Altering Enzymatic Activity: Recruitment of Carboxypeptidase Activity into an RTEM β-Lactamase/Penicillin-Binding Protein 5 Chimera

The D-Ala-D-Ala carboxypeptidases/transpeptidases (penicillin-binding proteins, PBPs) share considerable structural homology with class A β-lactamases (EC 3.5.2.6), although these β-lactamases have no observable D-Ala-D-Ala carboxypeptidase activity. With the objective of recruiting such activity into a β-lactamase background, we have prepared a chimeric protein by inserting a 28-amino acid segment of PBP-5 of Escherichia coli in place of the corresponding region of the RTEM-1 β-lactamase. The segment thus inserted encompasses two residues conserved in both families: Ser-70, which forms the acyl-enzyme intermediate during β-lactam hydrolysis, and Lys-73, whose presence has been shown to be necessary for catalysis. This chimera involves changes of 18 residues and gives a protein that differs at 7% of the residues from the parent. Whereas RTEM β-lactamase has no D-Ala-D-Ala carboxypeptidase activity, that of the chimera is significant and is, in fact, about 1% the activity of PBP-5 on diacetyl-L-Lys-D-Ala-D-Ala; in terms of free energy of activation, the chimera stabilizes the transition state for the reaction to within about 2.7 kcal/mol of the stabilization achieved by PBP-5. Furthermore, the chimera catalyzes hydrolysis exclusively at the carboxyl-terminal amide bond which is the site of cleavage by D-Ala-D-Ala carboxypeptidase. Though containing all those residues that are conserved throughout class A β-Lactamases and are thought to be essential for β-lactamase activity, the chimera has considerably reduced activity ({approx} 10^-5) on penams such as penicillins and ampicillins as substrates. As a catalyst, the chimera shows an induction period of {approx} 30 min, reflecting a slow conformational rearrangement from an inactive precursor to the active enzyme

Moral hazard under commercial and universal banking

Many claims have been made about the potential benefits, and the potential costs, of adopting a system of universal banking in the United States. We evaluate these claims using a model where there is a moral hazard problem between banks and “borrowers,” a moral hazard problem between banks and a deposit insurer, and a costly state verification problem. Under conditions we describe, allowing banks to take equity positions in firms strengthens their ability to extract surplus, and exacerbates problems of moral hazard. The incentives of universal banks to take equity positions will often be strongest when these problems are most severe.Universal banks

Deposit insurance: a reconsideration

This paper undertakes a simple general equilibrium analysis of the consequences of deposit insurance programs, the way in which they are priced and the way in which they fund revenue shortfalls. We show that the central issue is how the government will make up any FDIC losses. Under one scheme for making up the losses, we show that FDIC policy is irrelevant: it does not matter what premium is charged, nor does it matter how big FDIC losses are. Under another scheme, all that matters is the magnitude of the losses. And there is no presumption that small losses are “good.” We also show that multiple equilibria can be observed and Pareto ranked. Some economies may be “trapped” in equilibria with inefficient financial systems. Our analysis provides counterexamples to the following propositions. (1) Actuarially fair pricing of deposit insurance is always desirable. (2) Implicit FDIC subsidization of banks through deposit insurance is always undesirable. (3) “Large” FDIC losses are necessarily symptomatic of a poorly designed deposit insurance system.Deposit insurance

Latent Facies Mapping from Binary Geological Data

This is the published version. Copyright University of Chicago PressMany geological observation sets contain discrete-state data, which can be encoded as binary patterns. When there are conditional relationships between the variables, latent class analysis may be applied to subdivide the total sample into latent facies associations, which have local independence in the probability sense. Probabilities of latent facies assignments can be mapped areally as continuous surfaces of implied geological facies. Latent class analysis is rooted in simple probabilityt heory and can be a useful technique in geological applications where observations are descriptive or weakly numerical. The method is illustrated by a latent facies mapping of the Morrison Formation (Upper Jurassic) in the subsurface of west Kansas

Dynamical Evolution of Volume Fractions in Multipressure Multiphase Flow Models

Compared to single-pressure models, multipressure multiphase flow models require additional closure relations to determine the individual pressures of the different phases. These relations are often taken to be evolution equations for the volume fractions. We present a rigorous theoretical framework for constructing such equations for compressible multiphase mixtures in terms of submodels for the relative volumetric expansion rates ∆Ei of the phases. These quantities are essentially the rates at which the phases dynamically expand or contract in response to pressure differences, and represent the general tendency of the volume fractions to relax toward values that produce local pressure equilibrium. We present a simple provisional model of this type in which ∆Ei is proportional to pressure differences divided by the time required for sound waves to traverse an appropriate characteristic length. It is shown that the resulting approach to pressure equilibrium is monotonic rather than oscillatory, and occurs instantaneously in the incompressible limit

Phase transition of a one-dimensional Ising model with distance-dependent connections

The critical behavior of Ising model on a one-dimensional network, which has long-range connections at distances $l>1$ with the probability $\Theta(l)\sim l^{-m}$, is studied by using Monte Carlo simulations. Through studying the Ising model on networks with different $m$ values, this paper discusses the impact of the global correlation, which decays with the increase of $m$, on the phase transition of the Ising model. Adding the analysis of the finite-size scaling of the order parameter $[]$, it is observed that in the whole range of $0<m<2$, a finite-temperature transition exists, and the critical exponents show consistence with mean-field values, which indicates a mean-field nature of the phase transition.Comment: 5 pages,8 figure

Erratum: Sustained expansion of NKT cells and antigen-specific T cells after injection of α-galactosyl-ceramide loaded mature dendritic cells in cancer patients

[No abstract available

Sustained expansion of NKT cells and antigen-specific T cells after injection of α-galactosyl-ceramide loaded mature dendritic cells in cancer patients

Natural killer T (NKT) cells are distinct glycolipid reactive innate lymphocytes that are implicated in the resistance to pathogens and tumors. Earlier attempts to mobilize NKT cells, specifically, in vivo in humans met with limited success. Here, we evaluated intravenous injection of monocyte-derived mature DCs that were loaded with a synthetic NKT cell ligand, α-galactosyl-ceramide (α-GalCer; KRN-7000) in five patients who had advanced cancer. Injection of α-GalCer-pulsed, but not unpulsed, dendritic cells (DCs) led to \u3e100-fold expansion of several subsets of NKT cells in all patients; these could be detected for up to 6 mo after vaccination. NKT activation was associated with an increase in serum levels of interleukin-12 p40 and IFN-γ inducible protein-10. In addition, there was an increase in memory CD8+ T cells specific for cytomegalovirus in vivo in response to α-GalCer-loaded DCs, but not unpulsed DCs. These data demonstrate the feasibility of sustained expansion of NKT cells in vivo in humans, including patients who have advanced cancer, and suggest that NKT activation might help to boost adaptive T cell immunity in vivo

Management of Agitation During the COVID-19 Pandemic

The coronavirus disease 2019 (COVID-19) pandemic caused by the coronavirus SARS-CoV-2 has radically altered delivery of care in emergency settings. Unprecedented hardship due to ongoing fears of exposure and threats to personal safety, along with societal measures enacted to curb disease transmission, have had broad psychosocial impact on patients and healthcare workers alike. These changes can significantly affect diagnosing and managing behavioral emergencies such as agitation in the emergency department. On behalf of the American Association for Emergency Psychiatry, we highlight unique considerations for patients with severe behavioral symptoms and staff members managing symptoms of agitation during COVID-19. Early detection and treatment of agitation, precautions to minimize staff hazards, coordination with security personnel and psychiatric services, and avoidance of coercive strategies that cause respiratory depression will help mitigate heightened risks to safety caused by this outbreak