Collision-Based Electrochemical Detection of Lysozyme Aggregation.

Proteins are utilized across many biomedical and pharmaceutical industries; therefore, methods for rapid and accurate monitoring of protein aggregation are needed to ensure proper product quality. Although these processes have been previously studied, it is difficult to comprehensively evaluate protein folding and aggregation by traditional characterization techniques such as atomic force microscopy (AFM), electron microscopy, or X-ray diffraction, which require sample pre-treatment and do not represent native state proteins in solution. Herein, we report early tracking of lysozyme (Lyz) aggregation states by using single-particle collision electrochemistry (SPCE) of silver nanoparticle (AgNP) redox probes.

G551D mutation impairs PKA-dependent activation of CFTR channel that can be restored by novel GOF mutations.

G551D is a major disease-associated gating mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) protein, an ATP- and phosphorylation-dependent chloride channel. G551D causes severe cystic fibrosis (CF) disease by disrupting ATP-dependent channel opening; however, whether G551D affects phosphorylation-dependent channel activation is unclear. Here, we use macropatch recording and Ussing chamber approaches to demonstrate that G551D impacts on phosphorylation-dependent activation of CFTR, and PKA-mediated phosphorylation regulates the interaction between the x-loop in nucleotide-binding domain 2 (NBD2) and cytosolic loop (CL) 1.

Nanoparticle Characterization Through Nano-Impact Electrochemistry: Tools and Methodology Development.

The field of nanomaterials has been expanding rapidly into many diverse applications within the last 20 years. With this growth, there is a significant need for new method development for the detection and characterization of nanomaterials. Understanding the physical properties of nanoscale entities and their associated reaction kinetics is crucial for monitoring their effect on environmental and human health, and in their use for practical applications.

Slowing ribosome velocity restores folding and function of mutant CFTR.

Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR), with approximately 90% of patients harboring at least one copy of the disease-associated variant F508del. We utilized a yeast phenomic system to identify genetic modifiers of F508del-CFTR biogenesis, from which ribosomal protein L12 (RPL12/uL11) emerged as a molecular target. In the present study, we investigated mechanism(s) by which suppression of RPL12 rescues F508del protein synthesis and activity.

Easy-to-Use Sensors for Field Monitoring of Copper Contamination in Water and Pesticide-Sprayed Plants.

Copper (Cu)-containing pesticides are commonly used in agriculture to control fungal and bacterial diseases, but the release of large quantities of Cu in water and soil can lead to harmful long-term consequences on the environment, organisms, and ecosystem health. Technology available to measure Cu accumulation in the field is too expensive and complicated for general population use. We describe a low-cost sensor with simplified user operation for measuring Cu content in environmental and agricultural samples at sensitivity levels comparable with a laboratory-based atomic absorption spectroscopy (AAS) method.

Early vs Delayed Weightbearing After Microfracture of Osteochondral Lesions of the Talus: A Prospective Randomized Trial.

Background: Osteochondral lesions of the talus (OLTs) are common injuries in young, active patients. Microfracture is an effective treatment for lesions less than 150 mm in size. Most commonly employed postoperative protocols involve delaying weightbearing for 6 to 8 weeks (DWB), though one study suggests that early weightbearing (EWB) may not be detrimental to patient outcomes.

Nanomaterial-functionalized Cellulose: Design, Characterization and Analytical Applications.

Cellulose-nanomaterial hybrid systems are promising platforms for the development of portable devices that can be used for fast and inexpensive analysis in the clinical, environmental and food monitoring fields. By combining the chemical and physical properties of the cellulosic network with the unique optical, electrical and catalytic functions of nanomaterials, it is possible to create versatile devices with engineered sensing functions. This review describes the most commonly used types of nanomaterials, their unique properties and assembly in hybrid structures in conjunction with cellulose paper and provides an overview of the most commonly used detection methodologies and their performance for selected applications.

Comparison of Calcaneal Exposure Through the Extensile Lateral and Sinus Tarsi Approaches.

Unlabelled: The purpose of this study was to compare the exposure of the posterior facet with the extensile lateral (EL) approach compared with the sinus tarsi (ST) approach. We hypothesized that the ST approach will provide a similar exposure of the posterior calcaneal facet. A total of 8 sequential ST then EL approaches were performed on cadavers.

Comparative Study of Assisted Ambulation and Perceived Exertion With the Wheeled Knee Walker and Axillary Crutches in Healthy Subjects.

Background: Functional limitations after lower extremity surgery often require the use of an assistive device for ambulation during rehabilitation and recovery. There are no known objective data evaluating the wheeled knee walker as an assistive device for protected ambulation. The purpose of this study was to compare assisted ambulation and perceived exertion with the wheeled knee walker and the axillary crutches in healthy participants.

Analysis of cystic fibrosis-associated P67L CFTR illustrates barriers to personalized therapeutics for orphan diseases.

Emerging knowledge indicates the difficulty in categorizing unusual cystic fibrosis (CF) mutations, with regard to both pathogenic mechanism and theratype. As case in point, we present data concerning P67L mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), a defect carried by a small number of individuals with CF and sometimes attributed to a channel conductance abnormality. Findings from our laboratory and others establish that P67L causes protein misfolding, disrupts maturation, confers gating defects, is thermally stable, and exhibits near normal conductance.

The Cystic Fibrosis Transmembrane Conductance Regulator Potentiator Ivacaftor Augments Mucociliary Clearance Abrogating Cystic Fibrosis Transmembrane Conductance Regulator Inhibition by Cigarette Smoke.

Acquired cystic fibrosis transmembrane conductance regulator (CFTR) dysfunction may contribute to chronic obstructive pulmonary disease pathogenesis and is a potential therapeutic target. We sought to determine the acute effects of cigarette smoke on ion transport and the mucociliary transport apparatus, their mechanistic basis, and whether deleterious effects could be reversed with the CFTR potentiator ivacaftor (VX-770). Primary human bronchial epithelial (HBE) cells and human bronchi were exposed to cigarette smoke extract (CSE) and/or ivacaftor.

Robust Stimulation of W1282X-CFTR Channel Activity by a Combination of Allosteric Modulators.

W1282X is a common nonsense mutation among cystic fibrosis patients that results in the production of a truncated Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) channel. Here we show that the channel activity of the W1282X-CFTR polypeptide is exceptionally low in excised membrane patches at normally saturating doses of ATP and PKA (single channel open probability (PO) < 0.01).

Channel Gating Regulation by the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) First Cytosolic Loop.

In this study, we present data indicating a robust and specific domain interaction between the cystic fibrosis transmembrane conductance regulator (CFTR) first cytosolic loop (CL1) and nucleotide binding domain 1 (NBD1) that allows ion transport to proceed in a regulated fashion. We used co-precipitation and ELISA to establish the molecular contact and showed that binding kinetics were not altered by the common clinical mutation F508del. Both intrinsic ATPase activity and CFTR channel gating were inhibited severely by CL1 peptide, suggesting that NBD1/CL1 binding is a crucial requirement for ATP hydrolysis and channel function.

Long-range coupling between the extracellular gates and the intracellular ATP binding domains of multidrug resistance protein pumps and cystic fibrosis transmembrane conductance regulator channels.

The ABCC transporter subfamily includes pumps, the long and short multidrug resistance proteins (MRPs), and an ATP-gated anion channel, the cystic fibrosis transmembrane conductance regulator (CFTR). We show that despite their thermodynamic differences, these ABCC transporter subtypes use broadly similar mechanisms to couple their extracellular gates to the ATP occupancies of their cytosolic nucleotide binding domains. A conserved extracellular phenylalanine at this gate was a prime location for producing gain of function (GOF) mutants of a long MRP in yeast (Ycf1p cadmium transporter), a short yeast MRP (Yor1p oligomycin exporter), and human CFTR channels.

Determining optimal ultrasound off time with micropulse longitudinal phacoemulsification.

Purpose: To evaluate the optimum off time for the most efficient removal of lens fragments using micropulse ultrasound (US).

Setting: John A. Moran Eye Center Laboratories, University of Utah, Salt Lake City, Utah, USA.

An electrostatic interaction at the tetrahelix bundle promotes phosphorylation-dependent cystic fibrosis transmembrane conductance regulator (CFTR) channel opening.

The CFTR channel is an essential mediator of electrolyte transport across epithelial tissues. CFTR opening is promoted by ATP binding and dimerization of its two nucleotide binding domains (NBDs). Phosphorylation of its R domain (e.

Determining optimal torsional ultrasound power for cataract surgery with automatic longitudinal pulses at maximum vacuum ex vivo.

Purpose: To determine the optimal longitudinal power settings for Infiniti OZil Intelligent Phaco (IP) at varying torsional amplitude settings; and to test the hypothesis that increasing longitudinal power is more important at lower torsional amplitudes to achieve efficient phacoemulsification.

Design: Laboratory investigation.

Methods: setting: John A.

Optimum on-time duty cycle for micropulse technology.

Purpose: To evaluate the optimum on time for the most efficient removal of lens fragments using micropulsed ultrasound (US).

Setting: John A. Moran Eye Center Laboratories, University of Utah, Salt Lake City, Utah, USA.

Conserved allosteric hot spots in the transmembrane domains of cystic fibrosis transmembrane conductance regulator (CFTR) channels and multidrug resistance protein (MRP) pumps.

ATP-binding cassette (ABC) transporters are an ancient family of transmembrane proteins that utilize ATPase activity to move substrates across cell membranes. The ABCC subfamily of the ABC transporters includes active drug exporters (the multidrug resistance proteins (MRPs)) and a unique ATP-gated ion channel (cystic fibrosis transmembrane conductance regulator (CFTR)). The CFTR channel shares gating principles with conventional ligand-gated ion channels, but the allosteric network that couples ATP binding at its nucleotide binding domains (NBDs) with conformational changes in its transmembrane helices (TMs) is poorly defined.

Phacoemulsification efficiency with a radiused phaco tip.

Purpose: To evaluate radiused and nonradiused phacoemulsification tips to determine which tip is more efficient in removal of lens fragments using 3 ultrasound (US) modalities.

Setting: John A. Moran Eye Center Laboratories, University of Utah, Salt Lake City, Utah, USA.

Can an integrated orthotic and rehabilitation program decrease pain and improve function after lower extremity trauma?

Background: Patients with severe lower extremity trauma have significant disability 2 years after injury that worsens by 7 years. Up to 15% seek late amputation. Recently, an energy-storing orthosis demonstrated improved function compared with standard orthoses; however, the effect when integrated with rehabilitation over time is unknown.

Comparison of functional outcomes following bridge synostosis with non-bone-bridging transtibial combat-related amputations.

Background: The prevalence of penetrating wartime trauma to the extremities has increased in recent military conflicts. Substantial controversy remains in the orthopaedic and prosthetic literature regarding which surgical technique should be performed to obtain the most functional transtibial amputation. We compared self-reported functional outcomes associated with two surgical techniques for transtibial amputation: bridge synostosis (modified Ertl) and non-bone-bridging (modified Burgess).

Converting nonhydrolyzable nucleotides to strong cystic fibrosis transmembrane conductance regulator (CFTR) agonists by gain of function (GOF) mutations.

Cystic fibrosis transmembrane conductance regulator (CFTR) is the only ligand-gated ion channel that hydrolyzes its agonist, ATP. CFTR gating has been argued to be tightly coupled to its enzymatic activity, but channels do open occasionally in the absence of ATP and are reversibly activated (albeit weakly) by nonhydrolyzable nucleotides. Why the latter only weakly activates CFTR is not understood.

Pathologic comparison of asymmetric or sulcus fixation of 3-piece intraocular lenses with square versus round anterior optic edges.

Objective: To evaluate the pathologic findings of 3-piece intraocular lenses (IOLs) with asymmetric or sulcus fixation in pseudophakic cadaver eyes, comparing IOLs with square or round edges on the anterior optic surface.

Design: Comparative case series with pathology.

Participants: A total of 661 pseudophakic cadaver eyes, obtained from eye banks within the United States, implanted with different IOLs.