Single-molecule kinetic locking allows fluorescence-free quantification of protein/nucleic-acid binding

Fluorescence-free micro-manipulation of nucleic acids (NA) allows the functional characterization of DNA/RNA processing proteins, without the interference of labels, but currently fails to detect and quantify their binding. To overcome this limitation, we developed a method based on single-molecule force spectroscopy, called kinetic locking, that allows a direct in vitro visualization of protein binding while avoiding any kind of chemical disturbance of the protein’s natural function. We validate kinetic locking by measuring accurately the hybridization energy of ultrashort nucleotides (5, 6, 7 bases) and use it to measure the dynamical interactions of Escherichia coli/E. coli RecQ helicase with its DNA substrate.

Characterization of Two Hydrogen Peroxide Resistant Peroxidases from Rhodococcus opacus 1CP

The dye-decolorizing peroxidases (DyP) are a family of heme-dependent enzymes present on a broad spectrum of microorganisms. While the natural function of these enzymes is not fully understood, their capacity to degrade highly contaminant pigments such as azo dyes or anthraquinones make them excellent candidates for applications in bioremediation and organic synthesis. In this work, two novel DyP peroxidases from the organism Rhodococcus opacus 1CP (DypA and DypB) were cloned and expressed in Escherichia coli. The enzymes were purified and biochemically characterized. The activities of the two DyPs via 2,2′-azino-bis [3-ethylbenzthiazoline-6-sulphonic acid] (ABTS) assay and against Reactive Blue 5 were assessed and optimized. Results showed varying trends for DypA and DypB. Remarkably, these enzymes presented a particularly high tolerance towards H2O2, retaining its activities at about 10 mM H2O2 for DypA and about 4.9 mM H2O2 for DypB.

Therapist Materials for the Natural Function of Emotions

The session covered in this chapter looks at the natural, adaptive, helpful function of different emotions. Though emotions are not necessarily serving an adaptive or helpful function when someone has an emotional disorder, it is useful to understand that every emotion does have a useful purpose in nature. Knowing the function of the emotion can help clients understand why they are having certain emotions at certain times. Emotions such as fear, sadness, anxiety, anger, guilt, and disgust are discussed. In this session, clients also explore the function of internal physical sensations, with the aim of understanding that while sensations may be uncomfortable, they are not dangerous.

The Natural Function of Emotions

During this session, clients learn about the natural and adaptive function (the evolutionary purpose) of emotions. This chapter looks at emotions such as fear, sadness, anxiety, anger, guilt, shame, and disgust. Because some of these emotions are unpleasant, people have a tendency to think of them as bad. Every emotion has a purpose, however, and can be helpful to understand their natural functions. The symptoms of eating disorders, anxiety, and depression are all perpetuated by difficulties coping with uncomfortable or distressing emotional experiences. Understanding how emotions can be adaptive and natural can help us to cope with them better, and to fear or avoid them less.

Ever-Expanding Frontiers of Ecological Destruction

This chapter explores the multiple forces that predispose capitalism to ever-increasing amounts of environmental damage. Sociologists discuss the treadmill of production, the degradation of the biosphere coming from increasing population growth and GDP requiring consuming ever-greater amounts of natural resources. A more refined argument is expanding frontiers of production. Modern economies require the incorporation of ever-greater amounts of physical space into capitalist production. Whenever an area is converted to commercial use, its natural function gets destroyed. The chapter then considers how capitalism is dependent on the four cheaps: cheap food, cheap energy, cheap natural resources, and cheap labor. All four are subject to expanding frontiers of production. However, the physical destruction of environments is not the only problem with expanding frontiers of production. The incorporation of new space into capitalist production means the dislocation of the population originally living in that space. Ultimately, landlessness leads to political volatility and warfare.

Lip and Perioral Trauma: Principles of Aesthetic and Functional Reconstruction

Successful management of lip and perioral trauma requires a nuanced understanding of anatomy and surgical techniques. Surgical correction is particularly challenging in instances of tissue loss, due to a narrow tolerance for aesthetic deformity and highly specialized functions of the perioral region, including facial expression, communication, and oral competence. Restoring continuity of the orbicularis oris musculature is critical for dynamic sphincter function of the upper and lower lips. Lip and perioral tissue symmetry are also critical for aesthetic balance, and failure to restore a natural appearance can adversely affect personal identity, with attendant psychological trauma. This discussion of lip and perioral trauma management encompasses lip and perioral anatomy, evaluation of injuries, reconstructive techniques, and prevention and management of complications. Perioral injuries are classified by size, depth, and extent of injury, and the corresponding reconstructive approaches are a function of complexity. These approaches proceed sequentially up rungs of the reconstructive ladder including primary repair, local flaps, grafting, regional flaps, as well as microvascular free tissue transfers. Procedures may be single stage or require multiple stages or subsequent refinement. Regardless of the defect size or location, the guiding principle of repair in the perioral region is restoring natural function and aesthetic appearance. This still-evolving area of facial plastic and reconstructive surgery lends itself to artistry and technical precision, offering opportunities for further innovation to improve the outcomes of patients with lip and perioral trauma.

Soluble Pecam-1 as a Biomarker in Periprosthetic Joint Infection

A reliable workup with regard to a single diagnostic marker indicating periprosthetic joint infection (PJI) with sufficient sensitivity and specificity is still missing. The immunologically reactive molecule Pecam-1 is shed from the T-cell surface upon activation via proinflammatory signaling, e.g., triggered by specific pathogens. We hypothesized that soluble Pecam-1 (sPecam-1) can hence function as a biomarker of PJI. Fifty-eight patients were prospectively enrolled and assigned to one of the respective treatment groups (native knees prior to surgery, aseptic, and septic total knee arthroplasty (TKA) revision surgeries). Via synovial sample acquisition and ELISA testing, a database on local sPecam-1 levels was established. We observed a significantly larger quantity of sPecam-1 in septic (n = 22) compared to aseptic TKA revision surgeries (n = 20, p ≤ 0.001). Furthermore, a significantly larger amount of sPecam-1 was found in septic and aseptic revisions compared to native joints (n = 16, p ≤ 0.001). Benchmarking it to the gold standard showed a high predictive power for the detection of PJI. Local sPecam-1 levels correlated to the infection status of the implant, and thus bear a strong potential to act as a biomarker of PJI. While a clear role of sPecam-1 in infection could be demonstrated, the underlying mechanism of the molecule’s natural function needs to be further unraveled.

Theta-modulated oscillatory transcranial direct current stimulation over posterior parietal cortex improves associative memory

Associative memory (AM) reflects the ability to remember and retrieve multiple pieces of information bound together thus enabling complex episodic experiences. Despite growing interest in the use of transcranial direct current stimulation (tDCS) for the modulation of AM, there are inconsistent evidence regarding its benefits. An alternative to standard constant tDCS could be the application of frequency-modulated tDCS protocols, that mimic natural function-relevant brain rhythms. Here, we show the effects of anodal tDCS oscillating in theta rhythm (5 Hz; 1.5 ± 0.1 mA) versus constant anodal tDCS and sham over left posterior parietal cortex on cued recall of face-word associations. In a crossover design, each participant completed AM assessment immediately following 20-min theta-oscillatory, constant, and sham tDCS, as well as 1 and 5 days after. Theta oscillatory tDCS increased initial AM performance in comparison to sham, and so did constant tDCS. On the group level, no differences between oscillatory and constant tDCS were observed, but individual-level analysis revealed that some participants responded to theta-oscillatory but not to constant tDCS, and vice versa, which could be attributed to their different physiological modes of action. This study shows the potential of oscillatory tDCS protocols for memory enhancement to produce strong and reliable memory-modulating effects which deserve to be investigated further.

The endless battle between phages and CRISPR-Cas systems in Streptococcus thermophilus

This review describes the contribution of basic research on phage-bacteria interactions to the understanding of CRISPR-Cas systems as well as its various applications. It focuses on the natural function of CRISPR-Cas system as an adaptive defense mechanism against mobile genetic elements such as bacteriophage genomes and plasmids. Some of the advances in the characterization of the type II-A CRISPR-Cas system of Streptococcus thermophilus and Streptococcus pyogenes later led to the development of the CRISPR-Cas9 genome editing technology. We mostly discuss the three stages of the CRISPR-Cas system in S. thermophilus, namely the adaptation stage, which is unique to this resistance mechanism, the CRISPR RNA biogenesis as well as the DNA cutting activity in the interference stage to protect bacteria against phages. Finally, we looked into applications of CRISPR-Cas in microbiology, including overcoming limitations in genome editing.