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Wilson disease (WD) is a metabolic disorder caused by inactivation of the copper-transporting ATPase 2 (ATP7B) and copper (Cu) overload in tissues. Excess Cu causes oxidative stress and pathologic changes with poorly understood mechanistic connections. In Atp7b −/− mice with established liver disease, Cu overload activates the stress-sensitive transcription factor Nrf2 (nuclear factor erythroid-derived 2-like 2). Nrf2 targets, especially sulfotransferase 1e1 (Sult1e1), are strongly induced and cause elevation of sulfated sterols, whereas oxysterols are decreased. This sterol misbalance results in inhibition of the liver X receptor (LXR) and up-regulation of LXR targets associated with inflammatory responses. Pharmacological inhibition of Sult1e1 partially reverses oxysterol misbalance and LXR inhibition. Contribution of this pathway to advanced hepatic WD was demonstrated by treating mice with an LXR agonist. Treatment decreased inflammation by reducing expression of proinflammatory molecules, diminished fibrosis by down-regulating the noncanonical transforming growth factor–β signaling pathway, and improved liver morphology and function. Thus, the identified pathway is an important driver of WD.

Transferrin receptor 1 (TfR1) performs a critical role in cellular iron uptake. Hepatocyte TfR1 is also proposed to influence systemic iron homeostasis by interacting with the hemochromatosis protein HFE to regulate hepcidin production. Here, we generated hepatocyte Tfrc knockout mice (Tfrcfl/fl;Alb-Cre+), either alone or together with Hfe knockout or β-thalassemia, to investigate the extent to which hepatocyte TfR1 function depends on HFE, whether hepatocyte TfR1 impacts hepcidin regulation by serum iron and erythropoietic signals, and its contribution to hepcidin suppression and iron overload in β-thalassemia. Compared with Tfrcfl/fl;Alb-Cre− controls, Tfrcfl/fl;Alb-Cre+ mice displayed reduced serum and liver iron; mildly reduced hematocrit, mean cell hemoglobin, and mean cell volume; increased erythropoietin and erythroferrone; and unchanged hepcidin levels that were inappropriately high relative to serum iron, liver iron, and erythroferrone levels. However, ablation of hepatocyte Tfrc had no impact on iron phenotype in Hfe knockout mice. Tfrcfl/fl;Alb-Cre+ mice also displayed a greater induction of hepcidin by serum iron compared with Tfrcfl/fl;Alb-Cre− controls. Finally, although acute erythropoietin injection similarly reduced hepcidin in Tfrcfl/fl;Alb-Cre+ and Tfrcfl/fl;Alb-Cre− mice, ablation of hepatocyte Tfrc in a mouse model of β-thalassemia intermedia ameliorated hepcidin deficiency and liver iron loading. Together, our data suggest that the major nonredundant function of hepatocyte TfR1 in iron homeostasis is to interact with HFE to regulate hepcidin. This regulatory pathway is modulated by serum iron and contributes to hepcidin suppression and iron overload in murine β-thalassemia.

Wilson disease (WD) is caused by inactivation of the copper transporter Atp7b and copper overload in tissues. Mice with Atp7b deleted either globally (systemic inactivation) or only in hepatocyte recapitulate various aspects of human disease. However, their phenotypes vary, and neither the common response to copper overload nor factors contributing to variability are well defined. Using metabolic, histologic, and proteome analyses in three Atp7b-deficient mouse strains, we show that global inactivation of Atp7b enhances and specifically modifies the hepatocyte response to Cu overload. The loss of Atp7b only in hepatocytes dysregulates lipid and nucleic acid metabolisms and increases the abundance of respiratory chain components and redox balancing enzymes. In global knockouts, independently of their background, the metabolism of lipid, nucleic acid, and amino acids is inhibited, respiratory chain components are down-regulated, inflammatory response and regulation of chromosomal replication are enhanced. Decrease in glucokinase and lathosterol oxidase and elevation of mucin-13 and S100A10 are observed in all Atp7b mutant strains and reflect the extent of liver injury. The magnitude of proteomic changes in Atp7b−/− animals inversely correlates with the metallothioneins levels rather than liver Cu content. These findings facilitate identification of WD-specific metabolic and proteomic changes for diagnostic and treatment.

Background and aims Bone morphogenetic proteins BMP2 and BMP6 play key roles in systemic iron homeostasis by regulating production of the iron hormone hepcidin. The homeostatic iron regulator (HFE) also regulates hepcidin through a mechanism that intersects with the BMP-mothers against decapentaplegic homolog 1/5/8 (SMAD1/5/8) pathway. However, the relative roles of BMP2 compared with BMP6 and whether HFE regulates hepcidin through a BMP2-dependent mechanism remain uncertain. Approach and results We therefore examined the iron phenotype of mice deficient for both Bmp2 and Bmp6 or both Bmp2 and Hfe compared with single knockout (KO) mice and littermate controls. Eight-week-old double endothelial Bmp6/Bmp2 KO mice exhibited a similar degree of hepcidin deficiency, serum iron overload, and tissue iron overload compared with single KO mice. Notably, dietary iron loading still induced liver SMAD5 phosphorylation and hepcidin in double Bmp6/endothelial Bmp2 KO mice, although no other BMP ligand mRNAs were increased in the livers of double KO mice, and only Bmp6 and Bmp2 mRNA were induced by dietary iron loading in wild-type mice. In contrast, double Hfe/endothelial Bmp2 KO mice exhibited reduced hepcidin and increased extrahepatic iron loading compared to single Hfe or endothelial Bmp2 KO mice. Liver phosphorylated SMAD5 and the SMAD1/5/8 target inhibitor of DNA binding 1 (Id1) mRNA were also reduced in double Hfe/endothelial Bmp2 KO compared with single endothelial Bmp2 KO female mice. Finally, hepcidin and Id1 mRNA induction by homodimeric BMP2, homodimeric BMP6, and heterodimeric BMP2/6 were blunted in Hfe KO primary hepatocytes. Conclusions These data suggest that BMP2 and BMP6 work collaboratively to regulate hepcidin expression, that BMP2-independent and BMP6-independent SMAD1/5/8 signaling contributes a nonredundant role to hepcidin regulation by iron, and that HFE regulates hepcidin at least in part through a BMP2-independent but SMAD1/5/8-dependent mechanism.

Copper misbalance has been linked to fat accumulation in animals and experimental systems; however, information about copper homeostasis in human obesity is limited. In this study, the copper status of obese individuals was evaluated by measuring their levels of copper and cuproproteins in serum, adipose and hepatic tissues. The analysis of serum trace elements showed significant positive and element-specific correlation between copper and BMI after controlling for gender, age, and ethnicity. Serum copper also positively correlated with leptin, insulin, and the leptin/BMI ratio. When compared to lean controls, obese patients had elevated circulating cuproproteins, such as semucarbazide-sensitive amine oxidase (SSAO) and ceruloplasmin, and higher SSAO activity and copper levels in visceral fat. Although hepatic steatosis reduces copper levels in the liver, obese patients with no or mild steatosis have higher copper content in the liver compared to lean controls. In conclusion, obese patients evaluated in this study had altered copper status. Strong positive correlations of copper levels with BMI and leptin suggest that copper and/or cuproproteins may be functionally linked to fat accumulation.

Wilson disease (WND) is caused by inactivation of the copper transporter ATP7B and copper accumulation in tissues. WND presentations vary from liver steatosis to inflammation, fibrosis, and liver failure. Diets influence the liver phenotype in WND, but findings are inconsistent. To better understand the impact of excess calories on liver phenotype in WND, the study compared C57BL/6J Atp7b(−/−) and C57BL/6J mice fed for 12 weeks with Western diet or normal chow. Serum and liver metabolites, body fat content, liver histology, hepatic proteome, and copper content were analyzed. Wild-type and Atp7b(−/−) livers showed striking similarities in their responses to Western diet, most notably down-regulation of cholesterol biosynthesis, altered nuclear receptor signaling, and changes in cytoskeleton. Western diet increased body fat content and induced liver steatosis in males and females regardless of genotype; however, the effects were less pronounced in Atp7b(−/−) mice compared with those in the wild type mice. Although hepatic copper remained elevated in Atp7b(−/−) mice, liver inflammation was reduced. The diet diminished signaling by Rho GTPases, integrin, IL8, and reversed changes in cell cycle machinery and cytoskeleton. Overall, high calories decreased inflammatory response in favor of steatosis without improving markers of cell viability. Similar changes of cellular pathways during steatosis development in wild-type and Atp7b(−/−) mice explain histologic overlap between WND and non-alcoholic fatty liver disease despite opposite copper changes in these disorders.

The comparative efficacy of organic sources on cropping behavior, nutrient dynamics, physico-chemical and biological properties of soil and fruit quality under rain-fed agroecosystem on “Silver King” nectarines was investigated. Bio-organic nutrients, namely vermi-compost (VC), biofertilizer (BF), farmyard manure (FYM), compost (comp), vermiwash (VW) and cow urine (CU) were evaluated in 11 different treatment combinations. The treatment application of VC at 25 kg tree−1, BF at 40 g tree−1, FYM at 30 kg tree−1, comp at 15 kg tree−1, VW1:10 and CU1:10 significantly improved plant growth and fruit quality characteristics of nectarine trees. This superior combination also enhanced physico-chemical and biological properties of the rhizosphere soil when compared to control as conventional chemical fertilizer application nitrogen, phosphorus and potassium (NPK) fertilizers. Different treatments of bio-organic sources changed pH of the soil to neutral. Available macronutrient contents of soil (viz. N, P, ...

Iron is an essential element for numerous fundamental biologic processes, but excess iron is toxic. Abnormalities in systemic iron balance are common in patients with chronic kidney disease and iron administration is a mainstay of anemia management in many patients. This review provides an overview of the essential role of iron in biology, the regulation of systemic and cellular iron homeostasis, how imbalances in iron homeostasis contribute to disease, and the implications for chronic kidney disease patients.

A failure of iron to appropriately regulate liver hepcidin production is central to the pathogenesis of hereditary hemochromatosis. SMAD1/5 transcription factors, activated by bone morphogenetic protein (BMP) signaling, are major regulators of hepcidin production in response to iron; however, the role of SMAD8 and the contribution of SMADs to hepcidin production by other systemic cues remain uncertain. Here, we generated hepatocyte Smad8 single (Smad8fl/fl ;Alb-Cre+ ), Smad1/5/8 triple (Smad158;Alb-Cre+ ), and littermate Smad1/5 double (Smad15;Alb-Cre+ ) knockout mice to investigate the role of SMAD8 in hepcidin and iron homeostasis regulation and liver injury. We found that Smad8;Alb-Cre+ mice exhibited no iron phenotype, whereas Smad158;Alb-Cre+ mice had greater iron overload than Smad15;Alb-Cre+ mice. In contrast to the sexual dimorphism reported for wild-type mice and other hemochromatosis models, hepcidin deficiency and extrahepatic iron loading were similarly severe in Smad15;Alb-Cre+ and Smad158;Alb-Cre+ female compared with male mice. Moreover, epidermal growth factor (EGF) failed to suppress hepcidin in Smad15;Alb-Cre+ hepatocytes. Conversely, hepcidin was still increased by lipopolysaccharide in Smad158;Alb-Cre+ mice, although lower basal hepcidin resulted in lower maximal hepcidin. Finally, unlike most mouse hemochromatosis models, Smad158;Alb-Cre+ developed liver injury and fibrosis at 8 weeks. Liver injury and fibrosis were prevented in Smad158;Alb-Cre+ mice by a low-iron diet and were minimal in iron-loaded Cre- mice. Conclusion: Hepatocyte Smad1/5/8 knockout mice are a model of hemochromatosis that encompasses liver injury and fibrosis seen in human disease. These mice reveal the redundant but critical role of SMAD8 in hepcidin and iron homeostasis regulation, establish a requirement for SMAD1/5/8 in hepcidin regulation by testosterone and EGF but not inflammation, and suggest a pathogenic role for both iron loading and SMAD1/5/8 deficiency in liver injury and fibrosis.

The bone morphogenetic protein (BMP)-SMAD signaling pathway is a key transcriptional regulator of hepcidin in response to tissue iron stores, serum iron, erythropoietic drive and inflammation to increase the iron supply when needed for erythropoiesis, but to prevent the toxicity of iron excess. Recently, BMP2 was reported to play a non-redundant role in hepcidin regulation in addition to BMP6. Here, we used a newly validated BMP2 ELISA assay and mice with a global or endothelial conditional knockout (CKO) of Bmp2 or Bmp6 to examine how BMP2 is regulated and functionally contributes to hepcidin regulation by its major stimuli. Erythropoietin (EPO) did not influence BMP2 expression in control mice, and still suppressed hepcidin in Bmp2 CKO mice. Lipopolysaccharide (LPS) reduced BMP2 expression in control mice, but still induced hepcidin in Bmp2 CKO mice. Chronic dietary iron loading that increased liver iron induced BMP2 expression, whereas acute oral iron gavage that increased serum iron without influencing liver iron did not impact BMP2. However, hepcidin was still induced by both iron loading methods in Bmp2 CKO mice, although the degree of hepcidin induction was blunted relative to control mice. Conversely, acute oral iron gavage failed to induce hepcidin in Bmp6(−/−) or CKO mice. Thus, BMP2 has at least a partially redundant role in hepcidin regulation by serum iron, tissue iron, inflammation and erythropoietic drive. In contrast, BMP6 is absolutely required for hepcidin regulation by serum iron.

The research has been focused for the systematic examination and assessment of cropping behavior, soil productivity and the promotion of soil biodiversity in pomegranate based multiple crop sequencing. The study also sought to monitor the agronomic performance, nutrient availability and microbial communities’ in the rhizosphere. The potential of five crop sequencing systems comprised ‘Pomegranate–Urd–Garlic’ (P–U–G), ‘Pomegranate–Urd–Pea’ (P–U–P), ‘Pomegranate–Turmeric’ (P–T), ‘Pomegranate–Broccoli’ (P–B) and ‘Pomegranate monoculture’ (Pmono) under rainfed ecosystem has been demonstrated. After intercrop sequencing cycles, the essential relevant elements significantly improved growth attributes, physical, chemical and biological attributes of the rhizosphere soil compared to monoculture cropping practice. Maximum moisture content at field capacity (McFc, 24.9%), water holding capacity (WHC, 33.3%) and soil organic carbon (OC, 7.16 g kg−1) was recorded in P–U–P intercrop sequencing. This superior sequencing also recorded the available macronutrient contents (N, P, K), exchangeable Ca and Mg of soil which were increased by 8.15%, 36.9%, 8.6%, 31.5% and 22%, respectively over Pmono. Diethylenetriaminepentaacetic acid (DTPA)—extractable micronutrient cations (Fe, Cu, Zn, Mn) improved by 15.6%, 23.2%, 28.7% and 16.5%, respectively. Microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), and the plate count (colony-forming units, CFUs) of Bacillusspecies, Pseudomonasspecies, total soil fungi, Azotobacter chroococcumand actinobacteria exhibited significant variability. MBC and MBN ranged from 172.3 to 255.9 mg kg−1 and 12.1 to 17.2 mg kg−1, respectively. Microbial biomass of Pseudomonas species (116.5%), Bacillusspecies (80.5%), total soil fungi (204.4%), A. chroococcum(219.6%) and actinobacteria (205.6%) improved significantly over Pmono. Correlation analysis of different attributes representing the chemical and biological properties of soil also resulted in a significant association at P < 0.05 attribute pairs. Principal component analysis (PCA) of soil quality attributes was also worked out to evaluate the differences induced among intercrop sequencing systems. The results inferred that PC4 accounted for 100% of the total variance in all intercrop sequencing. Therefore, no correlation was observed between the soil properties, and the first, second and third PC (PC1, PC2, and PC3). It is expedient from the studies that the intercrop sequence systems have effects on soil fertility indicators and has the implications for agricultural productivity.

The aim of this research was designed in order to optimize integrated plant nutrient supply (IPNS) through balanced fertilization of organic, inorganic and microbial inoculants in strawberry cv. Chandler. The potential efficiency of bio-organics used along with chemical fertilizers on cropping behaviour, physical-chemical and biological properties of rhizosphere soil, fruit yield, quality attributes and leaf nutrient content was investigated. The significant improvement in physico-chemical properties of the soil and nutrient uptake was recorded. The uninoculated control received farmyard manure (FYM) and inorganic nitrogen (N) recorded the highest cation exchange capacity (CEC) and soil organic carbon (OC) content. Highest available N and phosphorus (P) of soil were recorded in vermicompost and inorganic N applied in two and one split, respectively. The concentration of micronutrients cations viz., iron (Fe), zinc (Zn), manganese (Mn) in soil was higher in treatment received vermicompost and inorg...

This study aimed to understand the relationship for productivity related traits using correlations and path coefficient analysis of ‘Dashehari’ mango under rainfed agro-climatic zone conditions of north-west Himalayas of India. The study also identifies the relationship among agronomic and yield-related characteristics to know the direct and indirect effect of independent variables on managerial ability of the crop. Correlation and path coefficient analyses were carried out over two consecutive years (2004, 2005) seasons for plant growth (tree trunk), leaf nutrient concentration, and fruit yield across four districts of Himachal Pradesh. Based on yield performance, an optimum sample size of 20 trees from each orchard comprised of 10 trees each of two categories: high productive (>90 kg tree −1 ) and low productive ( −1 ) were selected using the probability proportional to size multistage sampling scheme. Both high as well as low productive orchards were analyzed for soil and leaf chemical properties. Simple linear correlation and path coefficient analyses were worked out between all possible combinations. The contribution of leaf nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), zinc (Zn), copper (Cu), manganese (Mn), and iron (Fe) on growth and fruit yield was studied. In high productive trees, path model showed that leaf N, Mg, Fe, Cu and Mn concentrations recorded the high magnitude of direct effect on fruit yield. However, leaf N concentration had a strong positive indirect effect through leaf Zn. Leaf Cu and Zn concentrations had a positive indirect effect through leaf Mg and leaf Zn on fruit yield. In low productive trees, leaf N concentration had a strong positive direct effect and a strong positive indirect effect via leaf Zn on fruit yield. Similarly, the direct effect of tree volume on fruit yield was positive, while, the maximum positive indirect effect was through leaf K. A residual value of 0.7308 and 0.7292 indicates the collective influence of the variables was to the magnitude of 0.2692 and 0.2708 to the extent of 26.9 and 27.1% toward fruit yield, and contributed 40.7 and 14.7% toward tree volume in high and low productive mango trees, respectively. The collective influence of the variables included on tree volume to the magnitude of 0.8521 and concluded that leaf nutrient content has been contributed to the extent of 14.7% toward tree volume.

The comparative efficacy of bio-organic nutrients on cropping behavior and fruit quality of kiwifruit was analyzed using farm yard manure (FYM), vermicompost (VC), biofertilizers (BF), green manure (GM), and vermiwash (VW). Among various treatments the combination of FYM at 15 kg/vine, GM, VC at 15kg/vine, BFat50-g/ vine and VW at 2kg/vine significantly improved cropping behavior. This superior combination also resulted in considerably greater amounts of leaf macro-and micronutrients: N (2.49%), P (0.26%), K (1.48%), iron (Fe: 208.0 mg/kg), copper (Cu: 17.8 mg/kg), zinc (Zn: 36.2 mg/kg), and manganese (Mn: 88.3 mg/kg),which might be responsible for better cropping behavior, productivity and nutrient profile for sustainable kiwi fruit production. It can be concluded that with use of various sources of bio-organic materials under organic farming regime, there will be sufficient improvement in fruit quality and plant nutrient contents.

Iron is an essential element for numerous fundamental biologic processes, but excess iron is toxic. Abnormalities in systemic iron balance are common in patients with chronic kidney disease and iron administration is a mainstay of anemia management in many patients. This review provides an overview of the essential role of iron in biology, the regulation of systemic and cellular iron homeostasis, how imbalances in iron homeostasis contribute to disease, and the implications for chronic kidney disease patients.

Iron accumulation and oxidative stress are associated with neurodegenerative disease. Labile iron is known to catalyze free radical generation and subsequent neuronal damage, whereas the role of oxidative stress in neuronal iron accumulation is less well understood. Here, we examined the effect of hydrogen peroxide (H2O2) treatment on cellular iron-uptake, -storage, and -release proteins in the neuroblastoma cell line SH-SY5Y. We found no detectable change in the iron-uptake proteins transferrin receptor-1 and divalent metal ion transporter. In contrast, H2O2 treatment resulted in significant degradation of the iron-exporter ferroportin (Fpn). A decrease in Fpn is expected to increase the labile iron pool (LIP), reducing the iron-regulatory protein (IRP)-iron-responsive element interaction and increasing the expression of ferritin-H (Ft-H) for iron storage. Instead, we detected IRP1 activation, presumably due to oxidative stress, and a decrease in Ft-H translation. A reduction in Ft-H mRNA was also observed, probably dependent on an antioxidant-response element present in the Ft-H enhancer. The decrease in Fpn and Ft-H upon H2O2 treatment led to a time-dependent increase in the cellular LIP. Our study reveals a complex regulation of neuronal iron-release and iron-storage components in response to H2O2 that may explain iron accumulation detected in neurodegenerative diseases associated with oxidative stress.