Ferritin Gene Research Articles

Targeting OGF/OGFR signal to mitigate doxorubicin-induced cardiotoxicity

Enkephalins are reportedly correlated with heart function. However, their regulation in the heart remains unexplored. This study revealed a substantial increase in circulating levels of opioid growth factor (OGF) (also known as methionine enkephalin) and myocardial expression levels of both OGF and its receptor (OGFR) in subjects treated with doxorubicin (Dox). Silencing OGFR through gene knockout or using adeno-associated virus serotype 9 carrying small hairpin RNA effectively alleviated Dox-induced cardiotoxicity (DIC) in mice. Conversely, OGF supplementation exacerbated DIC manifestations, which could be abolished by administration of the OGFR antagonist naltrexone (NTX). Mechanistically, the previously characterized OGF/OGFR/P21 axis was identified to facilitate DIC-related cardiomyocyte apoptosis. Additionally, OGFR was observed to dissociate STAT1 from the promoters of ferritin genes (FTH and FTL), thereby repressing their transcription and exacerbating DIC-related cardiomyocyte ferroptosis. To circumvent the compromised therapeutic effects of Dox on tumors owing to OGFR blockade, SiO2-based modifiable lipid nanoparticles were developed for heart-targeted delivery of NTX. The pretreatment of tumor-bearing mice with the assembled NTX nanodrug successfully provided cardioprotection against Dox toxicity without affecting Dox therapy in tumors. Taken together, this study provides a novel understanding of Dox cardiotoxicity and sheds light on the development of cardioprotectants for patients with tumors receiving Dox treatment.

Exploring the evolutionary landscape and structural resonances of ferritin with insights into functional significance in plant

The mineral iron plays a crucial role in facilitating the optimal functioning of numerous biological processes within the cellular environment. These processes involve the transportation of oxygen, energy production, immune system functioning, cognitive abilities, and muscle function. However, it is crucial to note that excessive levels of iron can result in oxidative damage within cells, primarily through Fenton reactions. Iron availability and toxicity present significant challenges that have been addressed through evolution. Ferritin is an essential protein that stores iron and is divided into different subfamilies, including DNA-binding proteins under starvation (Dps), bacterioferritin, and classical ferritin. Ferritin plays a critical role in maintaining cellular balance and protecting against oxidative damage. This study delves into ferritin's evolutionary dynamics across diverse taxa, emphasizing structural features and regulatory mechanisms. Insights into ferritin's evolution and functional diversity are gained through phylogenetic and structural analysis in bacterial Dps, bacterioferritin, and classical ferritin proteins. Additionally, the involvement of ferritin in plant stress responses and development is explored. Analysis of ferritin gene expression across various developmental stages and stress conditions provides insights into its regulatory roles. This comprehensive exploration enhances our understanding of ferritin's significance in plant biology, offering insights into its evolutionary history, structural diversity, and protective mechanisms against oxidative stress.

Identification and differentiation of rice ferritin gene in two different chromosomes in several local Indonesian rice varieties

Abstract. Permatasari SE, Salamah A. 2024. Identification and differentiation of rice ferritin gene in two different chromosomes in several local Indonesian rice varieties. Biodiversitas 25: 2547-2555. The presence of ferritin protein in rice plants indicates a regulatory response to environmental iron stress. It prevents the destructive effects of the chain reaction resulting from Fe2+ accumulation in the cell. This study aimed to detect and identify the location of the ferritin gene (OsFER) in eight local rice varieties in Indonesia. OsFER2, as the target gene was amplified using PCR, visualized by electrophoresis, and then sequenced. The sequencing results were analyzed using DNA Baser, BioEdit, and ClustalX2. The predicted proteins were visualized using the SWISS-MODEL server, Rice Genome Annotation Project Database, and chromosome map tools. The results show that all rice varieties studied have 100% alignment similarity and OsFER characteristics on chromosome 11 at LOC_Os11g01530 and chromosome 12 at LOC_Os12g01530, which have striking differences. The complete protein structure of the complex is found on chromosome 12, while only a portion of alpha-helix ferritin is found on chromosome 11. Differences in the position of the ferritin gene at different chromosomes impact the functional changes in the ferritin protein as an iron homeostasis key role. The two genes show different characteristics and are located at different genomic positions, suggesting a potential regulatory impact on the level of iron stress resistance in Indonesian rice varieties.

Study on Ferritin Gene Expression to Evaluate the Health of White Leg Shrimp (Litopenaeus vannamei) Postlarvae Due to Changes in Water Temperature, Salinity, and pH

The growth and survival of marine organisms are influenced by environmental factors such as water temperature, salinity, and pH. Unsuitable environmental conditions may negatively impact marine organisms. The white leg shrimp (Litopenaeus vannamei), a euryhaline organism highly adapted to salinity, is a valuable species for aquaculture. This study examined the effects of water temperature, salinity, and pH on the health of postlarvae L. vannamei. Stress levels within the organisms were analyzed through the expression of a biomarker gene. Ferritin was selected as the biomarker gene for analysis. The experimental animal samples used were the hepatopancreas of L. vannamei postlarvae. The analysis was performed by qRT-PCR. The results showed that the adaptation of L. vannamei postlarvae to temperature was dependent on salinity. Under low-salinity conditions (5 psu), ferritin expression increased at 25 °C and 30 °C after 48 h of exposure; however, it decreased after 72 h of exposure. Under normal salinity conditions (27 psu), ferritin expression increased from 24 h to 72 h at water temperatures of 25 °C and 30 °C. These results indicate that low-salinity conditions may enable L. vannamei postlarvae to rapidly adapt to high temperatures. In conclusion, L. vannamei postlarvae adapt more efficiently to high temperatures under low-salinity conditions than that under high-salinity conditions. The results of this study could beneficially impact L. vannamei farming.

Preparation and brain targeting effects study of recombinant human ferritin nanoparticles

Human heavy-chain ferritin is a naturally occurring protein with high stability and multifunctionality in biological systems. This study aims to utilize a prokaryotic expression system to produce recombinant human heavy-chain ferritin nanoparticles and investigate their targeting ability in brain tissue. The human heavy-chain ferritin gene was cloned into the prokaryotic expression vector pET28a and transformed into Escherichia coli BL21 (DE3) competent cells to explore optimal expression conditions. The recombinant protein was then purified to evaluate its immunoreactivity and characteristics. Additionally, the distribution of the administered protein in normal mice and its permeability in an in vitro blood-brain barrier (BBB) model were measured. The results demonstrate that the purified protein can self-assemble extracellularly into nano-cage structures of approximately 10 nm and is recognized by corresponding antibodies. The protein effectively penetrates the blood-brain barrier and exhibits slow clearance in mouse brain tissue, showing excellent permeability in the in vitro BBB model. This study highlights the stable expression of recombinant human heavy-chain ferritin using the Escherichia coli prokaryotic expression system, characterized by favorable nano-cage structures and biological activity. Its exceptional brain tissue targeting and slow metabolism lay an experimental foundation for its application in neuropharmaceutical delivery and vaccine development fields.

Plasma and tissue transferrin and ferritin, and gene expression of ferritin, transferrin, and transferrin receptors I and II in channel catfish Ictalurus punctatus fed diets with different concentrations of inorganic or organic iron.

Ferritin, transferrin, and transferrin receptors I and II play a vital role in iron metabolism, health, and indication of iron deficiency anaemia in fish. To evaluate the use of high-iron diets to prevent or reverse channel catfish (Ictalurus punctatus) anaemia of unknown causes, we investigated the expression of these iron-regulatory genes and proteins in channel catfish fed plant-based diets. Catfish fingerlings were fed five diets supplemented with 0 (basal), 125, and 250 mg/kg of either inorganic iron or organic iron for 2 weeks. Ferritin, transferrin, and transferrin receptor I and II mRNA and protein expression levels in fish tissues (liver, intestine, trunk kidney, and head kidney) and plasma were determined. Transferrin (iron transporter) and TfR (I and II) genes were generally highly expressed in fish fed the basal diet compared to those fed the iron-supplemented diets. In contrast, ferritin (iron storage) genes were more expressed in the trunk kidney of fish fed the iron-supplemented diets than in those fed the basal diet. Our results demonstrate that supplementing channel catfish plant-based diets with iron from either organic or inorganic iron sources affected the expression of the iron-regulatory genes and increased body iron status in the fish.

Leishmanicidal and immunomodulatory activity of Terminalia catappa in Leishmania amazonensisin vitro infection

Leishmaniases are infectious-parasitic diseases that impact public health around the world. Antileishmanial drugs presented toxicity and increase in parasitic resistance. Studies with natural products show an alternative to this effect, and several metabolites have demonstrated potential in the treatment of various diseases. Terminalia catappa is a plant species with promising pharmaceutical properties. The objective of this work was to evaluate the therapeutic potential of extracts and fractions of T. catappa on Leishmania amazonensis and investigate the immunomodulatory mechanisms associated with its action. In anti-Leishmania assays, the ethyl acetate fraction exhibited activity against promastigotes (IC50 86.07 ± 1.09 μg/mL) and low cytotoxicity (CC50 517.70 ± 1.68 μg/mL). The ethyl acetate fraction also inhibited the intracellular parasite (IC50 25.74 ± 1.08 μg/mL) with a selectivity index of 20.11. Treatment with T. catappa ethyl acetate fraction did not alter nitrite production by peritoneal macrophages stimulated with L. amazonensis, although there was a decrease in unstimulated macrophages treated at 50 μg/mL (p = 0.0048). The T. catappa ethyl acetate fraction at 100 μg/mL increased TNF-α levels (p = 0.0238) and downregulated HO-1 (p = 0.0030) and ferritin (p = 0.0002) gene expression in L. amazonensis-stimulated macrophages. Additionally, the total flavonoid and ellagic acid content for ethyl acetate fraction was 13.41 ± 1.86 mg QE/g and 79.25 mg/g, respectively. In conclusion, the T. catappa ethyl acetate fraction showed leishmanicidal activity against different forms of L. amazonensis and displayed immunomodulatory mechanisms, including TNF-α production and expression of pro and antioxidant genes.

Conventional and germinated pearl millet flour (Pennisetum glaucum (L.) R. Br.) improves iron metabolism and antioxidant capacity in Wistar rats

The study evaluated the effect of the consumption of pearl millet whole grain pre-cooked conventional, germinated and extruded flours on iron metabolism and antioxidant capacity in rats. The animals received iron-free standard diet (SD iron-free) for 28 days and subsequently, were separated into 4 groups: ferrous sulfate (SD + FS), conventional millet flour (SD + CM), germinated millet flour (SD + GM), and extruded millet flour (SD + EM), for 21 days. The SD iron free group presented increased ferroportin, hephaestin and ferritin gene expression and MDA concentration compared to the SD + FS group. The SD + GM group increased DcytB, ferroportin and hephaestin gene expression. The SD + EM group increased the concentration of transferrin compared to the SD + FS group and serum iron, compared to the other groups, while reducing the liver's total antioxidant capacity (TAC). The SD + CM group increased ferritin expression. All test groups increased the gene expression of DMT1 and hephaestin, as well as serum iron and reduced transferrin, Nrf2 and TNF gene expression, and MDA, NO, SOD and CAT levels in relation to the SD + FS group. In view of this, millet provides promising bioavailability of iron. Besides, germination stands out for improving iron metabolism in relation to extrusion and is similar to conventional millet in antioxidant capacity.

Effects of Chlorella fusca-supplemented diet on intestinal microbiota and gene expression related to metabolism, stress, and immune response in Chelon labrosus

Microalgae are emerging as promising alternatives to traditional fishmeal and crop ingredients in aquaculture diets. In this study, we aimed to characterize the intestinal microbiota and evaluate the transcription of genes related to metabolism, stress, and the immune system in various tissues of thick-lipped grey mullet (Chelon labrosus) fed a diet containing 15 % of microalga Chlorella fusca against a microalgae-free control diet for 90 days. After this period, the fish were challenged with Aeromonas hydrophila and polyinosinic–polycytidylic acid (poly I:C) to assess their immune response during 72 h. Microbiota analysis revealed that dietary inclusion of microalgae did not produce significant differences in intestine diversity compared to the control group. The dominant genera differed between the groups, with Pseudomonas and Mycoplasma most abundant in the fish fed with C. fusca, and Brevinema, Cetobacterium, and Pseudomonas predominant in the control group. Gene expression analysis indicated significant changes in hif3α, mhcII, ferritin, and mx genes in different organs of fish fed the microalgae-supplemented diet. Gene expression in the head kidney showed variations after A. hydrophila or poly I:C challenges, and higher peak values were observed in poly I:C-injected fish. Furthermore, c3 mRNA levels were significantly up-regulated in the fish fed with C. fusca at 24 h following A. hydrophila challenge. In conclusion, the inclusion of C. fusca in the fish diet modifies the composition of microbial species in the intestine but does not seem to affect its functionality. Regarding the influence on gene expression, the microalgae-supplemented diet effectively modulates the transcription of stress and immune-related genes, which may potentially enhance the fish resistance to stress and infections.

Heterologous Expression of a Ferritin Homologue Gene PpFer1 from Prunus persica Enhances Plant Tolerance to Iron Toxicity and H2O2 Stress in Arabidopsis thaliana.

In plants, ferritin proteins play an important role in iron (Fe) storage which contributes to plant growth and development. However, the biological functions of ferritins in fruit trees are essentially unknown. In this study, three Ferritin genes were isolated from 'Zhentong No. 3' peach, which were named PpFer1-PpFer3. The expression levels of these genes were different in distinct tissues/organs. Notably, PpFer1 was the most abundantly expressed Ferritin family gene in all tested tissues of 'Zhentong No. 3' peach; its expression levels were significantly enhanced throughout the entire peach seedling under Fe toxicity and H2O2 stress, particularly in the leaves. In addition, over-expression of PpFer1 was effective in rescuing the retarded growth of Arabidopsis fer1-2 knockout mutant, embodied in enhanced fresh weight, primary root length, lateral root numbers, total root length, total leaf chlorophyll, stomatal conductance (Gs), net photosynthetic rate (Pn), transpiration rate, and tissue Fe concentration. This study provides insights into understanding the molecular mechanisms of Fe storage and sequestration in perennial fruit trees.

Identification and functional regulation of three alternative splicing isoforms of the fthl27 gene in miiuy croaker, Miichthys miiuy

Alternative splicing is an important basic mechanism for eukaryotes to control gene expression. Different forms of alternative splicing may lead to the production of protein subtypes with different functions, leading to the expansion of protein diversity in organisms, affecting cell production and metabolism, and is even related to the occurrence of many diseases. Many studies have shown that ferritin is usually associated with inflammation, vascular proliferation, and tumors, which is the focus of immunological research. It not only plays a role in iron metabolism and storage in the body, but also plays an important regulatory role in pathways related to immune and inflammatory regulation. However, there are few studies on alternative splicing events of the ferritin gene nowadays. Therefore, this study identified three different splicing isoforms in its ferritin gene fthl27 of Miichthys miiuy through Sanger sequencing, qRT-PCR, and other experimental techniques, and we found that three different splicing isoforms of the ferritin gene fthl27 in M. Miiuy cells showed an upregulation trend after being stimulated by Lipopolysaccharide (LPS) and poly (I: C). The experiment also found that the three isoforms may have different regulatory effects on the expression of inflammatory factors and antiviral immune factors, playing an important role in the innate immune response of fish.

The Effect of Ferritin Level and Gene Expression of β-globin Promoter with β-thalassemia Patients in Al-Qadisiyah Governorate, Iraq

Abstract BACKGROUND: The genetic condition β-thalassemia causes a deficit in the β-globin chain. Goblins are produced under the supervision of at least nine different genes. Thalassemia can be distinguished from other disorders by changes in these genes, which can lead to issues with hemoglobin synthesis. A typical side effect of thalassemia syndromes is iron overload, which raises the risk of mortality and can cause organ damage. Blood ferritin levels as well as total iron of body reserves have a positive correlation when there is no inflammation. OBJECTIVE: The purpose of this study was to assess the ferritin level of an Iraqi patient and the relationship between β-thalassemia and gene expression of β-globin. MATERIALS AND METHODS: A case–control study included 60 samples with mean age (17.76 ± 0.88; 28 males and 32 females) which had been collected from patients who were diagnosed with β-thalassemia and 60 samples with mean age (22.7 ± 0.75; 29 males, 31 females) which were collected from apparently healthy individuals as a control group (CG). The procedure’s outcome is monitored using polymerase chain reaction and the Fluorecare instrument. RESULTS: Ferritin levels in thalassemia patients were higher than in CG patients. The β-globin expression in the thalassemia group was significantly lower than in the CG. The discovery of two essential sequences thymine-adenine-thymine-adenine and cytosine-adenine-thymine-adenine in the β-gene promoter that are crucial in the start of transcription can account for this downregulation. Changes made to these sequences decreased the affinity of transcription factors, which in turn restricted the transcription of the messenger ribonucleic acid. Examples of these transcription factors are erythroid Kruppel-like factor and specificity protein 1. CONCLUSION: Ferritin can be a useful indicator of severe iron overload. The results showed that the level of expression of β-globin was dramatically downregulated within the thalassemia group as compared with the CG future prospective of this study.

Multiple roles of wheat ferritin genes during stress treatment and TaFER5D-1 as a positive regulator in response to drought and salt tolerance

Ferritin not only regulates the plant's iron content but also plays a significant role in the plant's development and resistance to oxidative damage. However, the role of the FER family in wheat has not been systematically elucidated. In this study, 39 FERs identified from wheat and its ancestral species were clustered into two subgroups, and gene members from the same group contain relatively conservative protein models. The structural analyses indicated that the gene members from the same group contained relatively conserved protein models. The cis-acting elements and expression patterns analysis suggested that TaFERs might play an important role combating to abiotic and biotic stresses. In the transcriptional analysis, the TaFER5D-1 gene was found to be significantly up-regulated under drought and salt stresses and was, therefore, selected to further explore the biological functions Moreover, the GFP expression assay revealed the subcellular localization of TaFER5D-1 proteins in the chloroplast, nucleus, membrane and cytoplasm. Over-expression of TaFER5D-1 in transgenic Arabidopsis lines conferred greater tolerance to drought and salt stress. According to the qRT-PCR data, TaFER5D-1 gene over-expression increased the expression of genes related to root development (Atsweet-17 and AtRSL4), iron storage (AtVIT1 and AtYSL1), and stress response (AtGolS1 and AtCOR47). So it is speculated that TaFER5D-1 could improve stress tolerance by promoting root growth, iron storage, and stress-response ability. Thus, the current study provides insight into the role of TaFER genes in wheat.

Iron Availability Influences Protein Carbonylation in Arabidopsis thaliana Plants.

Protein carbonylation is an irreversible form of post-translational modification triggered by reactive oxygen species in animal and plant cells. It occurs either through the metal-catalyzed oxidation of Lys, Arg, Pro, and Thr side chains or the addition of α, β-unsaturated aldehydes and ketones to the side chains of Cys, Lys, and His. Recent genetic studies concerning plants pointed to an implication of protein carbonylation in gene regulation through phytohormones. However, for protein carbonylation to stand out as a signal transduction mechanism, such as phosphorylation and ubiquitination, it must be controlled in time and space by a still unknown trigger. In this study, we tested the hypothesis that the profile and extent of protein carbonylation are influenced by iron homeostasis in vivo. For this, we compared the profile and the contents of the carbonylated proteins in the Arabidopsis thaliana wild-type and mutant-deficient in three ferritin genes under normal and stress conditions. Additionally, we examined the proteins specifically carbonylated in wild-type seedlings exposed to iron-deficient conditions. Our results indicated that proteins were differentially carbonylated between the wild type and the triple ferritin mutant Fer1-3-4 in the leaves, stems, and flowers under normal growth conditions. The profile of the carbonylated proteins was also different between the wild type and the ferritin triple mutant exposed to heat stress, thus pointing to the influence of iron on the carbonylation of proteins. Consistent with this, the exposure of the seedlings to iron deficiency and iron excess greatly influenced the carbonylation of certain proteins involved in intracellular signal transduction, translation, and iron deficiency response. Overall, the study underlined the importance of iron homeostasis in the occurrence of protein carbonylation in vivo.

Screening of morphology-related genes based on predator-induced transcriptome sequencing and the functional analysis of Dagcut gene in Daphnia galeata.

High fish predation pressure can trigger "induced defense" in Daphnia species, resulting in phenotypic plasticity in morphology, behavior, or life-history traits. The molecular mechanisms of defense morphogenesis (e.g., the tail spine and helmet) in Daphnia remain unclear. In the present study, the tail spine, helmet, and body of Daphnia galeata under fish and non-fish kairomones conditions were collected for transcriptome analysis. A total of 24 candidate genes related to the morphological defense of D. galeata were identified, including 2 trypsin, one cuticle protein, 1 C1qDC protein, and 2 ferritin genes. The function of the Dagcut gene (D. galeata cuticle protein gene) in relation to tail spine morphology was assessed using RNA interference (RNAi). Compared with the EGFP (Enhanced green fluorescent protein) treatment, after RNAi, the expression levels of the Dagcut gene (D. galeata cuticle protein gene) showed a significant decrease. Correspondingly, the tail spines of the offspring produced by D. galeata after RNAi of the Dagcut gene appeared curved during the experiment. In whole-mount in situ hybridization, a clear signal site was detected on the tail spine of D. galeata before RNAi which disappeared after RNAi. Our results suggest that the Dagcut gene may play an important role in tail spine formation of D. galeata, and will provide a theoretical basis for studying the molecular mechanisms of the morphological plasticity in cladocera in the future.

Ferritin from Mycobacterium abscessus is involved in resistance to antibiotics and oxidative stress.

Mycobacterium abscessus subsp. massiliense (Mycma) is a rapidly growing Mycobacterium belonging to the M. abscessus complex that is often associated with lung and soft tissue infection outbreaks. Mycma is resistant to many antimicrobials, including those used for treating tuberculosis. Therefore, Mycma infections are difficult to treat and may lead to high infectious complication rates. Iron is essential for bacterial growth and establishment of infection. During infection, the host reduces iron concentrations as a defense mechanism. To counteract the host-induced iron deficiency, Mycma produces siderophores to capture iron. Mycma has two ferritins (encoded by mycma_0076 and mycma_0077) modulated by different iron concentrations, which allow the survival of this pathogen during iron scarcity. In this study, we constructed knockout (Mycma 0076KO) and complemented (Mycma 0076KOc) gene strains for mycma_0076 to understand the function of 0076 ferritin. Deletion of mycma_0076 in Mycma led to the transition in colony morphology from smooth to rough, alteration of the glycopeptidolipids spectra, increased permeability of the envelope, reduction in biofilm formation, increased susceptibility to antimicrobials and hydrogen peroxide-induced oxidative stress, and decreased internalization by macrophages. This study shows that Mycma_0076 ferritin in Mycma is involved in resistance to oxidative stress and antimicrobials, and alteration of cell envelope architecture. KEY POINTS: • Deletion of the mycma_0076 gene altered colony morphology to rough; • Mycma 0076KO changed GPL profile; • Absence of Mycma_0076 ferritin results in increased susceptibility to antimicrobials and oxidative stress in Mycma. Legend: a In wild-type M. abscessus subsp. massiliensestrain, iron is captured from the environment by carboxymycobactins and mycobactins (1). Iron-dependent regulator (IdeR) proteins bind to ferrous iron (Fe+2) in the bacterial cytoplasm leading to the activation of the IdeR-Fe+2 complex (2). The activated complex binds to the promoter regions of iron-dependent genes, called iron box, which in turn help in the recruitment of RNA polymerase to promote transcription of genes such as mycma_0076 and mycma_0077 ferritin genes (3). Mycma_0076 and Mycma_0077 ferritins bind to excess iron in the medium and promote Fe2+ oxidation into ferric iron (Fe3+) and store iron molecules to be released under iron scarcity conditions. (4) Genes related to biosynthesis and transport of glycopeptidolipids (GPL) are expressed normally and the cell envelope is composed of different GPL species (colored squares represented on the cell surface (GPLs). Consequently, WT Mycma present smooth colony phenotype (5). b In Mycma 0076KO strain, the lack of ferritin 0076 causes overexpression of mycma_0077 (6), but does not restore wild-type iron homeostasis and thus may result in free intracellular iron, even in the presence of miniferritins (MaDps). The excess iron potentiates oxidative stress (7) by generating hydroxyl radicals through Fenton Reaction. During this process, through an unknown mechanism, that could involve Lsr2 (8), the expression of GPL synthesis locus is regulated positively and/or negatively, resulting in alteration of GPL composition in the membrane (as represented by different colors of squares on the cell surface), resulting in a rough colony phenotype (9). The changes of GPL can increase cell wall permeability, contributing to antimicrobial susceptibility (10).

Effect of Air Exposure and Re-Submersion on the Histological Structure, Antioxidant Response, and Gene Expression of Procambarus Clarkii.

Air exposure is an important environmental stressor during the transportation and cultivation of Procambarus clarkii. We evaluated the effect of re-submersion for 24 h after dry transportation for 24 h on the histological structure, antioxidant activity, and gene expression of crayfish. The antioxidant parameters of catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), and lactate dehydrogenase (LDH), and the relative expression of CAT, SOD, HSP70, and ferritin genes were subsequently measured in the hepatopancreas and gills at both stages. Histopathology found that air exposure led to vacuolation of the hepatopancreas and disorderly arrangement of respiratory epithelial cells (REC) in the gills. The activities of catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), and lactic dehydrogenase (LDH) in the hepatopancreas and gills increased with short-term air exposure. The relative expression of genes (CAT, SOD, HSP70, and Ferritin) were induced after short-term air exposure. During re-submersion, MDA content and CAT and SOD activities in the hepatopancreas and gills were restored after 24 h, however, LDH activity and hepatopancreatic tissue damage were not repaired. Our results indicate that air exposure can cause oxidative damage to P. clarkii, and CAT and SOD can be used to determine the response of crayfish exposed to air, in addition to some damage that can be eliminated after re-submersion to a limited degree. This study provides foundational data that re-submersion can improve crayfish performance under hypoxic stress to a certain extent and will lead to the development of more effective transportation strategies and decrease economic losses in the future.

Proof of concept and early development stage of market-oriented high iron and zinc rice expressing dicot ferritin and rice nicotianamine synthase genes

Micronutrient deficiencies such as iron (Fe), zinc (Zn), and vitamin A, constitute a severe global public health phenomenon. Over half of preschool children and two-thirds of nonpregnant women of reproductive age worldwide have micronutrient deficiencies. Biofortification is a cost-effective strategy that comprises a meaningful and sustainable means of addressing this issue by delivering micronutrients through staple foods to populations with limited access to diverse diets and other nutritional interventions. Here, we report on the proof-of-concept and early development stage of a collection of biofortified rice events with a high density of Fe and Zn in polished grains that have been pursued further to advance development for product release. In total, eight constructs were developed specifically expressing dicot ferritins and the rice nicotianamine synthase 2 (OsNAS2) gene under different combinations of promoters. A large-scale transformation of these constructs to Bangladesh and Philippines commercial indica cultivars and subsequent molecular screening and confined field evaluations resulted in the identification of a pool of ten events with Fe and Zn concentrations in polished grains of up to 11 μg g−1 and up to 37 μg g−1, respectively. The latter has the potential to reduce the prevalence of inadequate Zn intake for women of childbearing age in Bangladesh and in the Philippines by 30% and 50%, respectively, compared to the current prevalence. To our knowledge, this is the first potential biotechnology public-sector product that adopts the product cycle phase-gated approach, routinely applied in the private sector.

Drought stress alters iron accumulation in Sorghum bicolor seeds

Sorghum ( Sorghum bicolor L.) is a cultivated food crop with iron-rich seeds that thrives in semi-arid areas. Although the mechanism of iron transport has been investigated in detail at the molecular level, the effects of drought stress on its regulation remain unclear. We cultivated sorghum in a chamber without (control) and with drought stress to determine iron accumulation in seeds. The drought stress treatment significantly increased the iron content by approximately 2-fold in seeds and decreased it by 0.6-fold in leaves. The results of RNA-sequencing leaves indicated that drought stress impaired the expression of genes involved in photosynthesis. This agreed with the finding that drought stress significantly reduced plant biomass. The expression of genes encoding vacuole iron transporter and ferritin were increased under drought stress. These expression profiles were confirmed by quantitative polymerase chain reaction, which indicated that expression of genes encoding the vacuole iron transporter and ferritin was significantly increased in leaves and seeds under drought stress. Furthermore, photosynthesis impaired by drought stress might trigger a disturbance in iron homeostasis. Therefore, increased vacuolar transporters and ferritin transcription might be important for safely storing excess iron. Our data suggested that maintaining iron homeostasis via vacuole iron transporters and ferritin enhances iron accumulation in seeds under drought stress. • Sorghum bicolor is drought stress-tolerant and has iron-enriched seeds • Drought stress enhanced iron accumulation in seeds • Drought stress affected genes associated with iron transport/homeostasis • Drought stress increased vacuole iron transporter and ferritin gene expression • Iron homeostasis might involve iron accumulation in drought-stressed seeds

A ferritin gene in the marine copepod Acartia tonsa as a highly sensitive biomonitor for nano-contamination

Toxicology is not only for eco-risk assessments, but also for the real-time environmental monitoring based on the quick response of specific biomarkers. Ferritin gene (ftn) is a potential biomarker involving in crucial protective responses in biota. However, little information is available concerning the ftn in marine copepod Acartia tonsa (A. tonsa), a model organism widely applied in toxicology assessments. Our study for the first time identified and characterized the ftn in A. tonsa, along with its time-dependent transcriptional response to the reproductive toxicity of two newly emerged nanomaterials. The full-length cDNA of ftn contains a 114-bp 5′-untranslated region (UTR), a 236-bp 3′-untranslated region, and a 510-bp open reading frame which encodes an 18.51 kDa polypeptide composed of 169 amino acids. The ftn sequence has an iron binding signature and a potential phosphorylation site, which is closely-related to the ftn of Calanus sinicus and Pseudodiaptomus annandalei genes at the phylogenetical level. The ftn showed a quick and highly sensitive response to nanomaterial exposures, even at no observed effect concentrations. In detail, after exposure to nickel nanomaterials (up to 17.0 mg/L), the ftn was significantly upregulated immediately at 0.5 h and peaked at 9.5-fold in adults within 48 h, along with a significant reduction of egg hatching rate. When exposed to CdSe/ZnS quantum dots (up to 135 mg/L), no significant change in egg productions or hatching rates was observed, while the expression of ftn still significantly increased to over 3.0-fold in the initial 48 h. After that, the upregulation of ftn induced by CdSe/ZnS quantum dots or nickel nanoparticles both gradually returned back within 96 h. These findings demonstrate the highly sensitive response of this new cloned ftn to nanomaterial exposures, and highlight the suitability of ftn in A. tonsa as a promising biomonitor for nano-contamination in marine environments.