Iron Content Research Articles

Iron parameters analysis in dogs with myxomatous mitral valve disease.

Myxomatous mitral valve disease (MMVD) is the most common acquired cardiovascular disease in small breed dogs. In contrast to human patients with heart failure (HF), iron deficiency (ID) prevalence in dogs with MMVD is weakly known. The study aimed to assess the usability of ID markers in serum and reticulocyte parameters from whole blood of dogs with MMVD to evaluate early ID symptoms. Sixty-eight dogs (43 male and 25 female) were included in the study. MMVD dogs were assigned according to the 2019 ACVIM guidelines for groups B1 (n = 9), B2 (n = 10), C (n = 27) and D (n = 10). Groups were also combined into B1 and B2 as non-symptomatic HF and C with D as symptomatic HF. Healthy controls were 12 dogs. Serum iron concentration below the reference range in dogs with MMVD was 12.5%. Other ID indices, such as %SAT, UIBC, and TIBC were similar in the MMVD groups and healthy controls (p > 0.05 for all parameters). Statistical comparison between control group and 4 groups of different stages of MMVD showed that significant differences occur only in serum transferrin. The assessment of ferritin and soluble transferrin receptors using Western Blotting did not show differences between control (n = 7) and MMVD (n = 33) dogs. Study has shown positive correlation between ID parameters and echocardiographic indices such as LA/Ao and LVIDdN, and some biochemical parameters. A significant increase in reticulocytes percentage, assessed manually, was observed in the HF group of animals (p = 0.027) compared to the control group. Studies have shown that ID parameters in serum are not significantly different in dogs with MMVD compared to healthy dogs. However, there is a clear correlation between atrial size and normalised left ventricular size to body size and some biochemical parameters, including ID parameters and therefore the severity of MMVD.

Analysis of Iron (Fe) Content in Breast Milk of Breastfeeding Mothers with Level Anemia Normal, Mild and Moderate

Analysis of Iron (Fe) Content in Breast Milk of Breastfeeding Mothers with Level Anemia Normal, Mild and Moderate

Knockdown of LncRNA Lcn2-204 alleviates sepsis-induced myocardial injury by regulation of iron overload and ferroptosis.

Ferroptosis is an iron-dependent programmed cell death form resulting from lipid peroxidation damage, it plays a key role in organ damage and tumor development from various causes. Sepsis leads to severe host response after infection with high mortality. The long non-coding RNAs (LncRNAs) are involved in different pathophysiological mechanisms of multiple diseases. Here, we used cecal ligation and puncture (CLP) operation to mimic sepsis induced myocardial injury (SIMI) in mouse model, and LncRNAs and mRNAs were profiled by Arraystar mouse LncRNA Array V3.0. Based on the microarray results, 552 LncRNAs and 520 mRNAs were differentially expressed in the sham and CLP groups, among them, LncRNA Lcn2-204 was the highest differentially expressed up-regulated LncRNA. Iron metabolism disorder was involved in SIMI by bioinformatics analysis, meanwhile, myocardial iron content and lipocalin-2 (Lcn2) protein expressions were increased. The CNC network comprised 137 positive interactions and 138 negative interactions. Bioinformatics analysis showed several iron-related terms were enriched and six genes (Scara5, Tfrc, Lcn2, Cp, Clic5, Ank1) were closely associated with iron metabolism. Then, we constructed knockdown LncRNA Lcn2-204 targeting myocardium and found that it ameliorated cardiac injury in mouse sepsis model through modulating iron overload and ferroptosis. In addition, we found that LncRNA Lcn2-204 was involved in the regulation of Lcn2 expression in septic myocardial injury. Based on these findings, we conclude that iron overload and ferroptosis are the key mechanisms leading to myocardial injury in sepsis, knockdown of LncRNA Lcn2-204 plays the cardioprotective effect through inhibition of iron overload, ferroptosis and Lcn2 expression. It may provide a novel therapeutic approach to ameliorate sepsis-induced myocardial injury.

Potential of metallurgical iron-containing solid waste-based catalysts as activator of persulfate for organic pollutants degradation.

The production of solid wastes in the metallurgical industry has significant implications for land resources and environmental pollution. To address this issue, it is crucial to explore the potential of recycling these solid wastes to reduce land occupation while protecting the environment and promoting resource utilization. Steel slag, red mud, copper slag and steel picking waste liquor are examples of solid wastes generated during the metallurgical process that possess high iron content and Fe species, making them excellent catalysts for persulfate-based advanced oxidation processes (PS-AOPs). This review elucidates the catalytic mechanisms and pathways of Fe2+ and Fe0 in the activation PS. Additionally, it underscores the potential of metallurgical iron-containing solid waste (MISW) as a catalyst for PS activation, offering a viable strategy for its high-value utilization. Lastly, the article provides an outlook towards future challenges and prospects for MISW in PS activation for the degradation of organic pollutants.

Cellular iron deposition patterns predict clinical subtypes of multiple system atrophy.

Multiple system atrophy (MSA) is a primary oligodendroglial synucleinopathy, characterized by elevated iron burden in early-affected subcortical nuclei. Although neurotoxic effects of brain iron deposition and its relationship with α-synuclein pathology have been demonstrated, the exact role of iron dysregulation in MSA pathogenesis is unknown. Therefore, advancing the understanding of iron dysregulation at the cellular level is critical, especially in relation to α-synuclein cytopathology. Iron burden in subcortical and brainstem regions were histologically mapped in human post-mortem brains of 4 MSA-parkinsonian (MSA-P), 4 MSA-cerebellar (MSA-C), and 1 MSA case with both parkinsonian and cerebellar features. We then performed the first cell type-specific evaluation of pathological iron deposition in α-synuclein-affected and -unaffected cells of the globus pallidus, putamen, and the substantia nigra, regions of highest iron concentration, using a combination of iron staining with immunolabelling. Selective regional and cellular vulnerability patterns of iron deposition were compared between disease subtypes. In 7 MSA cases, expression of key iron- and closely related oxygen-homeostatic genes were examined. MSA-P and MSA-C showed different patterns of regional iron burden across the pathology-related systems. We identified subcortical microglia to predominantly accumulate iron, which was more distinct in MSA-P. MSA-C showed relatively heterogenous iron accumulation, with greater or similar deposition in astroglia. Iron deposition was also found outside cellular bodies. Cellular iron burden associated with oligodendrocytic, and not neuronal, α-synuclein cytopathology. Gene expression analysis revealed dysregulation of oxygen homeostatic genes, rather than of cellular iron. Importantly, hierarchal cluster analysis revealed the pattern of cellular vulnerability to iron accumulation, distinctly to α-synuclein pathology load in the subtype-related systems, to distinguish MSA subtypes. Our comprehensive evaluation of iron deposition in MSA brains identified distinct regional, and for the first time, cellular distribution of iron deposition in MSA-P and MSA-C and revealed cellular vulnerability patterns to iron deposition as a novel neuropathological characteristic that predicts MSA clinical subtypes. Our findings suggest distinct iron-related pathomechanisms in MSA clinical subtypes that are therefore not a consequence of a uniform down-stream pathway to α-synuclein pathology, and inform current efforts in iron chelation therapies at the disease and cellular-specific levels.

ANO6 (TMEM16F) inhibits gastrointestinal stromal tumor growth and induces ferroptosis.

Herein, we elucidate the potential role of ANO6 (TMEM16F) in gastrointestinal stromal tumors (GISTs). ANO6 expression in GIST and adjacent normal tissues was determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. Cell proliferation, apoptosis, and pyroptosis were examined utilizing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, terminal deoxynucleotidyl transferase dUTP Nick-End Labeling staining, and flow cytometry. In addition, the total iron and Fe2+ levels were assessed. IL-18 and IL-1β levels were also evaluated. Lipid reactive oxygen species (ROS), cystine (Cys), glutathione (GSH), and glutathione peroxidase 4 (GPX4) levels were evaluated using appropriate kits. Ferroptotic markers, including Ptgs2, Chac1, SLC7A11, and SLC3A2, were analyzed by RT-qPCR, western blotting, and immunohistochemistry. ANO6 expression decreased in GIST tissues. ANO6-plasmid inhibits proliferation, induces apoptosis, and promotes pyroptosis in GIST-T1 and GIST-T1 IR cells. The ANO6-plasmid induced ferroptosis, as confirmed by enhanced lipid ROS levels, increased intracellular concentrations of total iron and Fe2+, promoted Ptgs2 and Chac1 expression, reduced Cys, GSH, and GPX4 levels, and downregulated SLC7A11 and SLC3A2 expression after in vitro and in vivo treatment with ANO6-plasmid. Moreover, the ANO6-plasmid inhibited GIST growth in vivo. Therefore, ANO6 may be a promising therapeutic target for blocking the development of GIST via the induction of apoptosis, pyroptosis, and ferroptosis.

Silicon extraction from x-ray amorphous soil constituents: a method comparison of alkaline extracting agents

The growing interest in amorphous silica (ASi) within the fields of soil science and ecology underscores the necessity for a reliable protocol to estimate ASi contents in soil. Alkaline wet chemical extraction methods are commonly employed for silicon (Si) extraction from operationally defined (x-ray) amorphous Si phases or short-range ordered mineral phases in soils and marine sediments. In our study we conducted a comparative analysis of four alkaline extraction methods (1% sodium carbonate, 0.5 M sodium carbonate, 0.2 M sodium hydroxide, and 0.1 M Tiron), assessing their extraction selectivity as well as effectiveness using soils artificially enriched with varying, defined amounts of ASi. While extraction effectiveness was evaluated by determining the recovery rate of initially added ASi, extraction selectivity was determined by measuring aluminum (Al) and iron (Fe) concentrations as indicators of the dissolution of non-target mineral phases. Microwave plasma atom emission spectrometry was used to analyze Al, Fe, and Si concentrations in the extracts. Our results indicate that extraction with 0.2 M sodium hydroxide yields the best outcomes in terms of both extraction effectiveness and selectivity. This more recent extraction technique is conducted at the most alkaline pH (13.3) of all four methods tested, but at ambient temperature (21°C) decreasing the dissolution of non-target mineral phases. Though, no wet-chemical extraction used on heterogeneous samples like soil is precisely selective, and thus able to quantify the target analyte only. Hence, data obtained by such procedures still need to be interpreted with caution considering all their limitations.

Epoxiconazole degradation in water samples: a comparative study of Fenton, photo-Fenton, solar photo-Fenton, and solar photolysis processes.

Epoxiconazole (EPO) is classified as a persistent organic pollutant due to its ability to persist in the environment for prolonged periods. Its degradation is pivotal in mitigating its environmental impact. This investigation focuses on assessing the degradation of EPO using various methodologies, namely Fenton, photo-Fenton, solar photo-Fenton, and solar photolysis, conducted in both Milli-Q water and groundwater. These experiments encompassed evaluations at both the standard pH typically used in photo-Fenton reactions and the natural pH levels inherent to the respective aqueous environments. Additionally, EPO degradation products were analyzed after a 60-min reaction. Notably, in systems utilizing groundwater, the inclusion of additional iron was unnecessary, as the naturally occurring iron content in the groundwater facilitated the intended processes. Specifically, in Milli-Q water, solar photo-Fenton demonstrated an EPO degradation efficiency of 97%. Furthermore, the substitution of Milli-Q water with groundwater in Fenton-like processes did not significantly affect the efficacy of EPO degradation. These findings underscore the potential of solar photo-Fenton as an economically viable and environmentally sustainable strategy for EPO degradation.

Baby Food Pouches, Baby-Led Weaning, and Iron Status in New Zealand Infants: An Observational Study

Iron deficiency in infants can impact development, and there are concerns that the use of baby food pouches and baby-led weaning may impair iron status. First Foods New Zealand (FFNZ) was an observational study of 625 New Zealand infants aged 6.9 to 10.1 months. Feeding methods were defined based on parental reports of infant feeding at “around 6 months of age”: “frequent” baby food pouch use (five+ times per week) and “full baby-led weaning” (the infant primarily self-feeds). Iron status was assessed using a venepuncture blood sample. The estimated prevalence of suboptimal iron status was 23%, but neither feeding method significantly predicted body iron concentrations nor the odds of iron sufficiency after controlling for potential confounding factors including infant formula intake. Adjusted ORs for iron sufficiency were 1.50 (95% CI: 0.67–3.39) for frequent pouch users compared to non-pouch users and 0.91 (95% CI: 0.45–1.87) for baby-led weaning compared to traditional spoon-feeding. Contrary to concerns, there was no evidence that baby food pouch use or baby-led weaning, as currently practiced in New Zealand, were associated with poorer iron status in this age group. However, notable levels of suboptimal iron status, regardless of the feeding method, emphasise the ongoing need for paying attention to infant iron nutrition.

Influence of ocean alkalinity enhancement with olivine or steel slag on a coastal plankton community in Tasmania

Abstract. Ocean alkalinity enhancement (OAE) aims to increase atmospheric CO2 sequestration in the oceans through the acceleration of chemical rock weathering. This could be achieved by grinding rocks containing alkaline minerals and adding the rock powder to the surface ocean where it dissolves and chemically locks CO2 in seawater as bicarbonate. However, CO2 sequestration during dissolution coincides with the release of potentially bioactive chemicals and may induce side effects. Here, we used 53 L microcosms to test how coastal plankton communities from Tasmania respond to OAE with olivine (mainly Mg2SiO4) or steel slag (mainly CaO and Ca(OH)2) as alkalinity sources. Three microcosms were left unperturbed and served as a control, three were enriched with olivine powder (1.9 g L−1), and three were enriched with steel slag powder (0.038 g L−1). Olivine and steel slag powders were of similar grain size. Olivine was added in a higher amount than the steel slag with the aim of compensating for the lower efficiency of olivine to deliver alkalinity over the 3-week experiment. Phytoplankton and zooplankton community responses as well as some biogeochemical parameters were monitored. Olivine and steel slag additions increased total alkalinity by 29 and 361 µmol kg−1, respectively, corresponding to a respective theoretical increase of 0.9 % and 14.8 % of the seawater storage capacity for atmospheric CO2. Olivine and steel slag released silicate nutrients into the seawater, but steel slag released considerably more and also significant amounts of phosphate. After 21 d, no significant difference was found in dissolved iron concentrations (>100 nmol L−1) in the treatments and the control. The slag addition increased dissolved manganese concentrations (771 nmol L−1), while olivine increased dissolved nickel concentrations (37 nmol L−1). There was no significant difference in total chlorophyll-a concentrations between the treatments and the control, likely due to nitrogen limitation of the phytoplankton community. However, flow cytometry results indicated an increase in the cellular abundance of several smaller (∼<20 µm) phytoplankton groups in the olivine treatment. The abundance of larger phytoplankton (∼>20 µm) decreased much more in the control than in the treatments after day 10. Furthermore, the maximum quantum yields of photosystem II (Fv/Fm) were higher in slag and olivine treatments, suggesting that mineral additions increased photosynthetic performance. The zooplankton community composition was also affected, with the most notable changes being observed in the dinoflagellate Noctiluca scintillans and the appendicularian Oikopleura sp. in the olivine treatment. Overall, the steel slag used here was more efficient for CO2 removal with OAE than the olivine over the 3-week timescale of the experiment. Furthermore, the steel slag appeared to induce less change in the plankton community than the olivine when comparing the CO2 removal potential of both minerals with the level of environmental impact that they caused.

Biofortification for Enhancement of Zinc (Zn) and Iron (Fe) Content in Wheat (Triticum aestivum L.): A Comprehensive Review

Micronutrient deficiency, often termed "hidden hunger," is a significant global health issue affecting over three billion people worldwide. Given that wheat is a primary staple grain in many developing countries, it is crucial to focus on enhancing its nutritional content, as it often lacks essential micronutrients. Biofortification offers a promising solution to this problem by increasing the levels of key nutrients in the edible parts of crops through both agronomic and genetic approaches. This article examines the potential of biofortification in wheat, targeting vital micronutrients like iron and zinc. By employing traditional breeding methods alongside modern genetic techniques such as genome sequencing, Quantitative Trait Locus (QTL) mapping, and Genome Wide Association Studies (GWAS), researchers aim to boost the bioavailability and concentration of these nutrients in wheat varieties. The genetic variability observed in wild wheat relatives plays a significant role in identifying traits that can enhance the nutritional profile of wheat. The benefits of biofortified wheat are numerous, especially for resource-limited consumers who rely heavily on cereal-based diets. Increased concentrations of iron and zinc in wheat could improve the health and well-being of these populations, helping to combat hidden hunger on a global scale. This review paper provides a comprehensive overview of the current state of agronomic and genetic strategies for wheat biofortification, with a specific focus on increasing zinc and iron content in wheat grains. It discusses the progress made in biofortification research and outlines the various breeding approaches and genetic tools used to enhance wheat's nutritional content. By leveraging these strategies, wheat biofortification has the potential to address micronutrient deficiencies and contribute to improved public health outcomes worldwide.

Electrochemical ocean iron fertilization and alkalinity enhancement approach toward CO2 sequestration

Achieving net-zero emissions by 2050 requires the development of effective negative emission techniques, including ocean-based approaches for CO2 sequestration. However, the implementation and testing of marine CO2 removal (mCDR) techniques such as ocean iron fertilization (OIF) or ocean alkalinity enhancement (OAE) face significant challenges. Herein, a novel self-operating electrochemical technology is presented that not only combines OIF and OAE, but also recovers hydrogen gas (H2) from seawater, hence offering a promising solution for achieving quantifiable and transparent large-scale mCDR. Experimental results show that the electrochemical OIF (EOIF) can not only increase the concentration of ferrous iron (Fe+2) by 0–0.5 mg/L, but also significantly increases the seawater pH by 8% (i.e., a 25% decrease in the hydrogen ions concentration). The release of iron (Fe+2/Fe+3) can be regulated by adjusting the magnitude of the electric current and its form (e.g., pulsed current and polarity reversal), as well as by optimizing the electrode material and geometry. In certain ocean regions, enhanced iron concentrations stimulate the naturally occurring biological carbon pump (BCP), leading to increased phytoplankton growth, CO2 uptake, and subsequent export of carbon to the deep ocean. Simultaneously, the system increases seawater alkalinity and the buffer capacity, enhancing CO2 solubility and storage in the shallow ocean through the solubility pump. The obtained measurements demonstrate the scalability of EOIF and its ability to operate using solar energy at a lower cost. Overall, the proposed EOIF technology offers a practical, effective, and sustainable solution for addressing climate change on a large scale.

Impact of foliar application of iron and zinc fertilizers on grain iron, zinc, and protein contents in bread wheat (Triticum aestivum L.)

Impact of foliar application of iron and zinc fertilizers on grain iron, zinc, and protein contents in bread wheat (Triticum aestivum L.)

Agronomic biofortification of basmati rice (Oryza sativa L.) through iron and boron under varying seedling densities

Agronomic biofortification of basmati rice (Oryza sativa L.) through iron and boron under varying seedling densities

Drosophila Evi5 is a critical regulator of intracellular iron transport via transferrin and ferritin interactions

Vesicular transport is essential for delivering cargo to intracellular destinations. Evi5 is a Rab11-GTPase-activating protein involved in endosome recycling. In humans, Evi5 is a high-risk locus for multiple sclerosis, a debilitating disease that also presents with excess iron in the CNS. In insects, the prothoracic gland (PG) requires entry of extracellular iron to synthesize steroidogenic enzyme cofactors. The mechanism of peripheral iron uptake in insect cells remains controversial. We show that Evi5-depletion in the Drosophila PG affected vesicle morphology and density, blocked endosome recycling and impaired trafficking of transferrin-1, thus disrupting heme synthesis due to reduced cellular iron concentrations. We show that ferritin delivers iron to the PG as well, and interacts physically with Evi5. Further, ferritin-injection rescued developmental delays associated with Evi5-depletion. To summarize, our findings show that Evi5 is critical for intracellular iron trafficking via transferrin-1 and ferritin, and implicate altered iron homeostasis in the etiology of multiple sclerosis.

Obesity, liver steatosis and metabolic syndrome: The hidden enemies in transfusion-dependent thalassaemia.

In their paper, the authors quantified liver iron concentration (LIC) and hepatic steatosis (HS) using MRI-T2* technology in transfusion-dependent thalassaemia (TDT) patients and healthy controls and found that the prevalence of HS among patients with TDT was 36.4%. In comparison with healthy controls, the hepatic fat fraction (FF) was significantly higher in the TDT population (p = 0.013). Active hepatitis C virus infection, body mass index (BMI) and LIC were independent predictors of HS. An inverse correlation between hepatic FF and high-density lipoprotein cholesterol (p = 0.042) and a significant association of high glycaemia level (p = 0.037) with higher hepatic FF and a significant relationship (p = 0.026) between HS and higher BMI (though in a 'lean' group of patients) in TDT patients indicated that 'metabolic syndrome' was present in this subset with TDT. The impact of metabolic syndrome on TDT, including cardiac disease unrelated to iron overload, needs further study. Commentary on: Ricchi etal. Liver steatosis in patients with transfusion-dependent thalassaemia. Br J Haematol 2024 (Online ahead of print). doi: 10.1111/bjh.19496.

Sulfated Nutrition Modifies Nutrient Content and Photosynthetic Pigment Concentration in Cabbage under Salt Stress

Negative effects of salt stress may be counteracted by adequate management of sulfated nutrition. Herein, we applied 3.50, 4.25, and 5.00 mM SO42− in a nutrient solution to counteract salt stress induced by 75 and 150 mM NaCl in cabbage cv. Royal. The increase in NaCl concentration from 75 to 150 mM reduced the contents of macronutrients and micronutrients in the shoot. When increasing from 3.50 to 4.25 mM SO42−, the contents of nitrogen (N), phosphorous (P), potassium (K), calcium (Ca), magnesium (Mg), and sulfur (S) in shoots were enhanced, at both concentrations of NaCl. Increasing from 3.50 to 4.25 mM SO42− enhanced iron (Fe), zinc (Zn), manganese (Mn), and sodium (Na) concentrations with 75 mM NaCl. With 150 mM NaCl, the increase from 3.50 to 4.25 mM SO42− enhanced the contents of Cu and Mn, but also those of Na. Chlorophylls a, b, and total decreased as the concentration of SO42− increased in plants treated with 150 mM NaCl. With 75 mM NaCl, carotenoid concentration had a positive relationship with SO42−. Hence, the 4.25 mM SO42− concentration increased the contents of macronutrients and micronutrients in the presence of 75 mM NaCl, while, with 150 mM NaCl, it improved the contents of macronutrients except K. The chlorophyll a/chlorophyll b ratio remained close to 3 when the plants were treated with 5.00 mM SO42−, regardless of NaCl. Similarly, this level of SO42− increased the concentration of carotenoids, which translated into reductions in the total chlorophyll/carotenoid ratios, indicating a protective effect of the photosynthetic apparatus. It is concluded that higher doses of sulfur favor the accumulation of nutrients and increase the concentration of carotenoids under salt stress.

Iron in Drinking Water and its Impact on Human Health –A Study in Selected Units of Jalalabad Cantonment

Background: Prevalence of diseases due to appearance of excessive amount of iron in drinking water is increasing day by day. Often people are unaware about the harmful effects of excess iron that may be present in drinking water. Sometimes they overlook minor symptoms. Adequate study was not conducted before to rule out actual scenario. The aim of this study is to represent the detrimental effect of excessive iron in drinking water as well as to create awareness to get rid of it. Methods: A cross-sectional study was carried out in 17 units of 17 Infantry Division, Jalalabad Cantonment, Sylhet covering the period of January 2019 to May 2019. Total 52 participants were selected purposively who had been suffering from various kinds of abdominal symptoms; their source of drinking water was marked. Water was collected from 5 different sources to see iron level. Again they were monitored after establishing Reverse Osmosis water filter. Iron level at drinking water was also tested each time at laboratory by Atomic Absorption spectrometry method. Informed written consent was taken from the patients. A semi-structured questionnaire was developed and data were collected by face to face interview. Analysis was done by using Statistical Package for the Social Sciences (SPSS version 25). Results: Among 52 respondents, maximum i.e. 25(48.07%) developed moderate to severe constipation followed by 16(30.76%) developed dyspepsia and while others i.e.11(21.15%) had both the symptoms. Symptoms were most severe among the persons who used to drink water having highest concentration of Iron (1.9 mg/L). After establishment of Reverse Osmosis water purifier the iron concentration in water reduced significantly and only 4(8%) had constipation and 6(12%) had dyspepsia. Conclusion: Iron is a natural constituent of drinking water. But considering the harmful impact of excessive iron present in drinking water to human health, regular monitoring should be ensured to identify the risk group. Bangladesh Armed Forces Med J Vol 56 No (2) December 2023, pp 58-64

CYB561 supports the neuroendocrine phenotype in castration-resistant prostate cancer.

Castration-resistant prostate cancer (CRPC) is associated with resistance to androgen deprivation therapy, and an increase in the population of neuroendocrine (NE) differentiated cells. It is hypothesized that NE differentiated cells secrete neuropeptides that support androgen-independent tumor growth and induce aggressiveness of adjacent proliferating tumor cells through a paracrine mechanism. The cytochrome b561 (CYB561) gene, which codes for a secretory vesicle transmembrane protein, is constitutively expressed in NE cells and highly expressed in CRPC. CYB561 is involved in the α-amidation-dependent activation of neuropeptides, and contributes to regulating iron metabolism which is often dysregulated in cancer. These findings led us to hypothesize that CYB561 may be a key player in the NE differentiation process that drives the progression and maintenance of the highly aggressive NE phenotype in CRPC. In our study, we found that CYB561 expression is upregulated in metastatic and NE prostate cancer (NEPC) tumors and cell lines compared to normal prostate epithelia, and that its expression is independent of androgen regulation. Knockdown of CYB561 in androgen-deprived LNCaP cells dampened NE differentiation potential and transdifferentiation-induced increase in iron levels. In NEPC PC-3 cells, depletion of CYB561 reduced the secretion of growth-promoting factors, lowered intracellular ferrous iron concentration, and mitigated the highly aggressive nature of these cells in complementary assays for cancer hallmarks. These findings demonstrate the role of CYB561 in facilitating transdifferentiation and maintenance of NE phenotype in CRPC through its involvement in neuropeptide biosynthesis and iron metabolism pathways.

Mechanism of acupotomy on chondrocyte ferroptosis in rabbits with knee osteoarthritis based on HSPA5/GPX4 signaling pathway

To observe the effect of acupotomy on heat shock protein A family member 5 (HSPA5)/glutathione peroxidase 4 (GPX4) signaling pathway in the chondrocytes of the rabbits with knee osteoarthritis (KOA) and explore the mechanism of acupotomy on chondrocyte ferroptosis in KOA. Twenty-seven New Zealand rabbits were randomly divided into a normal group, a model group and an acupotomy group, with 9 rabbits in each group. The left hind limb was fixed by the modified Videman method for 6 weeks to establish KOA model. After modeling, acupotomy was given in the acupotomy group, once a week and for consecutive 3 weeks. Using Lequesne MG score, the local symptoms, physical signs and functions of knee joint were evaluated. With HE staining and saffrane-solid green staining adopted, the morphology of chondrocytes and cartilage tissue was observed. Under transmission electron microscope, the mitochondrial structure of chondrocytes was observed. The iron content of cartilage tissue was detected by iron ion kit. The mitochondrial membrane potential (Δψm) and the reactive oxygen species (ROS) level in cartilage tissue were determined by flow cytometry, and the mitochondrial damage rate was calculated. The mRNA expression of HSPA5, GPX4, type Ⅱ collagen α1 chain (COL2A1), matrix metalloproteinases (MMP) 3 and MMP13 was detected by the real-time quantitative PCR; and the protein expression of HSPA5, GPX4, type Ⅱ collagen (COL-Ⅱ), MMP3 and MMP13 was detected by Western blot. The mean flourscence intensity of HSPA5 and GPX4 in cartilage tissue was determined by immunofluorescence. Before intervention, compared with the normal group, the Lequesne MG scores were increased in the model group and the acupotomy group (P<0.01). After intervention, the Lequesne MG score in the acupotomy group was decreased when compared with that in the model group. In comparison with that in the normal group, the number of chondrocytes was reduced and the cells were disarranged; the layers of cartilage structure were unclear, the tide lines disordered and blurred; the mitochondria were wrinkled and the mitochondrial crista decreased or even disappeared in the model group. Compared with the model group, the number of chondrocytes was increased, the layers of cartilage structure were clear, the tide lines recovered, the number of mitochondria elevated, with normal structure and more crista in the acupotomy group. The iron content of cartilage tissue was increased (P<0.01), the Δψm of chondrocytes was declined, the mitochondrial damage rate was increased (P<0.01), the average fluorescence intensity of ROS was increased (P<0.01); the mRNA and corresponding protein expression of HSPA5, GPX4 and COL2A1 was decreased (P<0.01), the mRNA and protein expression of MMP3 and MMP13 was increased (P<0.01) and the average fluorescence intensity of HSPA5, GPX4 was decreased (P<0.01) in the model group when compared with those in the normal group. Compared with the model group, the iron content in cartilage tissue was reduced (P<0.01), the Δψm of chondrocytes was increased, the mitochondrial damage rate was decreased (P<0.01), and the average fluorescence intensity of ROS was decreased (P<0.01); the mRNA and corresponding protein expression of HSPA5, GPX4 and COL2A1 was higher (P<0.01), and the mRNA and protein expression of MMP3 and MMP13 was lower, and the average fluorescence intensity of HSPA5, GPX4 was increased (P<0.01) in the acupotomy group. Acupotomy can alleviate cartilage injury of KOA rabbits, and its mechanism may be related to the regulation of HSPA5/GPX4 signaling pathway to maintain iron homeostasis in articular cartilage, thus inhibiting chondrocyte ferroptosis and relieving extracellular matrix degradation.