3D dynamics of bacteria wall entrapment at a water-air interface.

Swimming bacteria can be trapped for prolonged times at the surface of an impenetrable boundary. The subsequent surface confined motility is found to be very sensitive to the physico-chemical properties of the interfaces which determine the boundary conditions for the flow. The quantitative understanding of this complex dynamics requires detailed and systematic experimental data to validate theoretical models for both flagellar propulsion and interfacial dynamics. Using a combination of optical trapping and holographic imaging we study the 3D dynamics of wall entrapment of swimming bacteria that are sequentially released towards a surfactant-covered liquid-air interface. We find that an incompressible surfactant model for the interface quantitatively accounts for the observed normal and tangential speed of bacteria as they approach the boundary. Surprisingly we also find that, although bacteria circulate over the air phase in counterclockwise circular trajectories, typical of free-slip interfaces, the body axis is still tilted "nose down" as found for no-slip interfaces.

Microrheology of DNA hydrogel gelling and melting on cooling.

We present systematic characterisation by means of dynamic light scattering and particle tracking techniques of the viscosity and of the linear viscoelastic moduli, G'(ω) and G''(ω), for two different DNA hydrogels. These thermoreversible systems are composed of tetravalent DNA-made nanostars whose sticky sequence is designed to provide controlled interparticle bonding. While the first system forms a gel on cooling, the second one has been programmed to behave as a re-entrant gel, turning again to a fluid solution at low temperature. The frequency-dependent viscous and storage moduli and the viscosity reveal the different viscoelastic behavior of the two DNA hydrogels. Our results show how little variations in the design of the DNA sequences allow tuning of the mechanical response of these biocompatible all-DNA materials.

Vibration detection by observation of speckle patterns.

When laser light illuminates a rough surface it is scattered into a speckle pattern that is strongly dependent on the surface geometry. Here, we show that it is possible to sense surface vibrations by measuring signal variations from a single pixel detector that collects a small portion of the scattered light. By carefully tuning the probing laser beam size and the detector's aperture it was possible to record a good quality signal in the acoustic band. This approach eliminates the need for an interferometer and thus opens the door to the possibility of detecting vibrations at distances of few hundreds of meters.

Hologram transmission through multi-mode optical fibers.

We demonstrate that a structured light intensity pattern can be produced at the output of a multi-mode optical fiber by shaping the wavefront of the input beam with a spatial light modulator. We also find the useful property that, as in the case for free space propagation, output intensities can be easily superimposed by taking the argument of the complex superposition of corresponding phase-only holograms. An analytical expression is derived relating output intensities ratios to hologram weights in the superposition.

A virtual reality interface for the immersive manipulation of live microscopic systems.

For more than three centuries we have been watching and studying microscopic phenomena behind a microscope. We discovered that cells live in a physical environment whose predominant factors are no longer those of our scale and for which we lack a direct experience and consequently a deep intuition. Here we demonstrate a new instrument which, by integrating holographic and virtual reality technologies, allows the user to be completely immersed in a dynamic virtual world which is a simultaneous replica of a real system under the microscope. We use holographic microscopy for fast 3D imaging and real-time rendering on a virtual reality headset. At the same time, hand tracking data is used to dynamically generate holographic optical traps that can be used as virtual projections of the user hands to interactively grab and manipulate ensembles of microparticles or living motile cells.

Brownian fluctuations and hydrodynamics of a microhelix near a solid wall.

We combine two-photon lithography and optical tweezers to investigate the Brownian fluctuations and propeller characteristics of a microfabricated helix. From the analysis of mean squared displacements and time correlation functions we recover the components of the full mobility tensor. We find that Brownian motion displays correlations between angular and translational fluctuations from which we can directly measure the hydrodynamic coupling coefficient that is responsible for thrust generation. By varying the distance of the microhelices from a no-slip boundary we can systematically measure the effects of a nearby wall on the resistance matrix. Our results indicate that a rotated helix moves faster when a nearby no-slip boundary is present, providing a quantitative insight on thrust enhancement in confined geometries for both synthetic and biological microswimmers.

A transition to stable one-dimensional swimming enhances E. coli motility through narrow channels.

Living organisms often display adaptive strategies that allow them to move efficiently even in strong confinement. With one single degree of freedom, the angle of a rotating bundle of flagella, bacteria provide one of the simplest examples of locomotion in the living world. Here we show that a purely physical mechanism, depending on a hydrodynamic stability condition, is responsible for a confinement induced transition between two swimming states in E. coli. While in large channels bacteria always crash onto confining walls, when the cross section falls below a threshold, they leave the walls to move swiftly on a stable swimming trajectory along the channel axis. We investigate this phenomenon for individual cells that are guided through a sequence of micro-fabricated tunnels of decreasing cross section. Our results challenge current theoretical predictions and suggest effective design principles for microrobots by showing that motility based on helical propellers provides a robust swimming strategy for exploring narrow spaces.

An optical reaction micro-turbine.

To any energy flow there is an associated flow of momentum, so that recoil forces arise every time an object absorbs or deflects incoming energy. This same principle governs the operation of macroscopic turbines as well as that of microscopic turbines that use light as the working fluid. However, a controlled and precise redistribution of optical energy is not easy to achieve at the micron scale resulting in a low efficiency of power to torque conversion. Here we use direct laser writing to fabricate 3D light guiding structures, shaped as a garden sprinkler, that can precisely reroute input optical power into multiple output channels. The shape parameters are derived from a detailed theoretical analysis of losses in curved microfibers. These optical reaction micro-turbines can maximally exploit light's momentum to generate a strong, uniform and controllable torque.

Dynamic density shaping of photokinetic E. coli.

Many motile microorganisms react to environmental light cues with a variety of motility responses guiding cells towards better conditions for survival and growth. The use of spatial light modulators could help to elucidate the mechanisms of photo-movements while, at the same time, providing an efficient strategy to achieve spatial and temporal control of cell concentration. Here we demonstrate that millions of bacteria, genetically modified to swim smoothly with a light controllable speed, can be arranged into complex and reconfigurable density patterns using a digital light projector. We show that a homogeneous sea of freely swimming bacteria can be made to morph between complex shapes. We model non-local effects arising from memory in light response and show how these can be mitigated by a feedback control strategy resulting in the detailed reproduction of grayscale density images.

Light controlled 3D micromotors powered by bacteria.

Self-propelled bacteria can be integrated into synthetic micromachines and act as biological propellers. So far, proposed designs suffer from low reproducibility, large noise levels or lack of tunability. Here we demonstrate that fast, reliable and tunable bio-hybrid micromotors can be obtained by the self-assembly of synthetic structures with genetically engineered biological propellers. The synthetic components consist of 3D interconnected structures having a rotating unit that can capture individual bacteria into an array of microchambers so that cells contribute maximally to the applied torque. Bacterial cells are smooth swimmers expressing a light-driven proton pump that allows to optically control their swimming speed. Using a spatial light modulator, we can address individual motors with tunable light intensities allowing the dynamic control of their rotational speeds. Applying a real-time feedback control loop, we can also command a set of micromotors to rotate in unison with a prescribed angular speed.

Active dynamics of colloidal particles in time-varying laser speckle patterns.

Colloidal particles immersed in a dynamic speckle pattern experience an optical force that fluctuates both in space and time. The resulting dynamics presents many interesting analogies with a broad class of non-equilibrium systems like: active colloids, self propelled microorganisms, transport in dynamical intracellular environments. Here we show that the use of a spatial light modulator allows to generate light fields that fluctuate with controllable space and time correlations and a prescribed average intensity profile. In particular we generate ring-shaped random patterns that can confine a colloidal particle over a quasi one-dimensional random energy landscape. We find a mean square displacement that is diffusive at both short and long times, while a superdiffusive or subdiffusive behavior is observed at intermediate times depending on the value of the speckles correlation time. We propose two alternative models for the mean square displacement in the two limiting cases of a short or long speckles correlation time. A simple interpolation formula is shown to account for the full phenomenology observed in the mean square displacement across the entire range from fast to slow fluctuating speckles.

Delayed Diagnosis of Osteonecrosis of the Jaw (ONJ) Associated with Bevacizumab Therapy in Colorectal Cancer Patients: Report of Two Cases.

Medication-induced Osteonecrosis of the Jaw (MRONJ) has been reported not only after use of antiresorptive agents (bisphosphonates and denosumab), but also in cancer patients receiving antiangiogenic agents, alone or combined with antiresorptive drugs. We report two cases of MRONJ observed in colorectal cancer patients after bevacizumab therapy only. MRONJ was diagnosed, respectively, two and seven months after a tooth extraction; both the patients had received two courses of bevacizumab infusions (for a total of 29 and 10 administrations, respectively). We discuss if tooth extraction during or after antiangiogenic therapy could be a potential trigger of MRONJ, but also if an underlying bone disease not evident before oral surgery might be a possible cause. A careful drug history has to be registered by dental specialists in cancer patients before oral surgery and adequate imaging might be obtained to avoid a delayed diagnosis.

A GPU Simulation Tool for Training and Optimisation in 2D Digital X-Ray Imaging.

Conventional radiology is performed by means of digital detectors, with various types of technology and different performance in terms of efficiency and image quality. Following the arrival of a new digital detector in a radiology department, all the staff involved should adapt the procedure parameters to the properties of the detector, in order to achieve an optimal result in terms of correct diagnostic information and minimum radiation risks for the patient. The aim of this study was to develop and validate a software capable of simulating a digital X-ray imaging system, using graphics processing unit computing. All radiological image components were implemented in this application: an X-ray tube with primary beam, a virtual patient, noise, scatter radiation, a grid and a digital detector. Three different digital detectors (two digital radiography and a computed radiography systems) were implemented. In order to validate the software, we carried out a quantitative comparison of geometrical and anthropomorphic phantom simulated images with those acquired. In terms of average pixel values, the maximum differences were below 15%, while the noise values were in agreement with a maximum difference of 20%. The relative trends of contrast to noise ratio versus beam energy and intensity were well simulated. Total calculation times were below 3 seconds for clinical images with pixel size of actual dimensions less than 0.2 mm. The application proved to be efficient and realistic. Short calculation times and the accuracy of the results obtained make this software a useful tool for training operators and dose optimisation studies.

Vestibular surgical access to the palatine root of the superior first molar: "low-dose cone-beam" CT analysis of the pathway and its anatomic variations.

Apicoectomy of the superior first molar palatine root until recently involved relatively destructive and laborious surgery because of the palatal access route, which not only necessitates an extensive flap, but also the readying of procedures required to deal with a possible hemorrhage from the palatine artery. With the advent of operative microscopy, endodontic surgery has become more precise and less invasive and its technical potential has increased; a possible innovative surgical technique may entail vestibular root access. The possibility of using low-dose, low-cost computed tomography (CT) dedicated to the jaw to obtain anatomic information to plan apicoectomy via the vestibular approach was evaluated; 31 patients were referred to CT with the NewTom apparatus. In 43 superior first molars, the mean distance of the palatine root from the external vestibular cortex was measured, and the frequency that the maxillary sinus lateral recess lay between the roots was determined. The mean root apex-vestibular cortex distance was 9.73 mm. In 25% of cases the maxillary sinus recessus lay between vestibular and palatine roots. CT may play an important role in optimizing palatine root apicoectomy through vestibular access, with regard to precision and preventing complications, with relatively low biological and economic cost, also possibly contributing to the affirmation of this new surgical procedure.

A multi-mode fiber probe for holographic micromanipulation and microscopy.

Holographic tweezers have revolutionized the way we do experiments at the micron scale. The possibility of applying controlled force fields on simultaneously trapped micro-particles has allowed to directly probe interactions and mechanical properties of colloids, macromolecules and living cells. Holographic micromanipulation requires the careful shaping of a laser beam that is then focused by a microscope objective onto a micro-hologram in the sample volume. The same objective is used for imaging. That approach is therefore limited to in vitro samples contained in transparent cells that are easily accessed optically. Here we demonstrate that the complex light propagator of a real multimode fiber can be directly measured. That allows to transmit a micro-hologram through a 1 metre long (60 µm core) optical fiber and produce dynamic arrays of focused spots at the fiber output. We show that those spots can be used for interactive holographic micromanipulation of micron sized beads facing the fiber tip. Scanning a single spot across the output fiber we can employ the same fiber as a probe for scanning fluorescence microscopy. Our findings open the way towards the fabrication of endoscopic probes which could be capable of seeing and manipulating single cells deep into biological tissues.

Computerized tomographic findings in bisphosphonate-associated osteonecrosis of the jaw in patients with cancer.

OBJECTIVES: To study bisphosphonate-associated osteonecrosis of the jaw with respect to radiographic, demographic, and clinical features. STUDY DESIGN: Thirty-two patients with painful exposed bone in the jaws subsequent to treatment with bisphosphonates for various neoplastic diseases were examined with both dental panoramic radiograph and multislice spiral computerized tomography (CT). Each case was assessed for the following radiologic signs: structural alteration of trabecular bone, cortical bone erosion, osteosclerosis, small sequestrum, extensive sequestrum, and presence of periosteal new bone. The McNemar, chi2, and Kruskall-Wallis tests were performed to analyze the association between radiologic signs, demographic data, clinical aspects of the lesion, and type and duration of bisphosphonate therapy. Hierarchical cluster analysis was used to group patients into categories on the basis of CT signs and dental panoramic radiograph signs. The agreement between CT and dental panoramic radiograph clusters was analyzed by kappa index. RESULTS: Dental panoramic radiograph proved positive for features of osteonecrosis in 18 cases and CT in 30. Computerized tomography was far superior to dental panoramic radiograph in detecting all the radiologic signs. The cluster analysis applied to CT signs showed a classification of the 32 patients in 4 categories characterized by an increasing number of signs coherent with the clinical extension of the lesions. Structural alteration of trabecular bone and cortical bone erosion, a constant finding in the 30 positive CTs, could be sometimes differentiated only with difficulty from those due to recent extraction. Dental panoramic radiograph missed the correct diagnosis of sequestration in 15 cases. Intense periosteal reaction was often found. The most extensive maxillary lesions were associated with new bone formation in Schneider mucous membrane, never described previously, and oroantral communications. CONCLUSIONS: Dental panoramic radiograph were found to be of limited use in assessing bisphosphonate-associated osteonecrosis of the jaw in patients for whom CT imaging was subsequently ordered.

Vertical alveolar ridge augmentation by means of a titanium mesh and autogenous bone grafts.

The aim of this study is to evaluate a surgical protocol for vertical ridge augmentation in the maxilla and mandible using autogenous onlay bone graft associated with a titanium mesh. A group of 18 partially edentulous patients, presenting the need for vertical bone augmentation of at least 4 mm, were treated before implant placement. During the first surgery, an autogenous bone graft was harvested from either the mandibular ramus or the mental symphysis and secured by means of titanium screws. Particulate bone was added and a titanium micro-mesh was used to stabilize and protect the graft. After a mean interval of 4.6 months, meshes and screws were removed and 37 endosseous implants were successfully placed. The desired bone gain was reached in all patients. Mean vertical bone augmentation obtained was 4.8 mm (range 4-7 mm). No major complications were recorded at recipient or donor sites. Abutment connection was carried out 2-3 months after implant placement. No implant was lost. Clinical parameters and probing depth, after prosthetic reconstruction, demonstrated the presence of a healthy peri-implant mucosa. The preliminary results suggest that, by using the presented technique, patients can be successfully rehabilitated by means of implant-supported prosthesis 6-7 months after the first surgery, even in case of severely atrophied maxilla.

Precision of transfer of preoperative planning for oral implants based on cone-beam CT-scan images through a robotic drilling machine.

In this study, an image-guided system for oral implant placement was assessed. A specially designed mechanical tool has been elaborated to transfer the preoperative implant axis planned on 3-dimensional imagery into a surgical template by a numerically controlled drilling machine. The main drawback of image-guiding systems is the use of preoperative computed tomography, which is expensive and delivers high radiation doses. Therefore, in this study the image-guiding system was coupled with a cone-beam tomograph that significantly decreased both cost and radiation doses. Three edentulous models were used. To determine the accuracy of the system, the ability of a 1.8-mm diameter drill to enter a 2.0-mm diameter, 10-mm-long titanium tube inserted on the model with no contact was verified. Because the drill entered the tubes with no contact and went beyond the end of the tube, the transfer error was less than 0.2 mm for translation and less than 1.1 degrees for rotation. The method presented here is low cost and high precision compared to other technological solutions such as tracking. Further assessment in the surgical field should lead to daily use of this system for flapless surgery, to prepare a prosthesis prior to surgery for immediate loading, to reduce risk of injuring critical anatomical structures and to eliminate manual placement error.