Total Iron Levels Research Articles

Endothelin A receptor antagonist attenuated renal iron accumulation in iron overload heme oxygenase-1 knockout mice.

Progressive iron accumulation and renal impairment are prominent in both patients and mouse models of sickle cell disease (SCD). Endothelin A receptor (ETA) antagonism prevents this iron accumulation phenotype and reduces renal iron deposition in the proximal tubules of SCD mice. To better understand the mechanisms of iron metabolism in the kidney and the role of the ETA receptor in iron chelation and transport, we studied renal iron handling in a nonsickle cell iron overload model, heme oxygenase-1 (Hmox-1-/-) knockout mice. We found that Hmox-1-/- mice had elevated plasma endothelin-1 (ET-1), cortical ET-1 mRNA expression, and renal iron content compared with Hmox-1+/+ controls. The ETA receptor antagonist, ambrisentan, attenuated renal iron deposition, without any changes to anemia status in Hmox-1-/- mice. This was accompanied by reduced urinary iron excretion. Finally, ambrisentan had an important iron recycling effect by increasing the expression of the cellular iron exporter, ferroportin-1 (FPN-1), and circulating total iron levels in Hmox-1-/- mice. These findings suggest that the ET-1/ETA signaling pathway contributes to renal iron trafficking in a murine model of iron overload.

Measurement of Tissue Non-Heme Iron Content using a Bathophenanthroline-Based Colorimetric Assay.

Iron is an essential micronutrient. Both iron overload and deficiency are highly detrimental to humans, and tissue iron levels are finely regulated. The use of experimental animal models of iron overload or deficiency has been instrumental to advance knowledge of the mechanisms involved in the systemic and cellular regulation of iron homeostasis. The measurement of total iron levels in animal tissues is commonly performed with atomic absorption spectroscopy or with a colorimetric assay based on the reaction of non-heme iron with a bathophenanthroline reagent. For many years, the colorimetric assay has been used for the measurement of the non-heme iron content in a wide range of animal tissues. Unlike atomic absorption spectroscopy, it excludes the contribution of heme iron derived from hemoglobin contained in red blood cells. Moreover, it does not require sophisticated analytical skills or highly expensive equipment, and can thus be easily implemented in most laboratories. Finally, the colorimetric assay can be either cuvette-based or adapted to a microplate format, allowing higher sample throughput. The present work provides a well-established protocol that is suited for the detection of alterations in tissue iron levels in a variety of experimental animal models of iron overload or iron deficiency.

The macro- and microelement provision of preschool children with psychomotor developmental disorders. Literature review and authors’ research

Background. An integral part of a child’s physical and mental development is a balanced diet. Insufficient or inadequate for a child developing organism’s needs nutrition in preschool age leads to a delay in physical, cognitive, and mental development. The study was aimed to assess the macro-and micronutrient provision status of preschool children with psychomotor developmental delay, identify the presence of macro- and micronutrient deficiencies, define the connection between macro- and micronutrient levels in patient’s serum and psychomotor developmental disorders. Materials and methods. The study analyzed the results of biochemical blood tests of blood serum samples for total calcium, magnesium, and iron level in 30 preschool children with psychomotor developmental disorders. Results. Mental retardation was observed in 53.33 % of examined children, autism spectrum disorders — in 20 % of examined children; minimal brain dysfunction — in 13.33 %, attention deficit hyperactivity disorder (ADHD) — in 13.33 %. 93.33 % of examined children had a deficiency of total calcium, magnesium — in 63.33 %, iron — in 20 %. Ten percent of patients had a deficiency of both total calcium, magnesium, and iron; one child (3.33 %) had no deficits. The most common deficiency in almost all subgroups of examined children was a combined deficiency of total calcium and magnesium. Only in the minimal brain dysfunction subgroup, in 50 % of cases, there was a combined deficiency of total calcium and magnesium, and in other 50 % of cases — an isolated deficiency of total calcium. This study identified a reliable inverse correlation of moderate strength between the level of total calcium and psychomotor development disorders and between the level of magnesium and the hyperexcitability syndrome. This study detected a weak unreliable correlation between the levels of magnesium and neurological disorders; an inverse unreliable weak correlation was found between iron and psychomotor developmental disorders. Conclusions. The most common deficiency in almost all subgroups of examined children was a combined deficiency of total calcium and magnesium. This study identified a reliable inverse correlation of moderate strength between the level of total calcium and psychomotor developmental disorders and between the level of magnesium and the hyperexcitability syndrome. There is also a weak direct unreliable correlation between magnesium level and psychomotor disorders. An inverse unreliable weak correlation was detected between the concentration of iron and psychomotor disorders.

Evaluation of the Physico-Chemical Quality of Well Water in the Analamanga Region (Antananarivo), Madagascar

The study made in some sites of the Analamanga Region, which are the Fokontany of Mandroseza, Ampanefy, Isaingy, Antalata and Malaho, shows the deterioration of the physico-chemical quality of the water of well. Elements that can cause illnesses for the beneficiaries are detected. The result is marked firstly by the acidity of the well water in all the Fokontany studied, secondly, by the rise in the rates of ammonium and organic matter in the wells which are located in the vicinity of several houses and those of nitrites in samples from wells located in the slums, near fields and rice paddies. Finally, for the Fokontany of Isaingy which is characterized by a very high number of hearths compared to other Fokontany, the waters of the wells are turbid and contain high levels of organic matter, total iron and nitrites.

The effect of protein supplements on weight loss, body composition, protein status, and micronutrients post laparoscopic sleeve gastrectomy (LSG): A Randomised Controlled Trial (RCT)

BackgroundLow protein intake post-bariatric surgery can result in protein malnutrition, and muscle mass loss. Authors aim to assess the effect of protein supplements on weight loss, body composition, and micronutrient status following LSG. MethodsThis is a double-blinded RCT conducted between February/2017 to January/2018. Eligible post LSG patients were randomized into the intervention group who received daily protein supplements containing 20 g of protein and placebo group received zero protein supplements. Both groups received a standardized diet. Weight loss, body composition, and micronutrient status were analyzed at 1, 3, and 6 months. Results48 participants were included in the final analysis (intervention: 21 and placebo:27). Excess weight loss percentage (EWL%) at 6 months was comparable between both groups (69.44 ± 21.99% and 71.40 ± 19.27% respectively). No significant difference observed in the anthropometric parameters. There was an increase in muscle mass and a decrease in muscle mass loss in the intervention group throughout the study period. However, these changes were not statistically significant. There was a significant increase in total protein (P=0.027) and magnesium (P=0.008) in the intervention group at 3 months. Albumin and iron levels were significantly higher at 6 months in the intervention group (P=0.036 &P=0.028 respectively). Other micronutrients did not differ at any time point between both groups. ConclusionProtein supplements resulted in significant improvement in total protein, albumin, magnesium, and iron levels post LSG. Although not significant, protein supplements helped in maintaining the muscle mass and preventing muscle mass loss. Original articleThis RCT is an original article and provides a level 2 evidence.

Heme Scavenging with Apohemoglobin-Haptoglobin Complex in Beta Thalassemia Intermedia Improves Anemia and Iron Indices: A Novel Therapeutic Approach

Beta-thalassemia intermediate (TI) is a genetic disorder of hemoglobin (Hb) synthesis characterized by anemia, ineffective erythropoiesis and iron overload. TI treatment is complex and regular transfusion therapy may be needed for growth failure, skeletal deformity, exercise intolerance, or when Hb levels decline because of progressive splenomegaly. Iron overload can be seen in TI, and serum ferritin measurement may often underestimate the extent of iron overload. There are limited therapeutic options for management of anemia and iron overload in TI. In this study, we look at the potential use of an engineered scavenger protein complex (apoHb-Hp) as a strategy to treat anemia in TI using a C57BL/6 mouse model.Sixteen beta-thalassemic mice (C57BL/6 heterozygous for the Hbb β-globin gene deletion (Hbb td3th/BrjK) (beta-thalassemia, Jackson Laboratory)) were treated with 50 µL of apoHb-Hp complex at a concentration of 27.95 mg/mL for six weeks to simultaneously scavenge cell-free Hb and free heme. Animal weight and RBC parameters (Hb, hematocrit (Hct), red blood cell (RBC) count, red cell distribution width) were measured at baseline and at 6 weeks post treatment. Total iron levels, transferrin concentration and transferrin saturation were measured at the third and sixth week of treatment. At the end of the experiment, the spleen and liver weights were measured and markers of liver function (Alanine transaminase (ALT), Aspartate transaminase (AST), Alkaline phosphatase (ALP)) were assessed. Further, total iron content of the liver and spleen was quantified using a using Perls' staining for non‐heme ferric iron (Fe3+).The mean Hb at baseline for this cohort was 10.5 g/dL, Hct was 36% and RBC count was 7.6 10 6/ µL. After treatment with apoHb-Hp complex for 6 weeks, Hb levels improved to 11.3 g/dL and Hct to 38% vs. Hb of 10.1 g/dL and Hct of 35% for vehicle treated animals. The mean total RBC count at was 9.2 10 6/ µL following treatment with apoHb-Hp, and 7.7 10 6/ µL for vehicle treated animals at 6 weeks. Iron parameters were also improved, with lower mean serum iron levels in the apoHb-Hp treated group at 6 weeks when compared to vehicle control (90.5 µg/dL vs. 135.2 µg/dL, p < 0.05). Serum transferrin levels were 131.1 mg/dL vs. 90.3 mg/dL (p<0.05) for the apoHb-Hp and vehicle treated groups at 6 weeks respectively. Furthermore, when compared to control mice, apoHb-Hp mice had a significant reduction in both liver and spleen weights at 6 weeks- liver 5.7g vs. 5.2g (p<0.05) and spleen 3.8g vs. 3.4g (p<0.05) respectively. Finally, liver function tests also showed improvement following treatment with apoHb-Hp at 6 weeks, ALT 71.1 units/L vs. 85.5 units/L for the vehicle, AST levels 180.2 units/L vs. 217.6 for the vehicle (p <0.05) of, and ALP levels 247.3 units/L vs. 304.4 units/L (p <0.05).Our study demonstrates that apoHb-Hp can improve anemia and reduce iron toxicity in TI mice as well as improve liver and spleen parameters. Thus, apoHb-Hp may be a potential treatment strategy in this disease and merits further study. [Display omitted] DisclosuresGopal: Alexion: Speakers Bureau; GBT: Consultancy; Pharming: Consultancy; Rigel Pharmaceuticals: Other: Clinical Trial, Research Funding.

Soluble, Colloidal, and Particulate Iron Across the Hydrothermal Vent Mixing Zones in Broken Spur and Rainbow, Mid-Atlantic Ridge.

The slow-spreading Mid-Atlantic Ridge (MAR) forms geological heterogeneity throughout the ridge system by deep crustal faults and their resultant tectonic valleys, which results in the existence of different types of hydrothermal vent fields. Therefore, investigating MAR hydrothermal systems opens a gate to understanding the concentration ranges of ecosystem-limiting metals emanating from compositionally distinct fluids for both near-field chemosynthetic ecosystems and far-field transport into the ocean interiors. Here, we present novel data regarding onboard measured, size-fractionated soluble, colloidal, and particulate iron concentrations from the 2018 R/V L’Atalante – ROV Victor research expedition, during which samples were taken from the mixing zone of black smokers using a ROV-assisted plume sampling. Iron size fractionation (<20, 20–200, and >200nm) data were obtained from onboard sequential filtering, followed by measurement via ferrozine assay and spectrophotometric detection at 562nm. Our results showed the persistent presence of a nanoparticulate/colloidal phase (retained within 20–200nm filtrates) even in high-temperature samples. A significant fraction of this phase was retrievable only under treatment with HNO3 – a strong acid known to attack and dissolve pyrite nanocrystals. Upon mixing with colder bottom waters and removal of iron in the higher parts of the buoyant plume, the larger size fractions became dominant as the total iron levels decreased, but it was still possible to detect significant (micromolar) levels of nanoparticulate Fe even in samples collected 5m above the orifice in the rising plume. The coolest sample (<10°C) still contained more than 1μM of only nitric acid-leachable nanoparticle/colloidal, at least 200 times higher than a typical Fe concentration in the non-buoyant plume. Our results support previous reports of dissolved Fe in MAR vent plumes, and we propose that this recalcitrant Fe pool – surviving immediate precipitation – contributes to maintaining high hydrothermal iron fluxes to the deep ocean.

An experimental study on release mechanism of iron and manganese from sediments to the water column in reservoirs

Iron and manganese accumulation in drinking water reservoirs is a challenging issue and should be controlled to prevent their adverse effects on human health. Accumulation of these elements not only clogs pipeline systems but also causes stains on fixtures and laundry. Also, high concentrations of iron and manganese may lead to various health problems when ingested. This study focuses on the release mechanism of iron and manganese from sediments to the water column in reservoirs and investigates methods to prevent this release. Effects of hypoxia, hypolimnetic aeration, alkalinity of water, and thermal stratification on iron and manganese concentrations were investigated through laboratory experiments. Experiments done simulating the water column showed that hypoxia caused more dissolution of ferrous iron when compared with that of manganese. Accordingly, aeration of the water column in hypoxic conditions lead to a significant decrease in ferrous iron concentrations (in our case reaching zero). However, manganese and total iron levels were not affected by the aeration of the water column. Alkalinity level of the water column was observed to have a great effect on the solubility of iron and manganese. Concentrations of total Fe and total Mn measured for acidic (pH=5) conditions were considerably greater than concentrations measured at neutral conditions. As for alkaline (pH=11) conditions, the opposite was observed with measured concentrations of total Fe and total Mn being lower than the ones measured for neutral conditions. Thermal stratification had an enhancing effect on the solubility of both iron and manganese ions. While aeration of the stratified water column slightly decreased the concentrations of Total Fe and Mn, it had a greater impact on decreasing Fe2+ concentrations.

Association Between Helicobacter pylori Infection and Serum Iron Profile.

Introduction: Helicobacter pylori (H. pylori) infection is reported to be the most frequent cause of morbidity and mortality in cases of upper gastrointestinal (GI) diseases. There is paucity of research between the possible association of H. pylori and iron stores and iron deficiency anemia (IDA). In this study, we will determine if there is an association between serum total iron-binding capacity (TIBC), serum iron and ferritin levels, and H. pylori infection.Methods: This case-control study was conducted in the gastroenterology ward of a major hospital in Pakistan from December 2020 to April 2021. Three hundred patients diagnosed with H. pylori were enrolled along with 300 participants in the control group. H. pylori was confirmed or excluded with the help of Giemsa stained gastric biopsy specimens. Blood was sent to the laboratory to test for ferritin, serum iron, and TIBC. Each sample was drawn in the morning to avoid any fluctuations.Results: The mean serum iron level was significantly lower in participants with H. pylori infection compared to those who did not have H. pylori infection (110.72 ± 28.38 ug/dL vs. 162.5 ± 21.18 ug/dL; p-value: <0.0001). Serum ferritin level was significantly higher in participants with H. pylori infection (536.82 ± 117.0 ng/dL vs. 391.31 ± 101.54 ng/dL; p-value: <0.0001).Conclusion: In comparison with the control group, TIBC and serum iron levels were found to be lower in the case group.

Effects of empagliflozin on erythropoiesis in patients with type 2 diabetes: Data from a randomized, placebo-controlled study.

Sodium-glucose cotransporter-2 (SGLT2) inhibitors have been shown to significantly reduce hospitalization for heart failure (HHF) and cardiovascular (CV) mortality in various CV outcome trials in patients with and without type 2 diabetes mellitus (T2D). SGLT2 inhibition further increased haemoglobin and haematocrit levels by an as yet unknown mechanism, and this increase has been shown to be an independent predictor of the CV benefit of these agents, for example, in the EMPA-REG OUTCOME trial. The present analysis of the EMPA haemodynamic study examined the early and delayed effects of empagliflozin treatment on haemoglobin and haematocrit levels, in addition to measures of erythropoiesis and iron metabolism, to better understand the underlying mechanisms. In this prospective, placebo-controlled, double-blind, randomized, two-arm parallel, interventional and exploratory study, 44 patients with T2D were randomized into two groups and received empagliflozin 10 mg or placebo for a period of 3months in addition to their concomitant medication. Blood and urine was collected at baseline, on Day 1, on Day 3 and after 3months of treatment to investigate effects on haematological variables, erythropoietin concentrations and indices of iron stores. Baseline characteristics were comparable in the empagliflozin (n=20) and placebo (n=22) group. Empagliflozin led to a significant increase in urinary glucose excretion (baseline: 7.3 ± 22.7g/24h; Day 1: 48.4 ± 34.7g/24h; P < 0.001) as well as urinary volume (baseline: 1740 ± 601mL/24h; Day 1: 2112 ± 837 mL/24h; P=0.011) already after 1day and throughout the 3-month study period, while haematocrit and haemoglobin were only increased after 3months of treatment (haematocrit: baseline: 40.6% ± 4.6%; Month 3: 42.2% ± 4.8%, P < 0.001; haemoglobin: baseline: 136 ± 19g/L; Month 3: 142 ± 25g/L; P=0.008). In addition, after 3months, empagliflozin further increased red blood cell count (P < 0.001) and transferrin concentrations (P=0.063) and there was a trend toward increased erythropoietin levels (P=0.117), while ferritin (P=0.017), total iron (P=0.053) and transferrin saturation levels (P=0.030) decreased. Interestingly, the increase in urinary glucose excretion significantly correlated with the induction of erythropoietin in empagliflozin-treated patients at the 3-month timepoint (Spearman rho 0.64; P=0.008). Empagliflozin increased haemoglobin concentrations and haematocrit with a delayed time kinetic, which was most likely attributable to increased erythropoiesis with augmented iron utilization and not haemoconcentration. This might be attributable to reduced tubular glucose reabsorption in response to SGLT2 inhibition, possibly resulting in diminished cellular stress as a mechanism for increased renal erythropoietin secretion.

Regular Physical Exercise Modulates Iron Homeostasis in the 5xFAD Mouse Model of Alzheimer's Disease.

Dysregulation of brain iron metabolism is one of the pathological features of aging and Alzheimer’s disease (AD), a neurodegenerative disease characterized by progressive memory loss and cognitive impairment. While physical inactivity is one of the risk factors for AD and regular exercise improves cognitive function and reduces pathology associated with AD, the underlying mechanisms remain unclear. The purpose of the study is to explore the effect of regular physical exercise on modulation of iron homeostasis in the brain and periphery of the 5xFAD mouse model of AD. By using inductively coupled plasma mass spectrometry and a variety of biochemical techniques, we measured total iron content and level of proteins essential in iron homeostasis in the brain and skeletal muscles of sedentary and exercised mice. Long-term voluntary running induced redistribution of iron resulted in altered iron metabolism and trafficking in the brain and increased iron content in skeletal muscle. Exercise reduced levels of cortical hepcidin, a key regulator of iron homeostasis, coupled with interleukin-6 (IL-6) decrease in cortex and plasma. We propose that regular exercise induces a reduction of hepcidin in the brain, possibly via the IL-6/STAT3/JAK1 pathway. These findings indicate that regular exercise modulates iron homeostasis in both wild-type and AD mice.

Puerarin Inhibits Ferroptosis and Inflammation of Lung Injury Caused by Sepsis in LPS Induced Lung Epithelial Cells

Background: Ferroptosis is a new type of programmed cell death, which plays an important role in lung injury caused by sepsis. Studies have reported that Puerarin (Pue) can treat lung injury caused by sepsis in children, but whether it plays a role by regulating iron death has not been reported.Methods: LPS induced human alveolar epithelial cell A549 to form a model of lung injury caused by sepsis. MTT detected the effect of Pue on A549 cell viability and the effect of Pue on LPS-induced A549 cell viability. The effects of Pue on LPS-induced inflammatory cytokines TNF-α, IL-8, IL-1β in A549 cells were determined by ELISA assay. The expression level of MDA was detected by TBARS colorimetric quantitative detection kit. GSH kit was used to detect the expression of GSH in cells. The iron kit detected the total iron level and the expression level of ferric divalent ions in the cells. DCFH-DA fluorescent probe was used to detect ROS levels. Western blot was used to detect the expression of ferroptosis-related proteins in cells.Results: Pue alleviated LPS-induced injury and inflammatory response in A549 cells, and Pue reduced the expression of ROS, MDA and GSH in LPS-induced A549 cells. In addition, Pue reduced total iron levels and ferrous ion levels in LPS-induced A549 cells, and decreased the expression of iron ferroptosis-related proteins.Conclusion: Puerarin inhibited ferroptosis and inflammation of lung injury caused by sepsis in children in LPS induced lung epithelial cells.

The effect of underlying inflammation on iron metabolism, cardiovascular risk and renal function in patients with type 2 diabetes.

AimTo investigate the impact of inflammation on iron metabolism, cardiovascular risk and renal function in type 2 diabetes (T2D).MethodsA total of 50 patients with T2D were included in this study. The patients were stratified into two groups based on their levels of C‐reactive protein (CRP), namely normal and high levels (n = 25/group). All laboratory tests were measured using standardised methods.ResultsFasting plasma glucose levels were elevated in patients with high CRP when compared to those with normal levels (p = 0.0413). Total serum iron levels were lower in patients with high CRP levels (12.78 ± 3.50) when compared to those with normal levels (15.26 ± 4.64), p = 0.0381. However, ferritin and transferrin levels were comparable between the groups (p > 0.05). The mean cell volume (MCV) in the high CRP group was lower (87.66 ± 3.62) than the normal level group (90.79 ± 4.52), p = 0.0096, whilst the lipograms were similar (p > 0.05). The estimated glomerular filtration rate (eGFR) was lower in the high CRP group (98.06 ± 11.64) than the normal level group (104.7 ± 11.11), p = 0.046. Notably, CRP levels were negatively associated with serum iron levels (r = –0.38, p = 0.0061), MCV (r = –0.41, p = 0.0031), potassium (r = –0.37, p = 0.0086) and sodium levels (r = –0.28, p = 0.0471). Regression analyses showed that only CRP (β = –0.16, standard error [SE]: 0.06, p = 0.0125) and sodium (β = 0.51, SE: 0.25, p = 0.0434) levels contributed significantly to the prediction of serum iron levels.ConclusionUnderlying inflammation in T2D is associated with increased incidence of hypertension and reduced levels of serum iron, MCV and renal function. Although there was no apparent clinical anaemia or renal dysfunction in these patients, mitigating inflammation may be effective in circumventing the ultimate development of iron deficiency anaemia and chronic kidney disease in T2D.

Assessing impacts of sand mining on water quality in Toutsang locality and design of waste water purification system

This paper focuses on assessing and development of water purification system for addressing water quality effects associated with sand mining in Toutsang locality, West-Cameroon. The study approach involved field investigations, laboratory analysis and design of waste water treatment system. Water samples were randomly collected from the study into well-labeled containers and conveyed to Unité de Recherche d’Analyse des Sols et Chimie de l’Environnement, Faculté d’Agronomie et des Sciences Agricoles (FASA) at the University of Dschang for analysis. Results from the physico-chemical analysis show elevated levels of total suspended solids (TSS), magnesium (Mg), turbidity and iron (Fe) in water samples collected downstream of the River close to the mining site, which exceeds the World Health Organization (WHO) standards. Based on the results of the physico-chemical analysis, a waste water treatment comprising of sedimentation tanks is proposed to treat waste water from the sand mining site. The designed water treatment system can be used to treat contaminated water from sand mining sites for domestic use. The proposed model tank needs to be implemented to restore Toutsang Water quality for indoor purpose and household use. • Toutsang water quality is accessed through Physico-chemical analysis of Toutsang water samples. • Sand mining activity increases the level of some physico-chemical parameters downstream of Toutsang River close to the mining site as compared to the World Health Organization standards. • Toutsang River contents very important amount of fine sediments in suspension which is the origin of it brownish coloration. • We investigate Toutsang River hydromorphological parameters including water flow, water velocity and water depth. • We design a sedimentation tank for Toutsang River treatment in order for it to become clean and appropriate for indoor and household purposes.

Safety and efficacy of a feed additive consisting of ferrous lysinate sulfate for all animal species (Phytobiotics Futterzusatzstoffe GmbH).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of ferrous lysinate sulfate as nutritional feed additive for all animal species. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) Panel was assigned to this mandate. Based on the results of a tolerance study, the FEEDAP Panel concluded that ferrous lysinate sulfate is safe in chickens for fattening when used up to the current maximum authorised levels of total iron in feed; this conclusion was extrapolated to all animal species and categories, at the respective maximum authorised iron levels in complete feed. The use of ferrous lysinate sulfate in animal nutrition up to the maximum iron content in complete feed authorised in the EU poses no concern to the safety of consumers. The FEEDAP Panel concluded that ferrous lysinate sulfate poses a risk to users by inhalation; the additive is not a dermal irritant, but is irritant to eyes and a skin sensitiser. The FEEDAP Panel considered that the use of ferrous lysinate sulfate in animal nutrition would not pose a risk for the environment. Owing to the limitations in the study provided, the FEEDAP Panel could not conclude on the efficacy of the additive for chickens for fattening, and thus, on the efficacy of ferrous lysinate sulfate for all animal species and categories.

Dietary Supplementation of Carob and Whey Modulates Gut Morphology, Hemato-Biochemical Indices, and Antioxidant Biomarkers in Rabbits.

This study aimed to evaluate the influence of the carob powder (CP) and sweet whey powder (WhP) inclusion into weaning feed on the gut morphology, hemato-biochemical parameters, and antioxidant biomarkers. The addition of 10 g/kg (basal diet +10 g/kg of CP, of WhP) or the mixture (5 g/kg of CP and 5 g/kg of WhP) in the rabbit's standard diet was assessed. A total of 40 weaned New Zealand White rabbits (4 weeks old) was distributed according to body weight (BW) into four treatments (n = 10) and the feed additives were provided for 7 weeks. Tissue samples and blood were obtained after slaughter. Final BW, daily weight gain, feed conversion ratio, intestinal morphology, and carcass dressing were positively affected by CP-WhP treatments compared with the control diet. Also, CP-WhP treatments significantly increased total proteins, calcium and iron levels, fecal cholesterol excretion, total antioxidants capacity, superoxide dismutase, and catalase in different tissues and significantly decreased total cholesterol, triglycerides, and glucose in blood serum. These changes were associated with a diminution of blood tumor necrosis factor alpha, lipid peroxidation, and carbonyl proteins in rabbit tissues. Both the additives separately and especially in the mix may enhance productive performance, protein profile, gut function, immunity, and antioxidant activity, with reducing lipid peroxidation, essential inflammatory mediator, and protein-carbonyl residues of growing rabbits. These findings suggest that CP-WhP dietary supplementation provides novel insights into a variety of bioactive compound mixtures with different beneficial modes of actions.

Electroacupuncture Ameliorates Cerebral Ischemic Injury by Inhibiting Ferroptosis.

Background: Our previous study found that electroacupuncture (EA) can promote the recovery of neurological functions, reduce the volume of cerebral infarction, and protect the neurovascular unit in middle cerebral artery occlusion (MCAO) rats. Some studies have shown that ferroptosis is closely related to ischemic stroke; however, whether EA plays a protective role by regulating ferroptosis is unknown.Objective: We aimed to investigate the inhibitory effects of EA on ferroptosis in MCAO rats.Methods: We used 36 adult male Sprague–Dawley rats in this study. MCAO rats were established according to the Zea method and treated with EA at a continuous wave of 2/100 Hz and ~2–4 V for 30 min for 7 consecutive days. We analyzed the coordinated motor deficit and volume of cerebral infarction in vivo through 9.4-tesla magnetic resonance imaging. Then, the ischemic brain tissue was isolated and the levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and iron were determined. Western blotting and real-time quantitative PCR were performed to evaluate the expression of glutathione peroxidase 4 (GPX4), transferrin (Tf), transferrin receptor 1 (TfR1), and ferritin heavy chain 1 (FTH1). To confirm the results, we used a transmission electron microscope to observe the mitochondrial morphology.Results: EA intervention significantly decreased the oxidative stress level and inhibited ferroptosis. EA significantly improved coordinated motor deficit (P < 0.01) and decreased cerebral infarct volume (P < 0.01) in the EA + MCAO group, compared with the MCAO group. EA downregulated the level of MDA (P < 0.01) and total iron (P < 0.01) and upregulated the level of SOD (P < 0.01) and GSH (P < 0.01) in the EA + MCAO group, compared with the MCAO group. EA increased the levels of GPX4 and GPX4 mRNA (P < 0.01) and FTH1 and FTH1 mRNA (P < 0.05, P < 0.01), whereas it decreased the levels of Tf and Tf mRNA (P < 0.05, P < 0.01) and TfR1 and TfR1 mRNA (P < 0.01) in the EA + MCAO group, compared with the MCAO group. EA also promoted the recovery of mitochondrial morphology according to the mitochondrial classification system for the ischemic cerebral tissue.Conclusion: Our results indicate that EA can inhibit ferroptosis by regulating oxidative stress and iron-related proteins, thus conferring protection against MCAO in a rat model.

Monitoring Acid Mine Drainage’s Effects on Surface Water in the Kizel Coal Basin with Sentinel-2 Satellite Images

Mining in the Kizel coal basin (Perm Region, Russia) ceased 20 years ago; however, AMD with high levels of total iron (Fetotal) and trace elements still affects the rivers. In this study, we attempted to estimate inter-annual and seasonal variability of AMD-related surface water contamination using Sentinel-2 images from 2016 to 2019. The acid mine water index (AMWI), which is a normalized difference of spectral reflectance in the red and blue bands, was calculated from Sentinel-2 images. We compared the AMWI values with measured Fetotal concentrations in the surface water. A statistically significant (at a 0.05 significance level) Spearman’s rank correlation between AMWI and Fetotal concentration was found for 5 out of 7 surface water sampling points. We found that surface water contamination reaches a seasonal maximum in July, 1–1.5 months after the end of the snowmelt high water period. Excessive summer rainfalls also contributes to increased contamination, causing contamination to possibly spread more than 200 km from the AMD sources. In contrast, arid summer conditions were associated with a substantially decreased AMD discharge and Fetotal concentrations in the surface water. The main uncertainties in our results are associated with the effect of contaminated bottom and bank sediments and suspended sediments on the spectral characteristics of the water surface, and the relatively coarse (10 m) spatial resolution of Sentinel-2 images. However, despite the data and method limitations, our results show that Sentinel-2 images have substantial potential for monitoring AMD-related contamination of surface water.

Protocols for assessing antibacterial and water coagulation potential of Moringa oleifera seed powder

This research evaluated Moringa oleifera seed powder (MOSP) as an antibacterial agent, and a coagulant. In the former, clinical isolates of Salmonella sp., Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were used, and in the latter, river and stream water were used. Both the isolates and water samples were treated with MOSP at varying final concentrations of 0.001, 0.002, 0.004 and 0.017 g/ml.For the antibacterial assay, a dose of 0.017 g/ml of MOSP was effective on all three isolates with CFU/ml reduction of 99.4, 78.8 and 57.3% on Salmonella sp., E. coli and S. aureus respectively. An ANOVA confirmed this finding at P<0.05; 0.0014 between the treated and control samples. The water treatment assay also showed a reduction of total hardness, fluoride, phosphate, nitrate, total iron and manganese levels below the water quality standards.The MOSP could serve as a cost-effective product for process integration in raw water treatment systems in rural and urban settings. The study shows bioactivity of the seed powder of Moringa, and provides grounds to isolate the active component for commercialization and usage by the wider population with limited or no access to potable water.

Involvement of Hepcidin in Cognitive Damage Induced by Chronic Intermittent Hypoxia in Mice.

Obstructive sleep apnea (OSA) patients exhibit different degrees of cognitive impairment, which is related to the activation of reactive oxygen species (ROS) production by chronic intermittent hypoxia (CIH) and the deposition of iron in the brain. As a central regulator of iron homeostasis, whether hepcidin is involved in OSA-induced cognitive impairment has not been clarified. In order to simulate OSA, we established the mouse model by reducing the percentage of inspired O2 (FiO2) from 21% to 5%, 20 times/h for 8 h/day. We found hepcidin was rising during CIH, along with increasing iron levels and neuron loss. Then, we constructed a mouse with astrocyte-specific knockdown hepcidin gene (shHamp). During CIH exposure, the shHamp mice showed a lower level of total iron and neuronal iron in the hippocampus, via stabilizing ferroportin 1 (FPN1) and decreasing L-ferritin (FTL) levels, when compared with wild-type (WT) mice. Furthermore, the shHamp mice showed a decrease of ROS by downregulating the elevated NADPH oxidase (NOX2) and 4-hydroxynonenal (4-HNE) levels mediated by CIH. In addition, the shHamp mice presented improved cognitive deficit by improving synaptic plasticity and BDNF expression in the hippocampus when subjected to CIH. Therefore, our data revealed that highly expressed hepcidin might promote the degradation of FPN1, resulting in neuronal iron deposition, oxidative stress damage, reduced synaptic plasticity, and impaired cognitive performance during CIH exposure.