Administration Of Iron Dextran Research Articles

Management of a Right Tibial Proximal Third Open Comminuted Fracture: Surgical Disarticulation of Right Stifle Joint in a 1-Year-Old Hen Pigeon (Columba livia domestica)

Traumatic pelvic limb fractures in pet birds are rare and primarily result from trauma. Avian bones are fragile and prone to fragmentation due to various natural and anthropogenic events. This case report describes the diagnosis, anesthesia protocol, surgical management, post-operative care, and remarkable outcome of a right tibial comminuted fracture in a one-year-old hen pigeon (Columba livia domestica). A one-year-old hen pigeon was presented to the Veterinary Teaching Hospital, University of Jos, Nigeria, with a broken right pelvic limb after being attacked by a young lad. The attack occurred 30 minutes prior to presentation. Clinical examination revealed extensive tissue damage including an open wound with a complete fracture and bone fragment at the proximal third of the right tibia, accompanied by hemorrhages, tenderness, pale ocular mucous membranes, and lethargy. The impact of the attack was the primary cause of the comminuted fracture. Due to logistical constraints and the patient's critical condition, pre-operative radiographic imaging was not feasible. A definitive diagnosis of right tibial proximal third open comminuted fracture was readily made based on the gross tissue damage. Following standard aseptic protocol, stifle disarticulation was performed under local anesthesia to alleviate the bird's excruciating pain. Local anesthesia was achieved with 10mg lidocaine hydrochloride via local infiltration at the stifle. Due to the bird's critical condition, local anesthesia was preferred over general anesthesia. A 5mm stab skin incision was made distal to the right stifle, followed by blunt dissection of the musculature and disarticulation at the tibiofemoral joint. 5mm musculature distal to the stifle was transected and the skin was routinely sutured with size 3/0 silk suture. Post-operative care included injectable amoxicillin, piroxicam, and iron dextran administration, along with alternative-day wound dressing. Skin sutures were removed eight days post-surgery. The pigeon made a remarkable recovery, regaining balance and flying comfortably. Post-recovery imaging was not feasible as all arrangements made with the client following the bird's recovery ultimately fell through. To our knowledge, this case represents the first reported instance of a comminuted tibial fracture in a pigeon managed with stifle disarticulation. The successful outcome of this case proves stifle disarticulation to be an effective treatment option for complex fractures in birds, underscoring the importance of innovative strategies, prompt surgical intervention and addressing a significant gap in the literature. At the time of this report, the pet bird is currently nesting and brooding its eggs. Keywords: Hen pigeon, Comminuted tibial fracture, Proximal third, Stifle disarticulation.

Intravenous administration of iron dextran as a potential inducer for hemochromatosis: Development of an iron overload animal model.

Iron overload in transfusion-dependent thalassemia patients represents a significant public health challenge due to its high mortality rate and risks of severe complications. Therefore, developing safe and effective therapeutic modalities for managing iron overload is critical, as current animal models inadequately replicate human conditions. The aim of this study was to investigate the effects of intravenous iron dextran on hepatocyte morphology, liver iron concentration, and serum iron profile changes as a model for hemochromatosis. An experimental design with a post-test-only control group method was conducted using animal models. Fifty rats were used and divided into ten groups, nine received different intravenous doses of iron dextran: 10, 20, 30, 40, 50, 60, 80, 100, and 120 mg/kg body weight (BW) and a control group received no treatment. The results showed that intravenous iron dextran starting at a dose of 10 mg/kg BW caused significant changes in liver iron concentration while starting at 20 mg/kg BW significantly affected hepatocyte morphology, transferrin levels, unsaturated iron binding capacity, serum iron levels, and transferrin saturation. Intravenous iron dextran starting at 40 mg/kg BW resulted significant changes in the level of total iron binding capacity compared to control group. In conclusion, intravenous iron dextran significantly altered hepatocyte morphology, increased liver iron concentration, and modified the serum iron profile, reflecting changes that might be observed in patients with transfusion-dependent thalassemia.

Combined Injection for Control of Iron-Deficiency Anemia and Coccidiosis in Piglets Decreases Stress at Management Time.

The aim of the present study was to assess the short-term behavioral and physiological responses of piglets to different treatment protocols for the control of iron-deficiency anemia (IDA) and cystoisosporosis. Piglets were treated with either (1) an injection of iron combined with an oral application of toltrazuril (TLZ) by drenching or (2) a combination injection of TZL + gleptoferron; the behavior of the piglets was then evaluated. For this study, 288 piglets were divided into three experimental groups: 96 piglets were kept untreated (control group); 96 piglets received an oral administration of a generic TZL-based anticoccidial agent (20 mg/kg BW) along with intramuscular administration of iron dextran (200 mg/mL; 1 mL/piglet) at the same handling (oral + parenteral group, O + P); and 96 piglets received an intramuscular application of the combination product (parenteral group, P). For each treated piglet, the total handling time, flight reaction, and the intensity and frequency of vocalizations were determined using the methodology described by Scollo et al. (2020). Piglets in the O + P group were found to emit more screams during treatment administration than animals in the P group (21.05% vs. 8.42% of animals; p < 0.05). Piglets in the O + P group reacted worse to manipulation and oral administration because a higher percentage of animals continued to fidget even after handling (32.63% vs. 12.63%; p < 0.05). Differences in growth performance between the groups were not observed in our study (p > 0.05). In conclusion, the administration of a combination product reduced stress during administration, as indicated by reduced vocalizations and reactions to manipulation.

Exogenous iron caused osteocyte apoptosis, increased RANKL production, and stimulated bone resorption through oxidative stress in a murine model

Iron overload is a risk factor for osteoporosis due to its oxidative toxicity. Previous studies have demonstrated that an excessive amount of iron increases osteocyte apoptosis and receptor activator of nuclear factor κ-B ligand (RANKL) production, which stimulates osteoclast differentiation in vitro. However, the effects of exogenous iron supplementation-induced iron overload on osteocytes in vivo and its role in iron overload-induced bone loss are unknown. This work aimed to develop an iron overloaded murine model of C57BL/6 mice by intraperitoneal administration of iron dextran for two months. The iron levels in various organs, bone, and serum, as well as the microstructure and strength of bone, apoptosis of osteocytes, oxidative stress in bone tissue, and bone formation and resorption, were assessed. The results showed that 2 months of exogenous iron supplementation significantly increased iron levels in the liver, spleen, kidney, bone tissue, and serum. Iron overload negatively affected bone microstructure and strength. Osteocyte apoptosis and empty lacunae rate were elevated by exogenous iron. Iron overload upregulated RANKL expression but had no significant impact on osteoprotegerin (OPG) and sclerostin levels. Static and dynamic histologic analyses and serum biochemical assay showed that iron overload increased bone resorption without significantly affecting bone formation. Exogenous iron promoted oxidative stress in osteocytes in vivo and in vitro. Additional supplementation of iron chelator (deferoxamine) or N-acetyl-l-cysteine (NAC) partially alleviated bone loss, osteocyte apoptosis, osteoclast formation, and oxidative stress due to iron overload. These findings, in line with prior in vitro studies, suggest that exogenous iron supplementation induces osteoclastogenesis and osteoporosis by promoting osteocyte apoptosis and RANKL production via oxidative stress.

Myeloid Specific HO-1-SIRT1-p53 Nexus Protects Against Iron Induced Chronic Liver Injury in Sickle Cell Disease

Sickle cell disease (SCD) is an autosomal-recessive-genetic disorder that affects ~100,000 Americans and millions of people worldwide. People with SCD experience multiple complications, including acute pain episodes, stroke, and organ damage, with a median life expectancy of 42-47 years in the United States. Red blood cell transfusions are an important therapeutic intervention in SCD, however, chronically transfused SCD patients develop severe iron overload in the liver, heart, spleen, and endocrine organs with increased expression of inflammatory markers. In addition, there is evidence of abnormal iron metabolism in SCD with prior studies showing a low urinary hepcidin level in children with SCD and highly variable levels in adults. Thus, hepatic iron homeostasis in SCD both at baseline and upon repeated blood transfusion remains incompletely understood. In this study we evaluated iron homeostasis in Townes sickle cell mice at baseline and upon iron overload by iron dextran administration. We also evaluated the potential contribution of hepatic macrophages in hepatic iron homeostasis. Townes mice exhibited significant iron accumulation in the liver at baseline. We found significant reduction of hepcidin and increase in ferroportin and ferritin levels in the liver of sickle mice at baseline. However, upon hepatic iron overload (which was generated by intraperitoneal injections of iron dextran (1 g/kg) once a week for upto 10 weeks) the hepcidin ferroportin homeostasis was impaired and we found reduction of ferroportin and ferritin and increase in hepcidin in the sickle mouse liver. Immunohistochemically we could see a significant enrichment of iron loaded macrophages in the liver of SCD mice at baseline. Clodronate liposome mediated depletion of hepatic kupffer cells caused significant liver injury, hepatocyte death, systemic and hepatic iron accumulation and mortality in the mouse liver at baseline suggesting a protective role of hepatic kupffer cells in iron homeostasis. Mechanistic analysis further revealed protective mechanism through myeloid specific heme oxygenase 1 (HO-1)-sirtuin -1 (SIRT1)-p53 signaling that can potentially alleviate iron induced chronic liver injury in the SCD mice. At baseline, HO1 generated by kupffer cells stimulated SIRT1 activation in SCD mouse hepatocytes. Furthermore, HO-1 and SIRT1 prevented iron-induced senescence and hepatocyte cell death via regulating P53 signaling. Clodronate liposome-mediated kupffer cell loss drastically lowered HO-1 expression, which in turn reduced SIRT1 in SCD liver and accelerated senescence and cell death. In line with this finding, we also detected elevated levels of SIRT-1 in SCD patients’ blood serum. Work is currently underway to understand how modulation of HO-1-SIRT1-p53 signaling pathway can be used as a therapeutic option to combat iron induced chronic liver injury.

Intracellular Iron Overload Induces Metabolic Switching in Active Hematopoietic Stem Cells in Beta-Thalassemia

Beta-thalassemia (BThal) is a complex monogenic disorder, causing severe anemia, ineffective erythropoiesis and multi-organ secondary defects. Iron overload (IO), associated with ineffective erythropoiesis and therapeutic blood transfusions, is one of the main complications of the disease, leading to morbidities in various organs. Despite the improvement in chelation therapies, IO toxicity is still a relevant issue. Correction of BThal is achieved by transplantation of hematopoietic stem cells (HSC) from healthy donors or autologous HSC from patients upon gene therapy. The quality and the engraftment of HSC depend on the bone marrow (BM) microenvironment, thus niche-HSC crosstalk plays a crucial role for transplantation outcome. We provided the first demonstration of impaired function of HSC caused by an altered BM niche in BThal (Aprile et al., Blood 2020). HSC from Hbbth3/+ (th3) BThal mice showed more active cycling profile and response to stress. We reported that IO reduces the hematopoietic supportive capacity of BM mesenchymal stromal cells from patients and recent evidence highlighted that both IO and iron deficiency are sources of cellular stress detrimental to HSC maintenance. Thus, we hypothesized that accumulation of iron might directly affect HSC function in BThal, as a model of chronic IO. Transcriptome analysis of sorted HSC from th3 mice showed a positive enrichment of genes involved in iron homeostasis, such as TfR1, Steap3, Fth1 and Ftl1, suggesting iron uptake and storage. Consistently, BThal HSC displayed higher levels of intracellular free reactive iron, as compared to wild-type (wt) controls (1.7±0.2 calcein MFI fold to wt, p<0.0001). We found that iron specifically accumulates in the mitochondria causing mitochondrial defects, as reduced size and lower membrane potential (0.4±0.05 TMRE/MTG fold to wt, p<0.001). These data, along with the downregulation of mitochondrial biogenesis and mitophagy genes, indicate an accumulation of damaged mitochondria in BThal HSC. In vivo administration of iron dextran to wt mice generated intracellular IO in HSC and was sufficient to decrease the mitochondrial membrane potential at levels comparable to those of th3 HSC, thus suggesting a direct effect of iron on mitochondrial activity. Since mitochondria are the powerhouse of the cell, we wondered if the mitochondrial dysfunction of BThal HSC could affect their production of metabolic energy. Metabolism indeed control HSC function: active HSC are more cycling and, because of their higher energy request, change the metabolism from glycolysis to mitochondrial oxidative phosphorylation (OXPHOS). Interestingly, we found that ATP levels in th3 HSC are lower compared to controls (wt 9.8±2.6 vs. th3 3.7±0.3 ATP nM, p<0.01). In line with mitochondrial dysfunction, in vitro inhibition of OXPHOS did not affect ATP content of BThal HSC since they switch to glycolysis for their energy demand. Consistently, th3 HSC showed positive enrichment of glycolytic genes and higher glucose uptake (wt 1639±163 vs. th3 2401±85 MFI 2NBDG, p<0.05). To unravel the basis of mitochondrial damage caused by IO, we investigated the role of reactive oxygen species (ROS). Iron is one of the sources of ROS and consistently, th3 HSC showed increased activation of antioxidant genes and higher levels of mitochondrial ROS (wt 2507±756 vs. th3 5205±459 MFI MitoSOX, p<0.01). In vivo reduction of mitochondrial ROS by MitoQ rescued mitochondrial activity (th3 0.3±0.07 vs. th3+MitoQ 0.9±0.2 TMRE/MTG fold to wt, p<0.05), thus demonstrating that mitochondrial dysfunction is reversible in BThal. Ongoing experiments will assess the metabolite profile, the restoration of HSC metabolism and in vivo function upon MitoQ and will also test HSC from BThal patients for mitochondrial defects. Overall, our study revealed that the multisystemic complications of IO in BThal also affect HSC. Iron has a direct impact on HSC metabolism by inducing oxidative stress and mitochondrial dysfunction. Despite the active cycling state, BThal HSC switch their metabolism to glycolysis because of the mitochondrial defect. Targeting mitochondrial iron or ROS might provide new tools to preserve HSC function in the setting of autologous transplantation upon gene therapy/editing for BThal. Moreover, this research will add novel insight about the regulation of HSC biology linking iron to mitochondrial metabolism.

Ferroptosis contribute to hepatic stellate cell activation and liver fibrogenesis.

Ferroptosis is a widely known regulator of cell death in connection with the redox state as a consequence of the depletion of glutathione or accumulation of lipid peroxidation. Hepatic stellate cells (HSCs) play a pivotal role in the progression of hepatic fibrosis by increasing the production and secretion of the extracellular matrix. However, the role of ferroptosis in HSC activation and liver fibrogenesis has not been clearly elucidated. The ferroptosis inducer RAS-selective lethal 3 (RSL3) or erastin treatment in HSCs caused cell death. This effect was suppressed only after exposure to ferroptosis inhibitors. We observed induction of ferroptosis by RSL3 treatment in HSCs supported by decreased glutathione peroxidase 4, glutathione deficiency, reactive oxygen species generation, or lipid peroxidation. Interestingly, RSL3 treatment upregulated the expression of plasminogen activator inhibitor-1, a representative fibrogenic marker of HSCs. In addition, treatment with ferroptosis-inducing compounds increased c-JUN phosphorylation and activator protein 1 luciferase activity but did not alter Smad phosphorylation and Smad-binding element luciferase activity. Chronic administration of iron dextran to mice causes ferroptosis of liver in vivo. The expression of fibrosis markers, such as alpha-smooth muscle actin and plasminogen activator inhibitor-1, was increased in the livers of mice with iron accumulation. Hepatic injury accompanying liver fibrosis was observed based on the levels of alanine aminotransferase, aspartate aminotransferase, and hematoxylin and eosin staining. Furthermore, we found that both isolated primary hepatocyte and HSCs undergo ferroptosis. Consistently, cirrhotic liver tissue of patients indicated glutathione peroxidase 4 downregulation in fibrotic region. In conclusion, our results suggest that ferroptosis contribute to HSC activation and the progression of hepatic fibrosis.

Identification and characterization of warm temperature acclimation proteins (Wap65s) in rainbow trout (Oncorhynchus mykiss)

Hemopexin is a vital glycoprotein for processing excessive iron in blood and functions as an iron scavenger in mammals. Teleosts however, unlike mammals, have two known hemopexin paralogs called warm temperature acclimation-related 65 kDa protein (Wap65-1 and Wap65-2, collectively termed Wap65s). Although Wap65s in rainbow trout have been considered notable biomarkers with significantly higher and/or lower expression under conditions of stress or disease, the individual roles, similarities and differences between the two paralogs are not well known. The aim of this study was to gain an understanding of the characteristics and functions of trout Wap65s from the perspective of iron-metabolism, physiological roles, and relevant immunological responses. The expression of Wap65-1 and −2 in this study was determined in the face of challenges by Aeromonas salmonicida, infectious hematopoietic necrosis virus (IHNV), and iron-dextran. Immuno-histochemistry (IHC) was employed to localize the major cell types for Wap65-2 expression, and trout leukocytes were isolated and incubated with LPS and OxLDL for comprehending the immunological characteristics of Wap65-2. We demonstrate that Wap65-1 is expressed only in the liver but Wap65-2 is systemically expressed in most organs and tissues. Interestingly, Wap65-1 expression was not significantly changed under A. salmonicida and iron-dextran administration, but was significantly decreased under IHNV. In contrast, Wap65-2 was up-regulated in all challenged groups, however with different expression patterns in the blood and liver. These results suggested that the two paralogs may participate in different biological roles. IHC showed that Wap65-2 antibody had high affinity for leukocyte-like cells, and macrophages but not lymphocytes significantly increased expression under LPS and OxLDL stimulation. These results support the conclusion that trout Wap65-2, not Wap65–1 may have conventional hemopexin functions such as reported in mammals including effects on iron metabolism, inflammation, and acute-phase protein.

Influence of the application methods of iron/anticoccidial products on the behaviour and associated stress factors of suckling piglets

The aim of the present study was to evaluate the effect of different treatment protocols for the control of iron-deficiency anaemia and coccidiosis, one combination injection versus injection of iron and oral application of toltrazuril, on the general behaviour of piglets. Piglets were divided into three experimental treatment groups: 24 piglets were left untreated; (control group, C); 24 piglets received an oral administration of generic anticoccidial agent (20 mg/kg BW) plus intramuscular administration of iron dextran (200 mg/ml; 1 ml/piglet) in the same handling (oral + parenteral group, O+P) and 24 piglets received an intramuscular application of the combination product (parenteral group, P). As expected, the handling time was higher in the oral + parenteral group than in the parenteral group. Regarding the behaviour of piglets, the time spent suckling after treatment was variable in the control and oral + parenteral groups, while piglets from the parenteral group spent more time suckling, and hence, their time spent resting was decreased. A statistical trend of lower concentration of cortisol was observed in P compared to O+P after processing. Regarding productivity indices, the average daily gain of piglets during lactation and, subsequently, their weaning weight were higher in P compared to O+P, and similar to the control. In conclusion, administering a combination product injection decreases the time of administration compared to individual treatments.

Naringin is a promising natural compound for therapy of iron-overload disorders

Naringin has been shown to exhibit satisfying iron chelation capacity. Considering the side effects of routinely-used iron chelator (desferrioxamine, DFO), we decided to evaluate the iron chelation potency of naringin to discover whether or not it can be a promising natural substitute for treatment of excessive iron-related diseases. 35 mice were classified into five groups of 7 and subjected to iron dextran administration to induce the iron-overload condition. Iron-overloaded mice were then treated with normal saline (as control), naringin or DFO Morphology changes, and iron deposition in liver tissues were studied using H&E and Perl’s staining. The results revealed that naringin is more potent than DFO in removing excessive iron ions deposited in liver tissues, indicating that naringin is a promising natural compound for therapy of iron overload disorders.

Selenium or ozone: Effects on liver injury caused by experimental iron overload

AimsIron is an important metal ion as a biocatalyst on the other hand iron overload causes various diseases. Iron overload can result in fibrosis and hepatocellular carcinoma with various pathophysiological mechanisms, including oxidative damage in the liver. Therefore; in this study the effects of ozone and selenium -whose antioxidant properties are known- were evaluated in liver injury induced by iron overload. Materials and methodsIron overload model was provided by intraperitoneal administration of 88 mg/kg iron dextrate for 4 weeks. After iron dextran administration, ozone and selenium administrations were made for 3 weeks. From the obtained blood and tissue samples total oxidant status (TOS) and total antioxidant status (TAS) were determined and histopathological examination was performed in liver tissue samples. Key findingsIn rats with iron overload, the lowest mean serum TOS was observed in the selenium administration group. The highest tissue TOS means and the lowest tissue TAS means were determined in the group in which ozone and selenium were administrated together. When histopathological data were evaluated, the presence of increased apoptosis in the ozone group compared to the iron group (p = 0.019) and selenium group (p = 0.019) was noted. Similarly, increased periportal inflammation (p = 0.001) and fibrosis (p = 0.005) were observed in the ozone group compared to the selenium group. SignificanceIn iron-induced liver damage, ozone was thought to be effective by decreasing ROS, but contrary to expectations, it was observed that it may negatively affect the picture by showing synergistic effect. However, the effects of selenium on both serum and tissue levels are promising.

Elucidating the Role of IL6 in Stress Erythropoiesis and in the Development of Anemia Under Inflammatory Conditions

Anemia of inflammation, also known as anemia of chronic disease is the second most common anemia after iron deficiency anemia. The predominant regulators of AI are the cytokine-interleukin-6 (IL6) and the hormone hepcidin (Hamp). IL6 has been implicated in inducing expression of hepcidin. Published data from our lab have shown that lack of IL6 or hepcidin in knockout mouse models (IL6-KO and Hamp-KO) injected with the heat-killed pathogen Brucella abortus(BA) results in recovery from anemia but interestingly the pattern of the recovery was different in IL6-KO and Hamp-KO mice, suggesting that the two proteins contribute independently to AI. Here, we validated the independent role of IL6 and Hamp in AI by generating a double-knockout (DKO) mouse model lacking the expression of both. In the first few days following BA administration, we observed severe reduction in the total number of BM cells in each model followed by a slow recovery in erythroid and multilineage hematopoietic cells. The recovery, initially, was more sustained in the BA-treated-DKO model. In particular, in the first week, BA-treated-DKO mice showed an increased number of erythroblasts in the bone marrow (BM) and spleen as seen in comparison to IL6-KO and Hamp-KO. IL6-KO mice showed an intermediate recovery profile when compared to DKO and Hamp-KO, the last one showing the worst profile in the BM. Interestingly, when the reticulocyte count in the DKO mice was compared to that of IL6-KO and Hamp-KO mice, it showed a biphasic trend, with a significant increase in number during the 2nd week, followed by a significant reduction during the 3rd week. We hypothesized that the initial surge in reticulocyte count in DKO was due to lack of hepcidin, which increases iron availability to erythroid cells, and concurrent lack of IL6, which favors BM erythropoiesis in presence of inflammatory stimuli. However, we also speculated that the excess of iron (as NTBI), which accumulates during the first two weeks, leads to oxidative stress and erythroid cell death in presence of inflammatory cytokines, despite the absence of IL6. We also surmised that, during the second week, a second wave of inflammatory cytokines is triggered by the adaptive response in response to the BA that would explain the negative effect on erythropoiesis after the initial recovery. To assess this hypothesis, we utilized an inflammation panel to analyze the cytokine expression in WT animals treated with PBS or BA at 6 hours, 24 hours and then around ~2 weeks. The cytokine levels were normalized after 24 hours. However, around two weeks, we observed a novel surge of cytokines such as IFN-g and TNFa in the BA treated mice, indicating their role in innate (immediate effect; 6 hours) and adaptive immune response, which activated a second wave of inflammation (around 2 weeks, during the recovery of hematopoiesis in the BM). Interestingly, while we observed oxidative stress and defective erythropoiesis in the bone marrow, this was not seen in the spleen, where increased and extramedullary erythropoiesis sustained some level of RBC production. Since the BA-treated-IL6-KO did not show any major defect in the BM after two weeks, we challenged them with administration of iron dextran. Upon treatment, also the IL6-KO mice treated with both BA and iron dextran shown increased production of reactive oxygen species as well as a defect in bone marrow erythropoiesis, similarly as in DKO or Hamp-KO mice, thereby explaining the plausible reason of reduced erythropoiesis in the bone-marrow. Furthermore, to identify mechanisms leading to oxidative stress, we established an in-vitro culture system where primary murine bone marrow cells were cultured for 18-20 hours in presence of serum isolated after 6hrs from either PBS treated or BA treated C57BL/6 mice. With the help of confocal microscopy, we observed an increase in mitochondrial superoxide in the cells treated with BA serum; interestingly we have also seen a decrease in Ter 119 population in the cells cultured with BA treated serum implicating that the erythroid cells are dying. To further investigate the downstream players related to the death of erythroid progenitors we are currently investigating the role caspase 1 (a major regulator in pyroptosis) and Gata-1. In conclusion, this study is elucidating some of the mechanisms associated with the anemia triggered by inflammation with the potential to identify new targets and treatments. Disclosures Rivella: Disc medicine, Protagonist, LIPC, Meira GTx: Consultancy; Meira GTx, Ionis Pharmaceutical: Membership on an entity's Board of Directors or advisory committees.

Protective Effect of Astragaloside IV on Hepatic Injury Induced by Iron Overload.

Suitable content of iron is essential for human body, but iron overload is associated with many kinds of diseases including chronic liver damage. Recently, researchers find that iron overload promotes hepatocyte autophagy and apoptosis. However, the mechanism of iron overload in liver damage remains unclear. In this study, Lo2 cells were selected as the research object, iron dextran was a model drug, and astragaloside IV was a therapeutic drug to explore the role of iron overload. MTT assay and Annexin/PI double staining were used to measure cell viability and apoptosis. Ultrastructure was observed by transmission electron microscopy. The expression levels of apoptosis and autophagy-related proteins were determined by real-time PCR and Western Blot. The results showed that iron dextran could significantly inhibit Lo2 cell viability and increase the apoptosis rate, while astragaloside IV could reverse the inhibition of Lo2 cell viability and decrease the apoptosis rate. Transmission electron microscopy showed a significant increase in the number of autophagosomes after administration of iron dextran, and the application of astragaloside IV reduced the production of autophagosomes. LC3II/I was significantly upregulated in the model group but decreased in the astragaloside IV treatment group, and P62 showed the opposite trend. Iron dextran significantly upregulated the expression of Bax and downregulated Bcl2, while astragaloside IV reversed this trend. Finally, the inhibition of hepcidin caused by iron dextran was counteracted by astragaloside IV. In conclusion, the experimental results show that the iron overload model mainly induces excessive autophagy and apoptosis of hepatocytes, thus causing damage to hepatocytes, but astragaloside IV plays a certain therapeutic role in reversing this damage.

Hazardous effect of the maintenance dose of iron sucrose versus iron dextran on oxidative burst activity of neutrophils in patients on regular hemodialysis; a pilot study

Introduction: Intravenous (IV) iron which is a potent pro-oxidant may decrease the phagocytic function of neutrophils and subsequently leads to repeated bacterial infections in patients on chronic regular hemodialysis (HD).Objectives: We aimed to evaluate the effect of IV iron dextran versus iron sucrose on oxidative burst activity of neutrophils in HD patients.Patients and Methods: A crossover study was included 20 prevalent HD patients were randomly divided into two groups; 10 patients each. Each group received a single dose of 100 mg iron sucrose or iron dextran then shifted to the other type after 4 weeks as a washout period. Patients with evidence of acute infections, diabetes mellitus, chronic liver disease, active collagen disease or any other acquired immune deficiency diseases were excluded. Patients on immunosuppressive drugs or with hypersensitivity to iron therapy or evidence of iron overload (TSAT &gt; 50% and/or ferritin &gt; 800 ng/mL) were also excluded. Erythropoietin maintenance dose was given to all patients. Urea reduction ratio (URR %), C-reactive protein (CRP) titer, complete blood picture and iron study were conducted. Neutrophil oxidative burst test was performed by flow-cytometry with an estimation of stimulation index before and after IV iron dosage. Results: Twenty patients (9 males, 11 females; mean age 49.40 ± 9.02 years) on chronic HD for a mean time of 10 ± 7.32 years. After 100 mg of IV iron infusion in HD patients, a highly significant difference in oxidative burst test of neutrophil cells before and after administration of iron regardless of its type was detected (P &lt; 0.01). This change over time is not significantly different between two types of iron (P &gt; 0.05). However, a significant increase in CRP titer after administration of iron dextran was seen (P = 0.016). Conclusion: Maintenance dosage of either IV iron sucrose or iron dextran (100 mg) had a hazardous effect on the neutrophilic phagocytosis demonstrated by the significant rise in the oxidative burst index in HD patients.

Oral iron administration in suckling piglets – a review

Iron deficiency is presently a serious problem in suckling piglets on pig farms. The most often used method of anaemia prevention in piglets is parenteral administration of iron dextran. Oral iron represents an alternative to this method. The goal of this article is to review current knowledge on oral iron administration in suckling piglets. The substances that can be used for this purpose include iron dextran, iron salts, iron chelates, carbonyl iron, an iron polymaltose complex and iron microparticles. The different methods of oral iron administration in piglets are discussed.

Supplemental iron dextran injections: Influence on hemoglobin concentrations and piglet growth

This case study assessed effectiveness of protocols for iron dextran administration on hemoglobin (Hb) concentrations in pigs and evaluated the effect of supplemental iron dextran injections prior to weaning on subsequent body weights (BW). Whole blood samples and body weights were collected from piglets at 14 days of age in four farms and at 27 days of age in the fifth farm. For Farms 1 to 3, six piglets per litter were matched by BW to provide a pair of heavy weight (HW) piglets, medium weight (MW) piglets, and light weight (LW) piglets in each litter. For Farms 4 and 5, MW piglets were not included. One piglet from each pair was injected intramuscularly with 200 mg iron dextran immediately after blood collection (treatment pigs). The other piglet in each pair served as the control. At 3 weeks after weaning, pigs were weighed and whole blood samples were collected to determine Hb concentrations. At 14 days of age and after weaning, the results were inconsistent among the farms. Prior to recommending supplemental iron injections for pigs, one must evaluate the existing on-farm protocol for iron administration.

Total dose iron dextran infusion versus oral iron for treating iron deficiency anemia in pregnant women: a randomized controlled trial

Study objective: To test safety, efficacy, and cost-effectiveness of total dose infusion (TDI) of low molecular weight (LMW) iron dextran for treatment of iron deficiency anemia (IDA) during pregnancy in comparison to oral ferrous fumarate.Design: Prospective interventional randomized controlled trial (RCT). Design classification. Canadian Task Force II3.Setting: Antenatal clinic and causality unit of a tertiary care referral facility and University Hospital.Patients: A total 66 anemic pregnant women (hemoglobin level between 7–10 g/dl).Intervention: Administration of a LMW iron dextran as a TDI (group A) or Oral iron ferrous fumarate 60 mg elemental iron three times daily (group B) followed by remeasurement of hemoglobin after 4 weeks.Measures and main results: The main outcome measure was clinical and laboratory improvement of anemia after 4 weeks of starting the therapy. Both groups showed a significant clinical improvement of anemia 4 weeks post-therapy. However, the first improvement of symptoms was significantly faster in group A. Complete blood count (CBC) as well as all iron indices were improved in both groups after 4 weeks of therapy, but were significantly better in group A than B. Side effects in group B were mainly gastrointestinal (GIT) while one case of mild hypersensitivity to TDI and another one case of local reaction at the site of injection were reported in group A.Conclusions: It is concluded that despite being equally effective in improving both clinical and laboratory evidence of IDA, TDI allows iron restoration with a single dose faster than oral iron therapy with a reasonable safety profile. It is a good example of office one-stop therapy. Nevertheless, noninvasive selfusage at home is a clear advantage of the cheaper oral iron therapy which makes it the first choice for treating IDA in the second and third trimesters of pregnancy in tolerable cases.Trial registration: ClinicalTrials.gov identifier: NCT03212781.

Safety and efficacy of iron dextran as a feed additive for piglets.

The additive, ‘Iron dextran 10%’, is a liquid preparation containing iron dextran (25%, of which 10% is total iron), sodium chloride (1.5%), phenol (0.4%) and water (73.1%). Iron dextran 10% is considered safe for suckling piglets when given at an oral dose of 1 mL/kg body weight (bw) once in each of the first 2 weeks of life; this dose corresponds to 100 mg Fe/kg bw. The administration of iron dextran 10% to piglets deficient in vitamin E and/or selenium is considered a risk. The oral use of iron dextran 10% in suckling piglets does not pose any safety concerns to consumers, provided that the conditions identified as safe for the target animal are respected. Iron dextran 10% is considered a respiratory sensitiser and may be harmful if inhaled; however, exposure by inhalation is not expected; therefore, the risk is considered to be negligible. Iron dextran 10% is an irritant to skin and eyes; a risk by skin sensitisation cannot be excluded. The presence of phenol in the additive should be considered to pose a hazard for users. The use of the additive for suckling piglets at the proposed level does not pose a safety concern to the environment. The available studies with orally administered iron dextran indicate that the additive is a bioavailable source of iron for suckling piglets; however, the efficacy of the additive when given to newborn pigs as proposed via water for drinking (voluntary intake) has not been demonstrated. The Panel proposed some recommendations regarding the dextran used in the manufacture of the additive and the procedure for administering the additive to piglets.

Glycoside rich fraction from Spondias pinnata bark ameliorate iron overload induced oxidative stress and hepatic damage in Swiss albino mice.

BackgroundIron in the overloaded condition in liver promotes the overproduction of free radicals that lead to oxidative stress and ultimately hepatic damage. The present study was designed to evaluate the ameliorating potential from iron overloaded hepatotoxicity by the glycosidic fraction from Spondious pinnata bark (SPW1) along with its antioxidant property.MethodsThe fraction was tested for its in vitro antioxidant, free radical scavenging property and iron chelation potential via standard biochemical assays. Iron overload condition was generated by the intraperitoneal administration of iron dextran in mice. The levels of serum enzymes, antioxidant enzymes in liver, markers of hepatic damage, liver iron, and ferritin content were measured in response to the oral treatment of SPW1. Histopathology of the liver sections was performed for visual confirmation of the amelioration potential of SPW1.ResultsThe fraction exhibited excellent in vitro antioxidant as well as free radical scavenging potential against both reactive oxygen species and reactive nitrogen species. Administration of SPW1 significantly normalized the disturbed levels of antioxidant enzymes, liver iron, lipid peroxidation, liver fibrosis, serum enzyme and ferritin better than standard desirox which were also supported by the morphological study of the liver sections. Phytochemical analysis as well as HPLC study, confirmed that the fraction mainly consisted of glycosidic phenolics and flavonoids that attributed to its biological activities.ConclusionsThe above results suggested that beneficial effects of SPW1 on iron overload induced hepatotoxicity that can be considered as a possible candidate against iron overload diseases.

Catherine Pasquier Memorial Award Lecture (shared): Critical role of oxidative and nitrosative stress in pathophysiology: Inflammation, cell injury and cancer

The presence of chronic inflammation associated with sustained cell death and proliferation turnover is a potential tissue environment for cancer development. In this situation, oxidative and nitrosative stress are the major driving force playing a key role either as a preventing or inducer/promoter of tumorigenesis. During inflammatory response, activated macrophages release a great variety of protease that degraded extracellular matrix, inflammatory mediators that plays a relevant role in chemotaxis and cell signaling, as well as oxygen and nitrogen reactive species (ROS and RNS, respectively) actively involved in cell defense through oxidation of essential biological molecules such as DNA, carbohydrates, proteins and lipids. Iron-loosely bound plays a relevant role in promoting oxidative-dependent reactions leading to cell damage. In this sense, we have shown that locally iron overload induced an accumulation of neutrophils with increased NADPH-derived superoxide anion generation in exudates from rats submitted to carrageenan-induced granuloma model. The increase of lipid peroxidation products was associated with the presence of iron-loosely bound, reduction of antioxidant status in inflammatory exudates and blood, as well as induction of hepatic acute phase response. A similar pro-inflammatory and oxidant reaction was observed by intraperitoneal iron-dextran administration in animals with Mycobacterium butyricum-induced adjuvant arthritis. The administration of a diet devoid of polyunsaturated fatty acids reduced the substrate for lipid peroxidation, and consequently a decrease of conjugated diennes and thiobarbituric acid-reactive substances in plasma and hepatic microsomal fraction were observed in animals with adjuvant arthritis. The acute phase response induced by different inflammatory mediators in hepatocytes has been studied in detail. The induction of carrageenan-induced granuloma drastically reduced the expression of drug metabolizing enzymes (DME) (CYP3A1, CYP4A, CYP2B1, CYP2D and CYP2E1) and their related-activities in rat livers. Cytokines (IL-1α, IL-6 and TNF-α) exerted a specific timely-dependent regulation of DME expression (CYP1A1, CYP1A2 and CYP3A4) and secretion of acute phase reactants (albumin, ferritin, fibrinogen and ceruloplasmin) in cultured primary human hepatocytes. Although TNF-α induces acute phase response and liver injury in different experimental settings such as sepsis, we have also shown that TNF-α-dependent induction of nitric oxide synthase type II (NOS-2) hepatic expression during PGE1 pre-administration drastically reduces D-galactosamine (D-GalN)-induced liver injury in rats. The pre-administration of PGE1 also reduced cell death induced by D-GalN in primary culture of human and rat hepatocytes. However, the cytoprotective properties of PGE1 do not involve a reduction of D-GalN-induced ROS production. Nitric oxide (NO) has demonstrated to exert pro- and anti-apoptotic properties according to local concentration, target cell, as well as the presence of other ROS such as superoxide anion generating high reactive peroxynitrite. We have observed that moderate increase of PGE1-dependent NO prevented further potent NF-kB-dependent NOS-2 expression and apoptosis induced by D-GalN in cultured rat hepatocytes. However, the administration of PGE1 at advanced stages of cell injury or high doses of NO donor increased cell death in D-GalN-treated human and rat hepatocytes. The induction of apoptosis by D-GalN was associated with ROS production, mitochondrial dysfunction and post-translation (S-nitrosylation and Tyrosine nitration) modifications of proteins involved in unfolded protein response, enzymatic antioxidants, cell metabolism and death in cultured human hepatocytes. In consequences, the administration of antioxidants such as N-acetylcysteine, Q10 and MnTBAB reduced ROS generation, mitochondrial dysfunction and apoptosis induced by D-GalN and glycochenodeoxycholate (GCDCA) in cultured human hepatocytes. The cytoprotective properties of α-tocopherol appears to mostly related to gene regulation (NOS-2, PPAR-α and carnitine palmitoyl transferase) rather than to its antioxidant activity in D-GalN-induced cell death in hepatocytes. The reduction of cell death by α-tocopherol was also associated with reduction of CYP7A1, Na+-taurocholate co-transporting polypeptide (NTCP), and heme oxygenase-1 expression, as well as increased NOS-2 expression and NO production in GCDCA-treated hepatocytes. α-Tocopherol and NO donors increased NTCP cysteine S-nitrosylation and Tyrosine nitration, and reduced Taurocholic acid uptake in hepatocytes. The pro-apoptotic properties of high NO donor concentration have been successfully applied to exert antitumoral activity in liver cancer cells. In this sense, the administration of NO donor or NOS-3 overexpression induces p53-dependent cell death receptor expression and apoptosis in hepatoma cell lines. The first generation adenovirus specifically designed to induce NOS-3 overexpression in liver cancer cells induced oxidative and nitrosative stress, DNA damage, p53, cell death receptors expression and their S-nitrosylation, and apoptosis that was correlated to a reduction of cell proliferation and growth of tumors orthotopically implanted in fibrotic livers. The S-nitrosylation of cell death receptors (TNF-α, CD95 and TRAIL-R1) plays a critical role on the shift from extrinsic to intrinsic cell death pathways during antitumoral molecular therapy (Sorafenib Tosylate) in liver cancer cells. In conclusion, new data are emerging that elucidate the extraordinary role that plays ROS- and RNS-dependent posttranslational target modifications in regulating intracellular cell signaling.