Langerhans cells are precommitted to immune tolerance induction.

Antigen-dependent interactions between T lymphocytes and dendritic cells (DCs) can produce two distinct outcomes: tolerance and immunity. It is generally considered that all DC subsets are capable of supporting both tolerogenic and immunogenic responses, depending on their exposure to activating signals. Here, we tested whether epidermal Langerhans cells (LCs) can support immunogenic responses in vivo in the absence of antigen presentation by other DC subsets.

High-resolution episcopic microscopy data-based measurements of the arteries of mouse embryos: evaluation of significance and reproducibility under routine conditions.

Defining the role of genes in the genesis of congenital cardiovascular defects involves comparisons of the diameters of arteries measured in wild-type and genetically engineered mouse embryos. This study aims at evaluating the significance and reproducibility of measurements of the diameters of the great intrathoracic arteries of mouse embryos, as produced under routine conditions, by employing a recently suggested measuring method. Using high-resolution episcopic microscopy, we generated digital volume data of 60 mouse embryos (voxel size 1.

Analysis of behavior and trafficking of dendritic cells within the brain during toxoplasmic encephalitis.

Under normal conditions the immune system has limited access to the brain; however, during toxoplasmic encephalitis (TE), large numbers of T cells and APCs accumulate within this site. A combination of real time imaging, transgenic reporter mice, and recombinant parasites allowed a comprehensive analysis of CD11c+ cells during TE. These studies reveal that the CNS CD11c+ cells consist of a mixture of microglia and dendritic cells (DCs) with distinct behavior associated with their ability to interact with parasites or effector T cells.

Imaging heart development using high-resolution episcopic microscopy.

Development of the heart in vertebrate embryos is a complex process in which the organ is continually remodelled as chambers are formed, valves sculpted and connections established to the developing vascular system. Investigating the genetic programmes driving these changes and the environmental factors that may influence them is critical for our understanding of congenital heart disease. A recurrent challenge in this work is how to integrate studies as diverse as those of cardiac gene function and regulation with an appreciation of the localised interactions between cardiac tissues not to mention the manner in which both may be affected by cardiac function itself.

Towards online multiresolution community detection in large-scale networks.

The investigation of community structure in networks has aroused great interest in multiple disciplines. One of the challenges is to find local communities from a starting vertex in a network without global information about the entire network. Many existing methods tend to be accurate depending on a priori assumptions of network properties and predefined parameters.

[Fast track approach in radical cystectomy].

The origins of the fast track concept in the field of elective colon surgery can be traced back to the beginning of the 1990s. The first studies performed by Kehlet et al. sparked interest in this new form of patient management among physicians and hospital administrators.

Visualizing the neutrophil response to sterile tissue injury in mouse dermis reveals a three-phase cascade of events.

Neutrophil granulocytes traffic into sites of organ injury in which they may not only participate in tissue repair and pathogen clearance but may also contribute to collateral cell damage through the release of noxious mediators. The dynamics and mechanisms of neutrophil migration in the extravascular space toward loci of tissue damage are not well understood. Here, we have used intravital multi-photon microscopy to dissect the behavior of neutrophils in response to tissue injury in the dermis of mice.

Does open reduction of the developmental dislocated hip increase the risk of osteonecrosis?

Background: Osteonecrosis (ON) of the femoral head is one of the main complications associated with treatment of developmental dysplasia of the hips (DDH). The reported rates of ON vary widely between 6% and 48%, suggesting varying factors in these studies influence the rate. Several studies suggest open reduction combined with femoral shortening provides protection against ON.

Ret finger protein inhibits muscle differentiation by modulating serum response factor and enhancer of polycomb1.

Skeletal myogenesis is precisely regulated by multiple transcription factors. Previously, we demonstrated that enhancer of polycomb 1 (Epc1) induces skeletal muscle differentiation by potentiating serum response factor (SRF)-dependent muscle gene activation. Here, we report that an interacting partner of Epc1, ret finger protein (RFP), blocks skeletal muscle differentiation.

Active immunotherapy combined with blockade of a coinhibitory pathway achieves regression of large tumor masses in cancer-prone mice.

Vaccines that aim to expand tumor-specific CD8(+) T cells have yielded disappointing results in cancer patients although they showed efficacy in transplantable tumor mouse models. Using a system that more faithfully mimics a progressing cancer and its immunoinhibitory microenvironment, we here show that in transgenic mice, which gradually develop adenocarcinomas due to expression of HPV-16 E7 within their thyroid, a highly immunogenic vaccine expressing E7 only induces low E7-specific CD8(+) T-cell responses, which fail to affect the size of the tumors. In contrast, the same type of vaccine expressing E7 fused to herpes simplex virus (HSV)-1 glycoprotein D (gD), an antagonist of the coinhibitory B- and T-lymphocyte attenuator (BTLA)/CD160-herpes virus entry mediator (HVEM) pathways, stimulates potent E7-specific CD8(+) T-cell responses, which can be augmented by repeated vaccination, resulting in initial regression of even large tumor masses in all mice with sustained regression in more than half of them.

Beyond the random coil: stochastic conformational switching in intrinsically disordered proteins.

Intrinsically disordered proteins (IDPs) participate in critical cellular functions that exploit the flexibility and rapid conformational fluctuations of their native state. Limited information about the native state of IDPs can be gained by the averaging over many heterogeneous molecules that is unavoidable in ensemble approaches. We used single molecule fluorescence to characterize native state conformational dynamics in five synaptic proteins confirmed to be disordered by other techniques.

Amplitude-integrated EEG pattern predicts further outcome in preterm infants.

Changes in EEG background activity are powerful but nonspecific markers of brain dysfunction. Early EEG and amplitude-integrated EEG (aEEG) pattern predict further neurodevelopmental outcome in term infants; however, sufficient data for prognostic value of aEEG in preterm infants are not available so far. The aim of the study was to evaluate whether aEEG predicts further outcome and to compare it to cerebral ultrasound assessment.

First experiences with a new adjustable plate for osteosynthesis of scaphoid nonunions.

Background: Plate osteosynthesis of the scaphoid, as reported earlier by Ender, has lost its importance in the past few years, after Herbert's introduction of the simple and successful technique of screw osteosynthesis. Only in rare cases does one encounter failed healing or instability of the fragments. Even with a vascularized bone chip, it is not always possible to achieve consolidation.

Cutaneous immunosurveillance by self-renewing dermal gammadelta T cells.

The presence of γδ T cell receptor (TCR)-expressing cells in the epidermis of mice, termed dendritic epidermal T cells (DETCs), is well established. Because of their strict epidermal localization, it is likely that DETCs primarily respond to epithelial stress, such as infections or the presence of transformed cells, whereas they may not participate directly in dermal immune responses. In this study, we describe a prominent population of resident dermal γδ T cells, which differ from DETCs in TCR usage, phenotype, and migratory behavior.

The influence of music on aEEG activity in neurologically healthy newborns ≥32 weeks' gestational age.

Aim: Music is increasingly being used in neonatal intensive care units to aid neurodevelopmental care. The aim of this pilot study was to examine the possible effects of music on quiet sleep (QS) in neurologically healthy newborns.

Methods: Twenty newborns ≥32 weeks' gestational age admitted to the neonatal intensive care unit at The Royal Children's Hospital in Melbourne for specialist consultation were randomly assigned to experimental or control groups.

Targeted induction of antigen expression within dendritic cells modulates antigen-specific immunity afforded by recombinant BCG.

The targeted modulation of antigen expression by recombinant vaccine vehicles would significantly aid development of effective immunotherapeutic strategies. In this report we demonstrate that the Mycobacterium tuberculosis hspX promoter can be used to regulate in vivo induction of antigens expressed by recombinant Bacille Calmette Guérin (rBCG). HspX promoter induction occurred rapidly upon entry of rBCG into cultured dendritic cells (DCs), as evidenced by GFP levels in DCs when infected with BCG:P(hspX)-GFP, in which P(hspX) controlled GFP expression.

Multidirectional volar fixed-angle plating using cancellous locking screws for distal radius fractures--evaluation of three screw configurations in an extra-articular fracture model.

Introduction: Volar fixed-angle plating is a popular treatment for unstable distal radius fractures. Despite the availability of plating systems for treating distal radius fractures, little is known about the mechanical properties of multidirectional fixed-angle plates. The aim of this study was to compare the primary fixation stability of three possible screw configurations in a distal extra-articular fracture model using a multidirectional fixed-angle plate with metaphyseal cancellous screws distally.

Four-color single-molecule fluorescence with noncovalent dye labeling to monitor dynamic multimolecular complexes.

To enable studies of conformational changes within multimolecular complexes, we present a simultaneous, four-color single molecule fluorescence methodology implemented with total internal reflection illumination and camera-based, wide-field detection. We further demonstrate labeling histidine-tagged proteins noncovalently with Tris-nitrilotriacetic acid (Tris-NTA)-conjugated dyes to achieve single molecule detection. We combine these methods to colocalize the mismatch repair protein MutSα on DNA while monitoring MutSα-induced DNA bending using Förster resonance energy transfer (FRET) and to monitor assembly of membrane-tethered SNARE protein complexes.

Advances in imaging the innate and adaptive immune response to Toxoplasma gondii.

Toxoplasma gondii is an intracellular protozoan parasite that infects a wide variety of warm-blooded hosts and can have devastating effects in the developing fetus as well as the immunocompromised host. An appreciation of how this organism interacts with the host immune system is crucial to understanding the pathogenesis of this disease. The last decade has been marked by the application of various imaging techniques, such as bioluminescent imaging as well as confocal and multiphoton microscopy to study toxoplasmosis.

Three-dimensional molecular modeling with single molecule FRET.

Single molecule fluorescence energy transfer experiments enable investigations of macromolecular conformation and folding by the introduction of fluorescent dyes at specific sites in the macromolecule. Multiple such experiments can be performed with different labeling site combinations in order to map complex conformational changes or interactions between multiple molecules. Distances that are derived from such experiments can be used for determination of the fluorophore positions by triangulation.

Results of dynamic treatment of fractures of the proximal phalanx of the hand.

Background: The treatment of fractures of the proximal phalanx in three-phalanx fingers has for a long time been the domain of conservative static treatment in a plaster cast. After removal of the plaster, there was usually limitation of mobility of the interphalangeal joints. Fractures of the proximal phalanx are managed with conservative functional treatment in our clinic.

Optimizing methods to recover absolute FRET efficiency from immobilized single molecules.

Microscopy-based fluorescence resonance energy transfer (FRET) experiments measure donor and acceptor intensities by isolating these signals with a series of optical elements. Because this filtering discards portions of the spectrum, the observed FRET efficiency is dependent on the set of filters in use. Similarly, observed FRET efficiency is also affected by differences in fluorophore quantum yield.

Cell-autonomous and environmental contributions to the interstitial migration of T cells.

A key to understanding the functioning of the immune system is to define the mechanisms that facilitate directed lymphocyte migration to and within tissues. The recent development of improved imaging technologies, most prominently multi-photon microscopy, has enabled the dynamic visualization of immune cells in real-time directly within intact tissues. Intravital imaging approaches have revealed high spontaneous migratory activity of T cells in secondary lymphoid organs and inflamed tissues.