250 MHz Rapid Scan Cross Loop Resonator.

A 25 mm diameter 250 MHz crossed-loop resonator was designed for rapid scan electron paramagnetic resonance imaging. It has a saddle coil for the driven resonator and a fine wire, loop gap resonator for the sample resonator. There is good separation of E and B fields and high isolation between the two resonators, permitting a wide range of sample types to be measured. Applications to imaging of nitroxide, trityl, and LiPc samples illustrate the utility of the resonator. Using this resonator and a trityl sample the signal-to-noise of a rapid scan absorption spectrum is about 20 times higher than for a first-derivative CW spectrum.

Background correction in rapid scan EPR spectroscopy.

In rapid scan EPR the rapidly-changing magnetic field induces a background signal that may be larger than the EPR signal. A method has been developed to correct for that background signal by acquiring two sets of data, denoted as scan 1 and scan 2. In scan 2 the external field B0 is reversed and the data acquisition trigger is offset by one half cycle of the scan field relative to the settings used in scan 1. For data acquired with a cross-loop resonator subtraction of scan 2 from scan 1 cancels the background and enhances the EPR signal. Experiments were performed at an EPR frequency of about 258 MHz, which is in the range that is commonly used for in vivo imaging. Samples include nitroxide radicals, a trityl radical, a dinitroxide, and a nitroxide in the presence of a magnetic field gradient. This method has the advantage that no assumption is made about the shape of the background signal, and it provides an approach to automating the background correction.

Tabletop 700 MHz electron paramagnetic resonance imaging spectrometer.

Low frequency electron paramagnetic resonance imaging is a powerful tool to non-invasively measure the physiological status of tumors. Here, we report on the design and functionality of a rapid scan and pulse table-top imaging spectrometer based around an arbitrary waveform generator and 25mm cross-loop resonator operating at 700 MHz. Two and four-dimensional rapid scan spectral-spatial images are presented. This table-top imager is a prototype for future pre-clinical imagers.

Rapid-scan EPR imaging.

In rapid-scan EPR the magnetic field or frequency is repeatedly scanned through the spectrum at rates that are much faster than in conventional continuous wave EPR. The signal is directly-detected with a mixer at the source frequency. Rapid-scan EPR is particularly advantageous when the scan rate through resonance is fast relative to electron spin relaxation rates. In such scans, there may be oscillations on the trailing edge of the spectrum. These oscillations can be removed by mathematical deconvolution to recover the slow-scan absorption spectrum. In cases of inhomogeneous broadening, the oscillations may interfere destructively to the extent that they are not visible. The deconvolution can be used even when it is not required, so spectra can be obtained in which some portions of the spectrum are in the rapid-scan regime and some are not. The technology developed for rapid-scan EPR can be applied generally so long as spectra are obtained in the linear response region. The detection of the full spectrum in each scan, the ability to use higher microwave power without saturation, and the noise filtering inherent in coherent averaging results in substantial improvement in signal-to-noise relative to conventional continuous wave spectroscopy, which is particularly advantageous for low-frequency EPR imaging. This overview describes the principles of rapid-scan EPR and the hardware used to generate the spectra. Examples are provided of its application to imaging of nitroxide radicals, diradicals, and spin-trapped radicals at a Larmor frequency of ca. 250MHz.

Resonators for In Vivo Imaging: Practical Experience.

Resonators for preclinical electron paramagnetic resonance imaging have been designed primarily for rodents and rabbits and have internal diameters between 16 and 51 mm. Lumped circuit resonators include loop-gap, Alderman-Grant, and saddle coil topologies and surface coils. Bimodal resonators are useful for isolating the detected signal from incident power and reducing dead time in pulse experiments. Resonators for continuous wave, rapid scan, and pulse experiments are described. Experience at the University of Chicago and University of Denver in design of resonators for in vivo imaging is summarized.

Systematic review and meta-analysis of randomised controlled trials on the effects of potassium supplements on serum potassium and creatinine.

OBJECTIVES: High potassium intake could prevent stroke, but supplementation is considered hazardous. We assessed the effect of oral potassium supplementation on serum or plasma potassium levels and renal function. SETTING: We updated a systematic review of the effects of potassium supplementation in randomised clinical trials carried out worldwide, published in 2013, extending it to July 2015. We followed the PRISMA guidelines. PARTICIPANTS: Any individual taking part in a potassium supplementation randomised clinical trial. Studies included met the following criteria: randomised clinical trials, potassium supplement given and circulating potassium levels reported. INTERVENTION: Oral potassium supplementation. PRIMARY OUTCOME MEASURES: Serum or plasma potassium and serum or plasma creatinine. RESULTS: A total of 20 trials (21 independent groups) were included (1216 participants from 12 different countries). All but 2 were controlled (placebo n=16, control n=2). Of these trials, 15 were crossover, 4 had a parallel group and 1 was sequential. The duration of supplementation varied from 2 to 24 weeks and the amount of potassium given from 22 to 140 mmol/day. In the pooled analysis, potassium supplementation caused a small but significant increase in circulating potassium levels (weighted mean difference (WMD) 0.14 mmol/L, 95% CI 0.09 to 0.19, p<1×10(-5)), not associated with dose or duration of treatment. The average increase in urinary potassium excretion was 45.75 mmol/24 hours, 95% CI 38.81 to 53.69, p<1×10(-5). Potassium supplementation did not cause any change in circulating creatinine levels (WMD 0.30 µmol/L, 95% CI -1.19 to 1.78, p=0.70). CONCLUSIONS: In short-term studies of relatively healthy persons, a moderate oral potassium supplement resulted in a small increase in circulating potassium levels and no change in renal function.

Simeprevir and sofosbuvir with or without ribavirin to treat recurrent genotype 1 hepatitis C virus infection after orthotopic liver transplantation.

Although combination simeprevir (SIM) plus sofosbuvir (SOF) is an approved regimen for genotype 1 chronic hepatitis C virus (HCV), data regarding its safety and efficacy in liver transplant recipients remain limited. A multicenter retrospective study was performed to determine the efficacy and tolerability of a 12-week regimen of SIM/SOF with or without ribavirin (RBV) in 56 consecutive liver transplant recipients in 2014; 79% of patients had genotype 1a, 14% had cirrhosis, and 73% were treatment experienced. Sustained virological response at 12 weeks (SVR12) was 88% by intention to treat analysis (95% confidence interval, 84%-90%). Four patients relapsed, but no on-treatment virological failures occurred. The Q80K polymorphism did not impact SVR12, but there was a trend toward decreased sustained virological response with advanced fibrosis (P = 0.18). HCV RNA was detectable at treatment week 4 in 21% of patients, and those who had detectable levels were less likely to achieve SVR12 (58% versus 95%; P = 0.003). Five patients had baseline Child-Pugh class B cirrhosis, and 2 of them died (1 following early discontinuation of therapy). An additional discontinuation resulted from a severe photosensitivity reaction in a patient on concomitant cyclosporine. Seven patients receiving RBV developed progressive anemia requiring intervention. Immunosuppression dose modifications were minimal. SIM/SOF for 12 weeks was effective and well tolerated by compensated liver transplant recipients especially when administered without concomitant RBV or cyclosporine. SIM/SOF appears to have a niche as the only 12-week RBV-free treatment regimen currently recommended by guidelines for compensated transplant recipients. However, 12 weeks may not be the optimal duration of therapy for those with detectable virus at week 4 or possibly for those with cirrhosis. These data require confirmation by prospective randomized clinical trials. Liver Transplantation 22 635-643 2016 AASLD.

Imaging disulfide dinitroxides at 250 MHz to monitor thiol redox status.

Measurement of thiol-disulfide redox status is crucial for characterization of tumor physiology. The electron paramagnetic resonance (EPR) spectra of disulfide-linked dinitroxides are readily distinguished from those of the corresponding monoradicals that are formed by cleavage of the disulfide linkage by free thiols. EPR spectra can thus be used to monitor the rate of cleavage and the thiol redox status. EPR spectra of (1)H,(14)N- and (2)H,(15)N-disulfide dinitroxides and the corresponding monoradicals resulting from cleavage by glutathione have been characterized at 250 MHz, 1.04 GHz, and 9 GHz and imaged by rapid-scan EPR at 250 MHz.

Improved sensitivity for imaging spin trapped hydroxyl radical at 250 MHz.

Radicals, including hydroxyl, superoxide, and nitric oxide, play key signaling roles in vivo. Reaction of these free radicals with a spin trap affords more stable paramagnetic nitroxides, but concentrations in vivo still are so low that detection by electron paramagnetic resonance (EPR) is challenging. Three innovative enabling technologies have been combined to substantially improve sensitivity for imaging spin-trapped radicals at 250 MHz. 1) Spin-trapped adducts of BMPO have lifetimes that are long enough to make imaging by EPR at 250 MHz feasible. 2) The signal-to-noise ratio of rapid-scan EPR is substantially higher than for conventional continuous-wave EPR. 3) An improved algorithm permits image reconstruction with a spectral dimension that encompasses the full 50 G spectrum of the BMPO-OH spin adduct without requiring the wide sweeps that would be needed for filtered backprojection. A 2D spectral-spatial image is shown for a phantom containing ca. 5 µM BMPO-OH.

The obesity epidemic: challenges, health initiatives, and implications for gastroenterologists.

Obesity is the next major epidemiologic challenge facing today's doctors, with the annual allocation of healthcare resources for the disease and related comorbidities projected to exceed $150 billion in the United States. The incidence of obesity has risen in the United States over the past 30 years; 60% of adults are currently either obese or overweight. Obesity is associated with a higher incidence of a number of diseases, including diabetes, cardiovascular disease, and cancer. Consumption of fast food, trans fatty acids (TFAs), and fructose-combined with increasing portion sizes and decreased physical activity-has been implicated as a potential contributing factor in the obesity crisis. The use of body mass index (BMI) alone is of limited utility for predicting adverse cardiovascular outcomes, but the utility of this measure may be strengthened when combined with waist circumference and other anthropomorphic measurements. Certain public health initiatives have helped to identify and reduce some of the factors contributing to obesity. In New York City and Denmark, for example, such initiatives have succeeded in passing legislation to reduce or remove TFAs from residents' diets. The obesity epidemic will likely change practice for gastroenterologists, as shifts will be seen in the incidence of obesity-related gastrointestinal disorders, disease severity, and the nature of comorbidities. The experience gained with previous epidemiologic problems such as smoking should help involved parties to expand needed health initiatives and increase the likelihood of preventing future generations from suffering the consequences of obesity.

Effect of bioactive glass crystallization on the conformation and bioactivity of adsorbed proteins.

Protein adsorption onto the surface of a biomaterial mediates cell adhesion and enhances tissue-implant integration. In a previous study, we demonstrated that crystallization of bioactive glass (BG) significantly increased the negative zeta potential and decreased serum protein adsorption onto the material surface. In this study, the conformation of protein adsorbed onto the surface of amorphous bioactive glass (ABG) and crystallized bioactive glass (CBG) was analyzed and correlated to bone marrow mesenchymal stem cell adhesion and spreading. ABG and CBG were immersed in three different protein solutions containing 10% fetal bovine serum, bovine serum albumin (BSA), and fibronectin (FN) for 4 h at 37 degrees C. Grazing angle Fourier transform infrared spectroscopy (GA-FTIR) demonstrated that the ratio of (amide I)/(amide II) functional groups of all proteins adsorbed onto ABG was greater than that for proteins adsorbed onto CBG. The Gaussian curve fitting analysis suggests that the significant expression of amide I, rich in charged and flexible unordered secondary structure of adsorbed FN, stimulated bone cell adhesion and spreading on the surface of ABG. CBG enforces protein conformation that exposes amide II, rich in neutral and stable beta-sheet structure and alpha-helix, which limited cell adhesion and spreading. Although ABG adsorbed significantly higher quantity of BSA than FN, GA-FTIR analyses showed that the ratio of amide I/amide II was significantly higher for adsorbed FN. Therefore, the intensity of amide I or amide II bands cannot be taken as a measure of the quantity of adsorbed protein.