Structural Characterization of the D179N and D179Y Variants of KPC-2 ß-Lactamase: Ω-Loop Destabilization as a Mechanism of Resistance to Ceftazidime-Avibactam.

Klebsiella pneumoniae carbapenemases (KPC-2 and KPC-3) present a global clinical threat, as these ß-lactamases confer resistance to carbapenems and oxyimino-cephalosporins. Recent clinically identified KPC variants with substitutions at Ambler position D179, located in the Ω loop, are resistant to the ß-lactam/ß-lactamase inhibitor combination ceftazidime-avibactam, but susceptible to meropenem-vaborbactam. To gain insights into ceftazidime-avibactam resistance conferred by D179N/Y variants of KPC-2, crystal structures of these variants were determined. The D179N KPC-2 structure revealed that the change of the carboxyl to an amide moiety at position 179 disrupted the salt bridge with R164 present in wild-type KPC-2. Additional interactions were disrupted in the Ω loop, causing a decrease in the melting temperature. Shifts originating from N179 were also transmitted toward the active site, including ∼1-Å shifts of the deacylation water and interacting residue N170. The structure of the D179Y KPC-2 ß-lactamase revealed more drastic changes, as this variant exhibited disorder of the Ω loop, with other flanking regions also being disordered. We postulate that the KPC-2 variants can accommodate ceftazidime because the Ω loop is displaced in D179Y or can be more readily displaced in D179N KPC-2. To understand why the ß-lactamase inhibitor vaborbactam is less affected by the D179 variants than avibactam, we determined the crystal structure of D179N KPC-2 in complex with vaborbactam, which revealed wild-type KPC-2-like vaborbactam-active site interactions. Overall, the structural results regarding KPC-2 D179 variants revealed various degrees of destabilization of the Ω loop that contribute to ceftazidime-avibactam resistance, possible substrate-assisted catalysis of ceftazidime, and meropenem and meropenem-vaborbactam susceptibility.

Spectroscopy of discrete vertically oriented single-crystals of n-type tetraazaterrylene: understanding the role of defects in molecular semiconductor photovoltaics.

Recent synthetic work has realized a novel (n-type) small-molecule acceptor, 7,8,15,16-tetra-aza-terrylene (TAT), single-crystals of which can be grown oriented along the c-axis crystallographic direction, and over-coated with pentacene to form a highly ordered donor/acceptor interface for use in organic photovoltaic devices. However, characterization of single TAT crystals reveals highly variable emission spectra and excited state dynamics - properties which strongly influence photovoltaic performance. Through the use of single-crystal widefield imaging, photoluminescence spectroscopy, time correlated single photon counting, and resonant Raman studies, we conclude that this variability is a result of long-lived low-energy trap-emission from packing defects. Interestingly, we also discovered that TAT crystals whose width exceeds ∼200 nm begin acting as waveguides and optical microcavity resonators for their own photoluminescence. Several strategies are proposed for leveraging the size-dependant optical properties of TAT pillars to further enhance device performance using this active layer design.

Polarization-driven stark shifts in quantum dot luminescence from single CdSe/oligo-PPV nanoparticles.

We demonstrate polarization-induced spectral shifts and associated linearly polarized absorption and emission in single CdSe/oligo-(phenylene vinylene) (CdSe/OPV) nanoparticles. A mechanism for these observations is presented in which charge separation from photoexcited ligands results in a significant Stark distortion of the quantum dot electron/hole wavefunctions. This distortion results in an induced linear polarization and an associated red shift in band-edge photoluminescence. These studies suggest the use of single quantum dots as local charge mobility probes.

Probing chemical reactions: evidence for exploration of an excited potential energy surface at thermal energies.

The reaction K + NaBr --> KBr + Na is probed during the reactive collision by a continuous wave laser tuned to frequencies not resonant with excitation in either reagents or products. Transient [K..Br..Na] absorbs a laser photon giving [K..Br..Na](*), which can decompose to Na(*) + KBr. Emission from excited Na(*) at the sodium D lines provides direct evidence of laser absorption during the reaction. Different excitation spectra were observed, depending on which sodium D line was monitored. This difference is explicable if, in the absence of the laser, the reaction flux partially bifurcates to a second potential energy surface during the reaction.

Laser-writable high-k dielectric for van der Waals nanoelectronics.

Similar to silicon-based semiconductor devices, van der Waals heterostructures require integration with high-k oxides. Here, we demonstrate a method to embed and pattern a multifunctional few-nanometer-thick high-k oxide within various van der Waals devices without degrading the properties of the neighboring two-dimensional materials. This transformation allows for the creation of several fundamental nanoelectronic and optoelectronic devices, including flexible Schottky barrier field-effect transistors, dual-gated graphene transistors, and vertical light-emitting/detecting tunneling transistors. Furthermore, upon dielectric breakdown, electrically conductive filaments are formed. This filamentation process can be used to electrically contact encapsulated conductive materials. Careful control of the filamentation process also allows for reversible switching memories. This nondestructive embedding of a high-k oxide within complex van der Waals heterostructures could play an important role in future flexible multifunctional van der Waals devices.

New routes to the functionalization patterning and manufacture of graphene-based materials for biomedical applications.

Graphene-based materials are being widely explored for a range of biomedical applications, from targeted drug delivery to biosensing, bioimaging and use for antibacterial treatments, to name but a few. In many such applications, it is not graphene itself that is used as the active agent, but one of its chemically functionalized forms. The type of chemical species used for functionalization will play a key role in determining the utility of any graphene-based device in any particular biomedical application, because this determines to a large part its physical, chemical, electrical and optical interactions. However, other factors will also be important in determining the eventual uptake of graphene-based biomedical technologies, in particular the ease and cost of manufacture of proposed device and system designs. In this work, we describe three novel routes for the chemical functionalization of graphene using oxygen, iron chloride and fluorine. We also introduce novel in situ methods for controlling and patterning such functionalization on the micro- and nanoscales. Our approaches are readily transferable to large-scale manufacturing, potentially paving the way for the eventual cost-effective production of functionalized graphene-based materials, devices and systems for a range of important biomedical applications.

Directional charge separation in isolated organic semiconductor crystalline nanowires.

One of the fundamental design paradigms in organic photovoltaic device engineering is based on the idea that charge separation is an extrinsically driven process requiring an interface for exciton fission. This idea has driven an enormous materials science engineering effort focused on construction of domain sizes commensurate with a nominal exciton diffusion length of order 10 nm. Here, we show that polarized optical excitation of isolated pristine crystalline nanowires of a small molecule n-type organic semiconductor, 7,8,15,16-tetraazaterrylene, generates a significant population of charge-separated polaron pairs along the π-stacking direction. Charge separation was signalled by pronounced power-law photoluminescence decay polarized along the same axis. In the transverse direction, we observed exponential decay associated with excitons localized on individual monomers. We propose that this effect derives from an intrinsic directional charge-transfer interaction that can ultimately be programmed by molecular packing geometry.

Birth dates of trigeminal motoneurons and metamorphic reorganization of the jaw myoneural system in frogs.

Drastic alterations in oral behavior characterize metamorphosis of anuran amphibians. Changes cascade through all components of the jaw apparatus from bone to muscle to nerve. In this investigation, tritiated thymidine autoradiography was used to determine the production schedule of the trigeminal motoneurons in the leopard frog, Rana pipiens. The time of origin of these neurons and their subsequent fate are of special interest given the breakdown of the larval jaw muscles and the de novo generation of adult muscle fibers during metamorphosis. Specifically, we wanted to learn whether trigeminal motoneurons are added, deleted, or reused during metamorphic climax. The entire complement of trigeminal motoneurons was produced over a 4-day span commencing at embryonic stage 13 and terminating at stage 20. Newly formed neurons are added to the primordial trigeminal nucleus in an orderly pattern. Firstborn neurons settle in the ventrorostral region of the nucleus; cells with progressively later birth dates were added in a posterodorsal direction. No additional trigeminal motoneurons are generated during larval maturation or at metamorphosis, thus indicating that the same population of neurons is present throughout the lifespan of the animal. From these observations we suggest that, during metamorphosis, the trigeminal motoneurons that supply the larval muscles switch their allegiance to the newly formed adult jaw muscles. This change of peripheral targets can be viewed as a respecification of the trigeminal motoneurons.

Maturation and recycling of trigeminal motoneurons in anuran larvae.

Development of the trigeminal motor system was analyzed in Rana pipiens larvae and adults. The aim of this investigation was to determine the postmetamorphic fate of the primary motoneurons that innervate the larval jaw muscles. Specifically, we wanted to ascertain whether these neurons were deleted in conjunction with their muscular targets during metamorphosis or reused to innervate the adult jaw muscles. Cell counts and horseradish peroxidase tracer were used to distinguish between these two possibilities. The number of trigeminal motoneurons was relatively constant in premetamorphic and prometamorphic larvae. A small reduction in the cellular complement of the motor nucleus occurred during metamorphic climax, but the majority (approximately equal to 90%) of the primary motoneurons were retained from the larval to the adult nervous system. The cell loss may represent motoneurons that innervated specific larval muscles that have no adult successors and thus the entire myoneural unit degenerates. Retrograde tracers indicated that all trigeminal motoneurons extended axons into the jaw muscles of both premetamorphic larvae and adult frogs. These observations provide further support for the recycling of the trigeminal motoneurons.

The relationship of economic factors and stress among employed, married women with children.

A convenience sample of 62 married women with children at home were measured on five economic variables and three stress-related variables: mother/marital role satisfaction, mother work role fulfillment and physical indicators of stress. The analysis reveals several significant relationships for the fulfillment and stress variables. Regression analysis also was used to model the relationship among economic and stress-related variables. The study suggests that financial health modules be integrated into stress management and other health enhancement programs for married, employed women.

Using storyboarding to determine components of wellness for university students.

Using a needs-assessment technique called storyboarding to determine characteristics of health promotion and wellness among university students is discussed. Storyboarding allows students to work together as they generate ideas and eliminates the territorial defensiveness that often occurs when new ideas are suggested. Sixteen graduate students in a health education teaching methods course demonstrated the use of the technique to identify characteristics of wellness for university students. In addition, they identified content areas and outlined key programs, services, personal habits, and skills useful for enhancing university students' wellness.

Health-related variables and academic performance among first-year college students: implications for sleep and other behaviors.

The authors analyzed the effect of several health behaviors and health-related variables on grade point averages of a random sample of 200 students living in on-campus residence halls at a large private university. The set of variables included exercise, eating, and sleep habits; mood states; perceived stress; time management; social support; spiritual or religious habits; number of hours worked per week; gender; and age. Of all the variables considered, sleep habits, particularly wake-up times, accounted for the largest amount of variance in grade point averages. Later wake-up times were associated with lower average grades. Variables associated with the 1st-year students' higher grade point averages were strength training and study of spiritually oriented material. The number of paid or volunteer hours worked per week was associated with lower average grades.

Spindle cell angiosarcoma of the oropharynx.

Angiosarcomas are uncommon malignant tumours of vascular endothelium. In the head and neck, the scalp is the commonest site of origin. A rare case of spindle-cell angiosarcoma of the oropharynx, treated by surgery and post-operative radiotherapy, is reported. We discuss the clinical presentation and histopathological diagnosis of this lesion.

Properties and applications of chemically functionalized graphene.

The vast and yet largely unexplored family of graphene materials has great potential for future electronic devices with novel functionalities. The ability to engineer the electrical and optical properties in graphene by chemically functionalizing it with a molecule or adatom is widening considerably the potential applications targeted by graphene. Indeed, functionalized graphene has been found to be the best known transparent conductor or a wide gap semiconductor. At the same time, understanding the mechanisms driving the functionalization of graphene with hydrogen is proving to be of fundamental interest for energy storage devices. Here we discuss recent advances on the properties and applications of chemically functionalized graphene.

Linear dipole behavior in single CdSe-oligo(phenylene vinylene) nanostructures.

We report on linearly polarized absorption and emission from individual (4.3 nm) CdSe quantum dots whose surfaces are coordinated with monodisperse oligo-phenylene vinylene ligands. Shown previously to suppress quantum dot blinking, we demonstrate here that the electronic interaction of photoexcited ligands with the quantum dot core is manifested as a strong polarization anisotropy in absorption (M = 0.5), as well as distinct linear dipole emission patterns from the quantum dot core. Further, there is a correlation between the quantum dot emission moment and polarization orientation corresponding to the absorption maxima that is manifested as fluctuations in emission moment orientation in the X-Y plane. The observed polarization effects can be switched off by tuning the excitation away from the ligand absorption band. We propose a mechanism based on exciton dissociation from the photoexcited ligand, followed by the pinning of electrons at the quantum dot surface. The resulting Stark interaction is sufficiently strong to break the 2D degeneracy of the emission moment within the dot, and may therefore account for the linear dipole emission character.

Fluorescence lifetimes and correlated photon statistics from single CdSe/oligo(phenylene vinylene) composite nanostructures.

We present measurements of fluorescence intensity trajectories and associated excited-state decay times from individual CdSe/oligo(phenylene vinylene) (CdSe-OPV) quantum dot nanostructures using time-tagged, time-resolved (TTTR) photon counting techniques. We find that fluorescence decay times for the quantum dot emitter in these composite systems are at least an order of magnitude shorter than ZnS-capped CdSe quantum dot systems. We show that both the blinking suppression and associated lifetime/count rate behavior can be described by a modified version of the diffusive reaction coordinate model which couples slow fluctuations in quantum dot electron (1Se, 1Pe) energies to Auger-assisted hole trapping processes, hence modifying both blinking statistics and excited-state decay rates.

Blinking suppression and intensity recurrences in single CdSe-oligo(phenylene vinylene) nanostructures: experiment and kinetic model.

We report time-resolved single molecule fluorescence imaging of individual CdSe quantum dots that are functionalized with oligomeric conjugated organic ligands. The fluorescence intensity trajectories from these composite nanostructures display both a strong degree of blinking suppression and intensity fluctuations with characteristic recurrence times on the order of 10-60 s. In addition, fluorescence decay rate measurements of individual hybrid nanostructures indicate significantly modified non-radiative quantum dot decay rates relative to conventional ZnS-capped CdSe quantum dots. We show that a modified diffusive reaction coordinate model with slow fluctuations in quantum dot electron energies (1S(e), 1P(e)) can reproduce the experimentally observed behaviour.

Glimpses into measurements of the microcirculation in skin.

There are instances when chronic disease states manifest as microvascular disturbances of the skin. Objective assessment of these disturbances would permit a better understanding of the causative factors and, in certain circumstances, influence decisions of clinical management. Recent advances in technology that permit such assessment non-invasively are evaluated in this review.

Effectiveness of antibiotic prophylaxis in preventing bacteriuria after multichannel urodynamic investigations: a blind, randomized study in 124 female patients.

One hundred twenty-four women with chronic, persistent lower urinary tract symptoms who had been scheduled for elective urodynamic investigations at Mount Sinai's Urodynamic Investigative Unit were divided into two blind, randomized groups, receiving either a placebo or prophylactic antibiotic. At the time of urodynamic testing, the rate of unsuspected urinary tract infection was 8.1%. There was no statistically significant decrease in postinstrumentation infection rate in the group who received prophylactic antibiotics. We conclude that, given in the fashion described in the study, prophylactic antibiotics are not effective in preventing urinary tract infections caused by urodynamic testing.

Generalizability theory: a unified approach to assessing the dependability (reliability) of measurements in the health sciences.

The reliability of health promotion program evaluation measures, behavioral and attitudinal measures, and clinical measures is a concern to many health educators. Classical reliability coefficients, such as Cronbach's alpha, apply to narrowly defined, prespecified measurement situations. Classical theory does not provide adequate reliability assessments for criterion-referenced measures, for measurement situations having multiple sources of error, or for aggregate-level variables. Generalizability theory can be used to assess the reliability of measures in these situations that are not adequately modeled by Classical theory. Additionally, Generalizability theory affords a broader view and a deeper understanding of the dependability of measurements and the role of different sources of error in the variability of measures.