Nicotianamine-chelated iron positively affects iron status, intestinal morphology and microbial populations in vivo (Gallus gallus)

Abstract Objectives Assessment and comparison of the effects of various concentrations of soluble extracts of quinoa fiber (Chenopodium quinoa Willd.) and quercetin-3-glucoside on the zinc and iron status, brush border membrane (BBM) functionality, intestinal morphology, and cecal bacterial populations in-vivo (Gallus gallus). Methods The study utilized Gallus gallus intra-amniotic feeding, a clinically validated method to assess the effects of quinoa, quercetin, and control using seven groups (no injection, 18 Ω H2O, 5% inulin, 1% quercetin 3-glucoside, 2.5% quinoa fiber, 5% quinoa fiber, 1% quercetin 3-glucoside + 5% quinoa fiber). Upon hatch, the cecum, duodenum, pectoral muscle, liver, and blood samples were collected for the estimation of the relative abundance of the gut microbiome, mRNA gene expression Zn and Fe-related transporter proteins and brush border membrane functionality and morphology, glycogen, relative expression of lipid-related genes and hemoglobin levels, respectively. Results The results demonstrated an increase (P < 0.05) in villi height, weight, and surface area in the groups administered with quercetin, and a dose-dependent increase was observed with quinoa soluble fiber treatment. Additionally, an increase in ferroportin and duodenal cytochrome B (DcytB) was observed in the group injected with both quinoa and quercetin. Similarly, zinc transporter 7 (ZnT7) and sucrose-isomaltase (SI) gene expression was upregulated in this group. Further, the administration of quinoa soluble fiber altered the composition and function of the cecal microbiome. Conclusions The evidence suggests that quinoa and quercetin have a synergistic effect, together they are found to improve BBM morphology and functionality, affect the intestinal microbiome, increase short-chain fatty acid production, and thereby improving mineral solubility. Quinoa fibers, a polyphenol-rich superfood, may help fight micronutrient deficiencies in target populations. Funding Sources N/A.

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Soluble Extracts from Chia Seed (Salvia hispanica L.) Affect Brush Border Membrane Functionality, Morphology and Intestinal Bacterial Populations In Vivo (Gallus gallus)

Nutrients ◽

10.3390/nu11102457 ◽

2019 ◽

Vol 11 (10) ◽

pp. 2457 ◽

Cited By ~ 5

Author(s):

Pereira da Silva ◽

Kolba ◽

Stampini Duarte Martino ◽

Hart ◽

Tako

Keyword(s):

Brush Border ◽

Brush Border Membrane ◽

Gallus Gallus ◽

Intestinal Morphology ◽

Salvia Hispanica ◽

Bacterial Populations ◽

Relative Expression ◽

Chia Seed ◽

Salvia Hispanica L

This study assessed and compared the effects of the intra-amniotic administration of various concentrations of soluble extracts from chia seed (Salvia hispanica L.) on the Fe and Zn status, brush border membrane functionality, intestinal morphology, and intestinal bacterial populations, in vivo. The hypothesis was that chia seed soluble extracts will affect the intestinal morphology, functionality and intestinal bacterial populations. By using the Gallus gallus model and the intra-amniotic administration approach, seven treatment groups (non-injected, 18 Ω H2O, 40 mg/mL inulin, non-injected, 5 mg/mL, 10 mg/mL, 25 mg/mL and 50 mg/mL of chia seed soluble extracts) were utilized. At hatch, the cecum, duodenum, liver, pectoral muscle and blood samples were collected for assessment of the relative abundance of the gut microﬂora, relative expression of Fe- and Zn-related genes and brush border membrane functionality and morphology, relative expression of lipids-related genes, glycogen, and hemoglobin levels, respectively. This study demonstrated that the intra-amniotic administration of chia seed soluble extracts increased (p < 0.05) the villus surface area, villus length, villus width and the number of goblet cells. Further, we observed an increase (p < 0.05) in zinc transporter 1 (ZnT1) and duodenal cytochrome b (Dcytb) proteins gene expression. Our results suggest that the dietary consumption of chia seeds may improve intestinal health and functionality and may indirectly improve iron and zinc intestinal absorption.

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Low Phytate Peas (Pisum sativum L.) Improve Iron Status, Gut Microbiome, and Brush Border Membrane Functionality In Vivo (Gallus gallus)

Nutrients ◽

10.3390/nu12092563 ◽

2020 ◽

Vol 12 (9) ◽

pp. 2563 ◽

Cited By ~ 1

Author(s):

Tom Warkentin ◽

Nikolai Kolba ◽

Elad Tako

Keyword(s):

Iron Deficiency ◽

Gut Microbiota ◽

Brush Border ◽

Brush Border Membrane ◽

Iron Status ◽

Gallus Gallus ◽

Intestinal Microbiome ◽

Physiological Status ◽

Dietary Iron

The inclusion of pulses in traditional wheat-based food products is increasing as the food industry and consumers are recognizing the nutritional benefits due to the high protein, antioxidant activity, and good source of dietary fiber of pulses. Iron deficiency is a significant global health challenge, affecting approximately 30% of the world’s population. Dietary iron deficiency is the foremost cause of anemia, a condition that harms cognitive development and increases maternal and infant mortality. This study intended to demonstrate the potential efficacy of low-phytate biofortified pea varieties on dietary iron (Fe) bioavailability, as well as on intestinal microbiome, energetic status, and brush border membrane (BBM) functionality in vivo (Gallus gallus). We hypothesized that the low-phytate biofortified peas would significantly improve Fe bioavailability, BBM functionality, and the prevalence of beneficial bacterial populations. A six-week efficacy feeding (n = 12) was conducted to compare four low-phytate biofortified pea diets with control pea diet (CDC Bronco), as well as a no-pea diet. During the feeding trial, hemoglobin (Hb), body-Hb Fe, feed intake, and body weight were monitored. Upon the completion of the study, hepatic Fe and ferritin, pectoral glycogen, duodenal gene expression, and cecum bacterial population analyses were conducted. The results indicated that certain low-phytate pea varieties provided greater Fe bioavailability and moderately improved Fe status, while they also had significant effects on gut microbiota and duodenal brush border membrane functionality. Our findings provide further evidence that the low-phytate pea varieties appear to improve Fe physiological status and gut microbiota in vivo, and they highlight the likelihood that this strategy can further improve the efficacy and safety of the crop biofortification and mineral bioavailability approach.

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Saffron (Crocus sativus L.) Flower Water Extract Disrupts the Cecal Microbiome, Brush Border Membrane Functionality, and Morphology In Vivo (Gallus gallus)

Nutrients ◽

10.3390/nu14010220 ◽

2022 ◽

Vol 14 (1) ◽

pp. 220

Author(s):

Nikita Agarwal ◽

Nikolai Kolba ◽

YeonJin Jung ◽

Jacquelyn Cheng ◽

Elad Tako

Keyword(s):

Gene Expression ◽

Iron Status ◽

Paneth Cell ◽

Water Extract ◽

Gallus Gallus ◽

Cell Number ◽

Crocus Sativus ◽

Dose Dependent ◽

Crocus Sativus L

Saffron (Crocus sativus L.) is known as the most expensive spice. C. sativus dried red stigmas, called threads, are used for culinary, cosmetic, and medicinal purposes. The rest of the flower is often discarded, but is now being used in teas, as coloring agents, and fodder. Previous studies have attributed antioxidant, anti-inflammatory, hepatoprotective, neuroprotective, anti-depressant, and anticancer properties to C. sativus floral bio-residues. The aim of this study is to assess C. sativus flower water extract (CFWE) for its effects on hemoglobin, brush boarder membrane (BBM) functionality, morphology, intestinal gene expression, and cecal microbiome in vivo (Gallus gallus), a clinically validated model. For this, Gallus gallus eggs were divided into six treatment groups (non-injected, 18 Ω H2O, 1% CFWE, 2% CFWE, 5% CFWE, and 10% CFWE) with n~10 for each group. On day 17 of incubation, 1 mL of the extracts/control were administered in the amnion of the eggs. The amniotic fluid along with the administered extracts are orally consumed by the developing embryo over the course of the next few days. On day 21, the hatchlings were euthanized, the blood, duodenum, and cecum were harvested for assessment. The results showed a significant dose-dependent decrease in hemoglobin concentration, villus surface area, goblet cell number, and diameter. Furthermore, we observed a significant increase in Paneth cell number and Mucin 2 (MUC2) gene expression proportional to the increase in CFWE concentration. Additionally, the cecum microbiome analysis revealed C. sativus flower water extract altered the bacterial populations. There was a significant dose-dependent reduction in Lactobacillus and Clostridium sp., suggesting an antibacterial effect of the extract on the gut in the given model. These results suggest that the dietary consumption of C. sativus flower may have negative effects on BBM functionality, morphology, mineral absorption, microbial populations, and iron status.

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Nicotianamine-Chelated Iron Positively Affects Iron Status, Intestinal Morphology and Microbial Populations In Vivo (Gallus Gallus)

Current Developments in Nutrition ◽

10.1093/cdn/nzab044_005 ◽

2021 ◽

Vol 5 (Supplement_2) ◽

pp. 574-574

Author(s):

Jesse Beasley ◽

Alex Johnson ◽

Elad Tako

Keyword(s):

Goblet Cell ◽

Gallus Gallus ◽

Cell Number ◽

Intestinal Morphology ◽

Short Term ◽

Gastrointestinal Health ◽

Short Term Exposure ◽

The Impact

Abstract Objectives Iron (Fe) fortification involves low dose delivery of bioavailable Fe fortificants to food products during manufacture (or point-of-use) and is an effective population-based strategy to combat human Fe deficiencies that affect over 2 billion people globally. Iron fortification of wheat flour is now mandatory in 75 countries worldwide, however, the tendency of Fe fortificants to oxidize and cause undesired organoleptic and sensory properties pose significant challenges. Nicotianamine (NA) is a natural chelator of Fe and zinc (Zn) in higher plants and NA-chelated Fe is highly bioavailable in vitro. In graminaceous plants NA serves as the biosynthetic precursor to 2′ -deoxymugineic acid (DMA), a related Fe chelator and enhancer of Fe bioavailability, and increased NA/DMA biosynthesis has proved an effective Fe biofortification strategy in several cereal crops. In this study we aimed to investigate the impact of NA-chelated Fe on Fe status and gastrointestinal health in vivo. Methods We utilized the versatile chicken (Gallus gallus) model to assess NA-chelated Fe when delivered to chickens through intraamniotic administration (short-term exposure) or over a period of six weeks as part of a biofortified wheat diet containing increased NA, Fe, Zn and DMA (long-term exposure). Results Following short-term exposure, chickens that received an NA-chelated Fe treatment demonstrated significantly increased blood serum Fe, intestinal goblet cell number, and abundance of Bifidobacterium relative to chickens that received control Fe treatments (either EDTA-chelated Fe or unchelated Fe). Following long-term exposure, chickens that consumed the biofortified diet had significantly increased liver Fe, liver and muscle glycogen, and intestinal goblet cell number, as well as significantly different microbial population diversities (including a greater abundance of Bifidobacterium and Lactobacillus) relative to chickens that consumed the control diet. Conclusions Taken together these results demonstrate numerous health benefits associated with NA-chelated Fe in a biofortified wheat diet and uncover novel impacts of NA-chelated Fe on gastrointestinal health and functionality. Funding Sources This work was supported by grants from the USDA-ARS and HarvestPlus.

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CBIO-10. REDUCED IRON EXPORT FUNCTIONS IN A CELL INTRINSIC MANNER TO DRIVE GLIOBLASTOMA GROWTH

Neuro-Oncology ◽

10.1093/neuonc/noaa215.070 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii17-ii17

Author(s):

Katie Troike ◽

Erin Mulkearns-Hubert ◽

Daniel Silver ◽

James Connor ◽

Justin Lathia

Keyword(s):

Tumor Cells ◽

Iron Uptake ◽

Iron Homeostasis ◽

Iron Status ◽

Tumor Grade ◽

Hfe Gene ◽

Iron Release ◽

Female Patients ◽

Cellular Iron

Abstract Glioblastoma (GBM), the most common primary malignant brain tumor in adults, is characterized by invasive growth and poor prognosis. Iron is a critical regulator of many cellular processes, and GBM tumor cells have been shown to modulate expression of iron-associated proteins to enhance iron uptake from the surrounding microenvironment, driving tumor initiation and growth. While iron uptake has been the central focus of previous investigations, additional mechanisms of iron regulation, such as compensatory iron efflux, have not been explored in the context of GBM. The hemochromatosis (HFE) gene encodes a transmembrane glycoprotein that aids in iron homeostasis by limiting cellular iron release, resulting in a sequestration phenotype. We find that HFE is upregulated in GBM tumors compared to non-tumor brain and that expression of HFE increases with tumor grade. Furthermore, HFE mRNA expression is associated with significantly reduced survival specifically in female patients with GBM. Based on these findings, we hypothesize that GBM tumor cells upregulate HFE expression to augment cellular iron loading and drive proliferation, ultimately leading to reduced survival of female patients. To test this hypothesis, we generated Hfe knockdown and overexpressing mouse glioma cell lines. We observed significant alterations in the expression of several iron handling genes with Hfe knockdown or overexpression, suggesting global disruption of iron homeostasis. Additionally, we show that knockdown of Hfe in these cells increases apoptosis and leads to a significant impairment of tumor growth in vivo. These findings support the hypothesis that Hfe is a critical regulator of cellular iron status and contributes to tumor aggression. Future work will include further exploration of the mechanisms that contribute to these phenotypes as well as interactions with the tumor microenvironment. Elucidating the mechanisms by which iron effulx contributes to GBM may inform the development of next-generation targeted therapies.

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The Combined Application of the Caco-2 Cell Bioassay Coupled with In Vivo (Gallus gallus) Feeding Trial Represents an Effective Approach to Predicting Fe Bioavailability in Humans

Nutrients ◽

10.3390/nu8110732 ◽

2016 ◽

Vol 8 (11) ◽

pp. 732 ◽

Cited By ~ 29

Author(s):

Elad Tako ◽

Haim Bar ◽

Raymond Glahn

Keyword(s):

Gallus Gallus ◽

Feeding Trial ◽

Combined Application

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The early endosomal protein Rab21 is critical for enterocyte functions and intestinal homeostasis

10.1101/2020.11.18.378984 ◽

2020 ◽

Author(s):

Sonya Nassari ◽

Dominique Lévesque ◽

François-Michel Boisvert ◽

Steve Jean

Keyword(s):

Membrane Trafficking ◽

Genetic Diseases ◽

Growth Factor Receptor ◽

Small Gtpase ◽

Intestinal Morphology ◽

Endosomal Trafficking ◽

Intestinal Homeostasis ◽

Compensatory Proliferation ◽

Epidermal Growth

ABSTRACTMembrane trafficking is defined as the vesicular transport of molecules into, out of, and throughout the cell. In intestinal enterocytes, defects in endocytic/recycling pathways result in impaired function and are linked to genetic diseases. However, how these trafficking pathways regulate intestinal tissue homeostasis is poorly understood. Using the Drosophila intestine as an in vivo model system, we investigated enterocyte-specific functions for the early endosomal trafficking machinery in gut homeostasis. We focused on the small GTPase Rab21, which regulates specific steps in early endosomal trafficking. Rab21-depleted guts showed severe abnormalities in intestinal morphology, with deregulated homeostasis associated with a gain in mitotic cells and increased cell death. Increases in both apoptosis and yorkie signaling were responsible for compensatory proliferation and tissue inflammation. Using a RNA interference screen, we identified specific regulators of autophagy and membrane trafficking that phenocopied Rab21 loss. We further showed that Rab21-induced hyperplasia was rescued by inhibition of epidermal growth factor receptor signaling, and identified improperly trafficked cargoes in Rab21-depleted enterocytes. Our data shed light on an important role for early endosomal trafficking, and particularly Rab21, in enterocyte-mediated intestinal homeostasis.

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