LEfSe method identifying the OTUs with the greatest differences in abundance in the BFe and SFe groups.

<p>(a) Taxonomic cladogram obtained using LEfSe analysis of the 16S rRNA sequences. Treatment groups are indicated by the different colors; (b) Computed LDA scores of the relative abundance difference between the BFe and SFe groups. Comparison of the relative abundance at the (c) phylum; (d) order; (e) family; and (f) genus levels in the BFe and SFe groups.</p

Family and genus level cecal microbiota shifts between the BFe and SFe treatment groups.

<p>(a) Family level changes in the BFe and SFe groups as measured at the end of the study (day 42); (b) Genus level changes in the BFe and SFe groups as measured at the end of the study (day 42).</p

Observed alterations in the metabolic capacity of the cecal microbiota in the BFe group compared to the SFe group.

<p><b>Relative abundance of differentially–enriched KEGG microbial metabolic pathways in cecal microbiota, including</b> a) Unclassified; b) Organismal Systems; c) Human Diseases; d) Genetic Information Processing; e) Environmental Information Processing; and f) Cellular Processes. Treatment groups are indicated by the different colors, and FDR-corrected P values are displayed on the y-axis.</p

Duodenal mRNA gene expression of Fe-related proteins collected on day 42¹.

<p>¹ Changes in mRNA expression are shown relative to expression of 18S rRNA in arbitrary units (AU, * <i>P</i> < 0.05).</p

Ferritin concentration in <i>Caco-2</i> cells exposed to samples of beans only (whole bean), additional meal plan components and bean-based diets¹^-².

<p>¹Caco-2 bioassay procedures and preparation of the digested samples are described in the materials and methods sections.</p><p>²Cells were exposed to only MEM (minimal essential media) without added</p><p>food digests and Fe (n = 6).</p><p>^a-g Within a column, means without a common letter are significantly different</p><p>(p < 0.05).</p><p>Ferritin concentration in <i>Caco-2</i> cells exposed to samples of beans only (whole bean), additional meal plan components and bean-based diets<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0138479#t002fn001" target="_blank">¹</a>^-<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0138479#t002fn002" target="_blank">²</a>.</p

Ferritin protein and Fe concentration in the liver.

<p>¹Atomic mass for iron is 55.8g/mol</p><p>^a-b Within a column, means without a common letter are significantly different (p < 0.05).</p><p>Ferritin protein and Fe concentration in the liver.</p

Fe-related parameters assessed during the study.

<p>(1A) Blood hemoglobin concentration (g/L), (1B) Total body Hb-Fe (mg), (1C) Hemoglobin maintenance efficiency (%). * <i>P</i> < 0.05 between treatment groups.</p

Linoleic Acid:Dihomo-γ-Linolenic Acid Ratio Predicts the Efficacy of Zn-Biofortified Wheat in Chicken (Gallus gallus)

The amount of Zn absorbed from Zn-biofortified wheat material has been determined using an <i>in vivo</i> model of Zn absorption. The erythrocyte linoleic:dihomo -γ-linolenic acid (LA:DGLA) ratio was used as a biomarker of Zn status. Two groups of chickens (<i>n</i> = 15) were fed different diets: a high-Zn (46.5 μg Zn g^–1) and a low-Zn wheat-based diet (32.8 μg Zn g^–1). Dietary Zn intakes, body weight, serum Zn, and the erythrocyte fatty acid profile were measured, and tissues were taken for gene expression analysis. Serum Zn concentrations were greater in the high Zn group (<i>p</i> < 0.05). Duodenal mRNA expression of various Zn transporters demonstrated expression upregulation in the birds fed a low Zn diet (<i>n</i> = 15, <i>p</i> < 0.05). The LA:DGLA ratio was higher in the birds fed the low Zn diet (<i>p</i> < 0.05). The higher amount of Zn in the biofortified wheat resulted in a greater Zn uptake

Alterations in the Gut (<i>Gallus gallus</i>) Microbiota Following the Consumption of Zinc Biofortified Wheat (<i>Triticum aestivum</i>)‑Based Diet

The structure and function of cecal microbiota following the consumption of a zinc (Zn) biofortified wheat diet was evaluated in a well-studied animal model of human nutrition (<i>Gallus gallus</i>) during a six-week efficacy trial. Using 16S rRNA gene sequencing, a significant increase in β- but not α-microbial diversity was observed in the animals receiving the Zn biofortified wheat diet, relative to the control. No significant taxonomic differences were found between the two groups. Linear discriminant analysis revealed a group of metagenomic biomarkers that delineated the Zn replete versus Zn deficient phenotypes, such that enrichment of lactic acid bacteria and concomitant increases in Zn-dependent bacterial metabolic pathways were observed in the Zn biofortified group, and expansion of mucin-degraders and specific bacterial groups able to participate in maintaining host Zn homeostasis were observed in the control group. Additionally, the <i>Ruminococcus</i> genus appeared to be a key player in delineating the Zn replete microbiota from the control group, as it strongly predicts host Zn adequacy. Our data demonstrate that the gut microbiome associated with Zn biofortified wheat ingestion is unique and may influence host Zn status. Microbiota analysis in biofortification trials represents a crucial area for study as Zn biofortified diets are increasingly delivered on a population-wide scale

Metacridamides A and B, Macrocycles from Conidia of the Entomopathogenic Fungus <i>Metarhizium acridum</i>

<i>Metarhizium acridum</i>, an entomopathogenic fungus, has been commercialized and used successfully for biocontrol of grasshopper pests in Africa and Australia. Its conidia produce two novel 17-membered macrocycles, metacridamides A (<b>1</b>) and B (<b>2</b>), which consist of a Phe unit condensed with a nonaketide. Planar structures were elucidated by a combination of mass spectrometric and NMR techniques. Following hydrolysis of <b>1</b>, chiral amino acid analysis assigned the l-configuration to the Phe unit. A crystal structure established the absolute configuration of the eight remaining stereogenic centers in <b>1</b>. Metacridamide A (<b>1</b>) showed cytotoxicity to three cancer lines with IC₅₀'s of 6.2, 11.0, and 10.8 μM against Caco-2 (epithelial colorectal adenocarcinoma), MCF-7 (breast cancer), and HepG2/C3A (hepatoma) cell lines, respectively. In addition, metacridamide B (<b>2</b>) had an IC₅₀ of 18.2 μM against HepG2/C3A, although it was inactive at 100 μM against Caco-2 and MCF-7. Neither analogue showed antimicrobial, phytotoxic, or insecticidal activity