Enabling the characterization of the nonlinear electrokinetic properties of particles using low voltage.

Insulator-based electrokinetically driven microfluidic devices stimulated with direct current (DC) voltages are an attractive solution for particle separation, concentration, or isolation. However, to design successful particle manipulation protocols, it is mandatory to know the mobilities of electroosmosis, and linear and nonlinear electrophoresis of the microchannel/liquid/particle system. Several techniques exist to characterize the mobilities of electroosmosis and linear electrophoresis. However, only one method to characterize the mobility of nonlinear electrophoresis has been thoroughly assessed, which generally requires DC voltages larger than 1000 V and measuring particle velocity in a straight microchannel. Under such conditions, Joule heating, electrolysis, and the DC power source cost become a concern. Also, measuring particle velocity at high voltages is noisy, limiting characterization quality. Here we present a protocol-tested on 2 µm polystyrene particles-for characterizing the mobility of nonlinear electrophoresis of the liquid/particle system using a DC voltage of only 30 V and visual inspection of particle dynamics in a microchannel featuring insulating obstacles. Multiphysics numerical modelling was used to guide microchannel design and to correlate particle location during an experiment with electric field intensity. The method was validated against the conventional characterization protocol, exhibiting excellent agreement while significantly reducing measurement noise and experimental complexity.

Trends and challenges in microfluidic methods for protein manipulation-A review.

Proteins are important molecules involved in an immensely large number of biological processes. Being capable of manipulating proteins is critical for developing reliable and affordable techniques to analyze and/or detect them. Such techniques would enable the production of therapeutic agents for the treatment of diseases or other biotechnological applications (e.g., bioreactors or biocatalysis). Microfluidic technology represents a potential solution to protein manipulation challenges because of the diverse phenomena that can be exploited to achieve micro- and nanoparticle manipulation. In this review, we discuss recent contributions made in the field of protein manipulation in microfluidic systems using different physicochemical principles and techniques, some of which are miniaturized versions of already established macro-scale techniques.

In vitro reconstitution of herpes simplex virus 1 fusion identifies low pH as a fusion co-trigger.

Membrane fusion mediated by herpes simplex virus 1 (HSV-1) is a complex, multi-protein process that is receptor triggered and can occur both at the cell surface and in endosomes. To deconvolute this complexity, we reconstituted HSV-1 fusion with synthetic lipid vesicles in vitro. Using this simplified, controllable system, we discovered that HSV-1 fusion required not only a cognate host receptor but also low pH. On the target membrane side, efficient fusion required cholesterol, negatively charged lipids found in the endosomal membranes, and an optimal balance of lipid order and disorder. On the virion side, the four HSV-1 entry glycoproteins-gB, gD, gH, and gL-were sufficient for fusion. We propose that low pH is a biologically relevant co-trigger for HSV-1 fusion. The dependence of fusion on low pH and endosomal lipids could explain why HSV-1 enters most cell types by endocytosis. We hypothesize that under neutral pH conditions, other, yet undefined, cellular factors may serve as fusion co-triggers. The in vitro fusion system established here can be employed to systematically investigate HSV-1-mediated membrane fusion.IMPORTANCEHSV-1 causes lifelong, incurable infections and diseases ranging from mucocutaneous lesions to fatal encephalitis. Fusion of viral and host membranes is a critical step in HSV-1 infection of target cells that requires multiple factors on both the viral and host sides. Due to this complexity, many fundamental questions remain unanswered, such as the identity of the viral and host factors that are necessary and sufficient for HSV-1-mediated membrane fusion and the nature of the fusion trigger. Here, we developed a simplified in vitro fusion assay to examine the fusion requirements and identified low pH as a co-trigger for virus-mediated fusion in vitro. We hypothesize that low pH has a critical role in cell entry and, potentially, pathogenesis.

Amplification factor in DC insulator-based electrokinetic devices: a theoretical, numerical, and experimental approach to operation voltage reduction for particle trapping.

Insulator-based microfluidic devices are attractive for handling biological samples due to their simple fabrication, low-cost, and efficiency in particle manipulation. However, their widespread application is limited by the high operation voltages required to achieve particle trapping. We present a theoretical, numerical, and experimental study that demonstrates these voltages can be significantly reduced (to sub-100 V) in direct-current insulator-based electrokinetic (DC-iEK) devices for micron-sized particles. To achieve this, we introduce the concept of the amplification factor-the fold-increase in electric field magnitude due to the presence of an insulator constriction-and use it to compare the performance of different microchannel designs and to direct our design optimization process. To illustrate the effect of using constrictions with smooth and sharp features on the amplification factor, geometries with circular posts and semi-triangular posts were used. These were theoretically approximated in two different systems of coordinates (bipolar and elliptic), allowing us to provide, for the first time, explicit electric field amplification scaling laws. Finite element simulations were performed to approximate the 3D insulator geometries and provide a parametric study of the effect of changing different geometrical features. These simulations were used to predict particle trapping voltages for four different single-layer microfluidic devices using two particle suspensions (2 and 6.8 µm in size). The general agreement between our models demonstrates the feasibility of using the amplification factor, in combination with nonlinear electrokinetic theory, to meet the prerequisites for the development of portable DC-iEK microfluidic systems.

Toward low-voltage dielectrophoresis-based microfluidic systems: A review.

Dielectrophoretically driven microfluidic devices have demonstrated great applicability in biomedical engineering, diagnostic medicine, and biological research. One of the potential fields of application for this technology is in point-of-care (POC) devices, ideally allowing for portable, fully integrated, easy to use, low-cost diagnostic platforms. Two main approaches exist to induce dielectrophoresis (DEP) on suspended particles, that is, electrode-based DEP and insulator-based DEP, each featuring different advantages and disadvantages. However, a shared concern lies in the input voltage used to generate the electric field necessary for DEP to take place. Therefore, input voltage can determine portability of a microfluidic device. This review outlines the recent advances in reducing stimulation voltage requirements in DEP-driven microfluidics.

Direct and indirect effects of parenting practices on socio-moral approval of aggression in Polish young adults. Do all practices matter?

The purpose of this article was to determine the socialisation antecedents of socio-moral approval of aggression (SMAA). In Study 1, we assessed factorial structure and reliability of the SMAA with a sample of 355 students who reported on the extent to which they approved of six forms of aggressive behaviour and six justifications of aggression. Two-factor solutions were obtained with regard to forms and justifications of aggressive acts. Thus, approval of extreme and minor aggression was distinguished as well as legitimate and illegitimate justifications of aggression. In Study 2, we tested the path models of the socialisation antecedents that contributed to the high approval of minor and extreme aggressive acts as well as legitimate and illegitimate justifications of aggression. Data were collected from 173 undergraduate students. Path analyses showed that high levels of approval of extremely aggressive acts and of illegitimate justifications of aggression were preceded by a sequence of negative life events, beginning with frequent misbehaviour in childhood, corporal punishment used by parents and ending with delinquency in adolescence. The approval of minor aggression had little relation to socialisation factors apart from a detrimental effect of psychological aggression while approval of legitimate justifications of aggression had no socialisation antecedents.

The usefulness of distinguishing types of aggression by function.

Far from being a universally defined notion, aggression is a changing and multifaceted phenomenon encompassing various concepts. There is no consensus as to how different types of aggression should be classified: multiple ways of doing so using a variety of criteria exist in the scientific literature. Some scientists categorise aggressive acts according to how they are expressed, while others prefer to look at motive, function, purpose and objective. Despite the claim of some authors that distinguishing between different types of aggressive acts is not always productive, categorising these according to different purposes and objectives can be very useful, both for developing theory and because such an approach serves forensic practice as well as preventive and therapeutic interventions, as these focus on the propensities and personality of the individual. Furthermore, given that the main functional classifications analysed show a common tendency to dichotomise, it would seem appropriate for their terminology and some of their measurement instruments to be standardised.