Engineering a blue light inducible SpyCatcher system (BLISS) as a tool for protein photopatterning and optogenetics

The SpyTag-SpyCatcher protein conjugation system has recently exploded in popularity due to its fast kinetics and high yield under biologically favorable conditions in both in vitro and intracellular settings. We imagine we can further expand the utility of this system by introducing the ability to spatially and temporally control the conjugation event. Taking inspiration from photoreceptor proteins in nature, we designed a method to integrate light dependency into the protein conjugation reaction. The light-oxygen-voltage 2 domain of Avena sativa (AsLOV2) undergoes a dramatic conformational change in response to blue light. We have thus genetically fused the SpyTag into the AsLOV2 domain to create a Blue Light Inducible SpyCatcher System (BLISS). In this design (Figure 1), the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag. We screened several likely insertion points in the Jα-helix, and found a variant with desirable light switching behavior where after one hour of irradiation, the reaction is 80% complete, while only 10% of the AsLOV2-SpyTag protein reacted in the dark. This reaction can be quenched within minutes by returning the reaction to the dark. We demonstrated the spatial aspect of this light control mechanism through photopatterning proteins onto Ni-NTA coated slides. As our system is made solely from protein components, which can be genetically encoded, we can extend the same spatiotemporal control of proteins inside cells. We anticipate BLISS will be a strong tool for fabricating protein microassays, crafting biomaterial composition, as well as optically controlling enzyme activity and protein localization in cells. Please click Additional Files below to see the full abstract

Doppler cooling of gallium atoms: 2. Simulation in complex multilevel systems

This paper derives a general procedure for the numerical solution of the Lindblad equations that govern the coherences arising from multicoloured light interacting with a multilevel system. A systematic approach to finding the conservative and dissipative terms is derived and applied to the laser cooling of gallium. An improved numerical method is developed to solve the time-dependent master equation and results are presented for transient cooling processes. The method is significantly more robust, efficient and accurate than the standard method and can be applied to a broad range of atomic and molecular systems. Radiation pressure forces and the formation of dynamic dark-states are studied in the gallium isotope 66Ga.Comment: 15 pages, 8 figure

Mending the Levee: How Supernaturally Anchored Conceptions of the Person Impact on Trauma Perception and Healing among Children (Cases from Madagascar and Nepal)

When dealing with children and youth who experience distressing events, psychosocial diagnostics and healing programmes principally resort to biomedical models. Children are often viewed as individualised ‘victims’ suffering from trauma and ‘in need’ of outside help. Highlighting case studies from Madagascar and Nepal, this article argues that the biomedical approach to trauma would be strengthened by a concomitant analysis of social networks, including the perceived relations with the supernatural. The various tandems of family and kin relationships, the living and the dead, constitute not only a social ‘levee’ breached by distressing events, but also the locus around which social relations are rebuilt

Singularities of nn-fold integrals of the Ising class and the theory of elliptic curves

We introduce some multiple integrals that are expected to have the same singularities as the singularities of the $n$-particle contributions $\chi^{(n)}$ to the susceptibility of the square lattice Ising model. We find the Fuchsian linear differential equation satisfied by these multiple integrals for $n=1, 2, 3, 4$ and only modulo some primes for $n=5$ and $6$, thus providing a large set of (possible) new singularities of the $\chi^{(n)}$. We discuss the singularity structure for these multiple integrals by solving the Landau conditions. We find that the singularities of the associated ODEs identify (up to $n= 6$) with the leading pinch Landau singularities. The second remarkable obtained feature is that the singularities of the ODEs associated with the multiple integrals reduce to the singularities of the ODEs associated with a {\em finite number of one dimensional integrals}. Among the singularities found, we underline the fact that the quadratic polynomial condition $1+3 w +4 w^2 = 0$, that occurs in the linear differential equation of $\chi^{(3)}$, actually corresponds to a remarkable property of selected elliptic curves, namely the occurrence of complex multiplication. The interpretation of complex multiplication for elliptic curves as complex fixed points of the selected generators of the renormalization group, namely isogenies of elliptic curves, is sketched. Most of the other singularities occurring in our multiple integrals are not related to complex multiplication situations, suggesting an interpretation in terms of (motivic) mathematical structures beyond the theory of elliptic curves.Comment: 39 pages, 7 figure

Anaphylaxis

Anaphylaxis is an acute, potentially fatal systemic reaction with varied mechanisms and clinical presentations. Although prompt recognition and treatment of anaphylaxis are imperative, both patients and healthcare professionals often fail to recognize and diagnose early signs and symptoms of the condition. Clinical manifestations vary widely, however, the most common signs are cutaneous symptoms, including angioedema, urticaria, erythema and pruritus. Immediate intramuscular administration of epinephrine into the lateral thigh is first-line therapy, even if the diagnosis is uncertain. The mainstays of long-term management include specialist assessment, avoidance measures, and the provision of an epinephrine auto-injector and an individualized anaphylaxis action plan. This article provides an overview of the causes, clinical features, diagnosis and acute and long-term management of this serious allergic reaction

El autor piensa en público : David Lodge presenta su novela Pensamientos Ocultos'

Obra ressenyada: David LODGE, Pensamiento ocultosapplication/pd

Predictive processing and the varieties of psychological trauma

A recently popular framework in the cognitive sciences takes the human nervous system to be a hierarchically arranged Bayesian prediction machine. In this paper, we examine psychological trauma through the lens of this framework. We suggest that this can help us to understand the nature of trauma, and the different effects that different kinds of trauma can have. We end by exploring synergies between our approach and current theories of PTSD, and gesture toward future directions