Iron Overload Research Articles

EVALUATION OF THE RELATIONSHIP BETWEEN IRON LOAD, AGE, AND CARDIAC FUNCTION IN CHILDREN AND YOUNG ADULTS WITH THALASSEMIA MAJOR: A PULSED-WAVE DOPPLER AND TISSUE DOPPLER IMAGING STUDY

Objective: To determine the structural and functional cardiac differences in children and young adults with thalassemia major (TM) compared to healthy subjects using pulsed-wave Doppler and tissue Doppler imaging methods and determine the relationship between iron overload and these differences. Materials and Methods: We analyzed the data of pediatric and young adult TM patients (n = 44) aged 4–22 years and an age- and gender-matched control group (n = 40) in our hospital data system between Oct.01.2023 and Oct.01.2024. Height, weight, body mass index (BMI), systolic–diastolic blood pressure measurements, complete blood count, ferritin, cardiac T2* magnetic resonance imaging (MRI) values, and echocardiography results were recorded. In addition to comparisons between the two groups, correlation analysis was performed between ferritin–cardiac T2* MRI results and echocardiographic parameters and age in TM patients. Results: Our study showed growth retardation (low height standard deviation score (SDS), low weight SDS and low BMI SDS), dilatation of the left cavities (high left ventricular internal diameter end diastole (LVIDd)), increased left ventricular muscle mass (high left ventricular mass index (LVMI)), cardiac distinctive diastolic (restrictive pattern: left ventricular (LV) peak early diastolic flow (E)/peak late diastolic flow (A) and E/early diastolic myocardial peak flow (E') high), and subclinical systolic (LV peak systolic flow low and LV Tei index high) dysfunction. In addition, iron load (ferritin and cardiac T2* MRI) was correlated with LVMI, and cardiac diastolic and systolic function indicators. As age increased, ferritin value did not change, but cardiac T2* MRI value decreased and diastolic–systolic parameters worsened. Conclusion: Periodic cardiac T2* MRI and Doppler echocardiography examinations of patients with TM may detect subclinical myocardial dysfunction at an early stage, thus providing a window of opportunity for intervention.

Thalassemia in Pakistan: Addressing a Genetic Health Crisis

Thalassemia, a genetic disorder usually caused by the defective genes. These genes are responsible for production of hemoglobin, a protein necessary for the transfer of oxygen in body through blood. Prevalence of thalassemia in Pakistan is concerning to a significant level. Global prevalence of this disease has affected millions of people and their lifestyles. Moreover, the disease leads to the burden on the diseased person’s family and the medical resources of the country, causing an alarming concern. The main issue of high prevalence of this disorder in Pakistan is associated with high percentage of cousin marriages. This cultural practice between the close relatives increases the chances of inheriting mutated genes into next generations. Each year more than 5000 children in Pakistan are born with disease. The rate of thalassemia is considerably higher in first cousin marriages (76.7%) than in second cousin marriages (23.3%) [1]. Furthermore, the inadequate spread of awareness among people and the lack of preventative measure by people is contributing to the increase in the number of thalassemia cases Thalassemic patients do require blood transfusion in an adequate to survive and live a normal life. Moreover, after transfusion they need financially draining therapies like iron chelation that is being required to manage the overload of iron in blood. The limited medical resources and poor infrastructure of hospitals and clinics in the rural areas can lead to the increase of complications then reducing the problems come with this disease. This kind of public healthcare system can affect the diseased individuals to level to develop life threatening problems such as stunted growth, and organ failures etc. To address this problem, a national program stating that genetic screening before marriage is necessary can lead to the reduction in the rate if this disease. Premarital screening for inherited diseases including Thalassemia is being carried out in middle east for last more than a decade. Some countries like Iran have overcome this problem to a great level by taking these kinds of initiatives, which shows that how important is it to detect the disease occurrence through premarital screening and counseling [2]. There should be campaigns for the public awareness highlighting the importance of genetic screening so that the cause of occurrence of this disorder can be decreased. Additionally, medical advancements have provided the hope. Despite being costly and scarce, for certain patients with thalassemia, bone marrow transplants (BMT) have proven to be a curative treatment, success rates for those who satisfy the right criteria, such as younger patients with matched related donors, range approximately between 80 and 95% [3]. Future advances in repairing genetic abnormalities at their origins may be possible thanks to emerging gene-editing technologies like CRISPR. To make these remedies available in Pakistan, however, significant funding for medical infrastructure and research is needed. Despite being avoidable, thalassemia will continue to take lives and put a burden on families and communities unless immediate action is taken. Pakistan can considerably lessen the impact of this genetic health disaster and clear the path for a healthier future by placing a high priority on education, early identification, and cutting-edge treatments.

Iron-responsive nanoparticle-loaded bilayer dissolving microneedles for selective and controlled transdermal delivery of deferasirox in β-thalassemia major treatment

Deferasirox (DFX) is widely used to manage β-thalassemia major (β-TM), but its oral administration is limited by low bioavailability and side effects. To address these challenges, we developed iron-responsive nanoparticles (NP-IR) of DFX using ferrocene as the iron-responsive material, incorporated into dissolving microneedles (DMN) for transdermal delivery. The NP-IR measured 276.67 ± 7.80 nm with an entrapment efficiency of 47.54 ± 3.68 %. FTIR analysis confirmed DFX incorporation, while reduced crystallinity suggested enhanced formulation. In vitro testing demonstrated controlled DFX release in the presence of iron, highlighting its targeted responsiveness. The DMN containing NP-IR, composed of polyvinyl pyrrolidone and polyvinyl alcohol, showed less than 10 % height reduction and successfully penetrated the fourth layer of Parafilm®, simulating human skin penetration. Ex vivo studies validated effective DFX delivery through rat skin with high iron selectivity, while in vivo experiments in an iron overload rat model revealed sustained, controlled release, outperforming oral administration and potentially ehancing DFX efficacy in β-TM treatment.

Integrating transcriptome and physiological analysis reveals the stress responses of rose petals to surface water in the iron-mining area

Mining activities in the iron area mining have caused serious pollution of surface water in the area, which in turn affects the normal growth and development of plants. As a restorative plant with heavy metal adsorption function, the stress response mechanism of rose to surface water in the iron-mining area remains unelucidated. In this study, we investigated the effects of surface water treatment on the phenotypic, physiological and molecular levels of the plant using rose as a material. The results showed that surface water in the Bayan Obo iron mining area caused darkening and soft rotting of rose petals, increased degree of petal damage, decreased the content of MDA (Malondialdehyde), increased the ratio of GSH/GSSG (glutathione/oxidized glutathione) and NADPH/NADP+(nicotinamide adenine dinucleotide phosphate/oxidized nicotinamide adenine dinucleotide phosphate), suggesting that the rose produces more GSH to prevent oxidative stress. Transcriptome analysis further revealed a large number of differentially expressed genes (DEGs) involved in the TCA (the tricarboxylic acid) cycle and the phenylpropanoid biosynthesis pathways under surface water stress, and these DEGs can promote the production of NADPH, which in turn positively regulates the content of GSH. Multiple peroxidase-related genes (RhPRXs) also exhibited a significant stress response in this process. In summary, our research results reflect that roses under surface water stress in the iron-mining area exhibit certain defense mechanisms at both physiological and transcriptional levels, and also provide a reference for cultivating effective rose resources for repairing pollution in mining areas and evaluating excessive iron pollution as a biological indicator.

Green Tea Epigallocatechin 3-Gallate Reduced Platelet Aggregation and Improved Anticoagulant Proteins in Patients with Transfusion-Dependent β-Thalassemia: A Randomized Placebo-Controlled Clinical Trial.

Patients with transfusion-dependent β-thalassemia (TDT) with iron overload have been linked to hypercoagulability and increased platelet (PLT) activation that causes thrombosis. Green tea extract (GTE) rich in epigallocatechin-3-gallate (EGCG) exerts iron-chelating and antithrombotic properties. The study aimed to assess the effects of GTE treatment on plasma coagulation state and PLT function in vitro and in patients with TDT. The subjects consumed a placebo or GTE tablets (50 mg and 2 × 50 mg EGCG equivalent) every day for two months. Blood was then collected from the treated patients for analyses of PLT numbers, agonist-induced PLT aggregation, and anti-coagulation proteins. In our findings indicate that the in vitro treatment of GTE (at least 1 mg EGCG equivalent) inhibited PLT aggregation in patients who were healthy and with thalassemia platelet-rich plasma (PRP), which was significant in the healthy PRP. Consistently, GTE treatment inhibited the PLT aggregation that had been ex vivo generated by collagen or ADP. In addition, consumption of GTE tablets greatly inhibited PLT aggregation and increased the plasma levels of proteins C and S, as well as the free protein S concentrations depending upon the time course, but not the GTE dosage. Moreover, plasma ferritin levels decreased in both green tea tablet groups in a time-dependent manner (p < 0.05 in the second month). In conclusion, EGCG-rich GTE diminished PLT aggregation in patients who were healthy and patients with thalassemia plasma. It also improved PLT aggregation and hypercoagulability in patients with TDT by increasing the antithrombotic activity of protein C and protein S. This would suggest an adjuvant of GTE could reduce the risk of thrombosis associated with iron overload.

Neobacillus driksii sp. nov. isolated from a Mars 2020 spacecraft assembly facility and genomic potential for lasso peptide production in Neobacillus.

The microbial surveillance of the Mars 2020 assembly cleanroom led to the isolation of novel N. driksii with potential applications in cleanroom environments, such as hospitals, pharmaceuticals, semiconductors, and aeronautical industries. N. driksii genomes were found to possess genes responsible for producing lasso peptides, which are crucial for antimicrobial defense, communication, and enzyme inhibition. Isolation of N. driksii from cleanrooms, Agave plants, and dryland wheat soils, suggested niche-specific ecology and resilience under various environmentally challenging conditions. The discovery of potent antimicrobial agents from novel N. driksii underscores the importance of genome mining and the isolation of rare microorganisms. Bioactive gene clusters potentially producing nicotianamine-like siderophores were found in N. driksii genomes. These siderophores can be used for bioremediation to remove heavy metals from contaminated environments, promote plant growth by aiding iron uptake in agriculture, and treat iron overload conditions in medical applications.

N-3 polyunsaturated fatty acids enhanced efficacy of cytarabine in iron-overloaded NALM-6 cells via apoptotic and oxidative pathways

Despite progress in treating acute lymphoblastic leukemia (ALL), the adverse effects of chemotherapy toxicity and iron overload from transfusions continue to affect patients' quality of life. Polyunsaturated fatty acids (PUFAs) exhibit both antitumor and anti-inflammatory properties in leukemia. This study investigated the influence of n-3 PUFA on the efficacy of cytarabine in cells with iron overload. Iron overload was induced in NALM-6 cells using ferric ammonium citrate (FAC) and quantified through atomic absorption spectroscopy (AAS). The impact of n-3 PUFA on NALM-6 cells' response to cytarabine was evaluated using MTT, lactate dehydrogenase (LDH), apoptosis, and cell cycle assays. Additionally, gene expression analyses were performed on apoptotic, anti-apoptotic, and inflammatory genes, along with oxidative stress markers such as reactive oxygen species (ROS) and malondialdehyde (MDA) levels. The administration of n-3 PUFA significantly enhanced the effectiveness of cytarabine in iron-overloaded NALM-6 cells, leading to increased LDH secretion, elevated apoptosis rates, and G1 phase cell cycle arrest. These effects were associated with the upregulation of apoptotic genes such as P53 and caspase-8, the downregulation of the anti-apoptotic gene Bcl2, and a decrease in the inflammatory gene TNF-α. Furthermore, there was a notable increase in ROS and MDA levels. Overall, n-3 PUFA treatment improved cytarabine's efficacy in iron-overloaded NALM-6 cells by activating apoptotic processes and oxidative stress pathways.

GPX4 Promoter Hypermethylation Induced by Ischemia/Reperfusion Injury Regulates Hepatocytic Ferroptosis.

Glutathione peroxidase 4 (GPX4) is a key factor in ferroptosis, which is involved in ischemia-reperfusion injury. However, little is known about its role in hepatic ischemia-reperfusion injury (HIRI). This study aimed to investigate the role of GPX4 methylation in ferroptosis during HIRI. For the in vitro experiments, an oxygen and glucose deprivation cell model was established. For the in vivo experiments, an ischemia-reperfusion model was created by subjecting mice to simulated HIRI. Ferroptosis occurrence, GPX4 promoter methylation, and global methylation levels were then assessed. Ferroptosis was observed in oxygen and glucose deprivation, characterized by a significant decrease in cellular viability (P < 0.05), an increase in lipid peroxidation (P < 0.01), iron overload (P < 0.05), and down-regulation of GPX4 (P < 0.05). This ferroptosis was exacerbated by GPX4 knockdown (P < 0.01) and mitigated by exogenous glutathione (P < 0.01). Similarly, ferroptosis was evident in mice subjected to HIRI, with a down-regulation of GPX4 mRNA and protein expression (all P < 0.01), and an upregulation of acyl-CoA synthetase long-chain family member 4 mRNA and protein (all P < 0.01), as well as prostaglandin-endoperoxide synthase 2 mRNA and protein expression (all P < 0.05). Methylation levels increased, evidenced by upregulation of DNA methylation transferase expression (P < 0.05) and down-regulation of Ten-eleven translocation family demethylases (P < 0.01), along with an upregulation of GPX4 promoter methylation. Ferroptosis may be the primary mode of cell death in hepatocytes following ischemia-reperfusion injury. The methylation of the GPX4 promoter and elevated levels of global hepatic methylation are involved in the regulation of ferroptosis.

Broadening horizons: the multifaceted role of ferroptosis in breast cancer.

Breast cancer poses a serious threat to women's health globally. Current radiotherapy and chemotherapy regimens can induce drug-resistance effects in cancer tissues, such as anti-apoptosis, anti-pyroptosis, and anti-necroptosis, leading to poor clinical outcomes in the treatment of breast cancer. Ferroptosis is a novel programmed cell death modality characterized by iron overload, excessive generation of reactive oxygen species, and membrane lipid peroxidation. The occurrence of ferroptosis results from the imbalance between intracellular peroxidation mechanisms (executive system) and antioxidant mechanisms (defensive system), specifically involving iron metabolism pathways, amino acid metabolism pathways, and lipid metabolism pathways. In recent years, it has been found that ferroptosis is associated with the progression of various diseases, including tumors, hypertension, diabetes, and Alzheimer's disease. Studies have confirmed that triggering ferroptosis in breast cancer cells can significantly inhibit cancer cell proliferation and invasion, and improve cancer cell sensitivity to radiotherapy and chemotherapy, making induction of ferroptosis a potential strategy for the treatment of breast cancer. This paper reviews the development of the concept of ferroptosis, the mechanisms of ferroptosis (including signaling pathways such as GSH-GPX4, FSP1-CoQ1, DHODH-CoQ10, and GCH1-BH4) in breast cancer disease, the latest research progress, and summarizes the research on ferroptosis in breast cancer disease within the framework of metabolism, reactive oxygen biology, and iron biology. The key regulatory factors and mechanisms of ferroptosis in breast cancer disease, as well as important concepts and significant open questions in the field of ferroptosis and related natural compounds, are introduced. It is hoped that future research will make further breakthroughs in the regulatory mechanisms of ferroptosis and the use of ferroptosis in treating breast cancer cells. Meanwhile, natural compounds may also become a new direction for potential drug development targeting ferroptosis in breast cancer treatment. This provides a theoretical basis and opens up a new pathway for research and the development of drugs for the prevention and treatment of breast cancer.

Mechanisms of METTL14-Mediated m6A Modification in Promoting Iron Overload-Induced Lipid Peroxidative Damage in Vascular Endothelial Cells to Aggravate Atherosclerosis.

Atherosclerosis (AS) is a chronic multifactorial disease with damage to vascular endothelial cells (VECs). This study sought to delve into the mechanism of methyltransferase-like 14 (METTL14) in iron overload-induced lipid peroxidative damage in AS. AS mouse model and cell model were established. Levels of METTL14/circRNA coded by the Arhgap12 (circARHGAP12)/Aspartate β-hydroxylase (ASPH) were determined. AS plaque area/lipid deposition/lipid metabolism in AS mice and iron overload in VECs were evaluated. N6-methyladenosine (m6A) level and METTL14 enrichment and human antigen R (HuR) in circARHGAP12 or ASPH were measured. The mRNA stability of circARHGAP12 or ASPH was analyzed. We observed that METTL14 was upregulated in AS mice. METTL14 downregulation reduced plaque area/lipid deposition/iron overload/peroxidative damage in AS mice. In cell models, METTL14 downregulation could VEC injury/iron overload/lipid peroxidative damage. Mechanically, METTL14 increased the stability and expression of circARHGAP12 through m6A modification, further stabilized ASPH mRNA, and promoted ASPH transcription by binding to HuR. Overexpression of circARHGAP12 or inhibition of ASPH averted the protective role of METTL14 downregulation against iron overload-induced peroxidative damage in AS. In conclusion, METTL14-mediated m6A modification upregulated circARHGAP12 and ASPH to aggravate overload-induced lipid peroxidative damage and facilitate AS progression.

ACSL4-mediated ZIP7-VDAC3 interaction regulates endoplasmic reticulum-mitochondria iron transfer in hepatocytes under PFOS exposure

Perfluorooctane sulfonate (PFOS) is a persistent organic pollutant with adverse health consequences. Our previous studies showed that PFOS caused an increase in mitochondrial iron and accelerated the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4), one classic executor in the ferroptosis pathway. As ACSL4 is located in the mitochondria-associated endoplasmic reticulum (ER) membranes, here, we intended to further explore the role of ACSL4 in the inter-organelle iron crosstalk between ER and mitochondria under PFOS exposure. We found that PFOS caused ER iron accumulation in mice liver and human hepatocytes L-02. Inhibition of solute carrier family 39 member 7 (SLC39A7/ZIP7), a potential ER iron efflux channel supposed by us, alleviated PFOS-induced mitochondrial iron overload and further elevated ER iron level. Knockdown of voltage-dependent anion channel 3 (VDAC3) or mitochondrial calcium uniporter (MCU), the respective potential mitochondrial iron influx channels in outer/inner mitochondrial membrane, reversed the mitochondrial iron overload and aggravated ER iron accumulation in the cells under PFOS treatment. ACSL4 interacted with both ZIP7 and VDAC3 in mice liver and L-02 cells after treatment with PFOS. Upon inhibition of ACSL4, the ZIP7-VDAC3 interaction was reduced, mitigating mitochondrial iron overload and exacerbating iron accumulation in ER. Inhibiting VDAC3 or ZIP7 reversed the overloaded cytosolic iron under PFOS treatment, however, we found no further decrease in cytosolic iron after simultaneous inhibiting VDAC3 and ZIP7 compared with respectively inhibiting VDAC3 or ZIP7 alone. Our study provides evidence and reveals the molecular mechanism underneath the ER-mitochondria iron crosstalk under PFOS exposure, providing new insights into and enriches the understanding of the iron network-regulating function of the ferroptosis executor ACSL4 and highlighting its role in PFOS toxicity.

A Retrospective Analysis of Demographics, Clinical Features, and Treatment Patterns in Sickle Cell Disease Patients at a Tertiary Healthcare Centre of North East India.

Background Sickle cell disease (SCD) is a hereditary disorder marked by abnormal hemoglobin (HbS), leading to chronic hemolytic anemia, vaso-occlusive crises (VOCs), and multi-organ complications. In India, the prevalence of SCD is highest among tribal populations in states like Madhya Pradesh, Maharashtra, Odisha, and Assam, with the disease burden exacerbated by limited healthcare access, especially in rural regions. This study provides a comprehensive analysis of the demographic profile, clinical features, and treatment patterns of SCD patients at a tertiary healthcare center in Upper Assam, where the prevalence of SCD is high among the tea tribe communities. Methods This retrospective observational study included 250 patients diagnosed with various SCD subtypes who presented with SCD-related complications at Assam Medical College and Hospital between January 2020 and December 2023. Data were obtained from medical records in the departments of medicine and pediatrics, covering demographic variables (age, gender, ethnicity), clinical characteristics (complications, hemoglobin levels, genotype, history of hospitalizations), and treatment details (frequency and type of blood transfusions, use of hydroxyurea and chelating agents). Descriptive statistics summarized demographic and clinical features, while chi-square tests and t-tests were used for bivariate analysis. Logistic regression identified factors associated with high transfusion requirements. Results The study population had a mean age of 17.2 years, with 54.4% male predominance. Most patients (87%) had sickle cell anemia (HbSS), while the remainder had other genotypes including HbSA, HbSE, and sickle cell thalassemia. The most common presenting symptoms were fever (61.2%) and bone/joint pain (48.4%), indicative of VOCs and frequent infections. Pallor (30%) and abdominal pain (25.6%) were also prominent. Half of the patients (125) received hydroxyurea, though its uptake was limited by availability and cost. A high transfusion burden was noted, with 72.4% of patients requiring between five to 12 transfusions annually. However, only 22.4% received chelation therapy to manage iron overload, reflecting the cost constraints in accessing these agents. Laboratory findings indicated a mean hemoglobin level of 7.19 g/dL and elevated serum ferritin levels due to repeated transfusions. The frequency of blood transfusions was higher compared to Western studies, emphasizing the need for more accessible disease-modifying therapies in resource-limited settings. Conclusions The findings of this study illustrate the significant clinical and transfusion burden experienced by SCD patients in Upper Assam. This population relies heavily on blood transfusions due to limited access to hydroxyurea and other advanced therapies. The study underscores a critical need for improved access to hydroxyurea, expanded availability of chelation therapy, and greater healthcare support for managing SCD-related complications in resource-limited settings. As India's National Sickle Cell Anemia Elimination Mission is implemented, regional studies such as this are essential for tailoring public health interventions to meet the specific needs of high-prevalence areas, especially among underserved tribal communities.

The role of mitochondria in iron overload-induced damage

Iron overload is a pathological condition characterized by the abnormal accumulation of iron within the body, which may result from excessive iron intake, disorders of iron metabolism, or specific disease states. This condition can lead to significant health complications and may pose life-threatening risks. The excessive accumulation of iron can induce cellular stress, adversely affecting the structure and function of mitochondria, thereby compromising overall organ function. Given the critical role of mitochondria in cellular metabolism and homeostasis, it is imperative to investigate how mitochondrial dysfunction induced by iron overload contributes to disease progression, as well as to explore mitochondrial-related pathways as potential therapeutic targets for various iron overload disorders. This review examines the mechanisms by which mitochondria are implicated in iron overload-induced damage, including increased oxidative stress, mitochondrial DNA damage, and disruptions in energy metabolism. Additionally, it addresses the relationship between these processes and various forms of programmed cell death, as well as alterations in mitochondrial dynamics. Furthermore, the review discusses strategies aimed at alleviating and mitigating the complications associated with iron overload in patients by targeting mitochondrial pathways.

Iron and Inflammatory Cytokines Synergistically Induce Colonic Epithelial Cell Ferroptosis in Colitis.

Inflammatory bowel disease (IBD) is an inflammatory disease that occurs to the intestinal tract. Many patients with IBD often develop anemia and often receive oral iron supplementation. Many of them develop non-compliance with oral iron therapy, but the mechanisms are not well understood. We interrogated whether colonic epithelial iron overload impacts cell viability and disease severity. We observed increased expression of iron importers and iron accumulation in mature colonocytes in dextran sulfate sodium (DSS)-induced acute colitis and in humans with active colitis. Administration of hepcidin increased epithelial iron overload and aggravated colonic inflammation in DSS-treated mice and IL10-/- mice. Hepcidin-induced iron accumulation increased colonic epithelial death, which was prevented by treatment with Trolox, a vitamin E analog and a scavenger of lipid peroxides. By using cultured Caco-2 cells, we showed that iron and inflammatory cytokines (TNF-α and IL-1β) induced a synergistic increase in the number of necrotic cells. We then showed that the combined treatment by hepcidin and cytokines increased labile iron content and lipid peroxidation in Caco-2 cells. Moreover, liproxstatin-1, a ferroptosis inhibitor, and deferoxamine, an iron chelator, both abolished the hepcidin/cytokines induced death of Caco-2 cells, suggesting ferroptosis. We further elucidated that inflammatory cytokines promote lipid peroxidation and ferroptosis by inducing NOX1-dependent exhaustion of reduced glutathione (GSH). Collectively, our findings demonstrate that the inflammatory context predisposes colonic epithelial cells to iron overload mediated ferroptosis, exacerbating colonic inflammation.

Mito-TEMPO Ameliorates Sodium Palmitate Induced Ferroptosis in MIN6 Cells through PINK1/Parkin-Mediated Mitophagy

ObjectiveMitochondrial reactive oxygen species (mtROS) could cause damage to pancreatic β-cells, rendering them susceptible to oxidative damage. Hence, investigating the potential of the mitochondriatargeted antioxidant (Mito-TEMPO) to protect pancreatic β-cells from ferroptosis by mitigating lipid peroxidation becomes crucial. MethodsMIN6 cells were cultured in vitro with 100 μmol/L sodium palmitate (SP) to simulate diabetes. FerroOrange was utilized for the detection of Fe2+ fluorescence staining, BODIPY581/591C11 for lipid reactive oxygen species, and MitoSox-Red for mtROS. Alterations in mitophagy levels were assessed through the co-localization of lysosomal and mitochondrial fluorescence. Western blotting was employed to quantify protein levels of Acsl4, GPX4, FSP1, FE, PINK1, Parkin, TOMM20, P62, and LC3. Subsequently, interventions were implemented using Mito-TEMPO and Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) to observe changes in ferroptosis and mitophagy within MIN6 cells. ResultsWe found that SP induced a dose-dependent increase in Fe2+ and lipid ROS in MIN6 cells while decreasing the expression levels of GPX4 and FSP1 proteins. Through bioinformatics analysis, it has been uncovered that mitophagy assumes a crucial role within the ferroptosis pathway associated with diabetes. Additionally, SP decreased the expression of mitophagy-related proteins PINK1 and Parkin, leading to mtROS overproduction. Conversely, Mito-TEMPO effectively eliminated mtROS while activating the mitophagy pathways involving PINK1 and Parkin, thereby reducing the occurrence of ferroptosis in MIN6 cells. CCCP also demonstrated efficacy in reducing ferroptosis in MIN6 cells. ConclusionIn summary, Mito-TEMPO proved effective in attenuating mtROS production and initiating mitophagy pathways mediated by PINK1 and Parkin in MIN6 cells. Consequently, this decreased iron overload and lipid peroxidation, ultimately safeguarding the cells from ferroptosis.

Impact of body fat composition on liver iron overload severity in hemochromatosis: a retrospective MRI analysis.

To evaluate the correlation between ectopic adipose tissue and iron overload severity in patients with hemochromatosis. A retrospective cohort of 52 patients who underwent liver iron concentration quantification from January 2015 to October 2023 using a 3.0T MRI scanner. R2* relaxation times and proton density fat fraction (PDFF) were assessed for the entire liver volume and a specific region of interest (ROI) placed in the right lobe. Total body fat (TF), subcutaneous fat (SCF), intermuscular fat (IMF), and visceral fat (VSF) percentages were calculated from a single axial slice at the level of the third lumbar vertebra. Additionally, ratios of IMF-to-VSF, IMF-to-SCF, and SCF-to-VSF were calculated. Standard iron laboratory parameters were collected at least one month prior to MRI. Pearson correlation coefficient was used for correlation analysis. The mean age of participants was 53.9 ± 19.6years. IMF positively correlated with R2* values in the ROI (p = 0.005, rs = 0.382) and entire liver (p = 0.016, rs = 0.332). Conversely, VSF negatively correlated with R2* values from the ROI (p = < 0.001, rs = -0.488) and entire liver (p = < 0.001, rs = -0.459). Positive correlations were also found between IMF-to-VSF and R2* of the ROI (p = 0.003, rs = 0.400) and whole liver (p = 0.008, rs = 0.364). Ferritin levels positively correlated with R2* values calculated from ROI (p = 0.002, rs = 0.417) and whole liver volume (p = 0.004, rs = 0.397). A positive correlation was noted between PDFF of the entire liver and TF (p = 0.024, rs = 0.313). The percentage of Intermuscular and visceral adipose tissues correlates with the severity of liver iron overload in hemochromatosis patients.

Syzygium aromaticum Extract Mitigates Doxorubicin-Induced Hepatotoxicity in Male Rats.

Doxorubicin (DOX), an anticancer drug, is used to treat several types of tumors, but it has detrimental side effects that restrict its therapeutic efficacy. One is the iron-dependent form of ferroptosis, which is characterized by elevated ROS production and iron overload. Syzygium aromaticum has a diverse range of biological and pharmaceutical actions due to their antioxidant properties. This study investigated the effect of S. aromaticum extract (SAE) on hepatotoxicity caused by DOX in rats. Phytochemical analysis was performed to assess compounds in SAE. The ADMETlab 2.0 web server was used to predict the pharmacokinetic properties of the most active components of SAE when DOX was injected into rats. Molecular docking studies were performed using AutoDock Vina. Forty male Sprague Dawley rats were divided into four groups of ten rats each (G1 was a negative control group, G2 was given 1/10 of SAE LD50 by oral gavage (340 mg/kg), G3 was given 4 mg/kg of DOX intraperitoneally (i.p.) once a week for a month, and G4 was administered DOX as in G3 and SAE as in G2). After a month, biochemical and histopathological investigations were performed. Rats given SAE had promising levels of phytochemicals, which could significantly ameliorate DOX-induced hepatotoxicity by restoring biochemical alterations, mitigating ferroptosis, and upregulating the NRF-2-SLC7A-11-GPX-4 signaling pathway. These findings suggest that SAE could potentially alleviate DOX-induced hepatotoxicity in rats.

Hemochromatosis neural archetype reveals iron disruption in motor circuits.

Our understanding of brain iron regulation and its disruption in disease is limited. Excess iron affects motor circuitry, contributing to Parkinson's disease (PD) risk. The molecular mechanisms regulating central iron levels, beyond a few well-known genes controlling peripheral iron, remain unclear. We generated scores based on the archetypal brain iron accumulation observed in magnetic resonance imaging scans of individuals with excessive dietary iron absorption and hemochromatosis risk. Genome-wide analysis revealed that this score is highly heritable, identifying loci associated with iron homeostasis, and driven by peripheral iron levels. Our score predicted gait abnormalities and showed a U-shaped relationship with PD risk, identifying individuals with threefold increased risk. These results establish a hormetic relationship between brain iron and PD risk, where central iron levels are strongly determined by genetics via peripheral iron. This framework combining forward and reverse genetics is a powerful study design to understand genomic drivers underlying high dimensional phenotypes.

Synchrotron Radiation Study of Fe-N-C Catalysts for Dissemination of Fuel Cells

Fuel cells are attracting worldwide attention as a technology for achieving carbon neutrality. However, existing fuel cells use large amounts of precious metals, which limits their widespread use. In our laboratory, we are studying fuel cells that do not use precious metals. We have synthesized iron-based complex catalysts using the method reported by Plamen Atanassov1 et al. Iron nitrate nonahydrate is mixed in aqueous solution with glucose, 2-methylimidazole, and zinc nitrate hexahydrate, followed by hydrothermal synthesis at 200°C for 24 hours. Hydrogen heat treatment is performed at 950 °C, followed by acid treatment with nitric acid to dissolve excess metallic iron. Then, ammonia heat treatment is performed at 950 °C to complete the Fe-N-C catalyst. In our laboratory, we have been working on various process improvements to reduce the amount of metallic iron to make the four-electron oxygen reduction reaction proceed more seamlessly. What is particularly noteworthy is that we have dramatically reduced the amount of iron nitrate dihydrate, a catalyst raw material, by 1/40.The results of the XAFS measurement of the reduced catalyst at SPring-8 BL14B1 showed that the prepared Fe-N-C catalyst formed a good complex, since the peak was like that of iron phthalocyanine (II), an iron complex, which was very different from the peak of metallic iron (Fe foil) (Figure 1). Through these process improvements, we succeeded in creating a catalyst that exceeds the performance of current platinum catalysts. The structure of the catalyst changes the ease of oxygen adsorption. The prepared catalysts were used to determine the catalyst fine structure by High-Energy-Resolution-Fluorescence-Detected (HERFD-XAFS)2 and Kβ X-ray emission spectroscopy (XES) measurements using synchrotron radiation at BL-11XU in SPring-8, where the spectral broadening due to the inner-shell lifetime width is suppressed.Using a half-cell with only a cathode, the prepared Fe-N-C catalyst was sprayed onto carbon paper, and a potentiostat was manipulated to measure the adsorbed species on the Fe surface at a specific potential.We used N2 and O2 gases and discussed the structure and valence of the catalyst by the peak transitions, focusing on Fe in the Fe-N-C catalyst, The structure of Fe was discussed based on the energy position transition for each potential.The structure of Fe-N-C oxide was discussed based on the energy of the Fe K-edge (7115eV) in high-resolution XAFS (Fig. 2).The catalysts were measured on Fe-N-C catalysts prepared in the laboratory and were a mixture of planar 4- and 6-coordinated catalysts. From these results, it was considered that the highly active iron complex catalyst we prepared was a hybrid of multiple complex structures. The structure and activity will be discussed in an oral presentation.References 1 R. Gokhale, L-K. Tsui, K. Roach, Y. Chen, M. M. Hossen, K. Artyushkova, F. Garzon, P. Atanassov, “Hydrothermal synthesis of platinum-Group-Metal-Free Catalysts: Structural Elucidation and Oxygen Reduction Catalysis”, ChemElectroChem, 5, 14, (2017), 1848-1853 2N. Yamamoto, D.Matsumura, Y. Hagihara, K.Tanaka, Y. Hasegawa, K. Ishii, H. Tanaka, “Investigation of hydrogen superoxide adsorption during ORR on Pt/C catalyst in acidic solution for PEFC by in-situ high energy resolution XAFS”, Jounal of Power Sources, 557, 15, (2023), 232508AcknowledgmentsThe authors would like to express their sincere appreciation and celebration to Professor Dr. P. Atanassov of University of California, Irvine for his contribution to the fuel cell research. The authors would like to acknowledge Dr. S. Kusano and Emeritus Professor Dr. J. Mizuki of Kwansei Gakuin University for their cooperation in establishing the in-situ XAFS measurement method. And they also thank Dr. H. Kishi, Dr. T. Sakamoto and Dr. K. Asazawa of Daihatsu Motor Co., Ltd. for development of electrochemical cells.The synchrotron radiation experiments were performed at the BL11XU in the SPring-8 with the approval of the Japan Synchrotron Radiation Research Institute (JASRI) (Proposal Numbers of 2022B-H08,2023A-H06,2023B-H08,2024A-H10)The SPring-8 experiment was also supported by the Advanced Research Infrastructure for Materials and Nanotechnology in Japan (ARIM Japan) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan (Grant No. JPMXP1222QSO108, JPMXP1223QS0006, JPMXP1223QS0108, JPMXP1224QS0010)This work was supported by JSPS KAKENHI Grant Number JP22H02188) Figure 1

Targeting inhibition of T3JAM reduces brain cell ferroptosis in rat following ischemia/reperfusion via a mechanism involving prevention of TLR4-mediated iron overload

Iron overload-dependent ferroptosis is believed to contribute to the brain injury of ischemia/reperfusion (I/R), whereas toll-like receptor 4 (TLR4) can exert pro-ferroptosis effect via inhibiting the glutathione peroxidase 4 (GPX4) level, but the mechanisms behind these phenomenon are not fully elucidated. Tumor necrosis factor receptor correlated factor 3-interaction Jun amino-terminal kinase [JNK]-activating modulator (T3JAM) can activate specific molecule and its downstream signaling pathways, including TLR4. This study aims to explore whether targeting T3JAM can reduce I/R-induced ferroptosis in brain via downregulating TLR4. A Sprague Dawley (SD) rat model of cerebral I/R injury was established by 2 h-ischemia plus 24 h-reperfusion, which displayed brain injury (increases in neurological deficit score and infarct volume) and upregulation of T3JAM and TLR4, concomitant with the increased ferroptosis, reflected by increases in the levels of transferrin receptor protein 1 (TfR1), total iron, Fe2+ and lipid peroxidation (LPO) while decreases in the levels of ferroportin (FPN) and GPX4. Consistently, similar results were achieved in the cultured HT22 cells subjected to 8h-oxygen-glucose deprivation plus 12 h-reoxygenation (OGD/R), and knockdown of T3JAM reversed these phenomena. Moreover, Telaprevir, an anti-hepatitis C virus (HCV) drug, could also provide beneficial effect on alleviating ischemic brain injury via inhibition of T3JAM. Based on these observations, we conclud that inhibition of T3JAM can reduce I/R-induced brain cell ferroptosis through downregulating TLR4 and that T3JAM could be a potential target for identifying novel or existing drugs (such as Telaprevir) to treat cerebral I/R injury.