Interaction between Alzheimer's Aβ(25–35) peptide and phospholipid bilayers: The role of cholesterol

AbstractThere is mounting evidence that the lipid matrix of neuronal cell membranes plays an important role in the accumulation of β-amyloid peptides into senile plaques, one of the hallmarks of Alzheimer's disease (AD). With the aim to clarify the molecular basis of the interaction between amyloid peptides and cellular membranes, we investigated the interaction between a cytotoxic fragment of Aβ(1–42), i.e., Aβ(25–35), and phospholipid bilayer membranes. These systems were studied by Electron Paramagnetic Resonance (EPR) spectroscopy, using phospholipids spin-labeled on the acyl chain. The effect of inclusion of charged phospholipids or/and cholesterol in the bilayer composition was considered in relation to the peptide/membrane interaction. The results show that Aβ(25–35) inserts in bilayers formed by the zwitterionic phospholipid dilauroyl phosphatidylcholine (DLPC), positioning between the outer part of the hydrophobic core and the external hydrophilic layer. This process is not significantly influenced by the inclusion of the anionic phospholipid phosphatidylglycerol (DLPG) in the bilayer, indicating the peptide insertion to be driven by hydrophobic rather than electrostatic interactions. Cholesterol plays a fundamental role in regulating the peptide/membrane association, inducing a membrane transition from a fluid-disordered to a fluid-ordered phase. At low cholesterol content, in the fluid-disordered phase, the insertion of the peptide in the membrane causes a displacement of cholesterol towards the more external part of the membrane. The crowding of cholesterol enhances its rigidifying effect on this region of the bilayer. Finally, the cholesterol-rich fluid-ordered membrane looses the ability to include Aβ(25–35)

Spatial steadiness of individual disorder modes upon controlled spectral tuning

Recent innovative applications in disordered photonics would strongly benefit from the possibility to achieve spectral tuning of the individual disorder localized photonic modes without affecting their spatial distributions. Here, we design and fabricate a two-dimensional disordered photonic system, made of a GaAs slab patterned with randomly distributed circular air scattering centers, supporting localized light modes with very small modal volume. The photoluminescence of InAs quantum dots embedded in the slab is used as a probe for near field experiments and gives direct access to the electric field intensity distribution of the localized random modes. We demonstrate that laser assisted oxidation of the GaAs slab performed by near field illumination can be used for a gentle tuning of the individual random modes without modifying the subtle balance leading to light localization given by multiple scattering

X-ray phase contrast microscopy at 300 nm resolution with laboratory sources.

We report the performance of an X-ray phase contrast microscope for laboratory sources with 300 nm spatial resolution. The microscope is based on a commercial X-ray microfocus source equipped with a planar X-ray waveguide able to produce a sub-micrometer x-ray beam in one dimension. Phase contrast images of representative samples are reported. The achieved contrast and resolution is discussed for different configurations. The proposed approach could represent a simple, inexpensive, solution for sub-micrometer resolution imaging with small laboratory setups

Organs on chip approach: A tool to evaluate cancer-immune cells interactions

In this paper we discuss the applicability of numerical descriptors and statistical physics concepts to characterize complex biological systems observed at microscopic level through organ on chip approach. To this end, we employ data collected on a micro uidic platform in which leukocytes can move through suitably built channels toward their target. Leukocyte behavior is recorded by standard time lapse imaging. In particular, we analyze three groups of human peripheral blood mononuclear cells (PBMC): heterozygous mutants (in which only one copy of the FPR1 gene is normal), homozygous mutants (in which both alleles encoding FPR1 are loss-of-function variants) and cells from ‘wild type’ donors (with normal expression of FPR1). We characterize the migration of these cells providing a quantitative con rmation of the essential role of FPR1 in cancer chemotherapy response. Indeed wild type PBMC perform biased random walks toward chemotherapy-treated cancer cells establishing persistent interactions with them. Conversely, heterozygous mutants present a weaker bias in their motion and homozygous mutants perform rather uncorrelated random walks, both failing to engage with their targets. We next focus on wild type cells and study the interactions of leukocytes with cancerous cells developing a novel heuristic procedure, inspired by Lyapunov stability in dynamical systems

From Petri Dishes to Organ on Chip Platform: The Increasing Importance of Machine Learning and Image Analysis

The increasing interest for microfluidic devices in medicine and biology has opened the way to new time-lapse microscopy era where the amount of images and their acquisition time will become crucial. In this optic, new data analysis algorithms have to be developed in order to extract novel features of cell behavior and cell–cell interactions. In this brief article, we emphasize the potential strength of a new paradigm arising in the integration of microfluidic devices (i.e., organ on chip), time-lapse microscopy analysis, and machine learning approaches. Some snapshots of previous case studies in the context of immunotherapy are included as proof of concepts of the proposed strategies while a visionary description concludes the work foreseeing future research and applicative scenarios

3D Microfluidic model for evaluating immunotherapy efficacy by tracking dendritic cell behaviour toward tumor cells

Immunotherapy efficacy relies on the crosstalk within the tumor microenvironment between cancer and dendritic cells (DCs) resulting in the induction of a potent and effective antitumor response. DCs have the specific role of recognizing cancer cells, taking up tumor antigens (Ags) and then migrating to lymph nodes for Ag (cross)-presentation to naïve T cells. Interferon-α-conditioned DCs (IFN-DCs) exhibit marked phagocytic activity and the special ability of inducing Ag-specific T-cell response. Here, we have developed a novel microfluidic platform recreating tightly interconnected cancer and immune systems with specific 3D environmental properties, for tracking human DC behaviour toward tumor cells. By combining our microfluidic platform with advanced microscopy and a revised cell tracking analysis algorithm, it was possible to evaluate the guided efficient motion of IFN-DCs toward drug-treated cancer cells and the succeeding phagocytosis events. Overall, this platform allowed the dissection of IFN-DC-cancer cell interactions within 3D tumor spaces, with the discovery of major underlying factors such as CXCR4 involvement and underscored its potential as an innovative tool to assess the efficacy of immunotherapeutic approaches

Near-field speckle imaging of light localization in disordered photonic

Optical localization in strongly disordered photonic media is an attractive topic for proposing novel cavity-like structures. Light interference can produce random modes confined within small volumes, whose spatial distribution in the near-field is predicted to show hot spots at the nanoscale. However, these near-field speckles have not yet been experimentally investigated due to the lack of a high spatial resolution imaging techniques. Here, we study a system where the disorder is induced by random drilling air holes in a GaAs suspended membrane with internal InAs quantum dots. We perform deep-subwavelength near-field experiments in the telecom window to directly image the spatial distribution of the electric field intensity of disordered-induced localized optical modes. We retrieve the near-field speckle patterns that extend over few micrometers and show several single speckles of the order of λ/10 size. The results are compared with the numerical calculations and with the recent findings in the literature of disordered media. Notably, the hot spots of random modes are found in proximity of the air holes of the disordered system

A multidisciplinary study using in vivo tumor models and microfluidic cell-on-chip approach to explore the cross-talk between cancer and immune cells

A full elucidation of events occurring inside the cancer microenvironment is fundamental for the optimization of more effective therapies. In the present study, the cross-talk between cancer and immune cells was examined by employing mice deficient (KO) in interferon regulatory factor (IRF)-8, a transcription factor essential for induction of competent immune responses. The in vivo results showed that IRF-8 KO mice were highly permissive to B16.F10 melanoma growth and metastasis due to failure of their immune cells to exert proper immunosurveillance. These events were found to be dependent on soluble factors released by cells of the immune system capable of shaping the malignant phenotype of melanoma cells. An on-chip model was then generated to further explore the reciprocal interactions between the B16.F10 and immune cells. B16.F10 and immune cells were co-cultured in a microfluidic device composed of three culturing chambers suitably inter-connected by an array of microchannels; mutual interactions were then followed using time-lapse microscopy. It was observed that WT immune cells migrated through the microchannels towards the B16.F10 cells, establishing tight interactions that in turn limited tumor spread. In contrast, IRF-8 KO immune cells poorly interacted with the melanoma cells, resulting in a more invasive behavior of the B16.F10 cells. These results suggest that IRF-8 expression plays a key role in the cross-talk between melanoma and immune cells, and under-score the value of cell-on-chip approaches as useful in vitro tools to reconstruct complex in vivo microenvironments on a microscale level to explore cell interactions such as those occurring within a cancer immunoenvironment

Broadband enhancement of light-matter interaction in photonic crystal cavities integrating site-controlled quantum dots

The fabrication of integrated quantum dot (QD)-optical microcavity systems is a requisite step for the realization of a wide range of nanophotonic experiments (and applications) that exploit the ability of QDs to emit nonclassical light, e.g., single photons. Thanks to their similar to 20-nm positioning accuracy and to their proven potential for single-photon operation, the QDs obtained by spatially selective hydrogen irradiation of dilute-nitride semiconductors-such as Ga(AsN) and Ga(PN)-are uniquely suited for integration with photonic nanodevices. In the present work, we demonstrate the ability to deterministically integrate single, site-controlled Ga(AsN)/Ga(AsN):H QDs within a photonic crystal (PhC) cavity. The properties of the fabricated QD-PhC cavity systems are then probed by photon correlation-providing clear evidence of single-photon emission-and time-resolved microphotoluminescence spectroscopy. Detailed information on the dynamics of our integrated nanodevices can be inferred by comparing these experiments to the solutions of a rate-equations system, developed by taking into account all the main processes leading to the capture, relaxation, and recombination of carriers in and out of the QD. This allows us to follow the evolution of the relevant recombination rates in our system for varying energy detuning, Delta E, between the QD and the PhC cavity. When the QD exciton transition is nearly resonant with the cavity mode, a large (>tenfold) enhancement of the spontaneous emission rate is observed, in substantial agreement with Jaynes-Cummings (JC) theory. For intermediate detunings (Delta E similar to 1.5-3.5 meV), on the other hand, the observed enhancement is significantly larger than that predicted by JC theory, due to the important role played by acoustic phonons in mediating the QD-PhC cavity coupling in a solid-state environment. Apart from its fundamental interest, the observation of such phonon-mediated, broadband enhancement of light-matter interaction significantly relaxes the requirements for the realization of a large variety of cavity QED-based experiments and applications. These include many photonic devices for which the use of site-controlled Ga(AsN)/Ga(AsN):H QDs would be inherently advantageous, such as those based on the coupling between more than one QD and a single cavity mode (e.g., few-QD nanolasers and QD solids)

Tuning of photonic crystal cavities by controlled removal of locally infiltrated water

We present a spectral tuning mechanism of photonic crystal microcavities based on microfluidics. The microinfiltration with water of one or few cavity holes and its subsequent controlled evaporation allow us to tune the cavity resonances in a spectral range larger than 20 nm, with subnanometer accuracy, and we also observe that the addition of water in the microcavity region improves its quality factor Q. (C) 2009 American Institute of Physics. [doi:10.1063/1.3247894