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Head of Institute: Prof. Oren Froy

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Publications

2019
Achmon, Y. ; Dowdy, F. R. ; Simmons, C. W. ; Zohar-Perez, C. ; Rabinovitz, Z. ; Nussinovitch, A. Degradation and bioavailability of dried alginate hydrocolloid capsules in simulated soil system. Journal of Applied Polymer Science 2019, 136. Publisher's VersionAbstract
Hydrocolloid capsules are common chemical carriers used in many types of applications in foods, biotechnology, and agriculture. Alginate microbeads and macrobeads are some of the more prevalent types of hydrocolloid capsules. Most studies to date have focused on alginate carrier applications but only a few have looked at their bioavailability after use. In this study, alginate carriers were subjected to simulated field conditions and their biodegradation in the soil was evaluated by respiration measurements, visualization, and volatile solids reduction. Using respiration rate, the degradation rate was calculated at 32 ± 3.1% (w/w) after 2 months. The visually estimated volume and volatile solids reduction gave degradation rates of 40 ± 8.6% (v/v) and 22.5 ± 2.5% (w/w), respectively. Moreover, water-loss calculations suggested that the carriers can serve as a stand-alone soil amendment for water retention. These findings emphasize the importance of studying hydrocolloid bioavailability in the soil and alginate carrier suitability for future applications. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 48142. © 2019 Wiley Periodicals, Inc.
Blaychfeld-Magnazi, M. ; Reshef, N. ; Zornitzki, T. ; Madar, Z. ; Knobler, H. The effect of a low-carbohydrate high-fat diet and ethnicity on daily glucose profile in type 2 diabetes determined by continuous glucose monitoring. European Journal of Nutrition 2019. Publisher's VersionAbstract
{Background and aims: Nutrition is an integral part of type 2 diabetes (T2DM) treatment, but the optimal macronutrient composition is still debated and previous studies have not addressed the role of ethnicity in dietary response. The current study aims were to compare the effect of short-term glycemic response to low-carbohydrate high-fat (LC-HF) diet vs. high-carbohydrate low-fat (HC-LF) diet using continuous glucose monitoring (CGM) and to evaluate the response of individuals with T2DM of Yemenite (Y-DM) and non-Yemenite origin (NY-DM). Methods: Twenty T2DM males, ten Y-DM and ten NY-DM underwent meal tolerance test and indexes of insulin resistance and secretion were calculated. Subsequently, patients were connected to CGM to assess daily glycemic control and glucose variability in response to isocaloric HC-LF or LC-HF diet, receiving each diet for 2 days by providing prepared meals. Daily glucose levels, area under the glucose curve (G-AUC) and parameters of glucose variability [standard deviation (SD), mean amplitude of glycemic excursions (MAGE) and mean absolute glucose (MAG)] were evaluated. Results: The LC-HF resulted in a significantly lower G-AUC (p < 0.001) and in lower variability parameters (p < 0.001) vs. the HC-LF diet. However, Y-DM showed less reduction in glucose variability indices upon diet-switching vs. NY-DM; MAGE decreased, respectively, by 69% vs. 89%
Nakonechny, F. ; Avisar, D. ; Ludmer, Z. ; Brauner, N. ; Ullmann, A. Application of Partially Miscible Solvent System for an Efficient Extraction of Organic Pollutants from Contaminated Sludge. Water, Air, and Soil Pollution 2019, 230. Publisher's VersionAbstract
A systematic study of extraction of various organic pollutants from highly contaminated solid media (e.g., sludge) by applying a phase transition of a benign partially miscible solvent system composed of water/ethyl acetate/ethanol was conducted. This solvent system possesses an upper critical solution temperature of about 55 °C. The efficiency of the phase transition extraction (PTE) process is found to be higher and much faster compared to those obtained without phase transition (at ambient temperature). The influence of various operating conditions on the process efficiency was investigated. The performance of the phase transition extraction when applied on contaminated sludge is much better than the extraction with ethyl acetate only, although the latter is shown to be a very efficient solvent for extracting various organic contaminants (e.g., pharmaceuticals, persistent organic pollutants) from aqueous solutions. The efficiency of phase transition extraction from the aqueous solution was somewhat lower than that achieved with ethyl acetate, but it shows a clear advantage in the presence of detergents, as emulsion formation is prevented. © 2019, Springer Nature Switzerland AG.
Dolev, N. ; Katz, Z. ; Ludmer, Z. ; Ullmann, A. ; Brauner, N. ; Goikhman, R. New insights into chelator recycling by a chelating resin: From molecular mechanisms to applicability. Chemosphere 2019, 215, 800-806. Publisher's VersionAbstract
As part of the project of developing a “green” and highly feasible soil remediation process, recycling an eco-friendly chelating agent, glycine, using Chelex-100 chelating resin, was studied. Two model complexes, copper and nickel glycinates, were tested under various conditions, including equivalent viscosity but different temperature conditions. Two similar complexes demonstrated very different reactivity towards Chelex-100. An in-depth study led to the discovery of unusual metal-dependent mechanisms of the complex-to-resin metal transfer. Particularly, nickel transfer proceeds via a dissociative mechanism, whereas copper transfer does not require pre-dissociation of the complexes, and proceeds via the associative ligand-exchange mechanism. Both processes result in the recovery of the used chelator. The glycine solution was applied on the spiked soil, then recovered on Chelex-100 resin and successfully reused, thus demonstrating a proof of the concept. These findings contribute to the science, strategies, and methodology of both water purification and chelator recycling fields. © 2018 Elsevier Ltd
Gertler, A. ; Solomon, G. Pegylated Human Leptin D23L Mutant - Preparation and biological activity in vitro and in vivo in male ob /ob mice. Endocrinology 2019, 160, 891-898. Publisher's VersionAbstract
Recombinant monomeric human leptin (hLEP) and its D23L mutant were prepared in Escherichia coli and pegylated at their N-terminus using 20-kDa methoxy pegylated (PEG)-propionylaldehyde. As determined by both SDS-PAGE and size-exclusion chromatography, the pegylated proteins consisted of >90% monopegylated and <10% double-pegylated species. Circular dichroism spectra showed that their secondary structure, characteristic of all four α-helix bundle cytokines, was not affected by either the D23L mutation or pegylation. Because of the D23L mutation, affinity for hLEP receptor increased 25- and 40-fold for the pegylated and nonpegylated mutant, respectively. However, whereas the proliferation-promoting activity in vitro of nonmutated and mutated nonpegylated hLEP was identical, that of the respective pegylated mutant was approximately sixfold higher compared with the pegylated nonmutated hLEP. This difference was also seen in vivo. Both pegylated hLEPs at all doses significantly decreased body weight and food consumption, as compared with the vehicle-treated control. Once-daily administration of pegylated hLEP D23L at doses of 0.1, 0.3, and 1 mg/kg for 14 consecutive days in ob/ob mice resulted in significantly decreased body weight and food consumption as compared with respective pegylated hLEP-treated animals, with the biggest difference observed at 0.1 mg/kg. Repeated administration of either pegylated hLEP D23L or pegylated hLEP significantly decreased blood glucose levels compared with the control before glucose challenge and after oral glucose tolerance test, but with no difference between the two treatments. The pegylated hLEP D23L mutant seems to be a more potent reagent suitable for in vivo studies than the pegylated nonmutated hLEP. © Copyright 2019 Endocrine Society.
Ramos-Nieves, J. M. ; Giesy, S. L. ; Schwark, W. S. ; Gertler, A. ; Boisclair, Y. R. Technical note: Effects of pegylation and route of administration on leptin kinetics in newborn lambs. Journal of Animal Science 2019, 97, 3768-3775. Publisher's VersionAbstract
Chronic energy insufficiency resulting from inadequate feed intake or increased nutrient demand reduces plasma leptin in ruminants. Treatment of energy-deficient ruminants with exogenous leptin has identified some physiological consequences of reduced plasma leptin, but their full complement remains unknown. Additional leptin-dependent responses could be identified by using strategies that interfere with leptin signaling such as administration of leptin mutants that act as competitive antagonists. The effectiveness of these antagonists depends on their fold excess over endogenous leptin, and this condition can be achieved under in vivo conditions by extending the half-life (t1/2) of the antagonist by addition of a polyethylene glycol (PEG) molecule (pegylation). Use of this approach in ruminants, however, is limited by the lack of information on the t1/2 of native and pegylated leptin and on the most effective route of administration. To answer these questions, newborn lambs (n = 3) were injected with an intravenous (i.v.) bolus of 150 µg of human leptin followed by blood sampling over the next 12 h. Analysis of the semilog plasma leptin concentration over time yielded a t1/2 of 43 ± 4.9 min; an i.v. bolus of 276 µg of bovine leptin yielded a comparable t1/2 (P > 0.05). Next, newborn lambs (n = 4) received a single dose of 229 μg/ kg of metabolic body weight (BW0.75) of pegylated super human leptin antagonist (PEG-SHLA) via the i.v. or subcutaneous (s.c.) route. Plasma PEG-SHLA concentration reached a peak of 1,528 ± 78 ng/mL after 1 min and a nadir of 71 ± 9 ng/mL after 24 h with the i.v. route versus a peak of 423 ± 43 ng/mL after 300 min and a nadir of 146 ± 22 ng/mL after 24 h for the s.c. route; the t1/2 of PEG-SHLA was 394 ± 29 min for the i.v. route and 433 ± 58 min for the s.c. route. Finally, plasma concentration of PEG-SHLA was modeled when given either i.v. or s.c. at a dose of 229 μg/kg BW0.75 every 12 h. Once a steady state was reached, peak and lowest concentrations PEG-SHLA over the 12-h windows were 2,269 and 403 ng/mL for the i.v. route and 814 and 555 ng/mL for the s.c. route. Weighted PEG-SHLA concentrations over the 12-h period were 1,455 and 713 ng/mL for the i.v. and s.c. route, translating into 364- and 178-fold excess over endogenous plasma leptin. These data confirm the effectiveness of pegylation in extending the t1/2 of leptin antagonists in newborn lambs and in increasing their circulation in fold excess over endogenous leptin. © The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved.
Kohay, H. ; Bilkis, I. ; Mishael, Y. Effect of polycation charge density on polymer conformation at the clay surface and consequently on pharmaceutical binding. Journal of Colloid and Interface Science 2019, 552, 517-527. Publisher's VersionAbstract
Polycation conformation upon adsorption to a negatively charged surface can be modulated by its charge density. At high charge density monomers directly interact with the surface in a ‘trains’ conformation and as charge density decreases a high degree of monomers dangle into solution in a ‘loops and tails’ conformation. In this study, the conformations of polycations upon adsorption to montmorillonite, as a function of polycation charge (20, 50 and 100% of the monomers, denoted as P-Q20, P-Q50 and P-Q100) were characterized and in accordance with their conformation, the adsorption of non-ionic and anionic molecules by the composite was tested. As expected, the adsorption of the nonionic pharmaceuticals increased to a composite with a ‘loops and tails’ conformation, due to both conformation and chemical properties. On the other hand, the anionic molecules, gemfibrozil and diclofenac, preferably adsorbed to composites with higher charge density (Q50 or Q100 composites). However, they showed different tendency toward the composites, i.e. higher adsorption of diclofenac by Q100 composite vs. higher adsorption of gemfibrozil by Q50 composite. To elucidate the differences in adsorption between these two pharmaceuticals, density functional theory calculations were employed. Both gemfibrozil and diclofenac were found to be better stabilized by methyl pyridinium sites (prevail in Q100 composite). The preferable adsorption of gemfibrozil by Q50 composite was explained by the presence of ‘loops and tails’ conformation enabling additional adsorption sites and diverse monomer-target molecule orientations. © 2019
Thawabteh, A. ; Lelario, F. ; Scrano, L. ; Bufo, S. A. ; Nowak, S. ; Behrens, M. ; Di Pizio, A. ; Niv, M. Y. ; Karaman, R. Bitterless guaifenesin prodrugs—design, synthesis, characterization, in vitro kinetics, and bitterness studies. Chemical Biology & Drug Design 2019, 93, 262-271. Publisher's VersionAbstract
Abstract A respected number of drugs suffer from bitter taste which results in patient incompliance. With the aim of solving the bitterness of guaifenesin, dimethyl maleate, maleate, glutarate, succinate, and dimethyl succinate prodrugs were designed and synthesized. Molecular orbital methods were utilized for the design of the ester prodrugs. The density functional theory (DFT) calculations revealed that the hydrolysis efficiency of the synthesized prodrugs is significantly sensitive to the pattern of substitution on C=C bond and distance between the nucleophile and the electrophile. The hydrolysis of the prodrugs was largely affected by the pH of the medium. The experimental t1/2 for the hydrolysis of guaifenesin dimaleate ester prodrugs in 1N HCl was the least and for guaifenesin dimethyl succinate was the highest. Functional heterologous expression of TAS2R14, a broadly tuned bitter taste receptor responding to guaifenesin, and experiments using these prodrugs revealed that, while some of the prodrugs still activated the receptor similarly or even stronger than the parent substance, succinate derivatization resulted in the complete loss of receptor responses. The predicted binding modes of guaifenesin and its prodrugs to the TAS2R14 homology model suggest that the decreased activity of the succinate derivatives may be caused by a clash with Phe247.
Di Pizio, A. ; Ben Shoshan-Galeczki, Y. ; Hayes, J. E. ; Niv, M. Y. Bitter and sweet tasting molecules: It's complicated. Neuroscience Letters 2019, 700, 56 - 63. Publisher's VersionAbstract
“Bitter” and “sweet” are frequently framed in opposition, both functionally and metaphorically, in regard to affective responses, emotion, and nutrition. This oppositional relationship is complicated by the fact that some molecules are simultaneously bitter and sweet. In some cases, a small chemical modification, or a chirality switch, flips the taste from sweet to bitter. Molecules humans describe as bitter are recognized by a 25-member subfamily of class A G-protein coupled receptors (GPCRs) known as TAS2Rs. Molecules humans describe as sweet are recognized by a TAS1R2/TAS1R3 heterodimer of class C GPCRs. Here we characterize the chemical space of bitter and sweet molecules: the majority of bitter compounds show higher hydrophobicity compared to sweet compounds, while sweet molecules have a wider range of sizes. Importantly, recent evidence indicates that TAS1Rs and TAS2Rs are not limited to the oral cavity; moreover, some bitterants are pharmacologically promiscuous, with the hERG potassium channel, cytochrome P450 enzymes, and carbonic anhydrases as common off-targets. Further focus on polypharmacology may unravel new physiological roles for tastant molecules.
Kolitsida, P. ; Zhou, J. ; Rackiewicz, M. ; Nolic, V. ; Dengjel, J. ; Abeliovich, H. Phosphorylation of mitochondrial matrix proteins regulates their selective mitophagic degradation. Proceedings of the National Academy of Sciences 2019. Publisher's VersionAbstract
Mitochondrial dysfunction underlies many age-related human pathologies. In normal cells, defective mitochondria are often degraded by mitophagy, a process in which these mitochondria are engulfed in autophagosomes and sent for degradation in the lysosome/vacuole. Surprisingly, studies on mitophagy in diverse eukaryotic organisms reveal an unexpected dimension of protein-level selectivity, wherein individual protein species exhibit divergent rates of mitophagic degradation. In this report, we show that this surprising intramitochondrial selectivity can be generated by differential phosphorylation of individual mitochondrial protein species, and we identify mitochondrial phosphatases and kinases that contribute to this regulation. By identifying a mechanism that regulates the intramitochondrial selectivity of mitophagic degradation, our findings open the door to potential manipulation of the quality-control process in the future.Mitophagy is an important quality-control mechanism in eukaryotic cells, and defects in mitophagy correlate with aging phenomena and neurodegenerative disorders. It is known that different mitochondrial matrix proteins undergo mitophagy with very different rates but, to date, the mechanism underlying this selectivity at the individual protein level has remained obscure. We now present evidence indicating that protein phosphorylation within the mitochondrial matrix plays a mechanistic role in regulating selective mitophagic degradation in yeast via involvement of the Aup1 mitochondrial protein phosphatase, as well as 2 known matrix-localized protein kinases, Pkp1 and Pkp2. By focusing on a specific matrix phosphoprotein reporter, we also demonstrate that phospho-mimetic and nonphosphorylatable point mutations at known phosphosites in the reporter increased or decreased its tendency to undergo mitophagy. Finally, we show that phosphorylation of the reporter protein is dynamically regulated during mitophagy in an Aup1-dependent manner. Our results indicate that structural determinants on a mitochondrial matrix protein can govern its mitophagic fate, and that protein phosphorylation regulates these determinants.
Wang, S. ; Duan, Y. ; Yan, Y. ; Adar, C. ; Braslavsky, I. ; Chen, B. ; Huang, T. ; Qiu, S. ; Li, X. ; Inglis, B. M. ; et al. Improvement of sperm cryo-survival of cynomolgus macaque (Macaca fascicularis) by commercial egg-yolk–free freezing medium with type III antifreeze protein. Animal Reproduction Science 2019, 210, 106177. Publisher's VersionAbstract
When nonhuman primate sperm undergoes cryopreservation in an egg yolk medium there is an increased risk that the egg yolk might adversely affect the sperm due to containing of avian pathogens. Although commercial egg-yolk-free medium for human sperm cryopreservation has been used for macaque sperm, the cryo-survival remains less than optimal. The present study, therefore, was conducted to determine the optimal concentration of antifreeze protein (AFP) III supplemented in a commercial egg-yolk-free medium for cynomolgus macaque (Macaca fascicularis) sperm cryo-survival. The function of frozen-thawed sperm was evaluated by post-thaw sperm motility, acrosome integrity, and mitochondrial function. Results indicate that the sperm motilities were greater when 0.1, 1, and 10 μg/ml of AFP III were supplemented into the sperm freezing medium (P < 0.05). In addition, the mitochondrial membrane potential was greater in the sperm cryopreserved with the medium that was supplemented with 0.1 μg/ml of AFP III (P < 0.05). The addition of AFP III at any of the concentrations, however, did not have any cryoprotection effect on the sperm acrosome, and the greatest concentrations of AFP III at 100 and 200 μg/ml had detrimental effects on acrosomal integrity (P < 0.05). Results of the present study indicated the methods used are effective for the cryopreservation of cynomolgus monkey sperm while reducing associated health risks due to avian pathogens being present in egg yolk-based extenders.
Yehuda, A. ; Slamti, L. ; Malach, E. ; Lereclus, D. ; Hayouka, Z. Elucidating the Hot Spot Residues of Quorum Sensing Peptidic Autoinducer PapR by Multiple Amino Acid Replacements. Front Microbiol 2019, 10, 1246.Abstract
The quorum sensing (QS) system of , an opportunistic human pathogen, utilizes the autoinducing PapR peptide signal that mediates the activation of the pleiotropic virulence regulator PlcR. A set of synthetic 7-mer PapR-derived peptides (PapR; ADLPFEF) have been shown to inhibit efficiently the PlcR regulon activity and the production of virulence factors, reflected by a loss in hemolytic activity without affecting bacterial growth. Interestingly, these first potent synthetic inhibitors involved D-amino acid or alanine replacements of three amino acids; proline, glutamic acid, and phenylalanine of the heptapeptide PapR. To better understand the role of these three positions in PlcR activity, we report herein the second generation design, synthesis, and characterization of PapR-derived combinations, alternate double and triple alanine and D-amino acids replacement at these positions. Our findings generate a new set of non-native PapR-derived peptides that inhibit the PlcR regulon activity and the production of virulence factors. Using the amino acids substitution strategy, we revealed the role of proline and glutamic acid on PlcR regulon activation. Moreover, we demonstrated that the D-Glutamic acid substitution was crucial for the design of stronger PlcR antagonists. These peptides represent potent synthetic inhibitors of QS and constitute new and readily accessible chemical tools for the study of the PlcR system. Our method might be applied to other quorum sensing systems to design new anti-virulence agents.
Amso, Z. ; Hayouka, Z. Antimicrobial random peptide cocktails: a new approach to fight pathogenic bacteria. Chem Commun (Camb) 2019, 55, 2007-2014.Abstract
Antibiotic resistance in bacteria has become a serious threat to public health, and therefore there is an urgent need to develop new classes of antimicrobial agents. Nowadays, natural antimicrobial peptides (AMPs) and their synthetic derivatives are considered as promising alternatives to traditional antibiotics. The broad molecular diversity of AMPs, in terms of sequences and structures, suggests that their activity does not depend on specific features of amino acid sequence or peptide conformation. We therefore selected two common properties of AMPs, (high percentage of hydrophobic and cationic amino acids), to develop a novel approach to synthesize random antimicrobial peptide mixtures (RPMs). Instead of incorporating a single amino acid at each coupling step, a mixture of hydrophobic and cationic amino acids in a defined proportion is coupled. This results in a mixture that contains up to 2n sequences, where n is the number of the coupling step, of random peptides with a defined composition, stereochemistry, and controlled chain length. We have discovered that RPMs of hydrophobic and cationic α-amino acids, such as phenylalanine and lysine, display strong and broad antimicrobial activity towards Gram-negative, Gram-positive, clinically isolated antibiotic resistant "superbugs", and several plant pathogenic bacteria. This review summarizes our efforts to explore the mode of action of RPMs and their potential as bioactive agents for multiple applications, including the prevention of biofilm formation and degradation of mature biofilm (related to human health), reduction of disease severity in plant bacterial disease models (related to crop protection), and inhibition of bacterial growth in milk (related to food preservation). All our findings illustrate the effectiveness of RPMs and their great potential for various applications.
Stern Bauer, T. ; Menagen, B. ; Avnir, D. ; Hayouka, Z. Random peptide mixtures entrapped within a copper-cuprite matrix: new antimicrobial agent against methicillin-resistant Staphylococcus aureus. Sci Rep 2019, 9 11215.Abstract
The emergence of global antibiotic resistance necessitates the urgent need to develop new and effective antimicrobial agents. Combination of two antimicrobial agents can potentially improve antimicrobial potency and mitigate the development of resistance. Therefore, we have utilized metal molecular doping methodology whereby antimicrobial random peptides mixture (RPMs) are entrapped in a bactericidal copper metal matrix. The copper/RPM composite exhibits greater antimicrobial activity toward methicillin-resistant Staphylococcus aureus (MRSA) than either copper or RPMs alone. Our findings indicate that this bactericidal antimicrobial biomaterial could be utilized to efficiently eradicate antibiotic-resistant pathogenic bacteria for health, agricultural and environmental applications.
Croote, D. ; Braslavsky, I. ; Quake, S. R. Addressing Complex Matrix Interference Improves Multiplex Food Allergen Detection by Targeted LC–MS/MS. Analytical ChemistryAnalytical Chemistry 2019. Publisher's VersionAbstract
The frequent use of precautionary food allergen labeling (PAL) such as “may contain” frustrates allergic individuals who rely on such labeling to determine whether a food is safe to consume. One technique to study whether foods contain allergens is targeted liquid chromatography-tandem mass spectrometry (LC–MS/MS) employing scheduled multiple reaction monitoring (MRM). However, the applicability of a single MRM method to many commercial foods is unknown as complex and heterogeneous interferences derived from the unique composition of each food matrix can hinder quantification of trace amounts of allergen contamination. We developed a freely available, open source software package MAtrix-Dependent Interference Correction (MADIC) to identify interference and applied it with a method targeting 14 allergens. Among 84 unique food products, we found patterns of allergen contamination such as wheat in grains, milk in chocolate-containing products, and soy in breads and corn flours. We also found additional instances of contamination in products with and without PAL as well as highly variable soy content in foods containing only soybean oil and/or soy lecithin. These results demonstrate the feasibility of applying LC–MS/MS to a variety of food products with sensitive detection of multiple allergens in spite of variable matrix interference.The frequent use of precautionary food allergen labeling (PAL) such as “may contain” frustrates allergic individuals who rely on such labeling to determine whether a food is safe to consume. One technique to study whether foods contain allergens is targeted liquid chromatography-tandem mass spectrometry (LC–MS/MS) employing scheduled multiple reaction monitoring (MRM). However, the applicability of a single MRM method to many commercial foods is unknown as complex and heterogeneous interferences derived from the unique composition of each food matrix can hinder quantification of trace amounts of allergen contamination. We developed a freely available, open source software package MAtrix-Dependent Interference Correction (MADIC) to identify interference and applied it with a method targeting 14 allergens. Among 84 unique food products, we found patterns of allergen contamination such as wheat in grains, milk in chocolate-containing products, and soy in breads and corn flours. We also found additional instances of contamination in products with and without PAL as well as highly variable soy content in foods containing only soybean oil and/or soy lecithin. These results demonstrate the feasibility of applying LC–MS/MS to a variety of food products with sensitive detection of multiple allergens in spite of variable matrix interference.
Abu Ahmad, N. ; Raizman, M. ; Weizmann, N. ; Wasek, B. ; Arning, E. ; Bottiglieri, T. ; Tirosh, O. ; Troen, A. M. Betaine attenuates pathology by stimulating lipid oxidation in liver and regulating phospholipid metabolism in brain of methionine-choline–deficient rats. The FASEB Journal 2019, fj.201802683R. Publisher's VersionAbstract
Methyl-donor deficiency is a risk factor for neurodegenerative diseases. Dietary deficiency of the methyl-donors methionine and choline [methionine-choline?deficient (MCD) diet] is a well-established model of nonalcoholic steatohepatitis (NASH), yet brain metabolism has not been studied in this model. We hypothesized that supplemental betaine would protect both the liver and brain in this model and that any benefit to the brain would be due to improved liver metabolism because betaine is a methyl-donor in liver methylation but is not metabolically active in the brain. We fed male Sprague-Dawley rats a control diet, MCD diet, or betaine-supplemented MCD (MCD+B) diet for 8 wk and collected blood and tissue. As expected, betaine prevented MCD diet?induced NASH. However, contrary to our prediction, it did not appear to do so by stimulating methylation; the MCD+B diet worsened hyperhomocysteinemia and depressed liver methylation potential 8-fold compared with the MCD diet. Instead, it significantly increased the expression of genes involved in ?-oxidation: fibroblast growth factor 21 and peroxisome proliferator?activated receptor α. In contrast to that of the liver, brain methylation potential was unaffected by diet. Nevertheless, several phospholipid (PL) subclasses involved in stabilizing brain membranes were decreased by the MCD diet, and these improved modestly with betaine. The protective effect of betaine is likely due to the stimulation of ?-oxidation in liver and the effects on PL metabolism in brain.?Abu Ahmad, N., Raizman, M., Weizmann, N., Wasek, B., Arning, E., Bottiglieri, T., Tirosh, O., Troen, A. M. Betaine attenuates pathology by stimulating lipid oxidation in liver and regulating phospholipid metabolism in brain of methionine-choline?deficient rats.Methyl-donor deficiency is a risk factor for neurodegenerative diseases. Dietary deficiency of the methyl-donors methionine and choline [methionine-choline?deficient (MCD) diet] is a well-established model of nonalcoholic steatohepatitis (NASH), yet brain metabolism has not been studied in this model. We hypothesized that supplemental betaine would protect both the liver and brain in this model and that any benefit to the brain would be due to improved liver metabolism because betaine is a methyl-donor in liver methylation but is not metabolically active in the brain. We fed male Sprague-Dawley rats a control diet, MCD diet, or betaine-supplemented MCD (MCD+B) diet for 8 wk and collected blood and tissue. As expected, betaine prevented MCD diet?induced NASH. However, contrary to our prediction, it did not appear to do so by stimulating methylation; the MCD+B diet worsened hyperhomocysteinemia and depressed liver methylation potential 8-fold compared with the MCD diet. Instead, it significantly increased the expression of genes involved in ?-oxidation: fibroblast growth factor 21 and peroxisome proliferator?activated receptor α. In contrast to that of the liver, brain methylation potential was unaffected by diet. Nevertheless, several phospholipid (PL) subclasses involved in stabilizing brain membranes were decreased by the MCD diet, and these improved modestly with betaine. The protective effect of betaine is likely due to the stimulation of ?-oxidation in liver and the effects on PL metabolism in brain.?Abu Ahmad, N., Raizman, M., Weizmann, N., Wasek, B., Arning, E., Bottiglieri, T., Tirosh, O., Troen, A. M. Betaine attenuates pathology by stimulating lipid oxidation in liver and regulating phospholipid metabolism in brain of methionine-choline?deficient rats.
Berkovich, B. - E. ; Stark, A. H. ; Eliakim, A. ; Nemet, D. ; Sinai, T. Rapid Weight Loss in Competitive Judo and Taekwondo Athletes: Attitudes and Practices of Coaches and Trainers. International Journal of Sport Nutrition and Exercise Metabolism 2019, 1 - 7. Publisher's VersionAbstract
Fasting, skipping meals, and dehydration are common methods of rapid weight loss used prior to competition in weight category sports. This study examines coaches? attitudes, perceptions, and practices regarding rapid weight loss among judo and taekwondo athletes. A convenience sample of experienced coaches and trainers (n?=?68) completed structured questionnaires. Participants in this study were 33.8?±?9.3 years old; 57 were males and 11 were females; and 59% were certified coaches, with 71% reporting over 20 years of involvement in sports and 68% having more than 10 years of teaching experience. The majority (90%) reported that they usually supervised athletes through the weight loss process. Interventions for weight loss began at 12.7?±?1.9 years of age, with a recommended precompetition weight loss duration of 16.2?±?8.2 days and an average reduction of 1.5?±?0.7 kg. The majority of the responders (92%) recommended that their athletes practice gradual weight loss methods using a combination of dehydration or increased physical activity (80.3%), sweat suits (50.8%), restricted fluid intake (39.3%), training in heated rooms (27%), and sauna (26.2%). Recommendations of spitting (27.8%) or using laxatives, diuretics, diet pills, or vomiting (21.3%) were also reported. Coaches and trainers often encouraged athletes to cut weight before competition. The methods recommended are potentially harmful with severe health risks, including compromised nutritional status and diminished athletic performance. This is of particular concern in young athletes who are still growing and developing physically. Enhancing knowledge and awareness for coaches, athletes, and parents regarding potential dangers, along with improved nutrition education, is critical for reducing the magnitude and misuse of rapid weight loss methods.Fasting, skipping meals, and dehydration are common methods of rapid weight loss used prior to competition in weight category sports. This study examines coaches? attitudes, perceptions, and practices regarding rapid weight loss among judo and taekwondo athletes. A convenience sample of experienced coaches and trainers (n?=?68) completed structured questionnaires. Participants in this study were 33.8?±?9.3 years old; 57 were males and 11 were females; and 59% were certified coaches, with 71% reporting over 20 years of involvement in sports and 68% having more than 10 years of teaching experience. The majority (90%) reported that they usually supervised athletes through the weight loss process. Interventions for weight loss began at 12.7?±?1.9 years of age, with a recommended precompetition weight loss duration of 16.2?±?8.2 days and an average reduction of 1.5?±?0.7 kg. The majority of the responders (92%) recommended that their athletes practice gradual weight loss methods using a combination of dehydration or increased physical activity (80.3%), sweat suits (50.8%), restricted fluid intake (39.3%), training in heated rooms (27%), and sauna (26.2%). Recommendations of spitting (27.8%) or using laxatives, diuretics, diet pills, or vomiting (21.3%) were also reported. Coaches and trainers often encouraged athletes to cut weight before competition. The methods recommended are potentially harmful with severe health risks, including compromised nutritional status and diminished athletic performance. This is of particular concern in young athletes who are still growing and developing physically. Enhancing knowledge and awareness for coaches, athletes, and parents regarding potential dangers, along with improved nutrition education, is critical for reducing the magnitude and misuse of rapid weight loss methods.
Shiff, Y. E. ; Reif, S. ; Marom, R. ; Shiff, K. ; Reifen, R. ; Golan-Gerstl, R. MiRNA-320a is less expressed and miRNA-148a more expressed in preterm human milk compared to term human milk. Journal of Functional Foods 2019, 57, 68 - 74. Publisher's VersionAbstract
ObjectivesTo investigate whether there is a difference in the profile of miRNAs between human milk (HM) from mothers of preterm versus HM from mothers of full-term infants. Second goal is to assess biological functions or implication related to those differences in miRNAs expression. Methods Four of the highly expressed miRNAs in milk were detected by qRT-PCR. Milk derived exosomes were incubated with cells. The expression of miRNAs and target gene were detected by qRT-PCR. Results MiRNA-320 was more highly expressed in the colostrum of fullterm than in preterm HM. The expression of MiRNA-148 was higher in preterm mother's milk than of full-term colostrum. MiRNA-320 and MIRNA-148a expression were upregulated in cells incubated with milk exosomes, which lead to a decrease in their target genes FASN1 and DNMT1 respectivilly. Conclusions Alterations in miRNAs expression in HM can affect biologic function in infants and may serve as a nutritional therapeutic target.
Curzon, A. Y. ; Chandrasekhar, K. ; Nashef, Y. K. ; Abbo, S. ; Bonfil, D. J. ; Reifen, R. ; Bar-el, S. ; Avneri, A. ; Ben-David, R. Distinguishing between Bread Wheat and Spelt Grains Using Molecular Markers and Spectroscopy. Journal of Agricultural and Food Chemistry 2019, 67, 3837 - 3841. Publisher's VersionAbstract
The increasing demand for spelt products requires the baking industry to develop accurate and efficient tools to differentiate between spelt and bread wheat grains. We subjected a 272-sample spelt-bread wheat set to several potential diagnostic methods. DNA markers for γ-gliadin-D (GAG56D), γ-gliadin-B (GAG56B), and the Q-gene were used, alongside phenotypic assessment of ease-of-threshing and near-infrared spectroscopy (NIRS). The GAG56B and GAG56D markers demonstrated low diagnostic power in comparison to the Q-gene genotyping, which showed full accordance with the threshing phenotype, providing a highly accurate distinction between bread wheat and spelt kernels. A highly reliable Q classification was based on a three-waveband NIR model [Kappa (0.97), R-square (0.93)], which suggested that this gene influences grain characteristics. Our data ruled out a protein concentration bias of the NIRS-based diagnosis. These findings highlight the Q gene and NIRS as important, valuable, but simple tools for distinguishing between bread wheat and spelt.The increasing demand for spelt products requires the baking industry to develop accurate and efficient tools to differentiate between spelt and bread wheat grains. We subjected a 272-sample spelt-bread wheat set to several potential diagnostic methods. DNA markers for γ-gliadin-D (GAG56D), γ-gliadin-B (GAG56B), and the Q-gene were used, alongside phenotypic assessment of ease-of-threshing and near-infrared spectroscopy (NIRS). The GAG56B and GAG56D markers demonstrated low diagnostic power in comparison to the Q-gene genotyping, which showed full accordance with the threshing phenotype, providing a highly accurate distinction between bread wheat and spelt kernels. A highly reliable Q classification was based on a three-waveband NIR model [Kappa (0.97), R-square (0.93)], which suggested that this gene influences grain characteristics. Our data ruled out a protein concentration bias of the NIRS-based diagnosis. These findings highlight the Q gene and NIRS as important, valuable, but simple tools for distinguishing between bread wheat and spelt.
Di Pizio, A. ; Waterloo, L. A. W. ; Brox, R. ; Löber, S. ; Weikert, D. ; Behrens, M. ; Gmeiner, P. ; Niv, M. Y. Rational design of agonists for bitter taste receptor TAS2R14: from modeling to bench and back. Cellular and Molecular Life Sciences 2019. Publisher's VersionAbstract
Human bitter taste receptors (TAS2Rs) are a subfamily of 25 G protein-coupled receptors that mediate bitter taste perception. TAS2R14 is the most broadly tuned bitter taste receptor, recognizing a range of chemically diverse agonists with micromolar-range potency. The receptor is expressed in several extra-oral tissues and is suggested to have physiological roles related to innate immune responses, male fertility, and cancer. Higher potency ligands are needed to investigate TAS2R14 function and to modulate it for future clinical applications. Here, a structure-based modeling approach is described for the design of TAS2R14 agonists beginning from flufenamic acid, an approved non-steroidal anti-inflammatory analgesic that activates TAS2R14 at sub-micromolar concentrations. Structure-based molecular modeling was integrated with experimental data to design new TAS2R14 agonists. Subsequent chemical synthesis and in vitro profiling resulted in new TAS2R14 agonists with improved potency compared to the lead. The integrated approach provides a validated and refined structural model of ligand–TAS2R14 interactions and a general framework for structure-based discovery in the absence of closely related experimental structures.