ICOS-Dependent and -Independent Functions of Memory CD4 T Cells in Allograft Rejection

Donor-reactive memory T cells undermine the survival of transplanted organs through multiple pathways. We have previously reported that memory CD4 T cells resist treatment with anti-CD154 antibody and donor-specific transfusion (DST/MR1) and promote cardiac allograft rejection via generation of effector CD4 T cells and alloantibody. We hypothesized that the helper functions of memory CD4 T cells are independent of T-cell costimulation through CD154 but instead are regulated by alternative costimulatory pathways. This study investigated how blocking ICOS/B7RP-1 interactions affects functions of donor-reactive memory CD4 T cells. Treatment with blocking anti-ICOS mAb synergized with DST/MR1 and prolonged mouse cardiac allograft survival despite the presence of donor-reactive memory CD4 T cells. While blocking ICOS did not diminish the expansion of preexisting memory CD4 T cells or the induction of allospecific effector T cells, it did inhibit recruitment of the activated memory and effector T cells into the graft. In addition, anti-ICOS mAb treatment in combination with DST/MR1 prevented help provided by memory CD4 T cells for production of donor-specific IgG antibody. These results demonstrate the potential efficacy of ICOS blockade in sensitized transplant patients and provide the foundation for rational use of ICOS blockade in combination with other graft-prolonging strategies.     

NMR‐based approach to the analysis of radiopharmaceuticals: radiochemical purity,specific activity,and radioactive concentration values by proton and tritium NMR spectroscopy

Compounds containing tritium are widely used across the drug discovery and development landscape. These materials are widely utilized because they can be efficiently synthesized and produced at high specific activity. Results from internally calibrated ³H and ¹H nuclear magnetic resonance (NMR) spectroscopy suggests that at least in some cases, this calibrated approach could supplement or potentially replace radio‐high‐performance liquid chromatography for radiochemical purity, dilution and scintillation counting for the measurement of radioactivity per volume, and liquid chromatography/mass spectrometry analysis for the determination of specific activity. In summary, the NMR‐derived values agreed with those from the standard approaches to within 1% to 9% for solution count and specific activity. Additionally, the NMR‐derived values for radiochemical purity deviated by less than 5%. A benefit of this method is that these values may be calculated at the same time that ³H NMR analysis provides the location and distribution of tritium atoms within the molecule. Presented and discussed here is the application of this method, advantages and disadvantages of the approach, and a rationale for utilizing internally calibrated ¹H and ³H NMR spectroscopy for specific activity, radioactive concentration, and radiochemical purity whenever acquiring ³H NMR for tritium location.     

Isolation of alpha-melanotropin from the pars intermedia of the larval amphibian, Ambystoma tigrinum

The effect of background adaptation on the steady-state levels of alpha-melanotropin in the pars intermedia of the larval amphibian. Ambystoma tigrinum, was investigated. Acid extracts of pars intermedia obtained from light-adapted and dark-adapted animals were analyzed by radioimmunoassay following Sephadex gel filtration chromatography, reverse-phase HPLC, and Sulfopropyl Sephadex cation-ion-exchange chromatography. For both background adaptation conditions similar results were obtained. The major form of alpha-melanotropin present in the pars intermedia has the following properties: (1) an apparent molecular mass of 1.5 kDa; (2) a net charge at pH 3.5 of +4; and (3) a retention time following reverse-phase HPLC similar to that of synthetic ACTH(1-13)amide. In dark-adapted animals a minor form of alpha-melanotropin which has a net charge of +3 at pH 3.5 was also detected. The latter form represented approximately 10% of the total alpha-melanotropin immunoreactivity in the pars intermedia of dark-adapted animals. These results strongly suggest that the predominant form of alpha-melanotropin in the pars intermedia of larval A. tigrinum is a nonacetylated ACTH(1-13)amide-like polypeptide.     

Thiocarbohydrazide-crosslinked chitosan as a bioinspired corrosion inhibitor for protection of stainless steel in 3.5% NaCl

A facile single-step synthesis was performed to cross-link chitosan with thiocarbohydrazide to yield thiocarbohydrazide-chitosan (TC-Cht) which was for the first time evaluated as an inhibitor for corrosion of stainless steel in 3.5% NaCl solution. A comprehensive electrochemical analysis employing electrochemical impedance spectroscopy (EIS), potentiodynamic polarization (PDP), and cyclic voltammetry (CV) was undertaken and showed that the TC-Cht acts by adsorption on the steel surface and exhibits mixed type behavior with predominantly cathodic nature. The adsorption of TC-Cht molecules on the surface of stainless steel followed the Langmuir isotherm. The TC-Cht showed a high inhibition efficiency of >94% at 500 mg L^?1 concentration. Surface investigation using SEM and EDX supported the inhibitor adsorption on the steel surface.     

Quantifying the Effects of Aging on Morphological and Cellular Properties of Human Female Pelvic Floor Muscles

Annals of Biomedical Engineering - Age-related pelvic floor muscle (PFM) dysfunction is a critical defect in the progression to pelvic floor disorders (PFDs). Despite dramatic prevalence of PFDs in...     

Lack of Amphetamine-Like Effects After Administration of Mefenorex in Normal Young Subjects

Mefenorex is an indirect sympathomimetic amine which acts as an anorectic drug and is used in combination with low diet to treat excess weight. The central nervous system (CNS) effects of mefenorex were assessed in a randomized, double-blind, three-way cross-over, placebo-controlled study involving nine healthy young male volunteers. They received either a single oral dose of mefenorex 80 mg (twice the recommended dose) or d-amphetamine sulfate 18 mg or a placebo at 1-week intervals. CNS pharmacodynamic measurements consisted of subjective evaluation (visual analogue scales and the Addiction Research Centre inventory (ARCI)), EEG, psychomotor performance and attention (tracking, simple and choice reaction times, tapping, continuous performance task, DSST, body sway) and memory (working memory and recall of a word list). d-Amphetamine produced a typical psychostimulant EEG profile (significant decrease in slow delta waves and increase in fast beta activities), significantly increased amphetamine, benzedrine and morphine–benzedrine scores of ARCI and significantly decreased body sway compared to placebo and mefenorex. A trend in favour of a stimulant effect occurred for all other parameters (particularly speed of reaction) and no changes of memory were noticed. In contrast, mefenorex did not produce an amphetamine-like EEG profile, neither significantly changed ARCI scores nor significantly modified psychomotor and memory performance compared to the placebo, although it induced a decrease in body sway. In conclusion, the present results indicate that a single oral dose of mefenorex, at twice the recommended daily dose, does not possess amphetamine-like subjective and EEG stimulant effects or sensations of well-being, often encountered with drugs of abuse liability potential, in a healthy young population.