Detection and controlled depletion of cancer cells using photothermal phase microscopy

We present a dual‐modality technique based on wide‐field photothermal (PT) interferometric phase imaging and simultaneous PT ablation to selectively deplete specific cell populations labelled by plasmonic nanoparticles. This combined technique utilizes the plasmonic reaction of gold nanoparticles under optical excitation to produce PT imaging contrast by inducing local phase changes when the excitation power is weak, or ablation of selected cells when increasing the excitation power. Controlling the entire process is carried out by dynamic quantitative phase imaging of all cells (labelled and unlabelled). We demonstrate our ability to detect and specifically ablate in vitro cancer cells over‐expressing epidermal growth factor receptors (EGFRs), labelled with plasmonic nanoparticles, in the presence of either EGFR under‐expressing cancer cells or white blood cells. The latter demonstration establishes an initial model for depletion of circulating tumour cells in blood. The proposed system is able to image in wide field the label‐free quantitative phase profile together with the PT phase profile of the sample, and provides the ability of both detection and selective cell ablation in a controlled environment.

Quantitative phase imaging with molecular specificity and specific cell depletion. ( a ) Label‐free quantitative phase profiles of mixed population of EGFR⁺/EGFR^– cancer cells. ( b ) When weak modulated PT excitation is applied, selective phase contrast is generated in the modulation frequency only for the EGFR⁺ cancer cells labelled with plasmonic nanoparticles. ( c ) When stronger modulated PT excitation is applied, selective ablation of the EGFR⁺ cancer cells labelled with plasmonic nanoparticles occurs. White scalebars represent 10 µm upon sample.     

The Interplay between Scientific Overlap and Cooperation and the Resulting Gain in Co-Authorship Interactions

Considering the importance of scientific interactions, understanding the principles that govern fruitful scientific research is crucial to policy makers and scientists alike. The outcome of an interaction is to a large extent dependent on the balancing of contradicting motivations accompanying the establishment of collaborations. Here, we assembled a dataset of nearly 20,000 publications authored by researchers affiliated with ten top universities. Based on this data collection, we estimated the extent of different interaction types between pairwise combinations of researchers. We explored the interplay between the overlap in scientific interests and the tendency to collaborate, and associated these estimates with measures of scientific quality and social accessibility aiming at studying the typical resulting gain of different interaction patterns. Our results show that scientists tend to collaborate more often with colleagues with whom they share moderate to high levels of mutual interests and knowledge while cooperative tendency declines at higher levels of research-interest overlap, suggesting fierce competition, and at the lower levels, suggesting communication gaps. Whereas the relative number of alliances dramatically differs across a gradient of research overlap, the scientific impact of the resulting articles remains similar. When considering social accessibility, we find that though collaborations between remote researchers are relatively rare, their quality is significantly higher than studies produced by close-circle scientists. Since current collaboration patterns do not necessarily overlap with gaining optimal scientific quality, these findings should encourage scientists to reconsider current collaboration strategies.     

Structural and mechanical analysis of tectorial membrane Tecta mutants

The tectorial membrane (TM) is an extracellular matrix of the cochlea whose prominent role in hearing has been demonstrated through mutation studies. The C1509G mutation of the Tecta gene, which encodes for the α-tectorin protein, leads to hearing loss. The heterozygote TM only attaches to the first row of outer hair cells (OHCs), and the homozygote TM does not attach to any OHCs. Here we measured the morphology and mechanical properties of wild-type, heterozygous, and homozygous Tecta TMs. Morphological analyses conducted with second- and third-harmonic imaging, scanning electron microscopy, and immunolabeling revealed marked changes in the collagen architecture and stereocilin-labeling patterns of the mutant TMs. The mechanical properties of the mutant TM were measured by force spectroscopy. Whereas the axial Young's modulus of the low-frequency (apical) region of Tecta mutant TM samples was similar to that of wild-type TMs, it significantly decreased in the basal region to a value approaching that found at the apex. Modeling simulations suggest that a reduced TM Young's modulus is likely to reduce OHC stereociliary deflection. These findings argue that the heterozygote C1509G mutation results in a lack of attachment of the TM to the OHCs, which in turn reduces both the overall number of OHCs that are involved in mechanotransduction and the degree of mechanotransduction exhibited by the OHCs that remain attached to the TM.     

Hydration of polyethylene glycol-grafted liposomes.

This study aimed to characterize the effect of polyethylene glycol of 2000 molecular weight (PEG2000) attached to a dialkylphosphatidic acid (dihexadecylphosphatidyl (DHP)-PEG2000) on the hydration and thermodynamic stability of lipid assemblies. Differential scanning calorimetry, densitometry, and ultrasound velocity and absorption measurements were used for thermodynamic and hydrational characterization. Using a differential scanning calorimetry technique we showed that each molecule of PEG2000 binds 136 +/- 4 molecules of water. For PEG2000 covalently attached to the lipid molecules organized in micelles, the water binding increases to 210 +/- 6 water molecules. This demonstrates that the two different structural configurations of the PEG2000, a random coil in the case of the free PEG and a brush in the case of DHP-PEG2000 micelles, differ in their hydration level. Ultrasound absorption changes in liposomes reflect mainly the heterophase fluctuations and packing defects in the lipid bilayer. The PEG-induced excess ultrasound absorption of the lipid bilayer at 7.7 MHz for PEG-lipid concentrations over 5 mol % indicates the increase in the relaxation time of the headgroup rotation due to PEG-PEG interactions. The adiabatic compressibility (calculated from ultrasound velocity and density) of the lipid bilayer of the liposome increases monotonically with PEG-lipid concentration up to approximately 7 mol %, reflecting release of water from the lipid headgroup region. Elimination of this water, induced by grafted PEG, leads to a decrease in bilayer defects and enhanced lateral packing of the phospholipid acyl chains. We assume that the dehydration of the lipid headgroup region in conjunction with the increase of the hydration of the outer layer by grafting PEG in brush configuration are responsible for increasing thermodynamic stability of the liposomes at 5-7 mol % of PEG-lipid. At higher PEG-lipid concentrations, compressibility and partial volume of the lipid phase of the samples decrease. This reflects the increase in hydration of the lipid headgroup region (up to five additional water molecules per lipid molecule for 12 mol % PEG-lipid) and the weakening of the bilayer packing due to the lateral repulsion of PEG chains.     

PTEN Negatively Regulates MAPK Signaling during Caenorhabditis elegans Vulval Development

Vulval development in Caenorhabditis elegans serves as an excellent model to examine the crosstalk between different conserved signaling pathways that are deregulated in human cancer. The concerted action of the RAS/MAPK, NOTCH, and WNT pathways determines an invariant pattern of cell fates in three vulval precursor cells. We have discovered a novel form of crosstalk between components of the Insulin and the RAS/MAPK pathways. The insulin receptor DAF-2 stimulates, while DAF-18 PTEN inhibits, RAS/MAPK signaling in the vulval precursor cells. Surprisingly, the inhibitory activity of DAF-18 PTEN on the RAS/MAPK pathway is partially independent of its PIP(3) lipid phosphatase activity and does not involve further downstream components of the insulin pathway, such as AKT and DAF-16 FOXO. Genetic and biochemical analyses indicate that DAF-18 negatively regulates vulval induction by inhibiting MAPK activation. Thus, mutations in the PTEN tumor suppressor gene may result in the simultaneous hyper-activation of two oncogenic signaling pathways.     

White Blood Cell Count and the Risk for Coronary Artery Disease in Young Adults

Background

The association between white blood cell (WBC) count and coronary artery disease (CAD) is unknown in young adults. Our objective was to assess the association between WBC count and its changes over time with CAD incidence in the Metabolic, Life-style and Nutrition Assessment in Young adults (MELANY) study, a cohort of Israeli army personnel.

Methods and Findings

29,120 apparently healthy young men (mean age; 31.2±5.5 years) with a normal baseline WBC count (3,000–12,000 cells/mm³) were followed during a mean follow up of 7.5±3.8 years for incidence of CAD. Participants were screened every 3–5 years using a stress test, and CAD was confirmed by coronary angiography. In a multivariate model adjusted for age, body mass index (BMI), LDL- and HDL-cholesterol, blood pressure, family history of CAD, physical activity, diabetes, triglycerides and smoking status, WBC levels (divided to quintiles) above 6,900 cells/mm³ (quintile 4) were associated with a 2.17-fold increase (95%CI = 1.18–3.97) in the risk for CAD as compared with men in quintile 1 (WBC≤5,400 cells/mm³). When modeled as a continuous variable, a WBC increment of 1000 cells/mm³ was associated with a 17.4% increase in CAD risk (HR 1.174; 95%CI = 1.067–1.290, p = 0.001). A decrease in the WBC level (within the normal range) during the follow-up period was associated with increased physical activity and decreased triglyceride levels as well as with reduced incidence of CAD.

Conclusions

WBC count is an independent risk factor for CAD in young adults at values well within the normal range. WBC count may assist in detecting subgroups of young men at either low or high risk for progression to CAD.     

The 3D structure of the tectorial membrane determined by second-harmonic imaging microscopy

The tectorial membrane (TM) is a highly hydrated non-cellular matrix situated over the sensory cells of the cochlea. It is widely accepted that the mechanical coupling, between the TM and outer hair cells stereocilia bundles, plays an important role in the cochlea energy transduction mechanism. Recently, we provided supporting evidence for the existence of mechanical coupling by demonstrating that the mechanical properties of the TM change along its longitudinal direction. Since the biochemical composition of the TM is similar throughout its entire length, it is likely that structural differences induce the observed material properties changes. Presently, however, the structure of the TM under physiological environments remains unknown. In this work, the 3D structure of native TM samples is shown by using two-photon second-harmonic imaging microscopy. We find that the collagen fibers at the basal region are arranged in a parallel orientation while being tilted in an angle with respect to the plane of the TM surface at the apical region. Moreover, we find an intensified marginal band at the basal OHC zone which forms a shell-like structure which engulfs the stereocilium imprints surface of the TM. In supports of our previous mechanical characterization, the analysis presented here provides a structural basis for the changes in TM's mechanical properties.