape changes but also the subsequent C5a-dependent chemotactic activity. In conclusion, the data suggest that the anaphylatoxin C5a regulates basic neutrophil cell processes by increasing the membrane elasticity and cell size as a consequence of actin-cytoskeleton polymerization and reorganization, transforming the neutrophil into a migratory cell able to invade the inflammatory site and subsequently clear pathogens and molecular debris.

The crucial role of neutrophil granulocytes in bone fracture healing.

Delayed bone fracture healing and the formation of non-unions represent an important clinical problem, particularly in polytrauma patients who suffer from posttraumatic systemic inflammation. However, the underlying pathomechanisms remain unclear. Neutrophil granulocytes are crucial effector cells in the systemic immune response and represent the most abundant immune cell population in the early fracture haematoma. Here we investigated the role of neutrophils in a mouse model of uncomplicated fracture healing and compromised fracture healing induced by an additional thoracic trauma. Twenty four hours before injury, 50 % of the mice were systemically treated with an anti-Ly-6G-antibody to reduce neutrophil numbers. In the isolated fracture model, Ly-6G-Ab treatment significantly increased the concentration of both pro- and anti-inflammatory cytokines, including interleukin (IL)-6 and IL-10, and chemokines, for example, C-X-C motif ligand 1 (CXCL1) and monocyte chemotactic protein-1 (MCP-1), in the fracture haematoma. Monocyte/macrophage recruitment was also significantly enhanced. After 21 d, bone regeneration was considerably impaired as demonstrated by significantly diminished bone content and impaired mechanical properties of the fracture callus. These results indicate that undisturbed neutrophil recruitment and function in the inflammatory phase after fracture is crucial to initiate downstream responses leading to bone regeneration. In the combined trauma model, the reduction of neutrophil numbers ameliorated pulmonary inflammation but did not provoke any significant effect on bone regeneration, suggesting that neutrophils may not play a crucial pathomechanistic role in compromised fracture healing induced by an additional thoracic trauma.

Directional superradiant emission from statistically independent incoherent nonclassical and classical sources.

Superradiance has been an outstanding problem in quantum optics since Dicke introduced the concept of enhanced directional spontaneous emission by an ensemble of identical two-level atoms. The effect is based on the correlated collective Dicke states which turn out to be highly entangled. Here we show that enhanced directional emission of spontaneous radiation can be produced also with statistically independent incoherent sources, via the measurement of higher-order correlation functions of the emitted radiation. Our analysis is applicable to a wide variety of quantum emitters, like trapped atoms, ions, quantum dots, or nitrogen-vacancy centers, and is also valid for incoherent classical emitters. This is experimentally confirmed with up to eight statistically independent thermal light sources. The arrangement to measure the higher-order correlation functions corresponds to a generalized Hanbury Brown-Twiss setup, demonstrating that the two phenomena, superradiance and the Hanbury Brown-Twiss effect, stem from the same interference phenomenon.

Quantum interference and entanglement of photons that do not overlap in time.

We discuss the possibility of quantum interferences and entanglement of photons that exist at different intervals of time, i.e., one photon being recorded before the other has been created. The corresponding two-photon correlation function is shown to violate Bell's inequalities.

Soft X-ray spectromicroscopy of phase-change microcapsules.

A synchrotron-based scanning transmission X-ray microscope (STXM) is used to investigate Micronal phase-change microcapsules. Prolonged X-ray illumination of the specimen leads to the breaking of the microcapsules' protective polymer shell and a partial separation of the core-shell species occurs. The paraffin wax and acrylic polymer components are characterized by carbon K-edge near X-ray edge absorption fine structure (NEXAFS) spectroscopy and components distribution mapping of the beam-damaged specimen is performed.

Mesenchymal stem cells in peripheral blood of severely injured patients.

PURPOSE: Mesenchymal stem cells (MSCs) are primarily stromal cells present in bone marrow and other tissues that are crucial for tissue regeneration and can be mobilized into peripheral blood after different types of organ damage. However, little is known about MSC appearance in blood in the setting of polytrauma. METHODS: We conducted a monocentered and longitudinal observational clinical study in 11 polytraumatized patients with an injury severity score (ISS) ≥ 24 to determine the numbers of MSCs in peripheral blood. Blood was collected from healthy volunteers and patients after polytrauma in the emergency room and 4, 12, 24, 48 h, 5 and 10 day later, and cells carrying MSC-surface markers (negative for CD45, positive for CD29, CD73, CD90, CD105, and CD166 in different combinations also employing the more stringent markers STRO1 and MSCA1) were detected and characterized using flow cytometry. Relative numbers of MSC-like cells were correlated with clinical parameters to evaluate if specific injury patterns had an influence on their presence in the blood cell pool. RESULTS: We were able to detect MSC marker-positive cells in both cohorts; however, the percentage of those cells present in the blood of patients during the first 10 day after injury was mostly similar to healthy volunteers, and significantly lowers starting at 4 h post trauma for one marker combination when compared to controls. Furthermore, the presence of a pelvis fracture was partly correlated with reduced relative numbers of MSC-like cells detectable in blood. CONCLUSIONS: Polytrauma in humans was associated with partly reduced relative numbers of MSC-like cells detected in peripheral blood in the time course after injury. Further studies need to define if this reduction was due to lower mobilization from the bone marrow or to active migration to the sites of injury.

Ultratag RBC kit for combined cardiac first-pass and multigated acquisition studies.

UNLABELLED: The authors developed a procedure to use the in vitro Ultratag (Mallinckrodt, St. Louis, MO) red blood cell (RBC) labeling kit for both first-pass (FP) and multigated acquisition (MUGA) studies with a high specific activity in a reduced volume (50 mCi/0.5 ml) and a high labeling efficiency that can be used with a single-crystal camera to yield a quality study. METHODS: A packed red blood cell (PRBC) bolus was created by two methods: (a) reducing the volume of the components of the Ultratag kit and (b) centrifuging the final dose volume. The labeling efficiency of each bolus was evaluated, each PRBC bolus was visually inspected for clots and percent hemolysis was assessed using a hemocytometer at 30 min, 1 hr and 2 hr postcentrifugation. RESULTS: Use of the first method, the 50% kit, provided the best results. However, the resulting volume from this kit only approached 1 ml, which is not clinically adequate for a first-pass study. In the second method, the total volume was centrifuged to form a PRBC bolus, which appeared to be stable in the syringe for at least 2 hr. A combined FP/MUGA study from a centrifuged 50% reduced kit was performed in one normal subject as a preliminary assessment of the clinical utility of this procedure. The image quality of the scan is diagnostically adequate. CONCLUSION: By using the in vitro Ultratag kit, a compact PRBC bolus was created that was stable in the syringe and could be reinjected safely into the patient for combined cardiac FP/MUGA studies.