Effects of Sources and Forms of Vitamin K3 on Its Storage Stability in Vitamin Premixes or Vitamin Trace Mineral Premixes

Six types of vitamin K3 (VK3); two sources (menadione sodium bisulfite, MSB; menadione nicotinamide bisulfite, MNB), and three different forms (crystal, micro-capsule, and micro-sphere) were used to determine the retention of VK3 in vitamin premixes (Experiment 1) or vitamin trace mineral (VTM) premixes (Experiment 2) after 1, 2, 3, and 6 months of storage. The retention of VK3 in vitamin premixes was evaluated at 25 °C/60% relative humidity or 40 °C/75% relative humidity in an incubator in Experiment 1 and in VTM premixes (choline chloride: 0 vs. 16,000 mg/kg) stored at room temperature in Experiment 2. The VK3 retention in vitamin premix or VTM premix decreased significantly with the extension of storage time (p < 0.05). In Experiment 1, the VK3 retention was higher in the 25 °C/60% incubator (56%) than in the 40 °C/75% incubator (28%). The MNB retention (52%) was higher than MSB retention (32%). The retention of VK3 in micro-capsules (43%) or micro-spheres (48%) was higher than the crystal form (35%) after six months of storage. In Experiment 2, there was no difference between the retention of MSB (49%) or MNB (47%). The retention of VK3 of micro-capsule (51%) or micro-sphere (54%) was higher than that of crystal form (40%). The VK3 retention was higher in the choline-free group (51%) than in the choline group (47%) after six months of storage. Finally, the predicted equations of VK3 retention with storage time in vitamin premixes or VTM premixes were established. The R2 of the prediction equations was ≥0.9005, indicating that time is an important factor in predicting VK3 retention. In conclusion, the higher temperature-relative humidity, choline had negative effects on VK3 retention during premix storage. MNB retention was higher than MSB during storage of vitamin premix. The encapsulated forms of VK3, micro-capsules and micro-spheres, could improve VK3 storage stability in vitamin premix and VTM premix.

New Insight Into Rumen Methanogen Function: a Barrier Blocking Trimethylamine From Entering the Host Body

Background: Trimethylamine (TMA) is the precursor of trimethylamine N-oxide (TMAO), which has been known to promote human cardiovascular disease. Methanomassiliicoccales (Mmc), TMA-utilizing methanogens, may function as a TMA barrier for the host in the rumen. This study aimed to investigate the role of Mmc in rumen TMA elimination and the effect of choline addition in the diet on the rumen TMA and TMAO concentrations in the plasma and milk of dairy cows.Results: Three experiments, 2 rumen in vitro fermentation trials and 1 dairy cow in vivo trial, were conducted. Four groups were set in Experiment 1: control, nitroglycerin (NG, a methanogen inhibitor), TMA (0.69 mg/mL), and TMA + NG. The methanogenic activity was completely inhibited in the NG group, and no methane production was observed in the NG and TMA + NG groups. The TMA content hardly reduced in the TMA + NG group (0.66 mg/mL) after 2 d of incubation; in contrast, it reduced by 47.2% in the TMA group. Methanogen 16S rRNA gene sequencing showed that the relative abundance of Mmc increased in the TMA group (P = 0.005), which was confirmed by real-time polymerase chain reaction testing. Two taxa (Group 9 sp. ISO4-G1 and Group 10 sp.) at the species level mainly contributed to the increase in the relative abundance of Mmc. Four groups were set in Experiment 2: control, NG, choline (choline chloride, 1.0 mg/mL), and choline + NG. Choline was completely degraded in 24 h, and the TMA content reached the peak point in the fermentation culture. The TMA content in the choline + NG group did not reduce after the occurrence of the peak point. In contrast, the TMA content in the choline group started to decrease after 24 h, corresponding to the rapid increase in methane production. Eight mid-lactating, rumen-fistulated Holstein cows were randomly assigned to the control (n = 4) or choline (n = 4) group in Experiment 3: In the choline group, the cows were gradually supplemented with 100–250 g/(cow·d) of choline chloride over 4 weeks. In the choline group, TMA accumulated in the rumen fluid, and the abundance of Mmc 16S rRNA gene and choline-degrading bacterial cutC gene increased in the rumen content (P < 0.050). The TMAO content in the plasma and milk of the dairy cows was approximately 10 times higher in the choline group than in the control at day 28.Conclusion: Rumen Mmc functioned as a TMA barrier for the host. The increase in dietary choline caused more TMA to enter the host body, resulting in higher TMAO deposition in the milk of the dairy cows. It should be noted that TMAO-rich milk might degrade food safety. Moreover, it should be discreet to mitigate rumen methane emission by inhibiting the rumen methanogen activities, which might destroy the rumen TMA barrier.

P4610Gut microbiome and atherosclerotic plaque instability: does choline consumption in red meat influence plaque stability?

Background Myocardial infarction is the major cause of deaths worldwide. Gut bacteria can process choline as abundant in red meat and subsequently converted by flavin containing monooxygenase in the liver to trimethylamin-N-oxide (TMAO) metabolite, which is strongly associated with cardiovascular events. Aim To investigate the gut microbiome and its association with atherosclerotic plaque instability. Methods Forty-eight Apolipoprotein E deficient mice were randomly divided into two groups and two time points, fed with a high fat diet (containing either 0.4% choline or 3% choline) at 12 weeks of age, for 7 weeks or for 14 weeks. All mice underwent Tandem Stenosis (TS) surgery to induce the development of unstable plaques. Stool samples were collected directly from the colon. Measurements of gut microbes were performed by AGRF diversity profiling. After bacterial genomic DNA isolation, 16S rRNA were sequenced by targeting 27F-519R (V1-V3) and 341F-806R (V3-V4) on the Illumina MiSeq platform. Vessel segments of TS were histologically processed and plaque composition of lipid, collagen, and intraplaque hemorrhage (marker of unstable plaques) were performed by a series of chemical staining and immunohistochemistry. Results Monocytes and granulocytes in mouse blood were significantly increased in the high choline group (p<0.05, unpaired t-test) after 7 weeks of high fat diet (21% fat, 0.15% Cholesterol, 3% Choline). Profiling of gut microbiota showed that Fimicutes were down regulated in the high choline group (p<0.05, unpaired t-test). Within Phylum Fimicutes, only Clostridia (class) Clostridiales (order) were significantly downregulated. Interestingly, histological analysis of TS segments showed that TER-119 (intraplaque haemorrhage marker) and CD42c (platelet marker) were significantly increased in the high choline group, indicating atherosclerotic plaques are more unstable and prone to rupture (p<0.05, unpaired t-test). Nevertheless, CD68 (Foam cells) in plaques, and total atherosclerotic plaque burden in the aortic sinus and aortic arch were not affected by the elevated levels of choline consumption. Conclusion Choline intake increases circulating monocytes and granulocyte numbers in the blood but not in the atherosclerotic plaque itself. Whereas the total plaque burden is not changed by an increased choline intake, the reduction of Fimicutes, Clostridia and Clostridiales seems to contribute to atherosclerotic plaque instability. Acknowledgement/Funding Heart Foundation 2018 Future Leader Fellowship (2018 FLF) ID: 102068 Chen

Prenatal Choline Supplementation Improves Child Color-location Memory Task Performance at 7 Y of Age (FS05-01-19)

Objectives To assess the effect of 3rd trimester maternal choline supplementation on child memory at 7 y of age. Animal studies have provided strong evidence that maternal choline supplementation improves offspring cognition, including memory, but few experimental studies have evaluated this intervention in humans. Methods Pregnant women (n = 26) were randomized to consume 480 mg (approx. the Adequate Intake [AI]) or 930 mg choline/d from gestational wk 27 until delivery as part of a controlled-feeding study. All food (providing 380 mg choline/d), choline supplements (100 mg or 550 mg/d), and prenatal vitamins were provided by the study. An ancillary follow-up was conducted to assess offspring cognitive functioning, including memory, at age 7 y (n = 20). Children performed a computer-based color-location memory task in which the location of dots on a cartoon figure were recalled after a retention interval (RI) of 1 or 8 s. The task increased in difficulty level (number of dots to be recalled; 1 to 5 dots) every 4 trials. Data were analyzed using mixed models; pairwise comparisons were corrected for multiplicity. Results Children in the 930 mg/d group passed more levels than those in the 480 mg/d group (P = .048; see Figure), indicative of superior memory span. While the choline group by RI interaction was non-significant (P = .31), contrasts revealed that the 930 (v. 480) mg/d group tended to pass more levels at the 1 s RI (P = .052), but not at the 8 s RI (P = .58). Similarly, the analysis of total dots correct revealed a choline group by RI interaction (P = .08) such that children in the 930 (v. 480) mg/d group recalled more dots correctly at the 1 s RI (P = .02), but not at the 8 s RI (P > .99). Conclusions Children whose mothers consumed 930 (v. 480) mg choline/d performed significantly better on a task of color-location memory at age 7 y, suggesting a long-term beneficial effect of prenatal choline supplementation on offspring cognition in humans. These findings indicate that the choline AI for pregnant women may not be sufficient for optimal offspring cognitive functioning and raise concerns about choline intake during pregnancy in North America, which on average is only approx. 70% of the AI. Funding Sources Funded by NIFA/USDA and the Balchem Corp. JEHN and CLB were supported by NIH Traineeships. CLB was supported by an Egg Nutrition Center Young Investigator Research Award for Early Exploration. Supporting Tables, Images and/or Graphs

Prenatal Choline Supplementation Improves Child Color-location Memory Task Performance at 7 Y of Age (FS05-01-19)

Objectives To assess the effect of 3rd trimester maternal choline supplementation on child memory at 7 y of age. Animal studies have provided strong evidence that maternal choline supplementation improves offspring cognition, including memory, but few experimental studies have evaluated this intervention in humans. Methods Pregnant women (n = 26) were randomized to consume 480 mg (approx. the Adequate Intake [AI]) or 930 mg choline/d from gestational wk 27 until delivery as part of a controlled-feeding study. All food (providing 380 mg choline/d), choline supplements (100 mg or 550 mg/d), and prenatal vitamins were provided by the study. An ancillary follow-up was conducted to assess offspring cognitive functioning, including memory, at age 7 y (n = 20). Children performed a computer-based color-location memory task in which the location of dots on a cartoon figure were recalled after a retention interval (RI) of 1 or 8 s. The task increased in difficulty level (number of dots to be recalled; 1 to 5 dots) every 4 trials. Data were analyzed using mixed models; pairwise comparisons were corrected for multiplicity. Results Children in the 930 mg/d group passed more levels than those in the 480 mg/d group (P = .048; see Figure), indicative of superior memory span. While the choline group by RI interaction was non-significant (P = .31), contrasts revealed that the 930 (v. 480) mg/d group tended to pass more levels at the 1 s RI (P = .052), but not at the 8 s RI (P = .58). Similarly, the analysis of total dots correct revealed a choline group by RI interaction (P = .08) such that children in the 930 (v. 480) mg/d group recalled more dots correctly at the 1 s RI (P = .02), but not at the 8 s RI (P > .99). Conclusions Children whose mothers consumed 930 (v. 480) mg choline/d performed significantly better on a task of color-location memory at age 7 y, suggesting a long-term beneficial effect of prenatal choline supplementation on offspring cognition in humans. These findings indicate that the choline AI for pregnant women may not be sufficient for optimal offspring cognitive functioning and raise concerns about choline intake during pregnancy in North America, which on average is only approx. 70% of the AI. Funding Sources Funded by NIFA/USDA and the Balchem Corp. JEHN and CLB were supported by NIH Traineeships. CLB was supported by an Egg Nutrition Center Young Investigator Research Award for Early Exploration. Supporting Tables, Images and/or Graphs

A New Mn-Salen Micellar Nanoreactor for Enantioselective Epoxidation of Alkenes in Water

A new chiral Mn-salen catalyst, functionalized with a long aliphatic chain and a choline group, able to act as surfactant catalyst for green epoxidation in water, is here described. This catalyst was employed with a commercial surfactant (CTABr) leading to a nanoreactor for the enantioselective epoxidation of some selected alkenes in water, using NaClO as oxidant. This is the first example of nanoreactor for enantioselective epoxidation of non-functionalized alkenes in water.

Effect dietary choline supplementation on egg quality and serumbiochemical profile in White Pekin Ducks

The aim of the present study was to evaluate the influence of choline in diet (a TMA precursor) on egg quality, serum biochemistry and presence of off flavour in eggs of White Pekin Ducks. 30 White Pekin ducks (45 weeks age) housed in individual cages were subjected to three dietary treatment groups and were. The dietary treatment groups consisted of two levels of choline as TMA precursor viz; 3g/kg diet, 6g/kg diet and a control diet devoid of choline. The results revealed highly significant (P<0.0001) effect of choline on yolk weight and HU than the control. However the other egg quality traits i.,e egg weight, albumen weight, shell weight, albumen index, yolk index, shape index, shell thickness were not influenced. Serum cholesterol and triglycerides were significantly (P<0.0001) reduced in high choline group (6g/kg diet) followed by low choline group (3g/kg diet) and control. The liver enzymes (SGPT and SGOT) were significantly lower (P<0.0001) in both the supplemented groups as compared to control. No significant effect was found for serum total protein and serum glucose levels. Presence of off flavour (fishy flavour) was more evident in the eggs of choline group (6g/kg diet) than the control group. It can be concluded that the presence of choline in the diet had minimal effect on the egg quality parameters of Pekin ducks except for yolk weight and albumen quality, however there was significant reduction in serum total cholesterol and triglycerides levels.

Effects of Pulsed Electric Fields on Phospholipid Molecular Conformation by Raman Spectra

In order to study the sterilization mechanism of pulsed electric fields（PEF） on microorganisms and the interaction between PEF conditions and cell membrane composed of phospholipids, the authors studied the phospholipid molecular conformation changes under various electric fields strength treatments using Raman spectra. It was demonstrated from the Raman spectra that there was little influence of PEF on the conformation of O-C-C-N+ backbone in the choline group of phospholipid. The intensity ratios of I1096/I1062 and I1096/I1126 represented the gauche/ trans ratioof C-C vibrations, but the first was increased directly, while the other one was increased firstly and then decreased with increasing electric fields strengths but the ratio was still higher than that of the control sample. I2885/I2850 represeted order/ disorder of chain-chain interactions which was decreased with increasing electric fields strengths. It was indicated that the intermolecular disordering in hydrocarbon chain of the lipid lattice was increased and the membrane fluidity was enhanced after PEF treatments.

Translocation of polysialic acid across model membranes: kinetic analysis and dynamic studies.

Transmembrane translocation of polyion homopolymers takes place in the case of polyanionic polysialic acid (polySia), polyanionic polynucleotides and polycationic polypeptides. The purpose of this work was to determine the role of membrane electrical parameters on the kinetics of polyion translocation, the influence of polysialic acid on ion adsorption on positively charged membrane surface and the dynamics of the phospholipid hydrocarbon chains and choline group by using 1H-NMR. The analysis of polyion translocation was performed by using the electrical equivalent circuit of the membrane for the initial membrane potential equal to zero. The changes in polysialic acid flux was up to 75% after 1 ms in comparison with the zero-time flux. Both a decrease of membrane conductance and an increase of polyion chain length resulted in the diminution of this effect. An increase of praseodymium ions adsorption to positively charged liposomes and an increase of the rate of segmental movement of the -CH2 and -CH3 groups, and the choline headgrup of lipid molecules, was observed in the presence of polySia. The results show that the direction of the vectorial polyion translocation depends both on the membrane electrical properties and the degree of polymerization of the polymer, and that polysialic acid can modulate the degree of ion adsorption and the dynamics of membrane lipids.

Lysophospholipids are natriuretic and diuretic in rats

The effects of lysophosphatidylcholine (LPC) and some of its analogues on renal sodium (UNaV) and water excretion (V) were examined in the pentobarbital sodium-anesthetized rat. LPC caused the increase in UNaV and V with a steep dose response. The natriuretic effect began within 20 min after a bolus injection and lasted for 7 h. The natriuresis and diuresis were not affected when the phospholipid was given as an LPC-albumin complex. Natriuretic activity was eliminated by substituting unsaturated bonds in the 1-acyl group and by removing the choline group on position 3. Natriuretic activity was not affected by substitution of 1-alkyl for 1-acyl groups and was stereospecific. These results demonstrate a unique, previously unrecognized effect of lysophospholipids and suggest that they may account for some of the previously demonstrated natriuretic activity of plasma.