Iron Injection Research Articles

Improving survival, growth rate, and animal welfare in piglets by avoiding teeth shortening and tail docking

Piglets are subjected to several painful procedures during their first week of life, including ear tagging, teeth clipping or grinding (although routinely prohibited in Europe), tail docking (although routinely prohibited in Europe), needle injections (vaccination and iron injection), and castration might be performed for male piglets. All these management practices cause pain and stress to the newborn piglets. The hypothesis of this experiment was that reducing painful interventions during the first week of life results in better zootechnical performance of the piglets, reduced piglet mortality, and that the overall welfare is improved. To investigate this, the 4 lightest piglets of the experimental group (EE) were not subjected to tail docking and teeth clipping or grinding. The 4 lightest piglets of the control group (CL) and the other piglets of the experimental group (EC) and the control group (CC) received treatments as the common practice. There were differences in behavior, but there were no differences regarding weight at weaning between CL and EE. However, mortality rate was higher in the lightest piglets, that is, CL (34.1%) and EE (23.0%), whereas mortality rate in the other piglets was much lower (9.0% for CC and 9.3% for EC). This information can be useful for a veterinarian to advise farmers on whether painful interventions should be performed.

Abstract 2156: Minocycline-induced Attenuation Of Iron Overload And Brain Injury Following Experimental Intracerebral Hemorrhage

Background and Purpose: Brain iron overload plays a detrimental role in brain injury after intracerebral hemorrhage (ICH). A recent study found that minocycline acts as an iron chelator and reduces iron-induced neuronal death in vitro. The present study investigated if minocycline reduces iron overload after ICH and iron-induced brain injury in vivo. Methods: This study was divided into three parts. (1) Male Sprague-Dawley rats with different sizes of ICH were euthanized 3 days later for serum total iron and brain edema determination. (2) Rats had an ICH treated with minocycline or vehicle. Rats were euthanized 1, 3 and 7 days later for serum iron, brain iron, and brain iron handling protein measurements. (3) Rats had a 50µl intracaudate injection of either saline, FeCl2, FeCl2+minocycline or FeCl2+macrophage/microglia inhibitory factor and were euthanized at one day later for measurements of brain edema, blood-brain barrier disruption and neuronal death. Results: After ICH, serum total iron and brain non-heme iron increased and these changes were reduced by minocycline treatment (e.g. serum total iron at day 3: 158±36 vs. 245±22 µg/dL in the vehicle-treated group, p<0.01). Minocycline also reduced ICH-induced upregulation of brain iron handling proteins and neuronal death. Intracaudate injection of iron caused brain edema, blood-brain barrier leakage and brain cell death, all of which were significantly reduced by co-injection with minocycline (p<0.05). Conclusions: The current study found that minocycline reduces iron overload after ICH and iron-induced brain injury. It is also well known minocycline is an inhibitor of microglial activation. Minocycline may be very useful for ICH patients because both iron accumulation and microglia activation contribute to brain damage following ICH.

Histopathologic Study of Livers From Chronic Iron Loaded C282Y Mice: Model of Hereditary Hemochromatosis?

Abstract 5288 Introduction:The C282Y mouse model of hemochromatosis was developed to facilitate study of hereditary hemochromatosis in humans. We studied the liver histologic findings in mice that were iron loaded over at least 10 weeks to determine if the pathological changes in the mouse liver mirror the chronic iron overload condition observed in humans with genetic hemochromatosis. Methods:C282Y and wild-type mice (C57BL) were iron loaded over a minimum of 10 weeks using weekly intraperitoneal injections of 400 mg/kg of iron dextran. Animals were sacrificed at least 2 weeks after receiving their last iron injection. Hematoxylin and eosin, Masson’s trichrome, and Perl’s Prussian blue stains were used to assess the amount and distribution of liver iron and the presence or absence of inflammation and fibrosis. The slides were evaluated by a veterinary pathologist (MAS) without knowledge of genetic or iron load status. Specific histopathologic characteristics were evaluated and graded as the mean of 10 40x high power fields. These included: Hepatocyte iron distribution graded as either periportal or diffuse, amount of iron deposition in cells was graded for hepatocytes and Kupffer cells (graded 0–5), iron deposition in endothelial cells (graded 0–4), amount of fibrosis (graded 0–4), presence or absence of inflammation, periportal lymphocyte infiltration (graded 0–3), and hematopoiesis (present, present but mild, or absent). Livers from 24 mice (3 +/+, 9 +/Y and 12 Y/Y) were studied. Results:When compared with animals that were not iron loaded, animals iron loaded with at least 4 g/kg of iron had increased cellular deposition of iron. Of the two mice that were not iron loaded, the +/Y mouse had no increase in tissue iron and the Y/Y mouse had 2+ iron in hepatocytes, Kupffer cells, and endothelial cells. The highest two categories of cellular iron were observed in the four animals loaded with the highest doses of iron (5.6–6.8 g/kg): grade 4–5 for hepatocytes and Kupffer cells, and grade 3–4 for endothelial cells. Mice loaded with 4 g/kg of iron (7 Y/Y, 8 +/Y and 3 +/+) had similar but generally lower levels of iron deposition: grade 3–5 in hepatocytes, grade 3–4 in Kupffer cells, and grade 3 in endothelial cells. The three genotypes could not be differentiated based on the amount of iron deposition. However, the distribution of iron in hepatocytes of the 3 wild type animals (+/+) was clearly periportal which differed from the diffuse distribution pattern observed in the Y/Y and +/Y mice. One +/Y mouse had a mild periportal distribution of hepatocyte iron. Iron-loaded animals in all 3 genetic groups had grade 1 or 2 fibrosis in the periportal area and around the iron laden Kupffer cell aggregates. None of animals had bridging fibrosis or cirrhosis. In the iron-loaded Y/Y and +/Y mice, hematopoiesis was present in close association with intrasinusoidal iron laden Kupffer cell aggregates. In the non-iron loaded mice hematopoiesis was rare in the Y/Y animal, absent in the +/Y mouse. Hematopoiesis was present but mild in all 3 of the +/+ mice as well as in 1 mouse of each other genotype, and was present in the remaining 17 iron-loaded mice. No hepatocyte necrosis or regenerative nodules were detected in any mouse. Discussion:Histologic evaluation of the C282Y mice showed that the deposition of iron in the liver increased with increased iron loads. The distribution of iron between Kupffer cells and hepatocytes was similar in all 3 genetic groups. The periportal iron distribution observed in humans with genetic hemochromatosis was not seen in either the Y/Y or +/Y genotypes, but was observed in the iron-loaded wild type +/+ mice. Liver histology with chronic intraperitoneal iron loading in the C282Y C57BL mice is comparable to that observed in humans with secondary iron overload and not those observed in humans with hereditary hemochromatosis. Why the liver histology in the iron loaded C282Y knock-in mouse does not mirror the changes observed in human livers from patients with genetic hemochromatosis is not clear. It may be the dosing regimen or the strain of the mouse or that additional genetic or environmental factors are required. Disclosures:No relevant conflicts of interest to declare.

Silencing of RhoA nucleotide exchange factor, ARHGEF3, reveals its unexpected role in iron uptake

AbstractGenomewide association meta-analysis studies have identified > 100 independent genetic loci associated with blood cell indices, including volume and count of platelets and erythrocytes. Although several of these loci encode known regulators of hematopoiesis, the mechanism by which most sequence variants exert their effect on blood cell formation remains elusive. An example is the Rho guanine nucleotide exchange factor, ARHGEF3, which was previously implicated by genomewide association meta-analysis studies in bone cell biology. Here, we report on the unexpected role of ARHGEF3 in regulation of iron uptake and erythroid cell maturation. Although early erythroid differentiation progressed normally, silencing of arhgef3 in Danio rerio resulted in microcytic and hypochromic anemia. This was rescued by intracellular supplementation of iron, showing that arhgef3-depleted erythroid cells are fully capable of hemoglobinization. Disruption of the arhgef3 target, RhoA, also produced severe anemia, which was, again, corrected by iron injection. Moreover, silencing of ARHGEF3 in erythromyeloblastoid cells K562 showed that the uptake of transferrin was severely impaired. Taken together, this is the first study to provide evidence for ARHGEF3 being a regulator of transferrin uptake in erythroid cells, through activation of RHOA.

Minocycline-Induced Attenuation of Iron Overload and Brain Injury After Experimental Intracerebral Hemorrhage

Brain iron overload plays a detrimental role in brain injury after intracerebral hemorrhage (ICH). A recent study found that minocycline acts as an iron chelator and reduces iron-induced neuronal death in vitro. The present study investigated if minocycline reduces iron overload after ICH and iron-induced brain injury in vivo. This study was divided into 4 parts: (1) rats with different sizes of ICH were euthanized 3 days later for serum total iron and brain edema determination; (2) rats had an ICH treated with minocycline or vehicle. Serum iron, brain iron, and brain iron handling proteins were measured; (3) rats had an intracaudate injection of saline, iron, iron+minocycline, or iron+macrophage/microglia inhibitory factor and were used for brain edema and neuronal death measurements; and (4) rats had an intracaudate injection of iron and were treated with minocycline. The brains were used for edema measurement. After ICH, serum total iron and brain nonheme iron increased and these changes were reduced by minocycline treatment. Minocycline also reduced ICH-induced upregulation of brain iron handling proteins and neuronal death. Intracaudate injection of iron caused brain edema, blood-brain barrier leakage, and brain cell death, all of which were significantly reduced by coinjection with minocycline. The current study found that minocycline reduces iron overload after ICH and iron-induced brain injury. It is also well known minocycline is an inhibitor of microglial activation. Minocycline may be very useful for patients with ICH because both iron accumulation and microglia activation contribute to brain damage after ICH.

Iron administration reduces airway hyperreactivity and eosinophilia in a mouse model of allergic asthma

The prevalence of allergic diseases has increased dramatically during the last four decades and is paralleled by a striking increase in iron intake by infants in affluent societies. Several studies have suggested a link between increased iron intake and the marked increase in prevalence of allergic diseases. We hypothesized that the increased iron intake by infants offers an explanation for the increased prevalence of allergic disease in industrialized societies during the past four decades. A well-established mouse model of ovalbumin (OVA)-driven allergic asthma was used to test the effects of differences in iron intake and systemic iron levels on the manifestations of allergic asthma. Surprisingly, iron supplementation resulted in a significant decrease in airway eosinophilia, while systemic iron injections lead to a significant suppression of both allergen-induced airway eosinophilia and hyperreactivity compared to placebo. In contrast, mice fed on an iron-deprived diet did not show any difference in developing experimentally induced allergic asthma when compared to those fed on an iron-sufficient control diet. In contrast to our hypothesis, airway manifestations of allergic asthma are suppressed by both increased levels of iron intake and systemic iron administrations in the mouse model.

Iron Enhances the Neurotoxicity of Amyloid β

Brain microbleeds often occur in Alzheimer's disease patients. Our previous studies have demonstrated that iron contributes to brain injury following intracerebral hemorrhage. This study investigated the effect of iron on amyloid β (Aβ)-mediated brain injury. There were two parts to this study. In first part, rats received an intracaudate injection of saline, iron, Aβ 25-35 or iron+Aβ 25-35. In the second part, rats received intracaudate injection of iron+Aβ and were treated with saline or cystamine, an inhibitor of transglutaminase. Rats were killed after 24 hours for brain edema measurement. DNA damage, neuronal death and tissue-type transglutaminase (tTG) expression were also examined. We found that brain water content in the ipsilateral caudate was higher (p<0.05) in rats injected with iron+Aβ than with iron, Aβ or saline. Combined iron+Aβ injection also resulted in more severe DNA damage (both single- and double-strand; p<0.01) and more Fluoro-Jade C staining (p<0.05). Expression of tTG increased markedly in the iron+Aβ group (p<0.05) and treatment with a tTG inhibitor reduced brain edema (p<0.05) and reduced degenerating neurons (124±25 vs. 249±50/mm(2) in vehicle-treated group, p<0.05). These results suggest that increased brain iron from microbleeds may exaggerate brain Aβ toxicity and that tTG is involved in the enhanced toxicity.

New rat models of iron sucrose-induced iron overload

The majority of murine models of iron sucrose-induced iron overload were carried out in adult subjects. This cannot reflect the high risk of iron overload in children who have an increased need for iron. In this study, we developed four experimental iron overload models in young rats using iron sucrose and evaluated different markers of iron overload, tissue oxidative stress and inflammation as its consequences. Iron overload was observed in all iron-treated rats, as evidenced by significant increases in serum iron indices, expression of liver hepcidin gene and total tissue iron content compared with control rats. We also showed that total tissue iron content was mainly associated with the dose of iron whereas serum iron indices depended essentially on the duration of iron administration. However, no differences in tissue inflammatory and antioxidant parameters from controls were observed. Furthermore, only rats exposed to daily iron injection at a dose of 75 mg/kg body weight for one week revealed a significant increase in lipid peroxidation in iron-treated rats compared with their controls. The present results suggest a correlation between iron overload levels and the dose of iron, as well as the duration and frequency of iron injection and confirm that iron sucrose may not play a crucial role in inflammation and oxidative stress. This study provides important information about iron sucrose-induced iron overload in rats and may be useful for iron sucrose therapy for iron deficiency anemia as well as for the prevention and diagnosis of iron sucrose-induced iron overload in pediatric patients.

Enhancement of Diesel Soot Combustion with Oxygen on Particulate Filters After Injection of Dicyclopentadienyl Iron (Ferrocene) in the Exhaust Pipe

Enhancement of Diesel Soot Combustion with Oxygen on Particulate Filters After Injection of Dicyclopentadienyl Iron (Ferrocene) in the Exhaust Pipe

Effects of the pyrones, maltol and ethyl maltol, on iron absorption from the rat small intestine

The pyrones, 3-hydroxy-2-methyl-4-pyrone (maltol) and 3-hydroxy-2-ethyl-4-pyrone (ethyl maltol) chelate iron with a high affinity and selectivity. The resulting 1:3 (metal-ligand) complexes, being neutral, are able to partition readily across cell membranes and thus may facilitate iron transport across the intestinal wall. Absorption of radioactive iron (59Fe) in the presence of these pyrones was investigated in male rats 1, 2, 4 and 6 h after intraduodenal administration of a 7 micrograms dose and compared with that of 59Fe given as the sulphate, gluconate, fumarate or complexed to EDTA. Total body absorption and distribution were calculated from the 59Fe content of various tissue samples. With all the iron preparations used, blood levels of 59Fe were highest 1 h after injection whilst the 59Fe content at the major site of deposition, i.e. the bone marrow, increased up to 6 h. No 59Fe was found in the urine. Total body absorption of 59Fe was significantly higher from the pyrones than from the other four preparations. Over the dose range 0.7-700 micrograms, the proportion of 59Fe absorbed from both iron maltol and iron sulphate decreased with increasing dose. Enhanced 59Fe uptake from maltol was evident at 0.7-70 micrograms but not at 700 micrograms suggesting that use of these pyrones will not result in iron overload. Absorption of 59Fe given into the stomach was slower in onset but was sustained longer presumably via a steady delivery of iron to the duodenum from the gastric reservoir.(ABSTRACT TRUNCATED AT 250 WORDS)

Self-managing anaemia at home

After being diagnosed with chronic kidney disease and anaemia, John Pearce was taught how to self- administer erythropoiesis-stimulating agents (ESAs) and iron injections so that he could treat himself at home—a vital timesaver and relief from the stresses of having to get to the hospital.

Haemoglobin Mass in an Anaemic Female Endurance Runner Before and After Iron Supplementation

Haemoglobin mass in a female endurance athlete was measured via carbon monoxide rebreathing upon diagnosis of iron-deficiency anemia (haemoglobin concentration = 8.8 g/dL, ferritin = 9.9 ng/mL) and regularly during treatment thereafter. Haemoglobin mass increased by 49% in the 2 wk following an intramuscular iron injection and continued to increase with oral iron supplementation for 15 wk. The presented case illustrates that haemoglobin mass is readily responsive to iron supplementation in a severely iron-deficient anemic athlete and that changes can be tracked efficiently using the CO-rebreathing method.

Comparison of oral versus parenteral iron supplementation on the health and productivity of piglets

The aim of this study was to compare oral iron supplementation in feed with a standard iron injection and assess the impact on the health and productivity of pigs. The...

Permanent tattoos after parenteral over the counter iron injections for anemia: A case report

Permanent tattoos after parenteral over the counter iron injections for anemia: A case report

Age-Dependent Retinal Iron Accumulation and Degeneration in Hepcidin Knockout Mice

Iron dysregulation can cause retinal disease, yet retinal iron regulatory mechanisms are incompletely understood. The peptide hormone hepcidin (Hepc) limits iron uptake from the intestine by triggering degradation of the iron transporter ferroportin (Fpn). Given that Hepc is expressed in the retina and Fpn is expressed in cells constituting the blood-retinal barrier, the authors tested whether the retina may produce Hepc to limit retinal iron import. Retinas of Hepc(-/-) mice were analyzed by histology, autofluorescence spectral analysis, atomic absorption spectrophotometry, Perls' iron stain, and immunofluorescence to assess iron-handling proteins. Retinal Hepc mRNA was evaluated through qPCR after intravitreal iron injection. Mechanisms of retinal Hepc upregulation were tested by Western blot analysis. A retinal capillary endothelial cell culture system was used to assess the effect of exogenous Hepc on Fpn. Hepc(-/-) mice experienced age-dependent increases in retinal iron followed by retinal degeneration with autofluorescent RPE, photoreceptor death, and subretinal neovascularization. Hepc(-/-) mice had increased Fpn immunoreactivity in vascular endothelial cells. Conversely, in cultured retinal capillary endothelial cells, exogenous Hepc decreased both Fpn levels and iron transport. The retina can sense increased iron levels, upregulating Hepc after phosphorylation of extracellular signal regulated kinases. These findings indicate that Hepc is essential for retinal iron regulation. In the absence of Hepc, retinal degeneration occurs. Increases in Hepc mRNA levels after intravitreal iron injection combined with Hepc-mediated decreases in iron export from cultured retinal capillary endothelial cells suggest that the retina may use Hepc for its tissue-specific iron regulation.

Effects of Nano Zero-Valent Iron on Oxidation−Reduction Potential

Oxidation-reduction potential (ORP) measurements have been widely used to assess the results of injection of nano zerovalent iron (nZVI) for groundwater remediation, but the significance of these measurements has never been established. Using rotating disk electrodes (RDE) in suspensions of nZVI, we found the electrode response to be highly complex but also a very sensitive probe for a range of fundamentally significant processes. The time dependence of the electrode response reflects both a primary effect (attachment of nZVI onto the electrode surface) and several secondary effects (esp., oxidation of iron and variations in dissolved H2 concentration). At nZVI concentrations above ∼200 mg/L, attachment of nZVI to the electrode is sufficient to give it the electrochemical characteristics of an Fe(0) electrode, making the electrode relatively insensitive to changes in solution chemistry. Lower nZVI concentrations give a proportional response in ORP, but much of this effect is mediated by the secondary effects noted above. Coating the nZVI with natural organic matter (NOM), or the organic polymers used to make stabile suspensions of nZVI, moderates its effect on ORP measurments. Our results provide the basis for interpretating ORP measurements used to characterize the results of injecting nZVI into groundwater.

Safety and Efficacy of Intravenous Iron Therapy in Postnatal Patients with Iron Deficiency Anemia

ABSTRACT Objective This prospective study was conducted to establish safety and efficacy of intravenous iron therapy in postnatal anemic patients. Results Around 217 anemic women were included in the study. Most of these patients (82.2%) were of 21 to 30 years of age having parity of 2 to 4 (53.9%). Almost 71% of patients had moderate anemia with Hb of 7 to 9 gm/dl. Around 208 patients (95.86%) required two to four injections of intravenous iron to build up Hb to optimum acceptable level. Only seven patients (3.22%) suffered from minor complications of local rash, while three (1.38%) had more severe allergic reactions but were managed successfully. Conclusion Intravenous iron is a safe and effective alternative in postnatal patients as it ensures iron therapy in low resource and noncompliant patients.

Acute Oxidative Stress following Intravenous Iron Injection in Patients on Chronic Hemodialysis: A Comparison of Iron-Sucrose and Iron-Dextran

Background/Aims: Intravascular iron release from parenteral iron formulations can catalyze various harmful oxidative reactions. These topics have mainly been investigated in vivo with iron-sucrose (IS). The aim of our study was to compare IS and iron-dextran (ID) regarding iron release and induction of oxidative stress. Methods: Twenty hemodialysis patients were enrolled. Plasma iron and oxidative stress parameters were measured before and 10 min after intravenous (IV) injection of 100 mg IS and ID, respectively. Results: Intravascular iron increased significantly more after IS. The mean ± SD changes in plasma iron were 314 ± 172% after IS versus 90 ± 44% after ID (p < 0.001). Comparable changes in non-transferrin bound iron were 86 ± 42 versus 45 ± 45%, respectively (p < 0.05). Plasma ascorbic free radical did not change following ID (–1.8 ± 11.2%) but increased by 29 ± 31.3% after IS (p < 0.01). Further, protein carbonyls increased following IS (p < 0.05) but not after ID. Conclusion: Increased oxidative stress was found only after IV injection with IS.

Early morbidity encountered in the dietary-related mouse model of Barrett's esophagus: a question of zinc?

Recently, a mouse model for Barrett's esophagus based on a zinc-deficient diet supplemented with deoxycholic bile acids has been published. The aim of this study was to attempt to reproduce these data and extend them by employing genetically modified mice and intraperitoneal iron supplementation. The study design encompassed six experimental groups (wild type, Apc-mutant and Smad4-mutant mice, with or without iron injections), with all animals fed with the zinc-deficient diet supplemented with deoxycholic bile acids. All treatments were started at 3-5 weeks of age (the majority [78%] at 5 weeks). Animals were scheduled for euthanasia at two distinct time points, namely at 3 and 6 months of age. All mice showed signs of considerable distress already 4 weeks after the start of the modified diets, and had to be euthanized before the first evaluation time point (mean age 9.3 weeks, range 5-15 weeks). No differences were observed between wild type and genetically modified mice, or between animals with or without iron supplementation. On histological examination, we could not detect any lesions (Barrett's esophagus-like or tumors) other than esophagitis. In the currently presented experimental settings, we were not able to reproduce the mouse model according to which Barrett's-like lesions could be detected in animals fed with the zinc-deficient diet supplemented with deoxycholic bile acids.

Depletion of luminal iron alters the gut microbiota and prevents Crohn's disease-like ileitis

BackgroundIron replacement therapy is a common treatment in patients with anaemia and Crohn's disease, but oral iron supplements are less tolerated. The pathogenesis of Crohn's disease is attributed to intestinal...