Concentrations Of Choline Chloride Research Articles

Choline chloride and amino acid solutions taste and hydration behavior with experimental thermodynamic properties and COSMO-PC-SAFT calculation

Aqueous amino acid solutions have been introduced as dietary supplements for both animals and humans. This study investigates the physicochemical properties of the solutions containing amino acids (l-glycine, d,l-alanine, l-proline), choline chloride, and water at temperature range of 288.15 to 318.15 K. The results show that increasing concentrations of amino acids and choline chloride lead to higher solution densities. Analysis of apparent molar volume (Vφ) and apparent molar isentropic compressibility (κφ) reveals that Vφ values increase with choline chloride concentration and temperature, indicating enhanced solute–solvent interactions, while κφ values decrease, suggesting increased solution compression. Thermodynamic analysis using the Redlich-Mayer model and COSMO-based modeling provides insights into molecular interactions. However, COSMO-based parameters show high average relative deviation percentage (ARD %) values, indicating poor predictive performance for the density of these systems. In contrast, the ePC-SAFT equation of state effectively predicts the densities, particularly for l-proline-based solutions, which show very low ARD % values, indicating high accuracy. The ePC-SAFT model also performs reasonably well for l-glycine solutions but shows poorer results for d,l-alanine-based solutions. The study also examines the sweetness and saltiness criteria (ASV and ASIC) of these solutions. The ASV values, which serve as a sweetness criterion, are higher than the ideal range of 0.5 < ASV < 0.7, suggesting an overly sweet taste. The ASIC values follow a similar trend, indicating increased saltiness. To achieve an appropriate grade of sweetness and saltiness, dilution to lower concentrations of the solution is recommended. Furthermore, the use of choline chloride is found to increase salt intake and enhance the taste of salt, which can be beneficial in amino acid supplements used in animal food.

Unraveling the interplay between choline chloride and collagen mimetic peptide with facile end modifications: A case study of Gly-Gly-(Pro-Hyp-Gly)8-Gly-Gly

Collagen mimetic peptides (CMPs) have garnered substantial interest in the field of regenerative medicine, tissue engineering, and biomaterials owing to their promising prospects for utilization within these domains. In the last few years, significant research efforts have been made to study how small CMPs can autonomously arrange themselves to create large scale collagen-like formations. These structures exhibit potential for advancing the field of next-generation biomaterials. In this investigation, we have synthesized a peptide named G-POG8, with the sequence Gly-Gly-(Pro-Hyp-Gly)8-Gly-Gly, and we have examined the behaviour of G-POG8 under different concentrations of choline chloride (ChCl) through various biophysical techniques. These methodologies include circular dichroism (CD), UV–vis, fluorescence and Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC) transmission electron microscopy (TEM), and dynamic light scattering (DLS). The findings indicate that the presence of Gly at both terminals of the CMP, G-POG8, results in a significant influence on the stability and aggregation behaviour of CMP triple helices. It is plausible that ChCl may have established favourable interactions with the terminal residues of G-POG8, potentially disrupting the end-to-end process of triple helix aggregation. This investigation lays the basis for the development of innovative biomaterials based on collagen, featuring improved properties, and opens up possibilities for cutting-edge biomedical applications.

Effect of source and amount of rumen-protected choline on hepatic metabolism during induction of fatty liver in dairy cows

Objectives were to determine the effect of supplementing increased amounts of rumen-protected choline (RPC) from sources with low (L, 28.8%) or high (H, 60.0%) concentration of choline chloride on hepatic metabolism when cows were subjected to feed restriction to develop fatty liver. It was hypothesized that increased supplementation of RPC reduces hepatic triacylglycerol and enhances glycogen concentrations. Pregnant, nonlactating multiparous Holstein cows (n = 110) at mean (± standard deviation) 232 ± 3.9 d of gestation were blocked by body condition (4.01 ± 0.52) and assigned to receive 0 (CON), 12.9 (L12.9 or H12.9), or 25.8 (L25.8 or H25.8) g/d of choline ion. Cows were fed for ad libitum intake on d 1 to 5 and restricted to 50% of the NEL required for maintenance and pregnancy from d 6 to 13. Intake of metabolizable methionine was maintained at 19 g/d during the feed restriction period by supplying rumen-protected methionine. Hepatic tissue was sampled on d 6 and 13 and analyzed for triacylglycerol, glycogen, and mRNA expression of genes involved in choline, glucose, and fatty acids metabolism, cell signaling, inflammation, autophagy, lipid droplet dynamics, lipophagy, and endoplasmic reticulum stress response. Blood was sampled and analyzed for concentrations of fatty acids, β-hydroxybutyrate (BHB), glucose, triacylglycerol, total cholesterol, and haptoglobin. Orthogonal contrasts evaluated the effect of supplementing RPC [CON vs. (1/4·L12.9 + 1/4·L25.8 + 1/4·H12.9 + 1/4·H25.8)], source of RPC [(1/2·L12.9 + 1/2·L25.8) vs. (1/2·H12.9 + 1/2·H25.8)], amount of RPC [(1/2·L12.9 + 1/2·H12.9) vs. (1/2·L25.8 + 1/2·H25.8)], and interaction between source and amount [(1/2·L12.9 + 1/2·H25.8) vs. (1/2·H12.9 + 1/2·L25.8)]. Least squares means and standard error of the means are presented in sequence as CON, L12.9, L25.8, H12.9, H25.8. Supplementation of RPC reduced hepatic triacylglycerol (9.3 vs. 6.6 vs. 5.1 vs. 6.6 vs. 6.0 ± 0.6% as-is) and increased glycogen contents (1.8 vs. 2.6 vs. 3.6 vs. 3.1 vs. 4.1 ± 0.2% as-is) on d 13 of the experiment. Feeding RPC reduced serum haptoglobin (136.6 vs. 85.6 vs. 80.6 vs. 82.8 vs. 81.2 ± 4.6 µg/mL) during the feed restriction period; however, blood concentrations of fatty acids, BHB, glucose, triacylglycerol, and total cholesterol did not differ among treatments. During feed restriction, supplementation of RPC enhanced the mRNA expression of genes related to choline metabolism (BHMT), uptake of fatty acids (CD36), and autophagy (ATG3), and reduced the expression of a transcript associated with endoplasmic reticulum stress response (ERN1). An increase in the amount of choline ion from 12.9 to 25.8 g/d enhanced the mRNA expression of genes associated with synthesis and assembly of lipoproteins (APOB100), and inflammation (TNFA), whereas it reduced the expression of genes linked to gluconeogenesis (PC), oxidation of fatty acids (ACADM, MMUT), ketogenesis (ACAT1), and synthesis of antioxidants (SOD1) on d 13 of the experiment. Feeding RPC, independent of the product used, promoted lipotropic effects that reduced hepatic lipidosis in dairy cows.

Choline supplementation regulates gut microbiome diversity, gut epithelial activity, and the cytokine gene expression in gilts.

Choline is an essential nutrient that is necessary for both fetal development and maintenance of neural function, while its effect on female ovarian development is largely unexplored. Our previous study demonstrated that choline supplementation promotes ovarian follicular development and ovulation, although its underlying mechanism was unclear. To uncover the potential regulation pathway, eighteen female Yorkshire × Landrace gilts were fed with either standard commercial diet (Control group, n = 9) or choline supplemented diet (Choline group, additional 500 mg/kg of control diet, n = 9) from day 90 of age to day 186. At day 186, feces samples were analyzed for effects on the gut microbiome using 16S ribosomal RNA gene V3-V4 region sequencing with Illumina MiSeq, serum samples were analyzed for trimethylamine (TMA) and trimethylamine-N-oxide (TMAO) using HILIC method, and jejunum tissues were analyzed for immune related gene expression using qRT-PCR. Our results show that choline supplementation did not alter the circulating level of TMA and TMAO (P > 0.05), but rather increased gut microbiome alpha diversity (P < 0.05). Beta diversity analysis results showed that the choline diet mainly increased the abundance of Firmicutes, Proteobacteria, and Actinobacteria, but decreased the abundance of Bacteroidetes, Spirochaetes, and Euryarchaeota at the phyla level. Meta-genomic analysis revealed that choline supplementation activated pathways in the gut microbiota associated with steroid hormone biosynthesis and degradation of infertility-causing environmental pollutants (bisphenol, xylene, and dioxins). To further verify the effect of choline on intestinal activity, a porcine intestine cell line (IPEC-J2) was treated with serial concentrations of choline chloride in vitro. Our data demonstrated that choline promoted the proliferation of IPEC-J2 while inhibiting the apoptotic activity. qRT-PCR results showed that choline significantly increased the expression level of Bcl2 in both IPEC-J2 cells and jejunum tissues. The expression of IL-22, a cytokine that has been shown to impact ovarian function, was increased by choline treatment in vitro. Our findings reveal the beneficial effect of choline supplementation on enhancing the gut microbiome composition and intestinal epithelial activity, and offer insights into how these changes may have contributed to the ovarian development-promoting effect we reported in our previous study.

Profiling the impact of choline chloride on the self-assembly of collagen mimetic peptide (Pro-Hyp-Gly)10

The promising applications of collagen in tissue engineering, regenerative medicine and biomaterials have intrigued great interest in development of collagen mimetic peptides (CMPs). In past decades, the large-scale collagen structures via the self-assembly of small CMPs have been extensively explored. Moreover, CMPs entail novel strategies in order to play a critical role in the discovery of new generation biomaterials. In this regard, we herein have synthesized a CMP, (Pro-Hyp-Gly)10 i.e. POG10. CMP has been examined in the presence of varying concentration of choline chloride (ChCl) by using various biophysical techniques such as circular dichroism (CD), UV–vis spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy (FTIR) spectroscopy, differential scanning calorimetry (DSC), transmission electron microscopy (TEM) and dynamic light scattering (DLS). For the first time, we have demonstrated the use of choline chloride (ChCl) to manipulate stability and aggregation of CMP triple helices in order to delineate the factors responsible for the stability of triple helices.

Enhanced Near‐Infrared Perovskite Light‐Emitting Devices by Introducing Choline Chloride Layer

AbstractThe existing interface defect states between perovskite and the transport layer is one of the dominant reasons to limit the performance of near‐infrared (NIR) perovskite light‐emitting devices (PeLEDs). Herein, a quaternary ammonium halide choline chloride layer is introduced to improve the wettability of the hole transport layer (HTL) and passivate defect states. As a consequence, the performance of the NIR PeLEDs is obviously enhanced. A maximum output power density of 4.7 mW cm−2 has been achieved by optimizing the concentration of choline chloride. Furthermore, the alternating current (AC) driving mode is introduced to reduce thermal degradation and charge accumulation in the interface region. It is demonstrated that the emission intensity as well as the stability of PeLEDs can be further improved.

Palmitate and pyruvate carbon flux in response to choline and methionine in bovine neonatal hepatocytes

Choline and methionine may serve unique functions to alter hepatic energy metabolism. Our objective was to trace carbon flux through pathways of oxidation and glucose metabolism in bovine hepatocytes exposed to increasing concentrations of choline chloride (CC) and d,l-methionine (DLM). Primary hepatocytes were isolated from 4 Holstein calves and maintained for 24 h before treatment with CC (0, 10, 100, 1000 μmol/L) and DLM (0, 100, 300 μmol/L) in a factorial design. After 21 h, [1-14C]C16:0 or [2-14C]pyruvate was added to measure complete and incomplete oxidation, and cellular glycogen. Reactive oxygen species (ROS), cellular triglyceride (TG), and glucose and ß-hydroxybutyrate (BHB) export were quantified. Exported very-low density lipoprotein particles were isolated for untargeted lipidomics and to quantify TG. Interactions between CC and DLM, and contrasts for CC (0 vs. [10, 100, 1000 μmol/L] and linear and quadratic contrast 10, 100, 1000 μmol/L) and DLM (0 vs. [100, 300 μmol/L] and 100 vs. 300 μmol/L) were evaluated. Presence of CC increased complete oxidation of [1-14C]C16:0 and decreased BHB export. Glucose export was decreased, but cellular glycogen was increased by the presence of CC and increasing CC. Presence of CC decreased ROS and marginally decreased cellular TG. No interactions between CC and DLM were detected for these outcomes. These data suggest a hepato-protective role for CC to limit ROS and cellular TG accumulation, and to alter hepatic energy metabolism to support complete oxidation of FA and glycogen storage regardless of Met supply.

Solubility of CO2 in Aqueous Solutions of Diethanolamine (DEA) and Choline Chloride

In this study, the solubility of CO2 in DEA in the presence and absence of choline chloride was reported at different temperatures of 276.15, 298.15, 313.15, and 333.15 K, and pressure range of 4-15 bar. The solubility of CO2 was evaluated using the pressure decay method in a batch isochoric stirred absorption cell. Also, the design of experiments performed with Qualitek-4 software using the Taguchi method. Henry’s law constants at three different temperatures were calculated from the correlation of solubility data. Results showed that increasing the initial pressure and DEA concentration and also decreasing the temperature, increases the solubility of CO2. Optimum operating conditions to maximize the amount of CO2 absorption including the temperature of 276.15 K (minimum level), the initial pressure of 15 bar (maximum level), the concentration of DEA of 40 wt.% (maximum level), and the concentration of choline chloride of 5 wt.% (middle level). Also, Qualitek-4 software predicted the amount of solubility at the optimum conditions which was 7.5% different from the measured value.

Urease Inhibitory Potential and Soil Ecotoxicity of Novel “Polyphenols–Deep Eutectic Solvents” Formulations

New formulations based on red grape pomace polyphenols and deep eutectic solvents (DES) have been here evaluated as inhibitors of urease of agricultural interest (jack bean urease, JBU). DES based on choline chloride (CHO) and betaine (BET), combined with ethylene glycol (EG), citric acid (CA), and urea (U), were used both as extracting and carrying agents for polyphenols, becoming active components of the formulations here obtained. Among the various DES combinations, U- and CA-based ones gave the best polyphenol extraction performances, 1.2–1.4-times higher than those of the hydroalcoholic mixture. Among the various DES–polyphenols formulations, the one composed by CHO–EG showed the best antioxidant potential and urease inhibition: 60–90% inhibition of the total JBU activity was achieved with a CHO concentration of 5–20 mM. Good results were also achieved with the BET–EG polyphenol formulation, which was able to inhibit ca. 50% of urease activity at 20 mM concentration of BET. Low phytotoxicity of DES a...

Glucose metabolism is differentially altered by choline and methionine in bovine neonatal hepatocytes.

Choline and methionine serve essential roles in the liver that may interact with glucose metabolism. Our objectives were to quantify glucose export, cellular glycogen, and expression of genes controlling oxidation and gluconeogenesis in primary bovine neonatal hepatocytes exposed to increasing concentrations of choline chloride (CC) and D,L-methionine (DLM) with or without fatty acids (FA). Primary hepatocytes isolated from 3 Holstein calves were maintained as monolayer cultures for 24 h before treatment with CC (61, 128, 2028, 4528 μmol/L) and DLM (16, 30, 100, 300 μmol/L) with or without a 1 mmol/L FA cocktail in a factorial design. After 24 h, media was harvested to quantify glucose, β-hydroxybutyrate (BHB), and cells harvested to quantify glycogen, DNA, and gene expression. No interactions between CC and DLM were detected. The potential two-way interaction between CC or DLM and FA was partitioned into three contrasts when P ≤ 0.20: linear without FA, linear with FA, difference of slope. Fatty acids did not affect glucose or cellular glycogen but increased pyruvate carboxylase (PC), cytosolic and mitochondrial phosphoenolpyruvate carboxykinase (PEPCKc, PEPCKm), and glucose-6-phosphatase (G6PC) expression. Increasing CC decreased glucose but increased cellular glycogen. Expression of PC and PEPCKc was increased by CC during FA treatment. Increasing DLM did not affect metabolites or PC expression, although PEPCKc was marginally decreased. Methionine did not affect G6PC, while CC had a marginal quadratic effect on G6PC. Oxidative and gluconeogenic enzymes appear to respond to FA in primary bovine neonatal hepatocytes. Increased PC and PEPCKc by CC during FA treatment suggest increased gluconeogenic capacity. Changes in G6PC may have shifted glucose-6-phosphate towards cellular glycogen; however, subsequent examination of G6PC protein is needed. Unaltered PC and marginally decreased PEPCKc suggest increased oxidative capacity with DLM, although BHB export was unaltered. The differential regulation supports unique effects of CC and DLM within bovine hepatocytes.

Digestible choline requirement of juvenile yellowtail kingfish (Seriola lalandi)

The present study was conducted to determine the digestible choline requirement of juvenile yellowtail kingfish (Seriola lalandi; YTK) in the presence of 2-amino-2-methyl-1-propanol (AMP), an inhibitor of choline biosynthesis. The second aim of this study was to determine if choline supplementation of a practical diet made from common raw materials can improve the growth performance of YTK and if choline supplementation dosages are affected by water temperature. Two eight-week experiments were conducted. The first was a dose-response experiment in which juvenile YTK (156.3 ± 15.3 g) were reared at 16 °C and fed five isonitrogenous and isoenergetic semi-purified diets containing 0.42 (Diet 1), 1.10 (Diet 2), 1.37 (Diet 3), 2.96 (Diet 4) or 6.05 g digestible choline kg−1 diet (Diet 5) by adding graded concentrations of choline chloride (CC). A sixth diet (Diet 6), comparable to Diet 4, was made without AMP to estimate the de novo synthesis of choline by YTK. The second was a factorial experiment in which juvenile YTK (157.3 ± 11.9 g) were reared at 16 °C and 24 °C and fed a practical diet supplemented with 0.0, 3.0 or 6.0 g of CC kg−1 which equates to a digestible choline concentration of 1.77, 3.54 and 4.66 g kg−1 diet, respectively. Results from experiment 1 indicated the breakpoint (broken-line regression model) in specific growth rate (SGR) and choline deposition rate occurred when digestible choline intake reached 26.1 mg kgBW−1 d−1 and 27.3 mg kgBW−1 d−1, respectively. On a dietary basis, the breakpoint in SGR and choline deposition rate occurred when diets provided 1.93 and 1.94 g digestible choline kg−1 diet, respectively. Experiment 2 results indicated there were no significant interactions between dietary choline and water temperature with respect to production indices such as SGR, food conversion ratio (FCR), whole body composition and nutrient retention. Choline deposition rate, SGR and FCR tended to be better in fish fed the practical diet supplemented with 3.0 g CC kg−1, but there was no additional advantage of raising the content to 6.0 g CC kg−1 diet. The present study confirms the necessity of adding CC to practical diets for YTK given that practical diets with no CC supplementation contain lower digestible choline than the estimated requirement established in experiment 1.

Effects of Excessive Choline Administration on Cardiovascular Risk Factors and Hepatic Cholesterol Metabolism in Female SD Rats

Choline has been considered to be linked to an increase of cardiovascular risk. It can be metabolized to trimethylamine by gut microbiota and further converted by hepatic flavin monooxygenase 3 (FMO3) to trimethylamine‐N‐oxide (TMAO) which is known to be a putative promoter of cardiovascular disease. However, the effects of excessive choline administration on the cardiovascular risk, especially in SD rats, have been little studied. The purpose of this study was to evaluate the effects of excessive choline administration on serum TMAO level, aortic wall thickness, and expression of hepatic genes responsible for cholesterol metabolism in female SD rats. Twenty female SD rats were fed AIN93‐G diet with or without various concentrations of choline chloride (1.5%, 2.25%, 3%) (w/w) in autoclaved water for 8 weeks. Aortic wall section of the rat was stained by hematoxylin and eosin. Serum TMAO and choline levels were determined by UPLC‐Synapt G2‐Si. Hepatic mRNA expression of FMO3, farnesoid X receptor (FXR), liver X receptor α (LXRα), sterol regulatory element‐binding protein 2 (SREBP‐2), cholesterol 7 alpha‐hydroxylase (CYP7A1), and low‐density lipoprotein receptor (LDLR) was determined by quantitative PCR. Serum TMAO level was significantly (p&lt;0.05) higher in the rat fed 1.5% choline‐containing water (1.5C) than control group (25.03 ± 14.27 vs. 0.34 ± 0.35 μM). However, serum choline levels were not significantly (p&gt;0.05) different (18.01 ± 7.15 vs. 23.13 ± 11.89 μM). Aortic wall of the 1.5C seemed to be thicker than the control group. Relative mRNA expression of CYP7A1 was down‐regulated in the 1.5C compared to the control group (0.32 ± 0.24 vs. 1.00 ± 0.56, p&lt;0.05), whereas that of FXR was up‐regulated (1.59 ± 0.60 vs. 1.00 ± 0.39, p&lt;0.05). However, mRNA expressions of hepatic FMO3, LDLR, LXRα, and SREBP‐2 were not significantly (p&gt;0.05) different. Excessive choline administration seemed to be able to affect cardiovascular system in female SD rats via production of TMAO and regulation of CYP7A1 and FXR. Additional research about secondary bile acid produced by gut bacteria is needed.Support or Funding InformationThis research has been supported by National Research Foundation of Korea (NRF‐ 2017R1D1A1B03028407) funded by the Ministry of Education of Korea.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Hypertonic Saline Suppresses NADPH Oxidase-Dependent Neutrophil Extracellular Trap Formation and Promotes Apoptosis.

Tonicity of saline (NaCl) is important in regulating cellular functions and homeostasis. Hypertonic saline is administered to treat many inflammatory diseases, including cystic fibrosis. Excess neutrophil extracellular trap (NET) formation, or NETosis, is associated with many pathological conditions including chronic inflammation. Despite the known therapeutic benefits of hypertonic saline, its underlying mechanisms are not clearly understood. Therefore, we aimed to elucidate the effects of hypertonic saline in modulating NETosis. For this purpose, we purified human neutrophils and induced NETosis using agonists such as diacylglycerol mimetic phorbol myristate acetate (PMA), Gram-negative bacterial cell wall component lipopolysaccharide (LPS), calcium ionophores (A23187 and ionomycin from Streptomyces conglobatus), and bacteria (Pseudomonas aeruginosa and Staphylococcus aureus). We then analyzed neutrophils and NETs using Sytox green assay, immunostaining of NET components and apoptosis markers, confocal microscopy, and pH sensing reagents. This study found that hypertonic NaCl suppresses nicotinamide adenine dinucleotide phosphate oxidase (NADPH2 or NOX2)-dependent NETosis induced by agonists PMA, Escherichia coli LPS (0111:B4 and O128:B12), and P. aeruginosa. Hypertonic saline also suppresses LPS- and PMA- induced reactive oxygen species production. It was determined that supplementing H2O2 reverses the suppressive effect of hypertonic saline on NOX2-dependent NETosis. Many of the aforementioned suppressive effects were observed in the presence of equimolar concentrations of choline chloride and osmolytes (d-mannitol and d-sorbitol). This suggests that the mechanism by which hypertonic saline suppresses NOX2-dependent NETosis is via neutrophil dehydration. Hypertonic NaCl does not significantly alter the intracellular pH of neutrophils. We found that hypertonic NaCl induces apoptosis while suppressing NOX2-dependent NETosis. In contrast, hypertonic solutions do not suppress NOX2-independent NETosis. Although hypertonic saline partially suppresses ionomycin-induced NETosis, it enhances A23187-induced NETosis, and it does not alter S. aureus-induced NETosis. Overall, this study determined that hypertonic saline suppresses NOX2-dependent NETosis induced by several agonists; in contrast, it has variable effects on neutrophil death induced by NOX2-independent NETosis agonists. These findings are important in understanding the regulation of NETosis and apoptosis in neutrophils.

Choline and methionine differentially alter methyl carbon metabolism in bovine neonatal hepatocytes.

Intersections in hepatic methyl group metabolism pathways highlights potential competition or compensation of methyl donors. The objective of this experiment was to examine the expression of genes related to methyl group transfer and lipid metabolism in response to increasing concentrations of choline chloride (CC) and DL-methionine (DLM) in primary neonatal hepatocytes that were or were not exposed to fatty acids (FA). Primary hepatocytes isolated from 4 neonatal Holstein calves were maintained as monolayer cultures for 24 h before treatment with CC (61, 128, 2028, and 4528 μmol/L) and DLM (16, 30, 100, 300 μmol/L), with or without a 1 mmol/L FA cocktail in a factorial arrangement. After 24 h of treatment, media was collected for quantification of reactive oxygen species (ROS) and very low-density lipoprotein (VLDL), and cell lysates were collected for quantification of gene expression. No interactions were detected between CC, DLM, or FA. Both CC and DLM decreased the expression of methionine adenosyltransferase 1A (MAT1A). Increasing CC did not alter betaine-homocysteine S-methyltranferase (BHMT) but did increase 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR) and methylenetetrahydrofolate reductase (MTHFR) expression. Increasing DLM decreased expression of BHMT and MTR, but did not affect MTHFR. Expression of both phosphatidylethanolamine N-methyltransferase (PEMT) and microsomal triglyceride transfer protein (MTTP) were decreased by increasing CC and DLM, while carnitine palmitoyltransferase 1A (CPT1A) was unaffected by either. Treatment with FA decreased the expression of MAT1A, MTR, MTHFR and tended to decrease PEMT but did not affect BHMT and MTTP. Treatment with FA increased CPT1A expression. Increasing CC increased secretion of VLDL and decreased the accumulation of ROS in media. Within neonatal bovine hepatocytes, choline and methionine differentially regulate methyl carbon pathways and suggest that choline may play a critical role in donating methyl groups to support methionine regeneration. Stimulating VLDL export and decreasing ROS accumulation suggests that increasing CC is hepato-protective.

Electrodeposition of Magnetic SmCo Films from Deep Eutectic Solvents and Choline Chloride-Ethylene Glycol Mixtures

Suitability of mixtures based on ethylene glycol and choline chloride is evaluated for the electrodeposition of magnetic SmCo alloys. Deep eutectic solvents (DES) are characterized by a wide electrochemical window, allowing the electrodeposition of metallic elements having a highly negative reduction potential. Electrodeposition of unconventional metals with reduction potential well below hydrogen evolution one is, in principle, feasible (e.g. Sm). Moreover, the limited presence of water during the growth of SmCo alloys may reduce oxidation and consequently improve the magnetic properties with respect to the employment of an aqueous bath. Deposits composition is strongly affected by process parameters and bath composition. Increasing choline chloride concentration and/or adding glycine to the solution allows to achieve higher Sm content in the deposit. Potentiostatic electrodeposition has been carried out in a conventional three electrodes cell, with potential interval selected from −0.7 V vs Ag wire to −0.95 V vs Ag wire, keeping the bath under stirring at 70°C. Magnetic SmCo films with composition from 0 to 50 wt% Sm have been obtained with coercivity up to 270 Oe.

Biosensor based on Butyrylcholinesterase for Detection of Carbofuran

Esterase enzymes play an important role in biology because they are responsible for the hydrolysis of choline esters. In their absence, the original state of the post synaptic membranes cannot be reestablished. Therefore, the aim of the work is to study the inhibiting action exerted by the group of compounds on these enzymes. Among these class of inhibiting compounds, pesticides are important because of the potential danger as a result of their large scale use in agriculture. Pesticides are generally determined using liquid or gas chromatography methods with various detection techniques. These methods are very sensitive and discriminating, however they require sample pretreatment such as extraction, preconcentration and clean up, which are skilled techniques and high cost treatment and also time consuming. In this study, acetyl cholinesterase and butyrylcholinesterase based biosensors have emerged as a promising tool for the detection and characterization of pesticides which are inhibitors of these enzymes. Although the physiological function of butyrylcholinesterase in comparison with acetyl cholinesterase is ambiguous, it has larger substrate specificity towards choline esters. Therefore, the development of a more selective electrode against choline, can lead to more sensitive determination of the inhibitor being investigated. Hence in the present work, a method based on inhibition of butyrylcholinesterase was attempted for quantification of carbofuran on the basis of cholinesterase inhibition. Butyrylcholinesterase with an activity of 10.2 units/mg was immobilized on a solid surface by cross linking with glutaraldehyde. The immobilized system was calibrated by correlating the inhibition of the butyrylcholinesterase activity with varying concentrations of the butyryl choline chloride and carbofuran. The sensing mechanism was investigated for its response to carbofuran concentrations ranging from 125 to 1,000 ppm. The effects of butyryl choline chloride concentration on the response of the sensing strip were also determined. Through this work, a sensing element made up of a carbon based electrode, containing the immobilized butyrylcholinesterase, has been described. This element has certain practical advantages like smaller size, user friendly, easy portability and disposability.

Statistical Approach for Media Optimization of Vitamin B12 Production by using newly isolated Rhizobium species with submerged fermentation

The present study deals with the improvement of the Vitamin B12 production by newly isolated Rhizobium sp. For this purpose various medium components were studied with Statistical approach to design fermentation medium. Eight variables were screened using Plackett Burman design. Further four selected variables were preceded with full factorial. Three significant variables from full factorial design such as mono sodium glutamate, cobalt nitrate and choline chloride concentration were optimized by using central composite design with response surface methodology. Using the optimal medium components (Choline chloride 6.25 g/l, Monosodium glutamate 1.50 g/l and Cobalt nitrate 0.38 g/l) the Vitamin B12 production was found five folds higher in the optimized medium as compared to the control medium.

A sensitive choline biosensor using Fe3O4 magnetic nanoparticles as peroxidase mimics

A sensitive choline biosensor using Fe(3)O(4) magnetic nanoparticles and a choline oxidase modified gold electrode was developed. Fe(3)O(4) magnetic nanoparticles as peroxidase mimics used in the choline biosensor can not only improve the sensitivity of the response signal, but also possess the favorable properties of stability, magnetic separation and easy preparation, etc. When using the reduction currents of square wave voltammetry as the detection signals, the interferences of ascorbic acid and uric acid to the choline biosensor can be reduced effectively. The reduction currents of the square wave voltammetry were increased with the logarithm values of the choline chloride concentration (from 10(-9) to 10(-2) M), the detection limit was estimated to be 0.1 nM (S/N = 3). This choline biosensor also exhibited favorable selectivity and stability in the determination of choline chloride.

Role of H 2O 2 on the kinetics of low-affinity high-capacity Na +-dependent alanine transport in SHR proximal tubular epithelial cells

The presence of high and low sodium affinity states for the Na +-dependent [ 14C]- l-alanine uptake in immortalized renal proximal tubular epithelial (PTE) cells was previously reported (Am. J. Physiol. 293 (2007) R538–R547). This study evaluated the role of H 2O 2 on the Na +-dependent [ 14C]- l-alanine uptake of ASCT2 in immortalized renal PTE cells from Wistar Kyoto rat (WKY) and spontaneously hypertensive rat (SHR). Na + dependence of [ 14C]- l-alanine uptake was investigated replacing NaCl with an equimolar concentration of choline chloride in vehicle- and apocynin-treated cells. Na + removal from the uptake solution abolished transport activity in both WKY and SHR PTE cells. Decreases in H 2O 2 levels in the extracellular medium significantly reduced Na +- K m and V max values of the low-affinity high-capacity component in SHR PTE cells, with no effect on the high-affinity low-capacity state of the Na +-dependent [ 14C]- l-alanine uptake. After removal of apocynin from the culture medium, H 2O 2 levels returned to basal values within 1 to 3 h in both WKY and SHR PTE cells and these were found stable for the next 24 h. Under these experimental conditions, the Na +- K m and V max of the high-affinity low-capacity state were unaffected and the low-affinity high-capacity component remained significantly decreased 1 day but not 4 days after apocynin removal. In conclusion, H 2O 2 in excess is required for the presence of a low-affinity high-capacity component for the Na +-dependent [ 14C]- l-alanine uptake in SHR PTE cells only. It is suggested that Na + binding in renal ASCT2 may be regulated by ROS in SHR PTE cells.

Mono-Enzyme Biosensor for the Detection of Organophosphorous Compounds

Organophosphorous compounds are important neuroactive molecules whose presents significant analytical challenges. New monoenzyme biosensor introduced here, is used for the measurement of the level of two organophosphorous compounds, paraoxon (POX) and ethyl parathion (EPA), in phosphate buffer. Prussian blue electrodepositionized on a graphite electrode served as a template for immobilization of monoenzyme layer. A monoenzyme layer containing choline oxidase was immobilized along with nafion and bovine serum albumin, by cross-linking with glutaraldehyde. The results revealed that the response of the proposed biosensor was linear between 5 μM and 80 μM in the presence of POX and EPA. Detection limits were 0.4 μM in both cases. Also, for the optimization of the biosensor performance, temperature and pH dependency, concentration of choline chloride as substrate of choline oxidase and incubation time were determined. The results showed that the optimum values for temperature, pH, substrate concentration and incubation time were room temperature, pH 7, 4 mM and 30 min, respectively.