DNA polymerases for whole genome amplification: considerations and future directions

In the same way that specialized DNA polymerases (DNAPs) replicate cellular and viral genomes, only a handful of dedicated proteins from various natural origins as well as engineered versions are appropriate for competent exponential amplification of whole genomes and metagenomes (WGA). Different applications have led to the development of diverse protocols, based on various DNAPs. Isothermal WGA is currently widely used due to the high performance of Φ29 DNA polymerase, but PCR-based methods are also available and can provide competent amplification of certain samples. Replication fidelity and processivity must be considered when selecting a suitable enzyme for WGA. However, other properties, such as thermostability, capacity to couple replication, and double helix unwinding, or the ability to maintain DNA replication opposite to damaged bases, are also very relevant for some applications. In this review, we provide an overview of the different properties of DNAPs widely used in WGA and discuss their limitations and future research directionsThis research was funded by MCIN/AEI/10.13039/501100011033 and ERDF A way of making Europe, grant PID2021-123403NB-I00 to M.R.-

The Dark Side of Goal Setting: The Role of Goals in Motivating Unethical Decision Making

A substantial literature has demonstrated that goal setting improves task performance (Locke & Latham, 1990). In this article we explore the proposition that challenging goals motivate not only constructive behavior, but also unethical behavior such as lying and cheating. We conducted eight scenario studies and an anagram experiment, and find support for our thesis. Respondents rated individuals with unmet goals as significantly more likely to engage in unethical behavior than similar individuals attempting to do their best or with met goals. Similarly, participants in the goal conditions in our experiment were significantly more likely to misrepresent their productivity in an anagram task than were participants in the do your best condition. This relationship was particularly strong when people had reward rather than mere goals, and when people were just short of reaching the goal. We explain our results in terms of the reference point adoption process consistent with Prospect Theory (Kahneman & Tversky, 1979), and identify specific contributions to goal setting theory and management practice

Artificial micro-swimmers in simulated natural environments

Microswimmers, such as bacteria, are known to show different behaviours depending on their local environment. They identify spatial chemical gradients to find nutrient rich areas (chemotaxis) and interact with shear flows to accumulate in high shear regions. Recently, artificial microswimmers have been developed which mimic their natural counterparts in many ways. One of the exciting topics in this field is to study these artificial motors in several natural settings like the ones bacteria interact with. In this Focus article, we summarize recent observations of artificial swimmers in chemical gradients, shear flows and other interesting natural environments simulated in the lab using microfluidics and nanotechnology

Underground parking structure built with deep foundations and vault precast elements in Spain

In many cases the only places available for the construction of a new car park are the existing streets or roads. These streets may also have important or historic buildings very close to the structure, which means that they cannot be disturbed in any way during the construction of the parking structure. In many cases the only places available for the construction of a new car park are the existing streets or roads. These streets may also have important or historic buildings very close to the structure, which means that they cannot be disturbed in any way during the construction of the parking structure

On Good Scholarship, Goal Setting, and Scholars Gone Wild

In this article, we define good scholarship, highlight our points of disagreement with Locke and Latham (2009), and call for further academic research to examine the full range of goal setting\u27s effects. We reiterate our original claim that goal setting, like a potent medication, can produce both beneficial effects and systematic, negative outcomes (Ordóñez, Schweitzer, Galinsky,& Bazerman, 2009), and as a result, it should be carefully prescribed and closely monitored

Investigating the Effects of Anger and Guilt on Unethical Behavior:A Dual-Process Approach

Although emotion has become one of the most popular research areas within organizational scholarship, few studies have considered its connection with unethical behavior. Using dual-process theory, we expand on the rationalist perspective within the field of behavioral ethics by considering the process through which two discrete emotions, anger and guilt, influence unethical behavior. Across two studies using different methodologies, we found that anger increases unethical behavior whereas guilt reduces unethical behavior. These effects were mediated by impulsive and deliberative processing. Overall, our results shed light on distinct mechanisms through which emotions can influence unethical behavior. Both theoretical and practical implications are discussed

Engineered viral DNA polymerase with enhanced DNA amplification capacity: a proof-of-concept of isothermal amplification of damaged DNA

© The Author(s) 2020.The development of whole genome amplification (WGA) and related methods, coupled with the dramatic growth of sequencing capacities, has changed the paradigm of genomic and genetic analyses. This has led to a continual requirement of improved DNA amplification protocols and the elaboration of new tailored methods. As key elements in WGA, identification and engineering of novel, faithful and processive DNA polymerases is a driving force in the field. We have engineered the B-family DNA polymerase of virus Bam35 with a C-terminal fusion of DNA-binding motifs. The new protein, named B35-HhH, shows faithful DNA replication in the presence of magnesium or an optimised combination of magnesium and manganese divalent cofactors, which enhances the replication of damaged DNA substrates. Overall, the newly generated variant displays improved amplification performance, sensitivity, translesion synthesis and resistance to salt, which are of great interest for several applications of isothermal DNA amplification. Further, rolling-circle amplification of abasic site-containing minicircles provides a proof-of-concept for using B35-HhH for processive amplification of damaged DNA samples.This work was funded by grants from Spanish Ministry of Science, Innovation and Universities [PGC2018-093726-B-100 AEI/FEDER UE to M.S. and PGC2018-093723-A-100 to M.R.R.] and a ComFuturo Grant from Fundación General CSIC [NewPols4Biotech to M.R-R.]. C.D. O. and A. L. were holders of a PhD fellowship [FPU16/02665 and FPU15/05797, respectively] from the Spanish Ministry of Science, Innovation and Universities. An institutional grant from Fundación Ramón Areces and Banco Santander to the Centro de Biología Molecular Severo Ochoa is also acknowledged. Tis paper is dedicated to the memory of Prof. Margarita Salas, an irreplaceable researcher and mentor

Equivalence of the Path Integral for Fermions in Cartesian and Spherical Coordinates

The path-integral calculation for the free energy of a spin-1/2 Dirac-fermion gas is performed in spherical polar coordinates for a flat spacetime geometry. Its equivalence with the Cartesian-coordinate representation is explicitly established. This evaluation involves a relevant limiting case of the fermionic path integral in a Schwarzschild background, whose near-horizon limit has been shown to be related to black hole thermodynamics.Comment: 16 page

Two-state theory of nonlinear Stochastic Resonance

An amenable, analytical two-state description of the nonlinear population dynamics of a noisy bistable system driven by a rectangular subthreshold signal is put forward. Explicit expressions for the driven population dynamics, the correlation function (its coherent and incoherent part), the signal-to-noise ratio (SNR) and the Stochastic Resonance (SR) gain are obtained. Within a suitably chosen range of parameter values this reduced description yields anomalous SR-gains exceeding unity and, simultaneously, gives rise to a non-monotonic behavior of the SNR vs. the noise strength. The analytical results agree well with those obtained from numerical solutions of the Langevin equation.Comment: 4 pages, 1 figur