cyc-DEP: Cyclic immunofluorescence profiling of particles collected using dielectrophoresis

Cancer is a highly heterogenous disease that requires precise detection tools and active surveillance methods. Liquid biopsy assays provide an agnostic way to follow the complex trajectory of cancer, providing better patient stratification tools for optimized treatment. Here, we present the development of a low-volume liquid biopsy assay called cyc-DEP (cyclic immunofluorescent imaging on dielectrophoretic chip) to profile biomarkers collected on a dielectrophoretic microfluidic chip platform. To enable on-chip cyclic imaging, we optimized a fluorophore quenching method and sequential rounds of on-chip staining with fluorescently conjugated primary antibodies. cyc-DEP allows for the quantification of a multiplex array of proteins using 25 µl of a patient plasma sample. We utilized nanoparticles from a prostate adenocarcinoma (LNCaP) cell line and a panel of six target proteins to develop our proof-of-concept technique. We then used cyc-DEP to quantify blood plasma levels of target proteins from healthy individuals, low-grade and high-grade prostate cancer patients (n = 3 each) in order to demonstrate that our platform is suitable for liquid biopsy analysis in its present form. To ensure accurate quantification of signal intensities and comparisons between different samples, we incorporated a signal intensity normalization method (fluorescent beads) and a custom signal intensity quantification algorithm that account for the distribution of signal across hundreds of collection regions on each chip. Our technique enabled a threefold improvement in multiplicity for detecting proteins associated with fluid samples, opening doors for early detection, and active surveillance through quantification of a multiplex array of biomarkers from low-volume liquid biopsies.     

Surfactant-free cellulose filaments stabilized oil in water emulsions

Cellulose - There has been significant interest over recent years in the production and application of sustainable and green materials. Among these, nanocellulose has incurred great interest...     

Time resolved measurements of the flow generated by suction feeding fish

The majority of aquatic vertebrates are suction feeders: by rapidly expanding the mouth cavity they generate a fluid flow outside of their head in order to draw prey into their mouth. In addition to the biological relevance, the generated flow field is interesting fluid mechanically as it incorporates high velocities, is localized in front of the mouth, and is unsteady, typically lasting between 10 and 50 ms. Using manometry and high-speed particle image velocimetry, this is the first study to quantify pressure within and outside the mouth of a feeding fish while simultaneously measuring the velocity field outside the mouth. Measurements with a high temporal (2 ms) and spatial (<1 mm) resolution were made for several feeding events of a single largemouth bass (Micropterus salmoides). General properties of the flow were evaluated, including the transient velocity field, its relationship to pressure within the mouth and pressure at the prey. We find that throughout the feeding event a relationship exists for the magnitude of fluid speed as a function of distance from the predator mouth that is based on scaling the velocity field according to the size of the mouth opening and the magnitude of fluid speed at the mouth. The velocity field is concentrated within an area extending approximately one mouth diameter from the fish and the generated pressure field is even more local to the mouth aperture. Although peak suction pressures measured inside the mouth were slightly larger than those that were predicted using the equations of motion, we find that these equations give a very accurate prediction of the timing of peak pressure, so long as the unsteady nature of the flow is included.     

Large-scale failure modes of dielectric elastomer actuators

Dielectric elastomer actuators (DEAs) show promise for robotic and mechatronic applications. However, to date, these actuators have experienced high rates of failure that have prevented their practical application. Here, large scale modes of failure of DEAs and their performance limits are studied. The objective is to provide design guidelines and bound the performance of DEAs that avoid failure. An idealized DEA is modeled and its failure is predicted as a function of film pre-stretch used during actuator fabrication, actuation voltage, and stretch rate. Three failure modes are considered: pull-in, dielectric strength, and material strength. Each failure mode is shown to dominate for different combinations of pre-stretch and stretch rate. High stretch rates lead to dielectric strength failure while low stretch rates lead to pull-in failure. Material strength failure is less important for most cases. Model predictions are validated experimentally using practical DEAs operating under load. This study suggests that DEAs cannot be operated reliably under load for long periods of time or low stretch rates due to pull-in failure limitations. To be reliable, DEAs must be used for short periods of time with high stretch rates.     

Genetics in primary care: a USA faculty development initiative

The Genetics in Primary Care (GPC) project is a USA national faculty development initiative with the goal of enhancing the training of medical students and primary care residents by developing primary care faculty expertise in genetics. Educational strategies were developed for the project by an executive committee with input from an advisory committee, comprising individuals with primary care, medical education and genetics expertise. These committees identified the key issues in genetics education for primary care as (1) considering inherited disease in the differential diagnosis of common disorders; (2) using appropriate counseling strategies for genetic testing and diagnosis, and (3) understanding the implications of a genetic diagnosis for family members. The group emphasized the importance of a primary care perspective, which suggests that the clinical utility of genetic information is greatest when it has the potential to improve health outcomes. The group also noted that clinical practice already incorporates the use of family history information, providing a basis for discussing the application of genetic concepts in primary care. Genetics and primary care experts agreed that educational efforts will be most successful if they are integrated into existing primary care teaching programs, and use a case-based teaching format that incorporates both clinical and social dimensions of genetic disorders. Three core clinical skills were identified: (1) interpreting family history; (2) recognizing the variable clinical utility of genetic information, and (3) acquiring cultural competency. Three areas of potential controversy were identified as well: (1) the role of nondirective counseling versus shared decision-making in discussions of genetic testing; (2) the intrinsic value of genetic information when it does not influence health outcomes, and (3) indications for a genetics referral. The project provides an opportunity for ongoing discussion about these important issues.     

Non-linear normal modes,invariance, and modal dynamics approximations of non-linear systems

Non-linear systems are here tackled in a manner directly inherited from linear ones, that is, by using proper normal modes of motion. These are defined in terms of invariant manifolds in the system's phase space, on which the uncoupled system dynamics can be studied. Two different methodologies which were previously developed to derive the non-linear normal modes of continuous systems — one based on a purely continuous approach, and one based on a discretized approach to which the theory developed for discrete systems can be applied-are simultaneously applied to the same study case-an Euler-Bernoulli beam constrained by a non-linear spring-and compared as regards accuracy and reliability. Numerical simulations of pure non-linear modal motions are performed using these approaches, and compared to simulations of equations obtained by a classical projection onto the linear modes. The invariance properties of the non-linear normal modes are demonstrated, and it is also found that, for a pure non-linear modal motion, the invariant manifold approach achieves the same accuracy as that obtained using several linear normal modes, but with significantly reduced computational cost. This is mainly due to the possibility of obtaining high-order accuracy in the dynamics by solving only one non-linear ordinary differential equation.     

Instantaneous and time-averaged flow fields of multiple vortices in the tip region of a ducted propulsor

The instantaneous and time-averaged flow fields in the tip region of a ducted marine propulsor are examined. In this flow, a primary tip-leakage vortex interacts with a secondary, co-rotating trailing edge vortex and other co- and counter-rotating vorticity found in the blade wake. Planar particle imaging velocimetry (PIV) is used to examine the flow in a plane approximately perpendicular to the mean axis of the primary vortex. An identification procedure is used to characterize multiple regions of compact vorticity in the flow fields as series of Gaussian vortices. Significant differences are found between the vortex properties from the time-averaged flow fields and the average vortex properties identified in the instantaneous flow fields. Variability in the vortical flow field results from spatial wandering of the vortices, correlated fluctuations of the vortex strength and core size, and both correlated and uncorrelated fluctuations in the relative positions of the vortices. This variability leads to pseudo-turbulent velocity fluctuations. Corrections for some of this variability are performed on the instantaneous flow fields. The resulting processed flow fields reveal a significant increase in flow variability in a region relatively far downstream of the blade trailing edge, a phenomenon that is masked through the process of simple averaging. This increased flow variability is also accompanied by the inception of discrete vortex cavitation bubbles, which is an unexpected result, since the mean flow pressures in the region of inception are much higher than the vapor pressure of the liquid. This suggests that unresolved fine-scale vortex interactions and stretching may be occurring in the region of increased flow variability.     

Palladium-Catalyzed N-Arylation of 3-Functionalized 2-Amino-4,5-dimethylpyrroles

A versatile class of 2-aminopyrroles containing various electron-withdrawing substituents at the 3-position have been N-arylated on the amine using a palladium-catalyzed cross-coupling reaction. Using this methodology, a pyrimidone-based tricyclic system has been prepared in just one step from a starting 2-aminopyrrole.

How Constant Momentum Acceleration Decouples Energy and Space Focusing in Distance–of–Flight and Time–of–Flight Mass Spectrometries

Resolution in time–of–flight mass spectrometry (TOFMS) is ordinarily limited by the initial energy and space distributions within an instrument’s acceleration region and by the length of the field–free flight zone. With gaseous ion sources, these distributions lead to systematic flight–time errors that cannot be simultaneously corrected with conventional static–field ion–focusing devices (i.e., an ion mirror). It is known that initial energy and space distributions produce non–linearly correlated errors in both ion velocity and exit time from the acceleration region. Here we reinvestigate an old acceleration technique, constant–momentum acceleration (CMA), to decouple the effects of initial energy and space distributions. In CMA, only initial ion energies (and not their positions) affect the velocity ions gain. Therefore, with CMA, the spatial distribution within the acceleration region can be manipulated without creating ion–velocity error. The velocity differences caused by a spread in initial ion energy can be corrected with an ion mirror. We discuss here the use of CMA and independent focusing of energy and space distributions for both distance–of–flight mass spectrometry (DOFMS) and TOFMS. Performance characteristics of our CMA–DOFMS and CMA–TOFMS instrument, fitted with a glow–discharge ionization source, are described. In CMA–DOFMS, resolving powers (FWHM) of greater than 1000 are achieved for atomic ions with a flight length of 285 mm. In CMA–TOFMS, only ions over a narrow range of m/z values can be energy–focused; however, the technique offers improved resolution for these focused ions, with resolving powers of greater than 2000 for a separation distance of 350 mm.