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Professor Bruce Ames demonstrated that nutritional recommendations should be adjusted in order to 'tune-up' metabolism and reduce mitochondria decay, a hallmark of aging and many disease processes. A major subset of tunable nutrients are the minerals, which despite being integral to every aspect of metabolism are often deficient in the typical Western diet. Mitochondria are particularly rich in minerals, where they function as essential cofactors for mitochondrial physiology and overall cellular health. Yet substantial knowledge gaps remain in our understanding of the form and function of these minerals needed for metabolic harmony. Some of the minerals have known activities in the mitochondria but with incomplete regulatory detail, whereas other minerals have no established mitochondrial function at all. A comprehensive metallome of the mitochondria is needed to fully understand the patterns and relationships of minerals within metabolic processes and cellular development. This brief overview serves to highlight the current progress towards understanding mineral homeostasis in the mitochondria and to encourage more research activity in key areas. Future work may likely reveal that adjusting the amounts of specific nutritional minerals has longevity benefits for human health.

Zinc deficiency continues to be a major concern for global public health. The zinc status of a target population is typically estimated by measuring circulating zinc levels, but the sampling procedures are not standardized and thus may result in analytical discrepancies. To examine this, we designed a study that controlled most of the technical parameters in order to focus on five pre-analytical variables reported to influence the measurement of zinc in blood samples, including (1) blood draw site (capillary or venous), (2) blood sample matrix (plasma or serum), (3) blood collection tube manufacturer (Becton, Dickinson and Company or Sarstedt AG & Co), (4) blood processing time (0, 4, or 24 hours), and (5) blood holding temperatures (4°C, 20°C, or 37°C). A diverse cohort of 60 healthy adults were recruited to provide sequential capillary and venous blood samples, which were carefully processed under a single chain of custody and measured for zinc content using inductively coupled plasma optical emission spectrometry. When comparing blood draw sites, the mean zinc content of capillary samples was 0.051 mg/L (8%) higher than venous blood from the same donors. When comparing blood sample matrices, the mean zinc content of serum samples was 0.034 mg/L (5%) higher than plasma samples from the same donors. When comparing blood collection tube manufacturer, the mean zinc content from venous blood samples did not differ between venders, but the mean zinc content from BD capillary plasma was 0.036 mg/L (6%) higher than Sarstedt capillary plasma from the same donors. When comparing processing times, the mean zinc content of plasma and serum samples was 5-12% higher in samples processed 4-24 hour after collection. When comparing holding temperatures, the mean zinc content of plasma and serum samples was 0.5-7% higher in samples temporarily held at 20°C or 37°C after collection. Thus even with the same donors and blood draws, significant differences in zinc content were observed with different draw sites, tube types, and processing procedures, demonstrating that key pre-analytic variables can have an impact on zinc measurement, and subsequent classification of zinc status. Minimizing these pre-analytical variables is important for generating best practice guidelines for assessment of zinc status.

Plasma zinc concentrations (PZC) have been shown to significantly increase during zinc supplementation. This study investigated the effects of daily preventive zinc supplementation on hair and nail zinc concentrations compared with a control group. In a randomized controlled trial, 6- to 23-month-old children (n = 3407) in Lao PDR were randomly assigned to one of four groups and followed for ~ 36 weeks: daily preventive zinc dispersible tablet (7 mg/d; PZ), daily micronutrient powder (10 mg zinc/d; MNP), therapeutic zinc supplements for diarrhea treatment (20 mg/d for 10 days; TZ), or daily placebo powder (Control). Plasma, hair, and nail zinc concentrations were assessed in a sub-sample of participants (n = 457) at baseline and endline. At baseline, 75% of children had low PZC (P P = 0.99). Nail zinc concentrations were marginally higher in the PZ (115.8 (111.6, 119.9) μg/g) and the MNP (117.8 (113.3, 122.3) μg/g) groups than in the TZ group (110.4 (106.0, 114.8) μg/g; P = 0.055) at endline. This study does not support the use of hair zinc as a biomarker of zinc exposure in young children. However, it provides some evidence that zinc concentrations in nails may respond to supplemental zinc interventions and supports the need for collecting additional data on this emerging biomarker.

Background: When consumed as whole grain, wheat has a high nutrient density that contributes to a healthy diet. However, products labeled as whole wheat can still contain a substantial amount of non-whole grain wheat, leading to confusion for consumers trying to maximize their whole grain intake. A biomarker of whole grain is needed to reveal the whole grain status within wheat-based foods.Objective: Wheat germ agglutinin (WGA), a lectin found predominantly in the germ tissue of wheat kernels, was evaluated as a biomarker of whole grain in commercial wheat products. Methods: The levels of WGA within wheat flour and pasta were assessed by an immunoblot method and compared to a whole grain standard. WGA content was also compared to other biomarkers including starch, minerals, phytate, and total protein content.Results: WGA content tightly correlated with the percentage of whole grain in pre-made mixtures of whole wheat and refined (white) flours. Several commercial flours labeled as whole wheat were then tested for WGA content and found to contain up to 40% less WGA compared to a whole grain standard. Several commercial pasta products labeled as whole wheat were also tested for WGA content and found to contain up to 90% less WGA compared to a whole grain standard. The discrepancies in WGA content were unlikely due to wheat varietial differences alone, as the WGA content measured in common varieties used in domestic wheat flour production varied less than 25%. Other wheat constituents including starch, mineral, phytate, and total protein were less consistent and did not discriminate between the commercial whole wheat flours and pasta products. Conclusions: The WGA content within wheat flour and pasta correlated with the levels of whole grain and identified discrepancies when tested in commercial wheat products compared to a whole wheat standard. WGA is a unique biomarker that could help identify which wheat products have the greatest amount of whole grain wheat.

In 2020, STEM training programs across the country were challenged to provide support to students during a nation-wide shutdown of research institutions in response to the COVID-19 pandemic. Five U.S. high school science internship programs funded by the Doris Duke Charitable Foundation, with a history of collaboration, developed innovative strategies for distance-learning (DL) opportunities during the pandemic. Forty under-represented high school and undergraduate students were paired with scientific mentors at one of the programs for a DL scientific internship. Summer training combined synchronous and asynchronous programming with research projects adapted for DL success. Ninety-five percent of students who participated were satisfied with the training experience, nearly identical to exit survey responses from 2019 when our programs were held in-person. More students were interested in pursuing a career in research at the end of the program and credited the DL experience with increasing interest in research careers. Some DL elements were ideal for underrepresented youth, including a more flexible schedule and elimination of cost and time for travel. While the lack of in-person instruction challenged our ability to create a strong student community, we found that preparation, communication, and flexibility were key elements to these successful DL programs. The increased emphasis on interpretation and analysis of data, rather than data collection, enhanced student learning. This manuscript highlights the changes made to our curricula, elements which were most successful, and recommends strategies for future distance-learning programming.

Lysosome function is compromised during aging and in many disease states. Interventions that promote lysosomal activity and acidification are thus of prime interest as treatments for longevity and health. Intracellular pH can be controlled by the exchange of protons for inorganic ions, and in cells from microbes to man, when potassium is restricted in the growth medium, the cytoplasm becomes acidified. Here we use a yeast model to show that potassium limited-cells exhibit hallmarks of increased acidity in the vacuole, the analog of the lysosome, and live long by a mechanism that requires the vacuolar machinery. The emerging picture is one in which potassium restriction shores up vacuolar acidity and function, conferring health benefits early in life and extending viability into old age. Against the backdrop of well-studied protein and carbohydrate restrictions that extend lifespan and healthspan, our work establishes a novel pro-longevity paradigm of inorganic nutrient limitation.

In the USA, western Washington (WWA) and the Alaska (AK) Interior are two regions where maritime and continental climates, high latitude and cropping systems necessitate early maturing spring wheat (Triticum aestivum L.). Both regions aim to increase the production of hard spring bread wheat for human consumption to support regional agriculture and food systems. The Nordic region of Europe has a history of breeding for early maturing spring wheat and also experiences long daylengths with mixed maritime and continental climates. Nordic wheat also carries wildtype (wt) NAM-B1, an allele associated with accelerated senescence and increased grain protein and micronutrient content, at a higher frequency than global germplasm. Time to senescence, yield, protein and mineral content were evaluated on 42 accessions of Nordic hard red spring wheat containing wt NAM-B1 over 2 years on experimental stations in WWA and the AK Interior. Significant variation was found by location and accession for time to senescence, suggesting potential parental lines for breeding programmes targeting early maturity. Additionally, multiple regression analysis showed that decreased time to senescence correlated negatively with grain yield and positively with grain protein, iron and zinc content. Breeding for early maturity in these regions will need to account for this potential trade-off in yield. Nordic wt NAM-B1 accessions with early senescence yet with yields similar to regional checks are reported. Collaboration among alternative wheat regions can aid in germplasm exchange and varietal development as shown here for the early maturing trait.

Micronutrient deficiencies are a significant public health problem affecting a large portion of the world population. Disproportionately affected populations, namely infants, young children, adolescents and women of reproductive age including pregnant women, are especially susceptible to the health consequences of insufficient micronutrient intakes. However, assessment of micronutrient deficiencies is not routinely included in population health surveys. This nutrition data gap hampers policy, program, and promotion efforts to prevent and treat micronutrient deficiencies. To address one of the barriers to micronutrient assessment, an expert group created a consensus of a target product profile (TPP) for a micronutrient assessment tool and associated blood collection device for use in population surveys. Experts in laboratory medicine, micronutrient assessment, population-based surveys, and product development reviewed proposed TPP standards and collaboratively established minimum and optimal characteristics. These experts defined the target population as infants and children from 6-59 months, adolescents and women of reproductive age 12-49 years including pregnant women. At minimum, the assessment tool should be a multiplex ELISA formatted for >1 analyte that uses a serum or plasma sample prepared from venous blood obtained by a phlebotomist with a 2-week training. Given the use case was specific for population surveys, experts agreed the minimum tool could be semi-quantitative, with analytical specificity of 99%. The TPP also considers the variable field environments for testing (e.g. storage conditions and time to results). The consensus TPP developed can be used to guide selection of existing technologies into population-based surveys, as well as future investment in product development. Partnerships focused on research and development, including industry, public sector, nonprofit, and academic institutions, can help advance the field and fill the micronutrient data gap.

BACKGROUND Nutritional deficiencies in children with cancer at time of diagnosis and during treatment may negatively affect disease outcome and increase treatment-related toxicity. Yet zinc, an essential nutrient important for supporting immune function and known for reducing diarrheal episodes, is rarely assessed in these children. PROCEDURES Fifty children (1 month to 18 years) with recently diagnosed cancer were enrolled in this study. An age and gender matched control group (n = 50) was also recruited. Plasma and urinary zinc, plasma copper, and C-reactive protein (CRP) levels were measured at baseline, 3, and 6 months following diagnosis. A retrospective review of the toxicity profile was performed in the cohort of children with cancer for the first 4 years after initial diagnosis. RESULTS CRP and plasma copper (both acute-phase reactants) were elevated in patients with cancer compared to controls at baseline, both p

Calcium-based renal calculi demonstrated significant heterogeneity in the structure, density, mineral composition, and material hardness not elucidated by routine clinical testing. Mineral density distributions within calcium oxalate stones revealed differential areas of low (590±80 mg/cc), medium (840±140 mg/cc), and high (1100±200 mg/cc) densities. Apatite stones also contained regions of low (700±200 mg/cc), medium (1100±200 mg/cc), and high (1400±140 mg/cc) densities within layers extending from single or multiple nucleation sites. Despite having lower average mineral density, calcium oxalate (CaOx) stones demonstrated higher material hardness compared to apatite stones, suggesting other chemical components might be involved in determining stone hardness properties. Carbon concentrated sites were identified between morphologic layers in CaOx stones and in stratified layers of apatite stones. Elemental analyses revealed numerous additional trace elements in both stone types. Despite the widespread assumption that stone mineral density is an indicator of susceptibility to lithotripsy, calcium stone mineral density estimates do not directly correlate with actual ex vivo stone hardness. Underlying stone heterogeneity in both structure and mineral density could explain why historical approaches have failed in accurately predicting response of stones to lithotripsy.

Elevated uric acid (UA) is a key risk factor for many disorders, including metabolic syndrome, gout and kidney stones. Despite frequent occurrence of these disorders, the genetic pathways influencing UA metabolism and the association with disease remain poorly understood. In humans, elevated UA levels resulted from the loss of the of the urate oxidase (Uro) gene around 15 million years ago. Therefore, we established a Drosophila melanogaster model with reduced expression of the orthologous Uro gene to study the pathogenesis arising from elevated UA. Reduced Uro expression in Drosophila resulted in elevated UA levels, accumulation of concretions in the excretory system, and shortening of lifespan when reared on diets containing high levels of yeast extract. Furthermore, high levels of dietary purines, but not protein or sugar, were sufficient to produce the same effects of shortened lifespan and concretion formation in the Drosophila model. The insulin-like signaling (ILS) pathway has been shown to respond to changes in nutrient status in several species. We observed that genetic suppression of ILS genes reduced both UA levels and concretion load in flies fed high levels of yeast extract. Further support for the role of the ILS pathway in modulating UA metabolism stems from a human candidate gene study identifying SNPs in the ILS genes AKT2 and FOXO3 being associated with serum UA levels or gout. Additionally, inhibition of the NADPH oxidase (NOX) gene rescued the reduced lifespan and concretion phenotypes in Uro knockdown flies. Thus, components of the ILS pathway and the downstream protein NOX represent potential therapeutic targets for treating UA associated pathologies, including gout and kidney stones, as well as extending human healthspan., Author summary Enzymatic purine degradation in humans ends with uric acid (UA). Multiple genetic and dietary factors raise UA levels above the norm, which is called hyperuricemia or hyperuricosuria when detected in the serum or urine, respectively. Clinical studies report a correlation between elevated UA and a plethora of chronic diseases including crystalopathies like UA kidney stones and gout, or metabolic and vascular disorders such as diabetes, obesity, and coronary artery disease. Here, we identified a regulatory role for the insulin-like signaling cascade affecting UA metabolism using a Drosophila melanogaster model. In the process we determined previously unrecognized potential drug targets to treat elevated UA levels and associated pathologies such as gout or UA kidney stones, with the potential additional benefit of extending human healthspan. Our work also establishes the fly as a model system to characterize the influence of genetic and dietary factors in gout or UA kidney stone development in a manner readily amenable for small-scale screening of drug interventions. The novelty of our findings, their broad impact, and relevance for multiple diseases opens up an important area of research to define mechanisms of UA accumulation.

SummaryElevated uric acid (UA) is a key factor for disorders, including gout or kidney stones and result from abrogated expression of Urate Oxidase (Uro) and diet. To understand the genetic pathways influencing UA metabolism we established a Drosophila melanogaster model with elevated UA using Uro knockdown. Reduced Uro expression resulted in the accumulation of UA concretions and diet-dependent shortening of lifespan. Inhibition of insulin-like signaling (ILS) pathway genes reduced UA and concretion load. In humans, SNPs in the ILS genes AKT2 and FOXO3 were associated with UA levels or gout, supporting a conserved role for ILS in modulating UA metabolism. Downstream of the ILS pathway UA pathogenicity was mediated partly by NADPH Oxidase, whose inhibition attenuated the reduced lifespan and concretion accumulation. Thus, genes in the ILS pathway represent potential therapeutic targets for treating UA associated pathologies, including gout and kidney stones.HighlightsIn Drosophila high uric acid (UA) levels shorten lifespan and cause UA aggregationConserved in flies and humans, the ILS pathway associates with UA pathologiesFoxO dampens concretion formation by reducing UA levels and ROS formationInhibition of NOX alleviates the lifespan attenuation and UA aggregation

© 2015 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. Urolithiasis affects around 10% of the US population with an increasing rate of prevalence, recurrence and penetrance. The causes for the formation of most urinary calculi remain poorly understood, but obtaining the chemical composition of these stones might help identify key aspects of this process and new targets for treatment. The majority of urinary stones are composed of calcium that is complexed in a crystalline matrix with organic and inorganic components. Surprisingly, mitigation of urolithiasis risk by altering calcium homeostasis has not been very effective. Thus, studies to identify other therapeutic stone-specific targets, using proteomics, metabolomics and microscopy techniques, have been conducted, revealing a high level of complexity. The data suggest that numerous metals other than calcium and many nonmetals are present within calculi at measurable levels and several have distinct distribution patterns. Manipulation of the levels of some of these elemental components of calcium-based stones has resulted in clinically beneficial changes in stone chemistry and rate of stone formation. The elementome—the full spectrum of elemental content—of calcium-based urinary calculi is emerging as a new concept in stone research that continues to provide important insights for improved understanding and prevention of urinary stone disease.

Metals such as copper, iron, manganese, and zinc are clearly required for proper metabolism and development, while imbalances can lead to systemic dysfunction and disease. As a result, organisms have evolved complex genetic systems for the regulation of metal levels, including import, export, and sequestration of metals within cells and sub-cellular compartments. The study of metal biology in insects has the potential to greatly expand our understanding of metal biology. The results of such studies might point to new possible therapeutic interventions for neurological and other human diseases, as well as new strategies for insect disease vector control. The articles collected in this Research Topic comprise review and original research on metal biology in insects.

This secondary data analysis addressed gaps in knowledge about effects of chronic water intake. Longitudinal data from the Adapt Study were used to describe effects of prescribing a sustained increase in water intake relative to baseline, for 4 weeks, on multiple indices of total body water (TBW) flux, regulation, distribution, and volume in five healthy, free‐living, young men, with mean total water intake initially below 2 L/day. Indices were measured weekly. Within‐person fixed effect models tested for significant changes in indices over time and associations between changes in indices. Agreement between indices was described. Mixed models tested if baseline between‐person differences in hydration indices modified changes in indices over time. Body water flux: The half‐life of water in the body decreased significantly. Body water regulation: Serum osmolality decreased significantly. Urine anti‐diuretic hormone, sodium, potassium, and osmolality decreased significantly. Plasma aldosterone and serum sodium increased significantly. Body water distribution: No significant changes were observed. Body water volume: Saliva osmolality decreased significantly. Body weight increased significantly by a mean ± SEM of 1.8% ± 0.5% from baseline over 4 weeks. Changes in indices were significantly inter‐correlated. Agreement between indices changed over 4 weeks. Baseline saliva osmolality significantly modified responses to chronic water intake. The results motivate hypotheses for future studies: Chronic TBW deficit occurs in healthy individuals under daily life conditions and increases chronic disease risk; Sustained higher water intake restores TBW through gradual isotonic retention of potassium and/or sodium; Saliva osmolality is a sensitive and specific index of chronic hydration status.

Cystinuria is an incompletely dominant disorder characterized by defective urinary cystine reabsorption that results in the formation of cystine-based urinary stones. Current treatment options are limited in their effectiveness at preventing stone recurrence and are often poorly tolerated. We report that the nutritional supplement α-lipoic acid inhibits cystine stone formation in the Slc3a1-/- mouse model of cystinuria by increasing the solubility of urinary cystine. These findings identify a novel therapeutic strategy for the clinical treatment of cystinuria.

Many late-onset proteotoxic diseases are accompanied by a disruption in homeostasis of metals (metallostasis) including iron, copper and zinc. Although aging is the most prominent risk factor for these disorders, the impact of aging on metallostasis and its role in proteotoxic disease remain poorly understood. Moreover, it is not clear whether a loss of metallostasis influences normal aging. We have investigated the role of metallostasis in longevity of Caenorhabditis elegans. We found that calcium, copper, iron, and manganese levels increase as a function of age, while potassium and phosphorus levels tend to decrease. Increased dietary iron significantly accelerated the age-related accumulation of insoluble protein, a molecular pathology of aging. Proteomic analysis revealed widespread effects of dietary iron in multiple organelles and tissues. Pharmacological interventions to block accumulation of specific metals attenuated many models of proteotoxicity and extended normal lifespan. Collectively, these results suggest that a loss of metallostasis with aging contributes to age-related protein aggregation.

Before initiating a mass zinc fortification program, this study assessed the prevalence of and risk factors for low zinc status among Cameroonian women and children. In a nationally representative survey, we randomly selected 30 clusters in each of 3 strata (North, South, and Yaounde/Douala) and 10 households per cluster, each with a woman aged 15-49 y and a child aged 12-59 mo (n = 1002 households). Twenty-four-hour dietary recalls (with duplicates in a subset) and anthropometric measurements were conducted, and non-fasting blood was collected to measure plasma zinc concentration (PZC) and markers of inflammation. PZC was adjusted for methodologic factors (time of collection and processing, and presence of inflammation). The prevalence of stunting was 33% (32% South; 46% North; 13% Yaounde/Douala). Among women, 82% had low adjusted PZC (

The loss of NPC1 protein function is the predominant cause of Niemann-Pick type C1 disease (NP-C1), a systemic and neurodegenerative disorder characterized by late-endosomal/lysosomal accumulation of cholesterol and other lipids. Limited evidence from post-mortem human tissues, an Npc1(-/-) mouse model, and cell culture studies also suggest failure of metal homeostasis in NP-C1. To investigate these findings, we performed a comprehensive transition metal analysis of cerebrospinal fluid (CSF), plasma and tissue samples from human NP-C1 patients and an Npc1(-/-) mouse model. NPC1 deficiency in the Npc1(-/-) mouse model resulted in a perturbation of transition metal homeostasis in the plasma and key organs (brain, liver, spleen, heart, lungs, and kidneys). Analysis of human patient CSF, plasma and post-mortem brain tissues also indicated disrupted metal homeostasis. There was a disparity in the direction of metal changes between the human and the Npc1(-/-) mouse samples, which may reflect species-specific metal metabolism. Nevertheless, common to both species is brain zinc accumulation. Furthermore, treatment with the glucosylceramide synthase inhibitor miglustat, the only drug shown in a controlled clinical trial to have some efficacy for NP-C1, did not correct the alterations in CSF and plasma transition metal and ceruloplasmin (CP) metabolism in NP-C1 patients. These findings highlight the importance of NPC1 function in metal homeostasis, and indicate that metal-targeting therapy may be of value as a treatment for NP-C.

Prolonged exposure to the flame retardants found in many household products and building materials is associated with adverse developmental, reproductive, and carcinogenic consequences. While these compounds have been studied in numerous epidemiological and animal models, less is known about the effects of flame retardant exposure on cell function. This study evaluated the toxicity of the commonly used fire retardant tris(1,3-dichloro-2-propyl)phosphate (TDCPP) in cell line derived from the kidney, a major tissue target of organohalogen toxicity. TDCPP inhibited cell growth at lower concentrations (IC50 27 μM), while cell viability and toxicity were affected at higher concentrations (IC50 171 μM and 168 μM, respectively). TDCPP inhibited protein synthesis and caused cell cycle arrest, but only at higher concentrations. Additionally, the antioxidant N-acetylcysteine (NAC) reduced cell toxicity in cells treated with TDCPP, suggesting that exposure to TDCPP increased oxidative stress in the cells. In summary, these data show that low concentrations of TDCPP result in cytostasis in a kidney cell line, whereas higher concentrations induce cell toxicity. Furthermore, TDCPP toxicity can be attenuated by NAC, suggesting that antioxidants may be effective countermeasures to some organohalogen exposures. Keywords: flame retardant, cytostasis, cell toxicity, antioxidant, cell cycle

Molecular pathways involved in dauer formation, an alternate larval stage that allows Caenorhabditis elegans to survive adverse environmental conditions during development, also modulate longevity and metabolism. The decision to proceed with reproductive development or undergo diapause depends on food abundance, population density, and temperature. In recent years, the chemical identities of pheromone signals that modulate dauer entry have been characterized. However, signals derived from bacteria, the major source of nutrients for C. elegans, remain poorly characterized. To systematically identify bacterial components that influence dauer formation and aging in C. elegans, we utilized the individual gene deletion mutants in E. coli (K12). We identified 56 diverse E. coli deletion mutants that enhance dauer formation in an insulin-like receptor mutant (daf-2) background. We describe the mechanism of action of a bacterial mutant cyaA, that is defective in the production of cyclic AMP, which extends lifespan and enhances dauer formation through the modulation of TGF-β (daf-7) signaling in C. elegans. Our results demonstrate the importance of bacterial components in influencing developmental decisions and lifespan in C. elegans. Furthermore, we demonstrate that C. elegans is a useful model to study bacterial-host interactions.

We determined the association between dietary zinc intake and incident calcium kidney stones in adolescents. We also examined the relationship between dietary zinc intake and urinary zinc excretion between cases and controls.We conducted a nested case-control study within a large pediatric health care system. Three 24-hour dietary recalls and spot urine chemistry analyses were obtained for 30 participants 12 to 18 years old with a first idiopathic calcium based kidney stone and 30 healthy controls matched for age, sex, race and month of enrollment. Conditional logistic regression models were used to estimate the association between daily zinc intake and incident calcium kidney stones, adjusting for dietary phytate, protein, calcium, sodium and oxalate. Multivariable linear regression was used to estimate the association between dietary and urine zinc, adjusting for urine creatinine and dietary phytate and calcium.Cases had lower daily zinc intake (8.1 mg) than controls (10 mg, p = 0.029). Daily zinc intake of boys and girls with calcium stones was 2 mg and 1.2 mg less, respectively, than the daily intake recommended by the Institute of Medicine. Odds of incident stones were reduced by 13% for every 1 mg increase in daily zinc intake (OR 0.87, 95% CI 0.75-0.99). There was an estimated 4.5 μg/dl increase in urine zinc for every 1 mg increase in dietary zinc (p = 0.009), with weak evidence of a smaller increase in urine zinc in cases than in controls (interaction p = 0.08).Decreased dietary zinc intake was independently associated with incident calcium nephrolithiasis in this population of adolescents.

Food fortification has been recommended to improve a population's micronutrient status. Biofortification techniques modestly elevate the zinc content of cereals, but few studies have reported a positive impact on functional indicators of zinc status.We determined the impact of a modest increase in dietary zinc that was similar to that provided by biofortification programs on whole-body and cellular indicators of zinc status.Eighteen men participated in a 6-wk controlled consumption study of a low-zinc, rice-based diet. The diet contained 6 mg Zn/d for 2 wk and was followed by 10 mg Zn/d for 4 wk. To reduce zinc absorption, phytate was added to the diet during the initial period. Indicators of zinc homeostasis, including total absorbed zinc (TAZ), the exchangeable zinc pool (EZP), plasma and cellular zinc concentrations, zinc transporter gene expression, and other metabolic indicators (i.e., DNA damage, inflammation, and oxidative stress), were measured before and after each dietary-zinc period.TAZ increased with increased dietary zinc, but plasma zinc concentrations and EZP size were unchanged. Erythrocyte and leukocyte zinc concentrations and zinc transporter expressions were not altered. However, leukocyte DNA strand breaks decreased with increased dietary zinc, and the level of proteins involved in DNA repair and antioxidant and immune functions were restored after the dietary-zinc increase.A moderate 4-mg/d increase in dietary zinc, similar to that which would be expected from zinc-biofortified crops, improves zinc absorption but does not alter plasma zinc. The repair of DNA strand breaks improves, as do serum protein concentrations that are associated with the DNA repair process. This trial was registered at clinicaltrials.gov as NCT02861352.

Disability or death due to intracerebral hemorrhage (ICH) is attributed to blood lysis, liberation of iron, and consequent oxidative stress. Iron chelators bind to free iron and prevent neuronal death induced by oxidative stress and disability due to ICH, but the mechanisms for this effect remain unclear. We show that the hypoxia-inducible factor prolyl hydroxylase domain (HIF-PHD) family of iron-dependent, oxygen-sensing enzymes are effectors of iron chelation. Molecular reduction of the three HIF-PHD enzyme isoforms in the mouse striatum improved functional recovery after ICH. A low-molecular-weight hydroxyquinoline inhibitor of the HIF-PHD enzymes, adaptaquin, reduced neuronal death and behavioral deficits after ICH in several rodent models without affecting total iron or zinc distribution in the brain. Unexpectedly, protection from oxidative death in vitro or from ICH in vivo by adaptaquin was associated with suppression of activity of the prodeath factor ATF4 rather than activation of an HIF-dependent prosurvival pathway. Together, these findings demonstrate that brain-specific inactivation of the HIF-PHD metalloenzymes with the blood-brain barrier–permeable inhibitor adaptaquin can improve functional outcomes after ICH in several rodent models.

Iron overload occurs frequently in thalassemia and other disorders that require regular blood transfusions. Excess iron is toxic owing to the generation of free radicals that lead to oxidation of biomolecules and tissue damage. In order to identify compounds that reduce oxidative injury from iron, we evaluated alpha-lipoic acid (LA), a multifunctional antioxidant, in iron-overloaded primary human fibroblasts (IMR-90). Oxidant stress was measured using dichlorodihydrofluorescein diacetate that is converted to the fluorescent dichlorofluorescein (DCF) upon oxidation. Exposure to ferric ammonium citrate (FAC) increased the iron-content of IMR-90 cells and caused a rise in oxidant appearance. The addition of LA improved the cellular redox status and attenuated the iron-mediated rise in oxidants in a dose-dependent manner. The R- and RS-enantiomers of LA demonstrated similar antioxidant activity. N-tert-butyl hydroxylamine (NtBHA) treated cells also exhibited a decrease in DCF fluorescence, but at a much higher concentration compared with LA. The combination acetyl-L-carnitine (ALCAR) and LA exhibited superior antioxidant effect at all dose levels. We conclude that LA is highly effective in reversing oxidative stress arising from iron overload and that its antioxidant efficacy is further enhanced in combination with ALCAR.

Chemotherapeutics used to treat prostate cancer are often from a class of drugs that target microtubule networks, such as paclitaxel. A previous report indicated that supplemental zinc sensitized prostate cancer cells to paclitaxel-induced apoptosis, suggesting that increased zinc levels might enhance paclitaxel efficacy. The effect of zinc deficiency on paclitaxel activity is not known though, so we tested this in two prostate cancer cell lines maintained under moderately zinc-deficient conditions. LNCaP and PC3 cell lines were used as models of early and late-stage prostate cancer, respectively. Cells cultured in reduced zinc levels did not demonstrate altered cell viability, growth rates, or intracellular zinc content. Additionally, zinc deficiency alone had no apparent effect on cell cycle kinetics or apoptosis levels. However, the IC50 for paclitaxel-induced cell cycle arrest increased in LNCaP cells from zinc-deficient compared to zinc-replete conditions. Consequently, paclitaxel-induced apoptosis was reduced in LNCaP cells from zinc-deficient compared to zinc-replete conditions. In PC3 cells, the effects of paclitaxel were independent of zinc status. Reduced extracellular zinc levels were shown to affect paclitaxel activity in a prostate cancer cell line. Given the prevalence of zinc deficiency, determining how chemotherapeutic action is modulated by zinc adequacy may have clinical importance.

Age-related macular degeneration (AMD) is the leading cause of severe visual impairment in the elderly in developed countries. AMD patients have elevated levels of iron within the retinal pigment epithelia (RPE), which may lead to oxidative damage to mitochondria, disruption of retinal metabolism, and vision impairment or loss. As a possible model for iron-induced AMD, we investigated the effects of excess iron in cultured human fetal RPE cells on oxidant levels and mitochondrial cytochrome c oxidase (complex IV) function and tested for protection by N-tert-butyl hydroxylamine (Nt-BHA), a known mitochondrial antioxidant. RPE exposure to ferric ammonium citrate resulted in a time- and dose-dependent increase in intracellular iron, which increased oxidant production and decreased glutathione (GSH) levels and mitochondrial complex IV activity. NtBHA addition to iron-overloaded RPE cells led to a reduction of intracellular iron content, oxidative stress, and partial restoration of complex IV activity and GSH content. NtBHA might be useful in AMD due to its potential to reduce oxidative stress, mitochondrial damage, and age-related iron accumulation, which may damage normal RPE function and lead to loss of vision.

Here we report the design, implementation, and initial use of an asymmetric steady-state continuous dual-nanospray ion source. This new source design consists of two independently controlled and continuously operating nanospray interfaces with funnel shaped counter electrodes. A steady-state ion mixing region combines the ions from the two sources into a single ion beam in the intermediate region after ion extraction from the nanospray sources but before the bulk of the pressure gradient of the vacuum interface. With this design we have achieved robust mixing of ions with no loss of duty cycle and remarkable ionization characteristics that appear entirely noncompetitive and potentially beneficial. This allows continuous introduction of internal mass calibration ions during a liquid chromatography-mass spectrometric analysis. This in turn allows for recalibration of individual spectra yielding sub part per million mass accuracy throughout the run. The steady-state approach presented here has several advantages over previous approaches. Since neither the voltage nor positions of the sprayers are changed, the nanospray has greater spray stability. The ions produced by the analyte sprayer are continuously sampled, as opposed to time-sharing which necessitates that the analyte ion stream be interrupted for some part of the duty cycle. There are no moving parts, no rapid changes to high voltages requiring additional control electronics, and no need for completely separate vacuum interfaces and the associated complexity. The sprayers are independently controlled and do not exhibit competition or mutual ionization suppression. This novel source has been implemented with a Bruker Apex II 9.4 T FTICR with a modified Apollo electrospray ion source as part of a nanoflow liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry analysis platform. Because of the low cost of implementation, the new source could potentially be applied to other forms of mass spectrometry, such as electrospray ionization-time-of-flight (ESI-TOF), which can benefit from internal mass calibration.

Background Lime-treated corn gruel ( atole) is a common weaning food in iron-deficient populations, especially in Mexico and Central America, and is a potential vehicle for fortification with iron. Objective The objective of this study was to screen promising iron compounds for use in the fortification of atole, using in vitro enzymatic digestion–dialysis techniques, while also considering their response to known iron absorption enhancers and inhibitors. Methods Atole, unaltered or preincubated with phytase, was fortified with iron (10 mg/L) from ferrous sulfate, ferrous bisglycinate, or ferrous fumarate, or with ferric chloride, ferric ammonium citrate, or ferric sodium ethylenediaminetetraacetic acid (NaFeEDTA), and submitted to in vitro digestion. Dialysis of calcium, copper, iron, phosphorus, and zinc (analyzed by inductively coupled plasma atomic emission spectrometry) was measured when atole was fortified with iron compounds alone or together with ascorbic acid or disodium ethyl- enediaminetetraacetic acid (Na2 EDTA). Results Iron dialyzability was higher with NaFeEDTA ( p < .05) than with all other iron compounds, which did not differ among themselves in iron dialyzability. Addition of ascorbic acid had no significant effect on iron dialysis, whereas Na2 EDTA enhanced iron dialyzability by 7 to 10 times in unaltered atole and 15 to 20 times in phytase-preincubated atole ( p < .05). Addition of Na2 EDTA always increased intrinsic zinc dialyzability, and most of the time this increase was significant. Phytase pretreatment generally increased mineral dialysis. Conclusions Adding EDTA (either as NaFeEDTA or as Na2 EDTA) to atole can increase the dialyzability of ferrous and ferric iron compounds and enhance the dialyzability of intrinsic zinc without any negative effects on calcium, phosphorus, or copper dialysis.

Zinc is an essential trace metal that has integral roles in numerous biological processes, including enzymatic function, protein structure, and cell signaling pathways. Both excess and deficiency of zinc can lead to detrimental effects on development and metabolism, resulting in abnormalities and disease. We altered the zinc balance within Caenorhabditis elegans to examine how changes in zinc burden affect longevity and healthspan in an invertebrate animal model. We found that increasing zinc levels in vivo with excess dietary zinc supplementation decreased the mean and maximum lifespan, whereas reducing zinc levels in vivo with a zinc-selective chelator increased the mean and maximum lifespan in C. elegans. We determined that the lifespan shortening effects of excess zinc required expression of DAF-16, HSF-1 and SKN-1 proteins, whereas the lifespan lengthening effects of the reduced zinc may be partially dependent upon this set of proteins. Furthermore, reducing zinc levels led to greater nuclear localization of DAF-16 and enhanced dauer formation compared to controls, suggesting that the lifespan effects of zinc are mediated in part by the insulin/IGF-1 pathway. Additionally, zinc status correlated with several markers of healthspan in worms, including proteostasis, locomotion and thermotolerance, with reduced zinc levels always associated with improvements in function. Taken together, these data support a role for zinc in regulating both development and lifespan in C. elegans, and that suggest that regulation of zinc homeostasis in the worm may be an example of antagonistic pleiotropy.

A coordinated network of zinc transporters and binding proteins tightly regulate cellular zinc levels. Canonical responses to zinc availability are thought to be mediated by changes in gene expression of key zinc transporters. We investigated the temporal relationships of actual zinc uptake with patterns of gene expression in membrane-bound zinc transporters in the human immortalized T lymphocyte Jurkat cell line. Cellular zinc levels were elevated or reduced with exogenous zinc sulfate or N,N,N',N-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), respectively. Excess zinc resulted in a rapid 44 % decrease in the rate of zinc uptake within 10 min. After 120 min, the expression of metallothionein (positive control) increased, as well as the zinc exporter, ZnT1; however, the expression of zinc importers did not change during this time period. Zinc chelation with TPEN resulted in a rapid twofold increase in the rate of zinc uptake within 10 min. After 120 min, the expression of ZnT1 decreased, while again the expression of zinc importers did not change. Overall, zinc transporter gene expression kinetics did not match actual changes in cellular zinc uptake with exogenous zinc or TPEN treatments. This suggests zinc transporter regulation may be the initial response to changes in zinc within Jurkat cells.

Background Patients with cystic fibrosis (CF) may be at risk for micronutrient depletion, particularly during periods of illness and infection. The purpose of this study was to investigate serum micronutrient status over time in adults with CF initially hospitalized with a pulmonary exacerbation. Materials and methods This was an ancillary study of a multicenter trial investigating the role of high-dose vitamin D supplementation in 24 adults with CF (mean age, 29.6 ± 7.3 years). We measured serum concentrations of copper (Cu), iron (Fe), calcium (Ca), magnesium (Mg), potassium (K), and sulfur (S) in subjects at the beginning of a pulmonary exacerbation and again at 3 months. Results Serum concentrations of Cu, Fe, and Ca were significantly lower at baseline compared with 3 months following the pulmonary exacerbation (Cu: baseline, 1.5 ± 0.6 vs 3 months, 1.6 ± 0.6 µg/mL, P = .027; Fe: 0.8 ± 0.3 vs 1.3 ± 1.1 µg/mL, P = .026; Ca: 9.7 ± 0.8 vs 10.8 ± 2.0 mg/dL, P = .024). Serum concentrations of K, Mg, and S did not change over time (K: baseline, 4.9 ± 0.3 vs 3 months, 5.1 ± 0.5 mEq/L; Mg: 1.8 ± 0.2 vs 2.0 ± 0.3 mg/dL; S: 1288.6 ± 343 vs 1309.9 ± 290 µg/mL; P > .05 for all). Conclusion Serum concentrations of Cu, Fe, and Ca increased significantly several months following recovery from acute pulmonary exacerbation in adults with CF. This may reflect decreased inflammation, improved food intake, and/or increased absorption following recovery.

© 2015 Killilea et al. One of the most common types of urinary stones formed in humans and some other mammals is composed of calcium oxalate in ordered hydrated crystals. Many studies have reported a range of metals other than calcium in human stones, but few have looked at stones from animal models such as the dog. Therefore, we determined the elemental profile of canine calcium oxalate urinary stones and compared it to reported values from human stones. The content of 19 elements spanning 7-orders of magnitude was quantified in calcium oxalate stones from 53 dogs. The elemental profile of the canine stones was highly overlapping with human stones, indicating similar inorganic composition. Correlation and cluster analysis was then performed on the elemental profile from canine stones to evaluate associations between the elements and test for potential subgrouping based on elemental content. No correlations were observed with the most abundant metal calcium. However, magnesium and sulfur content correlated with the mineral hydration form, while phosphorous and zinc content correlated with the neuter status of the dog. Inter-elemental correlation analysis indicated strong associations between barium, phosphorous, and zinc content. Additionally, cluster analysis revealed subgroups within the stones that were also based primarily on barium, phosphorous, and zinc. These data support the use of the dog as a model to study the effects of trace metal homeostasis in urinary stone disease.

Blood transfusion therapy is life-saving for patients with beta-thalassaemia and sickle cell disease (SCD), but often results in severe iron overload. This pilot study examined whether the biomarkers of tissue injury or inflammation differ in these two diseases. Plasma malondialdehyde (MDA) was significantly increased 1.8-fold in thalassaemia relative to control patients. In contrast, MDA in SCD was not significantly different from controls. In multivariate analysis, the strongest predictors of elevated MDA were liver iron concentration (P < 0.001) and specific diagnosis (P = 0.019). A significant 2-fold elevation of non-transferrin bound iron (NTBI) was observed in thalassaemia relative to SCD. NTBI was not a significant predictor of high MDA in multivariate analysis. SCD patients showed a significant 2.2-fold elevation of the inflammatory marker interleukin (IL)-6 relative to controls, and a 3.6- and 1.7-fold increase in IL-5 and IL-10 relative to thalassaemia. Although alpha-tocopherol was significantly decreased by at least 32% in both thalassaemia and SCD, indicating ongoing oxidant stress and antioxidant consumption, gamma-tocopherol, a nitric oxide-selective antioxidant, was increased 36% in SCD relative to thalassaemia. These results demonstrate that thalassaemia patients have increased MDA and circulating NTBI relative to SCD patients and lower levels of some cytokines and gamma-tocopherol. This supports the hypothesis that the biology of SCD may show increased inflammation and increased levels of protective antioxidants compared with thalassaemia.

Thalassemia (THAL) patients have a genetic anemia causing incomplete erythropoiesis and iron overload, possibly leading to cardiomyopathy and diabetes. Plasma zinc (Zn) and Copper (Cu) deficiency are also observed in some patients with THAL, however, these effects are not as well characterized. It has been suggested that oxidative stress can be caused by increased labile plasma iron (LPI), and decreased serum Zn and Cu levels, however this is unproven. LPI can initiate peroxidation releasing malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE). Thus, we hypothesized that that a functional Zn or Cu deficiency can increase oxidative stress and that changes in the levels of key iron trafficking proteins, such as soluble transferrin receptor (sTfR) or hemopexin (HPX) would also affect oxidative stress. 39 subjects with informed consent were enrolled (29 patients with THAL and 10 healthy controls). LPI was measured using dihydrorhodamine 123. Both MDA and 4-HNE were measured using N-methyl-2-phenylindole. Iron trafficking proteins were measured by MRM mass spectrometry. Trace mineral levels were assessed by ICP spectroscopy. Plasma levels of the iron trafficking proteins, transferrin, HPX and haptoglobin (HAP) were all decreased in THAL patients (P