Chemical Genetic Identification of PKC Epsilon Substrates in Mouse Brain.

PKC epsilon (PKCε) plays important roles in behavioral responses to alcohol and in anxiety-like behavior in rodents, making it a potential drug target for reducing alcohol consumption and anxiety. Identifying signals downstream of PKCε could reveal additional targets and strategies for interfering with PKCε signaling. We used a chemical genetic screen combined with mass spectrometry to identify direct substrates of PKCε in mouse brain and validated findings for 39 of them using peptide arrays and in vitro kinase assays. Prioritizing substrates with several public databases such as LINCS-L1000, STRING, GeneFriends, and GeneMAINA predicted interactions between these putative substrates and PKCε and identified substrates associated with alcohol-related behaviors, actions of benzodiazepines, and chronic stress. The 39 substrates could be broadly classified in three functional categories: cytoskeletal regulation, morphogenesis, and synaptic function. These results provide a list of brain PKCε substrates, many of which are novel, for future investigation to determine the role of PKCε signaling in alcohol responses, anxiety, responses to stress, and other related behaviors.

Brain-specific serine/threonine-protein kinase 1 is a substrate of protein kinase C epsilon involved in sex-specific ethanol and anxiety phenotypes.

Protein kinase C epsilon (PKCε) regulates behavioural responses to ethanol and plays a role in anxiety-like behaviour, but knowledge is limited on downstream substrates of PKCε that contribute to these behaviours. We recently identified brain-specific serine/threonine-protein kinase 1 (BRSK1) as a substrate of PKCε. Here, we test the hypothesis that BRSK1 mediates responses to ethanol and anxiety-like behaviours that are also PKCε dependent. We used in vitro kinase assays to further validate BRSK1 as a substrate of PKCε and used Brsk1-/- mice to assess the role of BRSK1 in ethanol- and anxiety-related behaviours and in physiological responses to ethanol. We found that BRSK1 is phosphorylated by PKCε at a residue identified in a chemical genetic screen of PKCε substrates in mouse brain. Like Prkce-/- mice, male and female Brsk1-/- mice were more sensitive than wild-type to the acute sedative-hypnotic effect of alcohol. Unlike Prkce-/- mice, Brsk1-/- mice responded like wild-type to ataxic doses of ethanol. Although in Prkce-/- mice ethanol consumption and reward are reduced in both sexes, they were reduced only in female Brsk1-/- mice. Ex vivo slice electrophysiology revealed that ethanol-induced facilitation of GABA release in the central amygdala was absent in male Brsk1-/- mice similar to findings in male Prkce-/- mice. Collectively, these results indicate that BRSK1 is a target of PKCε that mediates some PKCε-dependent responses to ethanol in a sex-specific manner and plays a role distinct from PKCε in anxiety-like behaviour.

Transcriptome Analysis of Listeria monocytogenes Exposed to Beef Fat Reveals Antimicrobial and Pathogenicity Attenuation Mechanisms.

Listeria monocytogenes is a deadly intracellular pathogen mostly associated with consumption of ready-to-eat foods. This study investigated the effectiveness of total beef fat (BF-T) from flaxseed-fed cattle and its fractions enriched with monounsaturated fatty acids (BF-MUFA) and polyunsaturated fatty acids (BF-PUFA), along with commercially available long-chain fatty acids (LC-FA), as natural antimicrobials against L. monocytogenes BF-T was ineffective at concentrations up to 6 mg/ml, while L. monocytogenes was susceptible to BF-MUFA and BF-PUFA, with MICs at pH 7 of 0.33 ± 0.21 mg/ml and 0.06 ± 0.03 mg/ml, respectively. The MIC of C14:0 was significantly lower than those of C16:0 and C18:0 (P < 0.05). Fatty acids c9-C16:1, C18:2n-6, and C18:3n-3 showed stronger inhibitory activity than c9-C18:1 and conjugated C18:2, with MICs of <1 mg/ml. Furthermore, global transcriptional analysis by transcriptome sequencing (RNA-seq) was performed to characterize the response of L. monocytogenes to selected fatty acids. Functional analysis indicated that antimicrobial LC-UFA repressed the expression of genes associated with nutrient transmembrane transport, energy generation, and oxidative stress resistance. On the other hand, upregulation of ribosome assembly and translation process is possibly associated with adaptive and repair mechanisms activated in response to LC-UFA. Virulence genes and genes involved in bile, acid, and osmotic stresses were largely downregulated, and more so for c9-C16:1, C18:2n-6, and C18:3n-3, likely through interaction with the master virulence regulator PrfA and the alternative sigma factor σBIMPORTANCEListeria monocytogenes is a bacterial pathogen known for its ability to survive and thrive under adverse environments and, as such, its control poses a significant challenge, especially with the trend of minimally processed and ready-to-eat foods. This work investigated the effectiveness of fatty acids from various sources as natural antimicrobials against L. monocytogenes and evaluated their potential role in L. monocytogenes pathogenicity modulation, using the strain ATCC 19111. The findings show that long-chain unsaturated fatty acids (LC-UFA), including unsaturated beef fat fractions from flaxseed-fed cattle, could have the potential to be used as effective antimicrobials for L. monocytogenes through controlling growth as well as virulence attenuation. This not only advances our understanding of the mode of action of LC-UFA against L. monocytogenes but also suggests the potential for use of beef fat or its fractions as natural antimicrobials for controlling foodborne pathogens.

Gene co-expression and alternative splicing analysis of key metabolic tissues to unravel the regulatory signatures of fatty acid composition in cattle.

Increasing the healthy/unhealthy fatty acid (FA) ratio in meat is one of the urgent tasks required to address consumer concerns. However, the regulatory mechanisms ultimately resulting in FA profiles vary among animals and remain largely unknown. In this study, using ~1.2 Tb high-quality RNA-Seq-based transcriptomic data of 188 samples from four key metabolic tissues (rumen, liver, muscle, and backfat) together with the contents of 49 FAs in backfat, the molecular regulatory mechanisms of these tissues contributing to FA formation in cattle were explored. Using this large dataset, the alternative splicing (AS) events, one of the transcriptional regulatory mechanisms in four tissues were identified. The highly conserved and absent AS events were detected in rumen tissue, which may contribute to its functional differences compared with the other three tissues. In addition, the healthy/unhealthy FA ratio related AS events, differential expressed (DE) genes, co-expressed genes, and their functions in four tissues were analysed. Eight key genes were identified from the integrated analysis of DE, co-expressed, and AS genes between animals with high and low healthy/unhealthy FA ratios. This study provides an applicable pipeline for AS events based on comprehensive RNA-Seq analysis and improves our understanding of the regulatory mechanism of FAs in beef cattle.

A Randomized, Placebo-controlled Trial of Fidaxomicin for Prophylaxis of Clostridium difficile-associated Diarrhea in Adults Undergoing Hematopoietic Stem Cell Transplantation.

Background: Clostridium difficile-associated diarrhea (CDAD) is common during hematopoietic stem-cell transplantation (HSCT) and is associated with increased morbidity and mortality. We evaluated fidaxomicin for prevention of CDAD in HSCT patients. Methods: In this double-blind study, subjects undergoing HSCT with fluoroquinolone prophylaxis stratified by transplant type (autologous/allogeneic) were randomized to once-daily oral fidaxomicin (200 mg) or a matching placebo. Dosing began within 2 days of starting conditioning or fluoroquinolone prophylaxis and continued until 7 days after neutrophil engraftment or completion of fluoroquinolone prophylaxis/clinically-indicated antimicrobials for up to 40 days. The primary endpoint was CDAD incidence through 30 days after study medication. The primary endpoint analysis counted confirmed CDAD, receipt of CDAD-effective medications (for any indication), and missing CDAD assessment (for any reason, including death) as failures; this composite analysis is referred to as "prophylaxis failure" to distinguish from the pre-specified sensitivity analysis, which counted only confirmed CDAD (by toxin immunoassay or nucleic acid amplification test) as failure. Results: Of 611 subjects enrolled, 600 were treated and analyzed. Prophylaxis failure was similar in fidaxomicin and placebo recipients (28.6% vs 30.8%; difference 2.2% [-5.1, 9.5], P = .278). However, most failures were due to non-CDAD events. Confirmed CDAD was lower in fidaxomicin vs placebo recipients (4.3% vs 10.7%; difference 6.4% [2.2, 10.6], P = .0014). Drug-related adverse events occurred in 15.0% of fidaxomicin recipients and 20.0% of placebo recipients. Conclusions: While no difference was demonstrated between arms in the primary analysis, results of the sensitivity analysis demonstrated that fidaxomicin significantly reduced the incidence of CDAD in HSCT recipients. Clinical Trials Registration: NCT01691248.

Crystal Structures of the Nuclear Receptor, Liver Receptor Homolog 1, Bound to Synthetic Agonists.

Liver receptor homolog 1 (NR5A2, LRH-1) is an orphan nuclear hormone receptor that regulates diverse biological processes, including metabolism, proliferation, and the resolution of endoplasmic reticulum stress. Although preclinical and cellular studies demonstrate that LRH-1 has great potential as a therapeutic target for metabolic diseases and cancer, development of LRH-1 modulators has been difficult. Recently, systematic modifications to one of the few known chemical scaffolds capable of activating LRH-1 failed to improve efficacy substantially. Moreover, mechanisms through which LRH-1 is activated by synthetic ligands are entirely unknown. Here, we use x-ray crystallography and other structural methods to explore conformational changes and receptor-ligand interactions associated with LRH-1 activation by a set of related agonists. Unlike phospholipid LRH-1 ligands, these agonists bind deep in the pocket and do not interact with residues near the mouth nor do they expand the pocket like phospholipids. Unexpectedly, two closely related agonists with similar efficacies (GSK8470 and RJW100) exhibit completely different binding modes. The dramatic repositioning is influenced by a differential ability to establish stable face-to-face π-π-stacking with the LRH-1 residue His-390, as well as by a novel polar interaction mediated by the RJW100 hydroxyl group. The differing binding modes result in distinct mechanisms of action for the two agonists. Finally, we identify a network of conserved water molecules near the ligand-binding site that are important for activation by both agonists. This work reveals a previously unappreciated complexity associated with LRH-1 agonist development and offers insights into rational design strategies.

Donor Lymphocyte Infusions Used to Treat Mixed-Chimeric and High-Risk Patient Populations in the Relapsed and Nonrelapsed Settings after Allogeneic Transplantation for Hematologic Malignancies Are Associated with High Five-Year Survival if Persistent Full Donor Chimerism Is Obtained or Maintained.

Mixed chimerism (MC), a persistent or increasing number of host cells after allogeneic hematopoietic stem cell transplantation (HSCT), is a predictor of disease relapse. Donor lymphocyte infusions (DLI) have the potential to enhance the graft-versus-malignancy (GVM) effect, reducing the risk of relapse in patients with MC. Hence, in addition to utilizing DLI in the relapsed setting, there is a motivation to pursue pre-emptive DLI for patients in complete remissions after HSCT. To assess the safety and efficacy of DLI, records of 86 patients who received DLI between 2003 and 2015 at a single institution were studied retrospectively. Patients who received DLI included 50 patients with relapsed/residual (RR) disease, 29 patients with emerging MC without detectable disease, and 7 patients in an "other" cohort who had neither RR disease nor emerging MC after HSCT. DLI were administered using a dose-escalation protocol. After DLI, 93% of MC patients converted to full donor chimerism (FDC). Nonrelapsed patients (MC and other) reported high overall survival (OS) at 1 and 5 years (83% at 1 year, 70% at 5 years for MC; 86% at 1 year, 69% at 5 years for other) and was statistically superior to 5-year OS for RR patients (nonrelapsed 69% versus RR 28%; P = .00032). Improved survival correlated with successful conversion to FDC after DLI for RR and MC cohorts: 71% 2-year OS for patients converted to FDC versus 13% for patients who failed to achieve FDC (P < .0001). DLI for nonrelapsed patients was associated with a superior 5-year progression-free survival (PFS) of 71% compared with 18% 5-year PFS in the RR group (P < .0001). Relapse/progressive disease was the most frequent cause of death (41%). Seven MC (24%), 2 other (29%), and 39 RR patients (78%) relapsed or did not respond after DLI. Overall, 6 patients (7%) died of graft-versus-host disease after DLI. Our results demonstrate a successful dose-escalation approach for nonrelapsed patients that correlated with high survival and a high rate of achieving FDC in MC and RR populations. DLI remain a viable option to boost the GVM effect in the relapsed setting and may pre-emptively protect against relapse in MC populations after HSCT.

Fabricating Nanometer-Thick Simultaneously Oleophobic/Hydrophilic Polymer Coatings via a Photochemical Approach.

The simultaneously oleophobic/hydrophilic coatings are highly desirable in antifogging, oil-water separation, and detergent-free cleaning. However, such coatings require special chemical structure, i.e., perfluorinated backbone and polar end-groups, and are too expensive for real-life application. Here, we have developed an UV-based photochemical approach to make nanometer-thick perfluoropolyethers without polar end-groups, which are not intrinsically simultaneously oleophobic/hydrophilic but cost-effective, become simultaneously oleophobic/hydrophilic. The contact angle, ellipsometry, and X-ray photoelectron spectroscopy (XPS) results indicated that the UV irradiation results in the covalent bonding between the polymer and the substrate, which renders more ordered packing of polymer chains and thus the appropriately small interchain distance. As a result, the small water molecules penetrate the polymer network while large oil molecules do not. As a result, the oil contact angle is larger than the water contact angle and the coating shows the simultaneous oleophobicity/hydrophilicity. Moreover, we also demonstrated that this nanometer-thick simultaneously oleophobic/hydrophilic coating has improved long-term antifogging performance and detergent-free cleaning capability and is mechanically robust. The photochemical approach established here potentially can be applied on many other polymers and greatly accelerate the development and application of simultaneously oleophobic/hydrophilic coatings.

A Microfluidic, Multi-Antibody Cell Capture Method to Evaluate Tumor Cells in Cerebrospinal Fluid in Patients With Suspected Leptomeningeal Metastases.

CONTEXT.­: Leptomeningeal disease (LMD) is a clinical sequela of central nervous system metastasis involving the cerebrospinal fluid (CSF), often seen in late-stage solid tumors. It has a grave prognosis without urgent treatment. Standard of care methodologies to diagnose LMD include CSF cytology, magnetic resonance imaging, and clinical evaluation. These methods offer limited sensitivity and specificity for the evaluation of LMD. Here, we describe the analytic performance characteristics of a microfluidic-based tumor cell enrichment and detection assay optimized to detect epithelial cells in CSF using both contrived samples as well as CSF from patients having suspected or confirmed LMD from carcinomas. OBJECTIVE.­: To demonstrate the feasibility of using a microfluidic, multi-antibody cell capture assay to identify and quantify tumor cells in CSF. DESIGN.­: An artificial CSF solution was spiked with 34 different human carcinoma cell lines at different concentrations and assayed for the ability to detect tumor cells to assess analytic accuracy. Two cell lines were selected to assess linearity, intra-assay precision, interinstrument precision, and sample stability. Clinical verification was performed on 65 CSF specimens from patients. Parameters assessed included the number of tumor cells, coefficient of variation percentage, and percentage of tumor cell capture (TCC). RESULTS.­: Among contrived samples, average tumor cell capture ranged from 50% to 82% (261 of 522; 436 of 531), and coefficients of variation ranged from 7% to 67%. The cell capture assay demonstrated a sensitivity of 92% and a specificity of 95% among clinical samples. CONCLUSIONS.­: This assay demonstrated the ability to detect and enumerate epithelial cells in contrived and clinical specimens in an accurate and reproducible fashion. The use of cell capture assays in CSF may be useful as a sensitive test for the diagnosis and longitudinal monitoring of LMD from solid tumors.

Separation of dietary omega-3 and omega-6 fatty acids in food by capillary electrophoresis.

A lower dietary omega-6/omega-3 (n-6/n-3) fatty acid ratio (<4) has been shown to be beneficial in preventing a number of chronic illnesses. Interest exists in developing more rapid and sensitive analytical methods for profiling fatty acid levels in foods. An aqueous CE method was developed for the simultaneous determination of 15 n-3 and n-6 relevant fatty acids. The effect of pH and concentration of buffer, type and concentration of organic modifier, and additive on the separation was investigated in order to determine the best conditions for the analysis. Baseline separations of the 15 fatty acids were achieved using 40 mM borate buffer at pH 9.50 containing 50 mM SDS, 10 mM ß-cyclodextrin, and 10% acetonitrile. The developed CE method has LODs of <5 mg/L and good linearity (R(2) > 0.980) for all fatty acids studied. The proposed method was successfully applied to the determination of n-3 and n-6 fatty acids in flax seed, Udo® oils and a selection of grass-fed and grain-fed beef muscle samples.

Influence of Production Factors on Beef Primal Tissue Composition.

This study used 1076 crossbred steers to evaluate the effects of calf-fed and yearling-fed beef production systems, implant strategies (with and without implants), and their interactions on the primal tissue composition (lean and fat components) of individual primal cuts using complete carcass dissection data. The results indicate that production system × implant interactions affected loin and rib primal weight percentages as well as marbling (p < 0.05) but did not affect the dissectible lean and fat contents of the individual primal cut (p > 0.05). Implants increased lean and decreased fat tissue contents of primal cut; however, the production system only affected lean content in the loin (p < 0.05) and fat content in the loin, round, and rib (p < 0.05). Redundancy analysis revealed a strong association between Angus breed percentage and marbling, as well as between Simmental breed percentage and multiple primal lean traits. Response surface regression models explained less variability in the tissue composition traits in calf-fed compared with yearling-fed animals, suggesting the need for further exploration using genomic studies.

Comparative effects of feeding citrus pulp and grape pomace on nutrient digestibility and utilization in steers.

Feedlot beef production in tropics and subtropics is being limited by shortage of feed ingredients due to prolonged droughts resulting in high prices of cereal grains and reduced profitability. To alleviate feed shortages and increase profitability for the feedlot industry, it is important to explore underutilized non-conventional resources such as fruit by-products. The current study compared the nutrient intake, digestibility and utilization effects of feeding either 150 g/kg of dried citrus pulp (DCP) or grape pomace (DGP) as alternative dietary fibre sources to 7 months-old Angus steers (281 ±â€¯15.7 kg) under feedlot conditions. Twenty-four steers were assigned to three dietary treatments (8 steers/treatment) in a completely randomised design. Each steer was treated as an experimental unit. Steers were adapted to diets for 21 d followed by 7 d of sample collection during which nutrient intake, digestibility, rumen fermentation parameters, nitrogen retention and microbial N supply were determined. All the data were analysed using the GLIMMIX procedures. Overall, steers fed DGP had the greatest dry matter intake (DM), organic matter (OM) crude protein (CP), ash free neutral detergent fibre (aNDFom), ether extract (EE) and starch intake followed by DCP and control (CON) diets (P ≤ .05). Apparent digestibilities of DM, OM and aNDFom were higher (P ≤ .05) for DCP diet compared to DGP and CON diets. Feeding DCP and DGP diets increased ruminal concentrations of total volatile fatty acids, acetate and isovalerate, and acetate to propionate ratio, and reduced propionate concentrations compared to the CON diet (P ≤ .05). The steers fed the CON diet had the greatest urinary excretions of allantoin, uric acid and total purine derivatives followed by those fed the DCP and DGP diets (P ≤ .05). Nitrogen intake, faecal nitrogen (N), N retention and N efficiency utilization were in the order of DGP > DCP > CON diets (P ≤ .05). Overall, feeding DGP as alternative fibre source to wheat bran improved nutrient intake, retention and efficiency of N utilization but reduced apparent nutrient digestibility compared to DCP. Current finding suggests that DGP may be a better fibre substitute for wheat bran in beef diets than DCP.

Polyunsaturated fatty acid, volatile and sensory profiles of beef from steers fed citrus pulp or grape pomace.

The present study compared the effects of feeding dried grape pomace (DGP) or citrus pulp (DCP) at 150 g/kg dry matter compared to a control diet on major polyunsaturated fatty acids (PUFA), volatile and sensory profiles of beef. Feeding DGP or DCP diets to Angus steers for 90 d increased the proportions of C18:2n-6, C20:4n-6, C18:3n-3, total conjugated linoleic acid (CLA), n-3 and n-6 PUFA in muscle. Control-fed beef had greater concentrations of C18:1n-9, total aldehydes, ketones, and alcohols compared to DCP and DGP. Feeding DGP and DCP diets produced less tender beef than control. Overall, finishing steers on diets containing DGP or DCP compared to control increased proportions of total CLA, n-3 and n-6 PUFA, and reduced concentrations of aldehydes, ketones, and alcohols, but did not affect beef sensory attributes except for a slight reduction in tenderness.

Pemmican, an endurance food: Past and present.

Pemmican is a meat product principally comprised of dried meat and fat that has served as a source of sustenance in cold, harsh climates and/or a means of preservation of meat for some Indigenous peoples, fur traders, polar explorers, military and police. Pemmican is acclaimed for its high nutrient density and long shelf life. However, for a meat product that has historically played a significant role and is often cited as the ultimate endurance food, there is a surprising paucity of scientific literature. The present study aims to review the literature to document the history of pemmican, its preparation, nutritional evaluation and additional use in the diet of animals. In view of food security and crisis situations in the world today, meat science may have a role to play in reviving, reformulating and potentially developing new processing strategies for a product like pemmican as a culturally appropriate food, with extended use as emergency provisions or for endurance athletes.

Evaluation of RNA quality and functional transcriptome of beef longissimus thoracis over time post-mortem.

Evaluating RNA quality and transcriptomic profile of beef muscle over time post-mortem may provide insight into RNA degradation and underlying biological and functional mechanisms that accompany biochemical changes occurring post-mortem during transformation of muscle to meat. RNA was extracted from longissimus thoracis (LT) sampled from British Continental crossbred heifer carcasses (n = 7) stored at 4°C in an abattoir drip cooler at 5 time points post-mortem, i.e., 45 min (0 h), 6 h, 24 h, 48 h, and 72 h. Following RNA-Sequencing, processed reads were aligned to the ARS-UCD1.2 bovine genome assembly. Subsequent differential expression (DE) analysis identified from 51 to 1434 upregulated and 27 to 2256 downregulated DE genes at individual time points compared to time 0 h, showing a trend for increasing counts of both upregulated and downregulated genes over time. Gene ontology and biological pathway term enrichment analyses on sets of DE genes revealed several processes and their timelines of activation/deactivation that accompanied or were involved with muscle transformation to meat. Although the quality of RNA in refrigerated LT remained high for several days post-mortem, the expression levels of several known biomarker genes for meat quality began to change from 24 h onwards. Therefore, to ensure accuracy of predictions on meat quality traits based on the expression levels of those biomarker genes in refrigerated beef muscle tissue, it is crucial that those expression measurements be made on RNA sampled within 24 h post-mortem. The present study also highlighted the need for more research on the roles of mitochondrial genes and non-coding genes in orchestrating muscle tissue processes after death, and how pre-mortem immune status might influence post-mortem meat quality.

Trans-10 18:1 in ruminant meats: A review.

Trans (t) fatty acids (TFA) from partially hydrogenated vegetable oils (i.e., industrial trans) have been phased out of foods in many countries due to their promotion of cardiovascular disease. This leaves ruminant-derived foods as the main source of TFA. Unlike industrial TFA where catalytic hydrogenation yields a broad distribution of isomers, ruminant TFA are enzymatically derived and can result in enrichment of specific isomers. Comparisons between industrial and ruminant TFA have often exonerated ruminant TFA due to their lack or at times positive effects on health. At extremes, however, ruminant-sourced foods can have either high levels of t10- or t11-18:1, and when considering enriched sources, t10-18:1 has properties similar to industrial TFA, whereas t11-18:1 can be converted to an isomer of conjugated linoleic acid (cis(c)9,t11-conjugated linoleic acid), both of which have potential positive health effects. Increased t10-18:1 in meat-producing ruminants has not been associated with negative effects on live animal production or meat quality. As such, reducing t10-18:1 has not been of immediate concern to ruminant meat producers, as there have been no economic consequences for its enrichment; nevertheless at high levels, it can compromise the nutritional quality of beef and lamb. In anticipation that regulations regarding TFA may focus more on t10-18:1 in beef and lamb, the present review will cover its production, analysis, biological effects, strategies for manipulation, and regulatory policy.

Enhancing the Nutritional Value of Red Meat through Genetic and Feeding Strategies.

Consumption of red meat contributes to the intake of many essential nutrients in the human diet including protein, essential fatty acids, and several vitamins and trace minerals, with high iron content, particularly in meats with high myoglobin content. Demand for red meat continues to increase worldwide, particularly in developing countries where food nutrient density is a concern. Dietary and genetic manipulation of livestock can influence the nutritional value of meat products, providing opportunities to enhance the nutritional value of meat. Studies have demonstrated that changes in livestock nutrition and breeding strategies can alter the nutritional value of red meat. Traditional breeding strategies, such as genetic selection, have influenced multiple carcass and meat quality attributes relevant to the nutritional value of meat including muscle and fat deposition. However, limited studies have combined both genetic and nutritional approaches. Future studies aiming to manipulate the composition of fresh meat should aim to balance potential impacts on product quality and consumer perception. Furthermore, the rapidly emerging fields of phenomics, nutrigenomics, and integrative approaches, such as livestock precision farming and systems biology, may help better understand the opportunities to improve the nutritional value of meat under both experimental and commercial conditions.

Effect of Feeding Barley, Corn, and a Barley/Corn Blend on Beef Composition and End-Product Palatability.

This study evaluated the relationship among palatability attributes, volatile compounds, and fatty acid (FA) profiles in meat from barley, corn, and blended (50:50, barley and corn) grain-fed steers. Multiple correspondence analysis with three dimensions (Dim) explained 62.2% of the total variability among samples. The Dim 1 and 2 (53.3%) separated pure from blended grain-fed beef samples. Blended grain beef was linked to a number of volatiles including (E,E)-2,4-decadienal, hexanal, 1-octen-3-ol, and 2,3-octanedione. In addition, blended grain-fed beef was linked to fat-like and rancid flavors, stale-cardboard, metallic, cruciferous, and fat-like aroma descriptors, and negative categories for flavor intensity (FI), off-flavor, and tenderness. A possible combination of linoleic and linolenic acids in the blended diet, lower rumen pH, and incomplete biohydrogenation of blended grain-fed polyunsaturates could have increased (p ≤ 0.05) long-chain n-6 fatty acids (LCFA) in blended grain-fed beef, leading to more accumulation of FA oxidation products in the blended than in barley and corn grain-fed meat samples. The Dim 3 (8.9%) allowed corn separation from barley grain beef. Barley grain-fed beef was mainly linked to alkanes and beef positive FI, whereas corn grain-fed beef was associated with pyrazines, in addition to aldehydes related to n-6 LCFA oxidation.

Randomized Placebo-Controlled Trial of Ferric Carboxymaltose in Heart Failure With Iron Deficiency: Rationale and Design.

BACKGROUND: Iron deficiency (ID) has a prevalence of ≈40% to 50% among patients in heart failure (HF) with reduced ejection fraction and is associated with worse prognosis. Several trials demonstrated that intravenous ferric carboxymaltose leads to early and sustained improvement in patient-reported outcomes and functional capacity in patients with HF with reduced ejection fraction with ID, yet morbidity and mortality data are limited. METHODS: The objective of the HEART-FID trial (Ferric Carboxymaltose in Heart Failure With Iron Deficiency) is to assess efficacy and safety of ferric carboxymaltose compared with placebo as treatment for symptomatic HF with reduced ejection fraction with ID. HEART-FID is a multicenter, randomized, double-blind, placebo-controlled trial enrolling ≈3014 patients at ≈300 international centers. Eligible patients are aged ≥18 years in stable chronic HF with New York Heart Association functional class II to IV symptoms, ejection fraction ≤40%, ID (ferritin <100 ng/mL or ferritin 100-300 ng/mL with a transferrin saturation <20%), and documented HF hospitalization or elevated N-terminal pro-brain natriuretic peptide. Consented patients are assigned to ferric carboxymaltose or placebo at baseline, with repeated visits/assessments every 6 months for additional study drug based on hemoglobin and iron indices for the trial duration. The primary end point is a hierarchical composite of death and HF hospitalization at 12 months and change from baseline to 6 months in the 6-minute walk test distance. CONCLUSIONS: The HEART-FID trial will inform clinical practice by clarifying the role of long-term treatment with intravenous ferric carboxymaltose, added to usual care, in ambulatory patients with symptomatic HF with reduced ejection fraction with ID. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT03037931.

Transportation of peripheral blood progenitor cell products: effect of ambient temperature.

BACKGROUND AIMS: Peripheral blood progenitor cell (PBPC) products are often transported at high cell concentrations (>200 × 109/L) over long distances, requiring >36 h transport time. METHODS: Fresh PBPC samples from eight healthy donors were studied with two viability assays for effects of temperature outside the transport container (ambient temperature). The Coleman 5272 container, routinely used by the National Marrow Donor Program (NMDP) with two -20°C gel packs, was compared with the Coleman 6216 container, which can hold four -20°C gel packs. RESULTS: The temperature inside the smaller transport container (5272) proved to be sensitive to ambient temperature, whereas the larger container (6216) was less sensitive. The viability of CD34(+) cells, and the survival of granulocyte-macrophage colony-forming units (GM-CFU), was more dependent on the ambient temperature for the smaller than for the larger container. CONCLUSIONS: PBPC products are most often transported at approximately 2-8°C. The inside temperature of the container currently used by the NMDP appears to be more sensitive to increases in temperature when exposed to higher ambient temperature for prolonged periods of time. Increasing the number of gel packs from two to four improves the stability of the temperature inside the container but would require a different container.