Sodium butyrate concentration

Cells were seeded at 1 to 2 X 10 cells/75cm2 flask with sodium butyrate concentrations from 0.5 to 5.0 mM and without butyrate in growth medium. After 8 days, the cells were harvested and protein was determined. Figure 1 shows total milligram protein/T-75cm2 flasks as plotted against sodium butyrate concentrations. The cell protein per flask of SKCO-1 decreased sharply with increased concentrations of butyrate when compared with control culture cells. With SW-480 and SW-620 culture cells, protein was decreased against butyrate concentrations, but the decrease was more pronounced with SW-620 cells. Cell protein of FHS and HT-29 cultures were unaffected (Fig. 1). 

Figure 1. Total milligrams of protein/ T-75 cm flasks vs. millimolar concentrations of sodium butyrate in growth medium. Total cell protein was determined after cells were incubated in growth medium with sodium butyrate for eight days at 37°C. O, FHS A, SKCO-1 %, FIT-29 , SW-480 SW-620.

The concentration of sodium butyrate was observed to have a differential effect on cell growth in colonic cell lines. After culturing for 8 days with 5 mM sodium butyrate, the cell protein per flask of the SCKO-1 line was decreased to less than 10% of the control cultures. In the SW-620 culture,cell protein per... 

A Cells exposed to medium containing 5mM sodium butyrate for times indicated. B After 48 hr, as in A, medium replaced with fresh medium without butyrate. C Cells exposed for 48 hr to medium containing the indicated concentrations of butyrate. Specific binding of, 2sI-choleragen determined as described in Ref. 5. (Data from Ref. 5J... 

Corn steep liquor (CSL), a byproduct of the com wet-milling process, was used in an immobilized cell continuous biofilm reactor to replace the expensive P2 medium ingredients. The use of CSL resulted in the production of 6.29 g/L of total acetone-butanol-ethanol (ABE) as compared with 6.86 g/L in a control experiment. These studies were performed at a dilution rate of 0.32 hr1. The productivities in the control and CSL experiment were 2.19 and 2.01 g/(Lh), respectively. Although the use of CSL resulted in a 10% decrease in productivity, it is viewed that its application would be economical compared to P2 medium. Hence, CSL may be used to replace the P2 medium. It was also demonstrated that inclusion of butyrate into the feed was beneficial to the butanol fermentation. A control experiment produced 4.77 g/L of total ABE, and the experiment with supplemented sodium butyrate produced 5.70 g/L of total ABE. The butanol concentration increased from 3.14 to 4.04 g/L. Inclusion of acetate in the feed medium of the immobilized cell biofilm reactor was not found to be beneficial for the ABE fermentation, as reported for the batch ABE fermentation. 

Next, an experiment was run in which 2.5 g/L of sodium butyrate was added to P2 medium to investigate whether it could be converted to butanol. A control experiment was run containing P2 medium. A separate control experiment was run before each experiment. This is essential because biomass accumulation in the reactor changes with time, thus affecting performance of the reactor (5). The reactor produced 4.77 g/L of total ABE, of which acetone, butanol, and ethanol were 1.51,3.14, and 0.12 g/L, respectively (Table 1). It resulted in a total ABE productivity of 1.53 g/(L-h) and a glucose utilization of 29.4% of that available in the feed of 59.1 g/L. The acid concentration in the effluent was 1.56 g/L. Following this, P2 medium was supplemented with sodium butyrate and the experiment was conducted at the same dilution rate. The reactor produced 1.55 g/L of acetone, 4.04 g/L of butanol, and 0.11 g/L of ethanol, for a total ABE concentration of 5.70 g/L, compared with 4.77 g/L in the control experiment. The productivity was 1.82 g/(L-h), compared with 1.53 g/(L-h) for the control experiment. These experiments suggested that butyrate was used by the culture to produce additional butanol. Note that 0.9 g/L of butanol was produced from 1.65 g/L of butyrate (2.5 g/L in feed, 0.85 g/L in effluent). The yield calculations do not include the amount of butyrate that was utilized by the culture. 

Sodium butyrate was employed at a final concentration of 20 mM. Palmltate and oleate were maintained as 1% stock solutions in 10% BRIJ 35. BRIJ 35 controls were negative. 

The cultured cells that we have been studying are immature and may therefore be models for the developing brain rather than the mature brain. The mouse neuroblastoma cells, however, can be induced to differentiate by exposure to 0.5mM sodium butyrate (Fig 7). This causes an increase in cytoplasm nucleus ratio, neurite outgrowth and induction of tyrosine hydroxylase and other enzymes. As shown in Fig 8, butyrate caused an increase in T3 uptake at 2 hours. The initial (l min) uptake was also increased. The apparent Ka of nuclear binding in intact cells with the finding that the apparent affinity of the nuclear receptor was decreased by butyrate but the Ka in isolated nuclei was not affected. The apparent inconsistency of an increased intracellular transport but a decreased intracellular free T/ concentration is unexplained. As described by others in developing rats and in cultured rat glial cells we also found that neuroblast differentiation was accompanied by an increase in nuclear receptor number. 

The HPLC data show that increasing Tween-80 from 0.07 to 0.20 M, while holding the PEG, sodium caprylate and ethyl butyrate concentrations constant, resulted in an increase in drug extracted from 7 to 22%. Using the same chemicals but increasing PEG concentration from 0.0 M to 0.5 M reduced by 60%, the amount of bupivacaine extracted. 

Ramachandran and Balasubramanian used the method to monitor structural alterations caused in water by added solutes. They used the spin probe 2,2,6,6-tetramethyl piperid-4-one TV oxide (TEMPO) to study structural alterations caused in water by the addition of urea and sodium butyrate. They studied the variation of hydrogen hyperflne linewidth as a function of the solute concentration and extracted information on the reorientation time [36]. They conclude that urea disrupts water structure continuously and this effect is significant at low molarities. With sodium butyrate they see evidence for two different environments, one attached to the solute and the other far away from it. 

Ethyl n-butyrate. Use a mixture of 88 g. (92 ml.) of n-butyric acid, 23 g. (29 ml.) of ethanol and 9 g. (5 ml.) of concentrated sulphuric acid. Reflux for 14 hours. Pour into excess of water, wash several times with water, followed by saturated sodium bicarbonate solution until all the acid is removed, and finally with water. Dry with anhydrous magnesium sulphate, and distU. The ethyl n-but3rrate passes over at 119 5-120-5°, Yield 40 g. An improved yield can be obtained by distilhng the reaction mixture through an efficient fractionating column until the temperature rises to 125°, and purifying the crude ester as detailed above under methyl acetate. 

Methylsuccinic acid has been prepared by the pyrolysis of tartaric acid from 1,2-dibromopropane or allyl halides by the action of potassium cyanide followed by hydrolysis by reduction of itaconic, citraconic, and mesaconic acids by hydrolysis of ketovalerolactonecarboxylic acid by decarboxylation of 1,1,2-propane tricarboxylic acid by oxidation of /3-methylcyclo-hexanone by fusion of gamboge with alkali by hydrog. nation and condensation of sodium lactate over nickel oxide from acetoacetic ester by successive alkylation with a methyl halide and a monohaloacetic ester by hydrolysis of oi-methyl-o -oxalosuccinic ester or a-methyl-a -acetosuccinic ester by action of hot, concentrated potassium hydroxide upon methyl-succinaldehyde dioxime from the ammonium salt of a-methyl-butyric acid by oxidation with. hydrogen peroxide from /9-methyllevulinic acid by oxidation with dilute nitric acid or hypobromite from /J-methyladipic acid and from the decomposition products of glyceric acid and pyruvic acid. The method described above is a modification of that of Higginbotham and Lapworth. ... 

In isolating higher-melting compounds prepared by this method (e.g., 7-naphthylbutyric acids), the layers are separated after cooling to 50-60°, benzene is used for the extraction, and the combined solution is clarified with Norite while still wet, concentrated somewhat, and allowed to cool for crystallization. In preparing methoxylated adds such as 7-anisyl- or 7-veratryl-butyric acid some demethylation occurs and a modification is necessary. The toluene layer and extracts are mixed with an excess of dilute sodium hydroxide and the organic solvents are... 

The deep red solution is cooled in an ice bath and neutralized by the addition, with shaking, of an ice-cold solution of 15 cc. of concentrated sulfuric acid in 200 cc. of water. The ether layer is separated, washed with water, and dried over sodium sulfate. The ether is removed by dropping the solution from a separatory funnel the stem of which extends to the bottom of an evacuated Claisen flask heated on the steam bath. The residue is a pale yellow oil consisting of a mixture of ethyl a-ethoxalyl-y-phenyl-butyrate and unchanged ethyl oxalate (Note 5). 

To a solution of 390 mg of 4-[3-methyl-trans-4a-cisoid-4a,5a-cis-5a-l,4a,5,5a,10b,10c-hexahydro-7-dioxino[5,4-a]cyclopenta[b]benzofuranyl] butyric acid in 5 ml of ethyl acetate cooled in an ice bath was added an excess of an ethereal solution of diazomethane, and after being stirred for 5 min the mixture was concentrated. The resulting oily material was dissolved in 3 ml of methanol, and to the solution was added 1 ml of 1 N hydrochloric acid and the mixture was stirred for 3 hours at room temperature. After concentration of the reaction mixture and addition of 1 ml of water, the mixture was extracted 3 times with each 5 ml of ethyl acetate. The combined layers of ethyl acetate were washed with 3 ml of water and 3 ml of aqueous saturated solution of sodium chloride, dried and concentrated to give 380 mg of crude crystals. The crude crystals were recrystallized from ethyl acetate-hexane to yield 200 mg of the pure captioned product (m.p. 56-57°C, yield 53%). 

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