MagA expression attenuates iron export activity in undifferentiated multipotent P19 cells

© 2019 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Magnetic resonance imaging (MRI) is a non-invasive imaging modality used in longitudinal cell tracking. Previous studies suggest that MagA, a putative iron transport protein from magnetotactic bacteria, is a useful gene-based magnetic resonance contrast agent. Hem-agglutinin-tagged MagA was stably expressed in undifferentiated embryonic mouse teratocarcinoma, multipotent P19 cells to provide a suitable model for tracking these cells during differentiation. Western blot and immunocytochemistry confirmed the expression and membrane localization of MagA in P19 cells. Surprisingly, elemental iron analysis using inductively-coupled plasma mass spectrometry revealed significant iron uptake in both parental and MagA-expressing P19 cells, cultured in the presence of iron-supplemented medium. Withdrawal of this extracellular iron supplement revealed unexpected iron export activity in P19 cells, which MagA expression attenuated. The influence of iron supplementation on parental and MagA-expressing cells was not reflected by longitudinal relaxation rates. Measurement of transverse relaxation rates (R2* and R2) reflected changes in total cellular iron content but did not clearly distinguish MagA-expressing cells from the parental cell type, despite significant differences in the uptake and retention of total cellular iron. Unlike other cell types, the reversible component R20 (R2* – R2) provided only a moderately strong correlation to amount of cellular iron, normalized to amount of protein. This is the first report to characterize MagA expression in a previously unrecognized iron exporting cell type. The interplay between contrast gene expression and systemic iron metabolism substantiates the potential for diverting cellular iron toward the formation of a novel iron compartment, however rudimentary when using a single magnetotactic bacterial gene expression system like magA. Since relatively few mammalian cells export iron, the P19 cell line provides a tractable model of ferroportin activity, suitable for magnetic resonance analysis of key iron-handling activities and their influence on gene-based MRI contrast

Review: Effects of Ractopamine Hydrochloride (Paylean) on welfare indicators for market weight pigs

This review summarizes the effects of ractopamine hydrochloride (RAC) dose (5, 7.5, 10, and 20 mg/kg) on market weight pig welfare indicators. Ractopamine hydrochloride (trade name Paylean) is a β-adrenergic agonist that was initially approved in the U.S. in 1999 at doses of 5 to 20 mg/kg to improve feed efficiency and carcass leanness. However, anecdotal reports suggested that RAC increased the rate of non-ambulatory (fatigued and injured) pigs at U.S. packing plants. This led to the addition of a caution statement to the Paylean label, and a series of research studies investigating the effects of RAC on pig welfare. Early research indicated that: (1) regardless of RAC administration, fatigued (non-ambulatory, non-injured) pigs are in a state of metabolic acidosis; (2) aggressive handling increases stress responsiveness at 20 mg/kg RAC, while 5 mg/kg reduces stress responsiveness to aggressive handling. Given this information, dosage range for Paylean was changed in 2006 to 5 to 10 mg/kg in market weight pigs. Subsequent research on RAC demonstrated that: (1) RAC has minimal effects on mortality, lameness, and home pen behavior; (2) RAC fed pigs demonstrated inconsistent prevalence and intensity of aggressive behaviors; (3) RAC fed pigs may be more difficult to handle at doses above 5 mg/kg; and (4) RAC fed pigs may have increased stress responsiveness and higher rates of non-ambulatory pigs when subjected to aggressive handling, especially when 20 mg/kg of RAC is fed

Induction of Biogenic Magnetization and Redox Control by a Component of the Target of Rapamycin Complex 1 Signaling Pathway

Most organisms are simply diamagnetic, while magnetotactic bacteria and migratory animals are among organisms that exploit magnetism. Biogenic magnetization not only is of fundamental interest, but also has industrial potential. However, the key factor(s) that enable biogenic magnetization in coordination with other cellular functions and metabolism remain unknown. To address the requirements for induction and the application of synthetic bio-magnetism, we explored the creation of magnetism in a simple model organism. Cell magnetization was first observed by attraction towards a magnet when normally diamagnetic yeast Saccharomyces cerevisiae were grown with ferric citrate. The magnetization was further enhanced by genetic modification of iron homeostasis and introduction of ferritin. The acquired magnetizable properties enabled the cells to be attracted to a magnet, and be trapped by a magnetic column. Superconducting quantum interference device (SQUID) magnetometry confirmed and quantitatively characterized the acquired paramagnetism. Electron microscopy and energy-dispersive X-ray spectroscopy showed electron-dense iron-containing aggregates within the magnetized cells. Magnetization-based screening of gene knockouts identified Tco89p, a component of TORC1 (Target of rapamycin complex 1), as important for magnetization; loss of TCO89 and treatment with rapamycin reduced magnetization in a TCO89-dependent manner. The TCO89 expression level positively correlated with magnetization, enabling inducible magnetization. Several carbon metabolism genes were also shown to affect magnetization. Redox mediators indicated that TCO89 alters the intracellular redox to an oxidized state in a dose-dependent manner. Taken together, we demonstrated that synthetic induction of magnetization is possible and that the key factors are local redox control through carbon metabolism and iron supply

Review: Effects of Ractopamine Hydrochloride (Paylean) on welfare indicators for market weight pigs

This review summarizes the effects of ractopamine hydrochloride (RAC) dose (5, 7.5, 10, and 20 mg/kg) on market weight pig welfare indicators. Ractopamine hydrochloride (trade name Paylean) is a β-adrenergic agonist that was initially approved in the U.S. in 1999 at doses of 5 to 20 mg/kg to improve feed efficiency and carcass leanness. However, anecdotal reports suggested that RAC increased the rate of non-ambulatory (fatigued and injured) pigs at U.S. packing plants. This led to the addition of a caution statement to the Paylean label, and a series of research studies investigating the effects of RAC on pig welfare. Early research indicated that: (1) regardless of RAC administration, fatigued (non-ambulatory, non-injured) pigs are in a state of metabolic acidosis; (2) aggressive handling increases stress responsiveness at 20 mg/kg RAC, while 5 mg/kg reduces stress responsiveness to aggressive handling. Given this information, dosage range for Paylean was changed in 2006 to 5 to 10 mg/kg in market weight pigs. Subsequent research on RAC demonstrated that: (1) RAC has minimal effects on mortality, lameness, and home pen behavior; (2) RAC fed pigs demonstrated inconsistent prevalence and intensity of aggressive behaviors; (3) RAC fed pigs may be more difficult to handle at doses above 5 mg/kg; and (4) RAC fed pigs may have increased stress responsiveness and higher rates of non-ambulatory pigs when subjected to aggressive handling, especially when 20 mg/kg of RAC is fed.This article is published as Ritter, M. J., A. K. Johnson, M. E. Benjamin, S. N. Carr, M. Ellis, L. Faucitano, T. Grandin et al. "Effects of Ractopamine Hydrochloride (Paylean) on welfare indicators for market weight pigs." Translational Animal Science 1, no. 4 (2017): 533-558. doi: 10.2527/tas2017.0060. Posted with permission.</p