Objective To modify the Supplemental Nutrition Assistance Program-Education curriculum based on formative research conducted at a homeless resource center. Methods Researchers interviewed 4 administrators/staff and 5 transitional housing residents. We used a content analysis approach to independently code transcripts, reconcile differences, and identify themes. We adapted the Create Better Health curriculum using these data. Results Curriculum adaptations included incorporating microwave-friendly recipes because of limited cooking facilities, providing strategies for making healthy food choices at facility-provided meals and convenience/dollar stores frequented by residents, and encouraging residents to exercise at their comfort levels because of common physical limitations. Classes located outside transitional housing units were planned for 60 minutes before and after mealtimes. Conclusions and Implications Adapting the Supplemental Nutrition Assistance Program-Education curriculum to meet the unique needs of transitional housing residents may enable healthy behavioral changes. Further research is required to evaluate the feasibility and efficacy of this revised curriculum.

To examine body shape perception in 218 adults without obesity or history of eating disorders during caloric restriction (CR). Comprehensive Assessment of Long-term Effects of Reducing Intake of Energy (CALERIE) is a 2-year, randomized clinical trial using a 2:1 assignment (CR, 25 CR is associated with reduced concern for body shape in men and women without obesity and with no history of eating disorders. Body shape perception among this sample was complex and influenced by multiple factors. Level I, randomized controlled trial.

Here the Human Pangenome Reference Consortium presents a first draft of the human pangenome reference. The pangenome contains 47 phased, diploid assemblies from a cohort of genetically diverse individuals 1 . These assemblies cover more than 99% of the expected sequence in each genome and are more than 99% accurate at the structural and base pair levels. Based on alignments of the assemblies, we generate a draft pangenome that captures known variants and haplotypes and reveals new alleles at structurally complex loci. We also add 119 million base pairs of euchromatic polymorphic sequences and 1,115 gene duplications relative to the existing reference GRCh38. Roughly 90 million of the additional base pairs are derived from structural variation. Using our draft pangenome to analyse short-read data reduced small variant discovery errors by 34% and increased the number of structural variants detected per haplotype by 104% compared with GRCh38-based workflows, which enabled the typing of the vast majority of structural variant alleles per sample.

ABSTRACT:Animals (grazing, working, or intrusion) in produce production areas may present a potential contamination source of foodborne pathogens on produce. Cattle grazing on native pecan production orchards, a common practice in the southern United States, provides an opportunity to study the impact of grazing practice and waiting periods on contamination rates of foodborne pathogens of tree nuts. Therefore, the objective of this study was to determine the prevalence of Salmonella and Shiga toxin-producing Escherichia coli (STEC) in native pecan production orchards as influenced by waiting periods between grazing cattle and pecan harvest. Soil (10 g), cattle feces (10 g), and in-shell pecans (25 g) were sampled from five cattle-grazed orchards in areas with cattle removed 2 or 4 months before harvest and not removed. Five nongrazing orchards were sampled at harvest for comparison. Detection and isolation of the pathogens were performed by enrichment, selective isolation, and multiplex PCR. Statistical analyses were performed using contingency tables with Pearson's chi-square test. The prevalence of STEC (36%) and Salmonella (29%) in cattle-grazed orchards was significantly higher than in nongrazed orchards (13 and 7%, respectively). STEC prevalence in cattle-grazed orchards was higher (38%) in areas with cattle at harvest than in fenced areas where cattle were removed 2 (29%) and 4 (27%) months before harvest. Salmonella prevalence was similar in areas without fencing (31%) and areas with cattle removed at 2 (22%) and 4 (30%) months before harvest. However, there were no significant differences (P > 0.05) in contamination rates between waiting periods for either pathogen, suggesting a limited impact of waiting periods on reducing the risk of contamination. HIGHLIGHTS:

The human reference genome is the most widely used resource in human genetics and is due for a major update. Its current structure is a linear composite of merged haplotypes from more than 20 people, with a single individual comprising most of the sequence. It contains biases and errors within a framework that does not represent global human genomic variation. A high-quality reference with global representation of common variants, including single-nucleotide variants, structural variants and functional elements, is needed. The Human Pangenome Reference Consortium aims to create a more sophisticated and complete human reference genome with a graph-based, telomere-to-telomere representation of global genomic diversity. Here we leverage innovations in technology, study design and global partnerships with the goal of constructing the highest-possible quality human pangenome reference. Our goal is to improve data representation and streamline analyses to enable routine assembly of complete diploid genomes. With attention to ethical frameworks, the human pangenome reference will contain a more accurate and diverse representation of global genomic variation, improve gene–disease association studies across populations, expand the scope of genomics research to the most repetitive and polymorphic regions of the genome, and serve as the ultimate genetic resource for future biomedical research and precision medicine.

Purpose: Very little research has investigated the effects of ultraendurance exercise on the bioenergetic status of muscle. The primary objective of this case study was to characterize the changes that occur in skeletal muscle mitochondria in response to a 100-km ultramarathon in monozygotic twins. A second objective was to determine whether mitochondrial function is altered by consuming a periodized lowcarbohydrate, high-fat diet during training compared with a high-carbohydrate diet. Methods: One pair of male monozygotic twins ran 100 kmon treadmills after 4 wk of training on either a high-carbohydrate or periodized low-carbohydrate, high-fat diet. Muscle biopsies were collected 4 wk before the run, as well as 4 and 52 h postrun. Blood draws were also performed immediately before as well as 4 and 52 h after the run. Results: Four hours postrun, respiratory capacity, citrate synthase activity, and mitochondrial complex protein content were decreased. Two days later, both twins showed signs of rapid recovery in several of these measures. Furthermore, blood levels of creatine phosphokinase, C-reactive protein, and aspartate transaminase were elevated 4 h after the run but partially recovered 2 d later. Conclusion: Although there were some differences between the twins, the primary finding is that there is significant mitochondrial impairment induced by running 100 km, which rapidly recovers within 2 d. These results provide ample rationale for future investigations of the effects of ultraendurance activity on mitochondrial function.

The primary objective of this case study was to investigate the changes in skeletal muscle mitochondria that occur in response to a 100-km ultramarathon in monozygotic twins. Secondarily, we sought to determine if periodically consuming a low-carbohydrate, high-fat diet (LCHFD) during training would affect these changes. To accomplish these objectives, one pair of male monozygotic twins ran 100 km side-by-side on treadmills. During 8 weeks of training leading up to this event, twin 1 (T1) and twin 2 (T2) consumed a calorie and macronutrient-matched, high-carbohydrate diet, with the exception of 2 consecutive days per week for the final 4 weeks where T2 consumed a calorie and protein-matched LCHFD. Training load was matched between the subjects, most of which they completed together. The subjects were extremely well-matched in VO2 max (61.2 and 60.3 ml/kg/min), BMI (20.4 and 20.6 kg/m2), and body mass (62 and 61.9 kg). During the 100-km run, food and fluid intake were matched, as well as the pace of the run. The run took 16 hours and 3 minutes to complete. Blood draws and muscle biopsies from the vastus lateralis were collected 4 weeks prior to the run, as well as 4- and 52-hours post-run. During the last 4 weeks of training, T2 lost an average of ~1 kg of body weight (BW) over the two days of LCHFD each week (comparing BW of 4 consecutive Mondays to Wednesdays, p=0.02), which was regained two days after reintroduction of carbohydrate, consistent with fluctuating glycogen levels. Though T2 trained with periodically low glycogen levels, respiratory-exchange ratio was no different from T1 throughout the 100-km run. Four hours post-run, state 3, uncoupled, and fatty acid-supported mitochondrial respiration were reduced in T1 by 22.3%, 34.1% and 25.7%, respectively, and by 28.4%, 49.4% and 39.9% in T2, which was associated with a decrease of 15-20% in citrate synthase (CS) activity. Both twins exhibited decreases in mitochondrial complex proteins, though this was more marked in T2. Blood CPK, CRP, and AST levels were also elevated post-run. Possibly the most novel finding from this study was that 52 hours post-run, respiratory capacity, mitochondrial proteins, CS activity and blood markers showed signs of rapid recovery. It is currently unclear by what mechanism mitochondrial respiration and content was reduced 4 hours post-run, or how it recovered in 2 days. Interestingly, maximal mitochondrial ROS production (glutamate, malate and succinate-supported) was decreased post-run at both timepoints but was increased compared to baseline with the addition of ADP (12.5 μM – 100 μM). This suggests that the mitochondria were less sensitive to ADP-induced reductions in ROS. Neither diet resulted in better preservation of mitochondrial function 4-hours post-run, though T2 showed some signs of supercompensation 52 hours-post run. This study is valuable because it provides evidence of an acute reduction in mitochondrial content and function following ultra-endurance activity, followed by rapid recovery. This study was approved by the Brigham Young University Institutional Review Board and conformed to the principles of human experimentation set by the Declaration of Helsinki.

Focal cortical dysplasia (FCD) is a common histopathologic finding in cortical specimens resected for refractory epilepsy. GABAergic neuronal abnormalities and K-Cl cotransporter type 2 (KCC2) immaturity may be contributing factors for FCD-related epilepsy. We examined surgical specimens from 12 cases diagnosed with FCD, and brain tissues without developmental abnormality obtained from 6 autopsy cases. We found that GABAergic neuronal density was abnormal in FCD with 2 distinct patterns. In 7 of 12 (58%) FCD subjects, the GABAergic neuron density in dysplastic regions and in neighboring nondysplastic regions was equally reduced, hence we call this a broad pattern. In the remaining cases, GABAergic neuron density was decreased in dysplastic regions but not in the neighboring nondysplastic regions; we designate this restricted pattern. The different patterns are not associated with pathologic subtypes of FCD. Intracytoplasmic retention of KCC2 is evident in dysmorphic neurons in the majority of FCD type II subjects (5/7) but not in FCD type I. Our study suggests that (1) broad GABAergic deficiency may reflect epileptic vulnerability outside the dysplastic area; and (2) abnormal distribution of KCC2 may contribute to seizure generation in patients with FCD type II but not in type I.

INTRODUCTION: Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy and usually associated with good prognosis. Typical sites for metastases include lymph nodes, lung and bone, while renal metastasis is rare. Here we present a case of primary papillary thyroid-like carcinoma of the kidney. Histologically and immunohistochemically, the tumor closely resembled PTC, but no abnormalities were found in the thyroid or cervical lymph nodes. CASE REPORT: 54-year-old Caucasian male non-smoker with history of GERD, and dyslipidemia was referred to ENT for evaluation of left tonsillar mass. PET CT showed hypermetabolic regions in the left base of tongue and lingual tonsil, and hypodensity in the lower pole of the left kidney. He underwent bilateral palatine tonsillectomy for invasive squamous cell carcinoma followed by adjuvant chemoradiation. MRI of the abdomen confirmed a 1.7cm left lower pole renal mass and he underwent laparoscopic partial nephrectomy. Pathological examination of the renal tumor was suggestive of papillary carcinoma of thyroid origin (immunohistochemical staining positive for TTF-1, PAX-8 and CK7, and negative for thyroglobulin, RCC, CK20 and racemase). Histologically, the tumor displayed nuclear inclusions, nuclear grooves and papillary architecture. RET oncogene amplification further supported the diagnosis of PTC. Neck ultrasound was negative for thyroid nodules or lymphadenopathy, and no residual hypermetabolic disease was seen on follow-up PET CT. Due to concern for metastatic PTC, the patient underwent total thyroidectomy with central lymph node dissection. Pathology showed only benign thyroid with lymphocytic thyroiditis. Post-thyroidectomy whole body scan showed no abnormal radiotracer uptake to suggest residual thyroid or disease. Serum thyroglobulin level remained undetectable with negative anti-TPO antibodies. DISCUSSION: This patient presented with an incidentally discovered renal tumor that histologically and immunohistochemically resembled PTC. Renal metastasis of PTC is very rare, with less than 20 cases reported, yet positive staining for TTF-1 is highly sensitive and specific for thyroid cells. The possible origins of the tumor include metastasis from an occult intra-thyroidal tumor, primary PTC that had undergone spontaneous resolution, or a primary renal tumor with thyroid-like characteristics. To our knowledge this is the second reported case of a primary papillary thyroid-like cancer of kidney.

Alzheimer's disease (AD), a progressive neurodegenerative disorder, is the most common untreatable form of dementia. Identifying molecular biomarkers that allow early detection remains a key challenge in the diagnosis, treatment, and prognostic evaluation of the disease. Here, we report a novel experimental and analytical model characterizing epigenetic alterations during AD onset and progression. We generated the first integrated base-resolution genome-wide maps of the distribution of 5-methyl-cytosine (5mC), 5-hydroxymethyl-cytosine (5hmC), and 5-formyl/carboxy-cytosine (5fC/caC) in normal and AD neurons. We identified 27 AD region-specific and 39 CpG site-specific epigenetic signatures that were independently validated across our familial and sporadic AD models, and in an independent clinical cohort. Thus, our work establishes a new model and strategy to study the epigenetic alterations underlying AD onset and progression and provides a set of highly reliable AD-specific epigenetic signatures that may have early diagnostic and prognostic implications.

We previously investigated the association between single nucleotide polymorphisms (SNPs) in genes related to obesity and inflammation and colorectal cancer in the CLUE II cohort. However, the relationships between these SNPs and colorectal adenomas have not been well evaluated. In a nested case‐control study of 135 incident adenoma cases and 269 matched controls in the CLUE II cohort (1989‐2000), we genotyped 17 candidate SNPs in 12 genes (PPARG, TCF7L2, ADIPOQ, LEP, IL10, CRP, TLR4, IL6, IL1B, IL8, TNF, RNASEL) and 19 tagSNPs in three genes (IL10, CRP, and TLR4). Conditional logistic regression was used to calculate odds ratios (OR) for adenomas (overall and by size, histology, location, number). Polymorphisms in the inflammatory‐related genes CRP, ADIPOQ, IL6, and TLR4 were observed to be associated with adenoma risk. At rs1205 in CRP, T (minor allele) carriers had a higher risk (OR 1.67, 95%CI 1.07‐2.60; reference: CC) of adenomas overall and adenomas with aggressive characteristics. At rs1201299 in ADIPOQ, the AC genotype had a higher risk (OR 1.58, 95%CI 1.00‐2.49) of adenomas, while the minor AA genotype had a borderline inverse association (OR 0.44, 95%CI 0.18‐1.08; reference: CC). At rs1800797 in IL6, the AA genotype had a borderline inverse association (OR 0.53, 95%CI 0.27‐1.05; reference: GG). Three TLR4 tagSNPs (rs10116253, rs1927911, rs7873784) were associated with adenomas among obese participants. None of these SNPs were associated with colorectal cancer in our prior study in CLUE II, possibly suggesting a different genetic etiology for early colorectal neoplasia.

Switchgrass (Panicum virgatum L.), as a model herbaceous crop species for bioenergy production, is targeted to improve biomass yield and feedstock quality. Plant height is a major component contributing to biomass yield. Accordingly, the objectives of this research were to analyze phenotypic variation for biomass and plant height and the association between them and to localize associated plant height QTLs. Two lowland switchgrass mapping populations, one selfed and another hybrid population established in the field at Perkins and Stillwater, Oklahoma, were deployed in the experiment for two years post establishment. Large genetic variation existed for plant biomass and height within the two populations. Plant height was positively correlated with biomass yield in the selfed population (r = 0.39, P<0.0001) and the hybrid population (r = 0.41, P<0.0001). In the selfed population, a joint analysis across all environments revealed 10 QTLs and separate analysis for each environment, collectively revealed 39 QTLs related to plant height. In the hybrid population, the joint analysis across overall environments revealed 35 QTLs and the separate analysis for each environment revealed 38 QTLs. The findings of this research contribute new information about the genetic control for plant height and will be useful for future plant breeding and genetic improvement programs in lowland switchgrass.

Previous studies demonstrated that live Mycobacterium leprae (M. leprae) infection promoted macrophage differentiation toward the M2 type, with elevated interleukin (IL)-10 production. The underlying mechanism is not entirely clear. In this study, we treated macrophages with primary M. leprae strains isolated from both lepromatous leprosy (L-lep) and tuberculoid leprosy (T-lep) patients. We found that infection by live M. leprae, regardless of the primary strain, resulted in M2 skewing in the infected macrophage. This skewing was associated with downregulated IRGM expression, a core organizer protein in the autophagy assembly and reduced autophagosome formation, and with lower annexin V staining and lower caspase 3 and caspase 9 activity. Moreover, live M. leprae-infected macrophages prevented efficient phagocytosis by uninfected bystander macrophages. As a result, the phagocytes secreted less pro-inflammatory cytokines, and preferentially primed anti-inflammatory T cell responses. Together, these results suggested that live M. leprae could employ a strain-independent mechanism to suppress inflammation, possibly involving the inhibition of autophagy and apoptosis in the infected macrophages.

Scab is the most damaging disease of pecan in the southeastern United States. Pecan trees can attain 44 m in height, so managing disease in the upper canopy is a problem. Fungicide is ordinarily applied using ground-based air-blast sprayers. Although mechanical hedge-pruning and topping of pecan is done for several reasons, improved management of scab is an important reason in the humid, wet Southeast. Resulting shoot growth on cut limbs of susceptible cultivars could lead to more severe scab. In three experiments over three years, we explored the effect of hedge-pruning trees to ∼12 to 14 m compared with non-hedge-pruned trees. All trees received fungicide treatments (air-blast sprays and ≤3 aerial applications). Hedge-pruning either had no effect, or increased or decreased scab severity only slightly on leaflets, immature, or mature fruit (a –9.95 to +14.63% difference in scab severity compared with the control). However, height in the canopy invariably had a large and significant effect on scab severity, and amounted to a 0.05 to 73.77% difference in severity between the lowest and highest sample in the canopy. Fruit weight depended on sample height, with fruit most often weighing less when collected at greater sample heights. A robust relationship between fruit weight and scab severity was found at the highest sample heights where scab was also most often severe (R 2 = 0.21 to 0.67, P < 0.0001). Hedge-pruning and topping pecan tree canopies to manage tree size will enable better fungicide coverage, reducing risk of a scab epidemic as more of the canopy is assured efficacious fungicide spray coverage.

During human brain development, multiple signaling pathways generate diverse cell types with varied regional identities. Here, we integrate single-cell RNA sequencing and clonal analyses to reveal lineage trees and molecular signals underlying early forebrain and mid/hindbrain cell differentiation from human embryonic stem cells (hESCs). Clustering single-cell transcriptomic data identified 41 distinct populations of progenitor, neuronal, and non-neural cells across our differentiation time course. Comparisons with primary mouse and human gene expression data demonstrated rostral and caudal progenitor and neuronal identities from early brain development. Bayesian analyses inferred a unified cell-type lineage tree that bifurcates between cortical and mid/hindbrain cell types. Two methods of clonal analyses confirmed these findings and further revealed the importance of Wnt/β-catenin signaling in controlling this lineage decision. Together, these findings provide a rich transcriptome-based lineage map for studying human brain development and modeling developmental disorders.

Lee Wheless1, Emily Kistner-Griffin1,2, Timothy J. Jorgensen3, Ingo Ruczinski4, Yvette Berthier-Schaad5,8, Bailey Kessing5, Judith Hoffman-Bolton6, Lesley Francis6, Yin Yao Shugart7, Paul T. Strickland8,9, W.H. Linda Kao8, Rhoda M. Alani10, Michael W. Smith11, and Anthony J. Alberg1,2,8 1Division of Epidemiology and Biostatistics, Department of Medicine, Medical University of South Carolina, Charleston, SC

BACKGROUND:The presence of population structure in a sample may confound the search for important genetic loci associated with disease. Our four samples in the Family Investigation of Nephropathy and Diabetes (FIND), European Americans, Mexican Americans, African Americans, and American Indians are part of a genome- wide association study in which population structure might be particularly important. We therefore decided to study in detail one component of this, individual genetic ancestry (IGA). From SNPs present on the Affymetrix 6.0 Human SNP array, we identified 3 sets of ancestry informative markers (AIMs), each maximized for the information in one the three contrasts among ancestral populations: Europeans (HAPMAP, CEU), Africans (HAPMAP, YRI and LWK), and Native Americans (full heritage Pima Indians). We estimate IGA and present an algorithm for their standard errors, compare IGA to principal components, emphasize the importance of balancing information in the ancestry informative markers (AIMs), and test the association of IGA with diabetic nephropathy in the combined sample. RESULTS:A fixed parental allele maximum likelihood algorithm was applied to the FIND to estimate IGA in four samples: 869 American Indians; 1385 African Americans; 1451 Mexican Americans; and 826 European Americans. When the information in the AIMs is unbalanced, the estimates are incorrect with large error. Individual genetic admixture is highly correlated with principle components for capturing population structure. It takes ~700 SNPs to reduce the average standard error of individual admixture below 0.01. When the samples are combined, the resulting population structure creates associations between IGA and diabetic nephropathy. CONCLUSIONS:The identified set of AIMs, which include American Indian parental allele frequencies, may be particularly useful for estimating genetic admixture in populations from the Americas. Failure to balance information in maximum likelihood, poly-ancestry models creates biased estimates of individual admixture with large error. This also occurs when estimating IGA using the Bayesian clustering method as implemented in the program STRUCTURE. Odds ratios for the associations of IGA with disease are consistent with what is known about the incidence and prevalence of diabetic nephropathy in these populations.

The U.S. Environmental Protection Agency (USEPA) has restricted concentrated animal feeding operation (CAFO) release of waste products into U.S. waters. These waste products must be disposed of using best management practices. Most of the waste is spread on cropland, but some operations have found other creative uses for waste products. Use of a phosphorus (P) reduction system to remove P from wastewater results in magnesium ammonium phosphate (MAP), a slowly soluble fertilizer. Using a P reduction system will not eliminate the need for land application of manure and wastewater, but it reduces the nutrient load in the waste that is applied thereby making compliance with regulations easier. In the first year of this study, MAP was compared to a controlled release fertilizer (CRF) with a similar nutrient element ratio on plant growth, fruit yield, nitrogen (N), P, potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), and zinc (Zn) concentration in tomato (Solanum lycopersicum L. 'Mountain Fresh Plus') plant parts. Plant growth and fruit production were similar with the two fertilizers, but the number of tomato culls was greater with either fertilizer than on control plants. Foliar N, P, Ca, and Mg concentration did not differ regardless of fertilizer treatment. Plants fertilized with CRF had a greater leaf K concentration than those fertilized with MAP, but foliar K concentration did not differ between fertilized and nonfertilized plants. Iron and Mn concentration in above-ground vegetative plant parts (stems and leaves) did not differ regardless of fertilizer treatment, but Zn concentration increased linearly as CRF increased. In the second year, MAP, each of the essential elements contained in MAP separately, and a hand mixture of each of these elements was tested for their effect on tomato plant growth, fruit yield, and tissue N, P, K, Ca, Mg, Fe, Mn, and Zn concentration and content. Magnesium ammonium phosphate and the hand mixture of fertilizer resulted in greater above-ground biomass excluding fruit stem weight and fruit yield than any of the individual nutrient treatments. Calcium sulfate resulted in a greater number and weight of tomatoes harvested than MAP. Nitrogen concentration did not differ among the fertilizer treatments for roots, stems, or leaves, but N content was greater in red fruit with the hand mix of fertilizer than with no fertilizer or with ammonium sulfate or Mg oxide. In immature green fruit at termination of the study, N content was greater with no fertilizer or Ca sulfate than with MAP or triple superphosphate (TSP). Phosphorus, K, and Ca concentrations did not differ among fertilizer treatments for any tissue tested. Magnesium concentration in green tomatoes differed among fertilizer treatments such that Mg concentration of green tomatoes from plants fertilized with TSP was greater than Mg concentration of green tomatoes fertilized with ammonium sulfate or Mg oxide. Phosphorus and K content of green fruit differed among fertilizer treatments with P and K concentration highest in green fruit from plants fertilized with Ca sulfate and lowest in green fruit from plants fertilized with MAP or TSP. Iron and Mn concentrations did not differ among fertilizer treatments for any tissue tested. Zinc concentration in leaves was greater when plants were fertilized with MAP, TSP, Ca sulfate, or Mg oxide than with ammonium sulfate. Zinc concentration of green fruit was greater when fertilized with MAP than with the hand mix, Ca sulfate or Mg oxide.Iron content was highest in green fruit from plants fertilized with TSP and lowest in plants fertilized with ammonium sulfate or control plants. Manganese content of leaves from control plants was greater than that of plants receiving ammonium sulfate while red fruit from plants fertilized with the hand mix had a greater Mn content than red fruit from any other treatment. Foliar Zn content was greater in plants fertilized with Ca sulfate than in those fertilized with the hand mix, ammonium sulfate, or TSP. In contrast, Zn content of red fruit fertilized with the hand mix was greater than for red fruit in any other treatment. Green fruit from control plants and those receiving MAP had a greater Zn content than plants fertilized with the hand mix, TSP, or Mg oxide. Fertilizer application increased Fe, Mn, and Zn content of several plant tissues. None of the labels of fertilizers applied stated that they contained micronutrients; however, small amounts of contamination were possible. Differing micronutrient contents of various plant tissues among fertilizer treatments were probably associated with other elements affecting plant growth or nutrient uptake. The nutrient elements present in the various fertilizers were not always the nutrient elements affected in the plants likely due to another element that may have limited plant growth or nutrient uptake. (C) 2016 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license.

BACKGROUND The cause of persistent injury-associated anemia is multifactorial and includes acute blood loss, an altered erythropoietin (EPO) response, dysregulation of iron homeostasis, and impaired erythropoiesis in the setting of chronic inflammation/stress. Hepcidin plays a key role in iron homeostasis and is regulated by anemia and inflammation. Erythropoietin is a main regulator of erythropoiesis induced by hypoxia. A unique rodent model of combined lung injury (LC)/hemorrhagic shock (HS) (LCHS)/chronic restraint stress (CS) was used to produce persistent injury-associated anemia to further investigate the roles of EPO, hepcidin, iron, ferritin, and the expression of EPO receptors (EPOr). METHODS Male Sprague-Dawley rats were randomly assigned into one of the four groups of rodent models: naive, CS alone, combined LCHS, or LCHS/CS. Plasma was used to evaluate levels of EPO, hepcidin, iron, and ferritin. RNA was isolated from bone marrow and lung tissue to evaluate expression of EPOr. Comparisons between models were performed by t tests followed by one-way analysis of variance. RESULTS After 7 days, only LCHS/CS was associated with persistent anemia despite significant elevation of plasma EPO. Combined LCHS and LCHS/CS led to a persistent decrease in EPOr expression in bone marrow on Day 7. The LCHS/CS significantly decreased plasma hepcidin levels by 75% on Day 1 and 84% on Day 7 compared to LCHS alone. Hepcidin plasma levels are inversely proportional to EPO plasma levels (Pearson R = −0.362, p < 0.05). CONCLUSION Tissue injury, hemorrhagic shock, and stress stimulate and maintain high levels of plasma EPO while hepcidin levels are decreased. In addition, bone marrow EPOr and plasma iron availability are significantly reduced following LCHS/CS. The combined deficit of reduced iron availability and reduced bone marrow EPOr expression may play a key role in the ineffective EPO response associated with persistent injury-associated anemia.