[USE OF LUMINESCENT INDICATORS IN THE VOLUMETRIC DETERMINATION OF VALERIC ACID IN VALERIAN TINCTURES].
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Polyhydroxyalkanoates (PHA) is a family of microbially synthesized polyesters consisting of various 3-hydroxyalkanoate monomers. Aeromonas hydrophila 4AK4 could be able to synthesize PHA copolymer consisting of 3-hydroxybutyrate (3-HB) and 3-hydroxyhexanoate (3-HHx). No data has been reported about the ability to synthesize the PHA with other monomers in A. hydrophila. In this study, propionic acid, valeric acid, heptanoic acid, nonanoic acid and undecanoic acid were used together with gluconate to find out whether A. hydrophila 4AK4 could synthesize the PHA consisting of odd carbon atom number monomers. The result showed that A. hydrophila 4AK4 could not growth when supplied with propionic acid, valeric acid, heptanoic acid and nonanoic acid and only undecanoic acid could be used to synthesize PHA. Wild type and recombinant A. hydrophila 4AK4 harboring phaA (beta-ketothiolase) and phaB (acetoacetyl-CoA reductase) were cultivated with undecanoic acid and glucose or undecanoic acid and gluconate served as carbon sources. PHA consisting of 3-HB and 3-hydroxyvalerate (3-HV) could be produced by both wild type and recombinant A. hydrophila 4AK4 and the latter could produce PHA with more 3-HB monomer. When the ratio of glucose or gluconate to undecanoic acid was 1:1, the cell dry weight (CDW) of A. hydrophila 4AK4 reached 1.14 g/L and PHA content was 60% of the CDW after cultivation for 24 h. When lauric acid and undecanoic acid were served as co-substrate, A. hydrophila 4AK4 could produce copolyester consisting of 3-HB, 3-HV and 3-HHx. Along with the increase of undecanoic acid proportion in the mixed carbon source, the 3-HV content of copolymer was increased while the 3-HB and 3-HHx content were decreased. In all cases, the CDW decreased along with the increase of undecanoic acid concentration, which indicated that undecanoic acid was not very good for A. hydrophila 4AK4 growth.
The synthesis and preliminary biological assays of a new monofunctional reagent to reversible derivatize monoclonal antibodies is described. This compound, comprising a 1,4-polyiol moiety, is cleavable by means of sodium periodate in mild conditions; moreover it also contains a phenolic residue suitable for 125I labelling and a carboxylic group for reaction with epsilon-lysyl amino group of antibodies. These features are suitable to study the monoclonal antibodies cell-internalization process and antigen expression on cell surface. The 125I labelled reagent has been coupled to the monoclonal antibody AR-3, an IgG1 directed toward the CAR-3 antigen widely expressed on human ovarian and colorectal adenocarcinomas. The cleavage capability is tested in different conditions both on reagent and on derivatized MAb. Cell labelling experiments are performed both on target and untarget cell lines.
The application of unfunctionized polymethacrylate resin (TSKgel G3000PWXL) as a stationary phase in liquid chromatography with UV detection for C1-C7 aliphatic monocarboxylic acids (formic acid, acetic acid, propionic acid, butyric acid, isovaleric acid, valeric acid, 3,3-dimethylbutyric acid, 4-methylvaleric acid, hexanoic acid, 2-methylhexanoic acid, 5-methylhexanoic acid and heptanoic acid) and C1-C7 aliphatic monoamines (methylamine, ethylamine, propylamine, isobutylamine, butylamine, isoamylamine, amylamine, 1,3-dimethylbutylamine, hexylamine, 2-heptylamine and heptylamine) was carried out. Using dilute sulfuric acid as the eluent, the TSKgel G3000PWXL, resin acted as an advanced stationary phase for these C1-C7 carboxylic acids. Excellent simultaneous separation and symmetrical peaks for these C1-C7 carboxylic acids were achieved on a TSKgel G3000PWXL column (150 mm x 6 mm i.d.) in 60 min with 0.25 mM sulfuric acid containing 1 mM 2-methylheptanoic acid at pH 3.3 as the eluent. Using dilute sodium hydroxide as the eluent, the TSKgel G3000PWXL resin also behaved as an advanced stationary phase for these C1-C7 amines. Excellent simultaneous separation and good peaks for these C1-C7 amines were achieved on the TSKgel G3000PWXL column in 60 min with 10 mM sodium hydroxide containing 0.5 mM 1-methylheptylamine at pH 11.9 as the eluent.
The application of unfunctionized polymethacrylate resin (TSKgel G3000PWXL) as a stationary phase in liquid chromatography with conductimetric detection for C1-C7 aliphatic monocarboxylic acids (formic acid, acetic acid, propionic acid, butyric acid, isovaleric acid, valeric acid, 3,3-dimethylbutyric acid, 4-methylvaleric acid, hexanoic acid, 2-methylhexanoic acid, 5-methylhexanoic acid and heptanoic acid) and C1-C7 aliphatic monoamines (methylamine, ethylamine, propylamine, isobutylamine, butylamine, isoamylamine, amylamine, 1,3-dimethylbutylamine, hexylamine, 2-heptylamine and heptylamine) was attempted with C8 aliphatic monocarboxylic acids (2-propylvaleric acid, 2-ethylhexanoic acid, 2-methylheptanoic acid and octanoic acid) and C8 aliphatic monoamines (1,5-dimethylhexylamine, 2-ethylhexylamine, 1-methylheptylamine and octylamine) as eluents, respectively. Using 1 mM 2-methylheptanoic acid at pH 4.0 as the eluent, excellent separation and relatively high sensitive detection for these C1-C7 carboxylic acids were achieved on a TSKgel G3000PWXL column (150 mm x 6 mm i.d.) in 60 min. Using 2 mM octylamine at pH 11.0 as the eluent, excellent separation and relatively high sensitive detection for these C1-C7 amines were also achieved on the TSKgel G3000PWXL column in 60 min.
A threonine overproducing mutant of Alcaligenes sp. SH-69 was isolated and its ability to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3HB-co-3HV), was investigated. The 3HV fraction in poly(3HB-co-3HV) produced from glucose as the sole carbon source exceeded 22 mol%, which is approximately six times higher than that achieved by the wild type under the same culture conditions. Furthermore, the addition of a relatively low concentration (10 mM) of propionic acid, valeric acid or levulinic acid to the glucose medium greatly increased the molar fraction of 3HV in the copolyester, to 38-77 mol%. The results suggest that metabolic engineering of the biosynthetic pathways supplying polyhydroxyalkanoate monomers, such as the threonine biosynthetic pathway, can lead to new poly(3HB-co-3HV)-producing strains.
4 sheep (wethers) and 4 bulls (130-140 kg live weight) received pellets of straw and concentrated feed of the charges 1-5 produced from the same wheat straw. The quota of wheat coarse meal in the pellets was, in the sequence 1 to 5:31.4; 29.7; 15.3; 11.5 and 7.8% (charge 5 contained in addition to this 7.4% dried sugar beet). The crude fibre content in the DM, in the same sequence, was as follows: 25.9; 28.1; 28.7; 31.7 and 34.4%. The total content of volatile fatty acids (VFA) in the rumen fluid of the sheep was between 96.6 and 97.8 mmol/l (all 5 charges) and in the rumen fluid of the bulls between 83.4 and 90.5 m mol/l. There were no significant differences between the groups. The molar acetic acid content after the feeding of charge 1 (poorer in crude fibre) was 67.4% (sheep) and 68.0% (bulls) and rose with the increasing crude fibre content in the case of charge 5 to 70.5% and 72.2% resp. The content of propionic acid did not rise in the pelleted rations with the growing cereal quota and, identically for sheep and bulls, amounted to 17.7 mol% and 17.8 mol% on an average of the 5 rations. The molar butyric acid content decreased significantly with both species with the growing crude fibre content of the pelleted charges. The sum of the VFA with branched chains (isobutyric acid, valeric acid and isovaleric acid) increased significantly in both species with the growing quota of cereal coarse meal in the pellet charges. In the bulls, however, the increase was distinctly steeper, which is probably connected with the significantly higher pH-value and a higher proteolysis rate in the rumen fluid of the bulls (charges 1 to 2) in comparison with the sheep.
Tetrachloroethene, also known as perchloroethylene, was reductively dechlorinated to trichloroethene and cis-1,2-dichloroethene by laboratory sulfate-reducing enrichment cultures. The causative organism or group was not identified. However, tetrachloroethene was dechlorinated to trichloroethene in 50 mM bromoethane-sulfonate-inhibited enrichments and to trichloroethene and cis-1,2-dichloroethene in 3 mM fluoroacetate-inhibited enrichments. Overall transformation varied from 92% tetrachloroethene removal in 13 days to 22% removal in 65 days, depending on conditions of the inoculum, inhibitor used, and auxilliary substrate used. Neither lactate, acetate, methanol, isobutyric acid, valeric acid, isovaleric acid, hexanoic acid, succinic acid, nor hydrogen appeared directly to support tetrachloroethene dechlorination, although lactate-fed inocula demonstrated longer-term dechlorinating capability.
The pathway for NH3 permeation across the apical membrane of MDCK cells was determined by measuring the effect of membrane fluidizing agents, protein reactive agents, and temperature on cellular NH3 influx. The rate of NH3 influx was calculated from the time course of increase in intracellular pH (pHi), measured with 2,7-biscarboxyethyl-5(6)-carboxyfluorescein, when MDCK cell monolayers were exposed to NH4Cl. The apical membrane NH3 permeability was 7.13 +/- 0.37 x 10(-3) cm/s (n = 12) at 37 degrees C and 1.23 +/- 0.07 x 10(-3) cm/s (n = 7) at 18 degrees C. In comparison, apical membrane permeability at 37 degrees C to the weak acids, valeric acid and acetic acid, were 1.39 +/- 0.11 x 10(-2) cm/s (n = 4) and 6.93 +/- 0.11 x 10(-3) cm/s (n = 4), respectively. The activation energy for NH3 permeation was 15.0 +/- 1.0 kcal/mol (17.5 degrees C-37.5 degrees C). In the presence of the membrane fluidizing agents, heptanol or chloroform, NH3 permeability increased in a dose-dependent manner. Heptanol (15 mM) significantly decreased the activation energy for NH3 permeation to 4.4 +/- 0.6 kcal/mol, P less than 0.001. The carboxyl reactive agent (1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide metho-p-toluensulfonic acid 1 mM), aminoreactive agents (4,4'-diisothiocyanostilbene-2,2'-disulfonic acid 50 microM; picrylsulphonic acid 1 mM), the sulphydryl reactive agent (p-chloromercuriphenylsulfonic acid 1 mM), and the nonspecific membrane protein cleaving agent pronase (1 mg/ml) had no effect on the NH3 influx. The results suggest that NH3 permeates the plasma membrane of MDCK cells via a lipid pathway.
The mechanisms of biotin reabsorption in rat kidney cortex were investigated using isolated brush-border membrane vesicles. An inwardly directed Na+ gradient specifically stimulated a transient biotin overshoot. Biotin transport was not affected by a valinomycin-induced K(+)-diffusion potential, and biotin(-)-Na+ stoichiometry was found to be 1:1. As a function of concentration, the uptake showed saturation in the presence of a Na+ gradient with an apparent Michaelis constant (Km) of 55 microM and Vmax of 217 pmol.mg protein-1.25 s-1. Desthiobiotin, 250 microM, norbiotin, bisnorbiotin, thioctic acid, valeric acid, probenecid, and nonanoic acid inhibited the transport of 30 microM biotin, whereas other biotin derivatives, as well as biocytin and organic acids found in the urine of biotinidase-deficient patients, did not. Preloading of the vesicles with biotin, desthiobiotin, norbiotin, and thioctic acid in the presence of Na+ increased initial uptake of biotin from the incubation medium (trans-stimulation). Our results indicate that biotin absorption in rat kidney fulfills the criteria for a specific carrier-mediated and electroneutral Na(+)-biotin- cotransport in a 1:1 ratio. The results are discussed in context with congenital biotinidase deficiency in humans.
OBJECTIVE: The objective of this study is to obtain the optimized separating conditions of formic acid, acetic acid, propionic acid, butyric acid, valeric acid, lactic acid, succinic acid from bacteria using reversed-phase high performance liquid chromatograph (RP-HPLC), and to set up a reliable progress in identifying the end-products of oral bacteria. METHODS: The conditions of separating seven short-chain fat acids were optimized using the method of gradient RP-HPLC. RESULTS: The study showed that the optimal chromatographic parameters were: 1.0 ml/min and 2.0 ml/min of flow rates, and the wavelength of ultraviolet detection was 217 nm. The composition of eluent: the ratio of phosphate buffer (pH 2.8) to methanol is 95:5 (v/v), gradient 5%-50%. The liner gradient wash will be performed in 10 minutes at room temperature. The average recovery rate was 92%, and RSD was 1.8%. CONCLUSION: The results of this paper will be helpful to determine acid composition of end products of oral bacteria.
The detection of volatile fatty acids (VFA) by gas chromatography of 85 purulent specimens from abscesses or pyogenic infections in cats, dogs, rodents, and ruminants was compared with the results of bacteriologic culturing, and proved to be a rapid means of presumptively diagnosing anaerobic infections. Of 83 bacteriologically positive specimens, 52 (61%) yielded obligate anaerobes and in 50 specimens, 1 or more VFA (butyric acid, isobutyric acid, valeric acid, isovaleric acid, caproic acid, or isocaproic acid) was detected. Forty-six specimens were positive for culturing of anaerobes and for detection of 1 or more of these VFA. By contrast, pus from infections caused by (facultative) aerobic microorganisms contained no VFA or only acetic and/or propionic acid.
OBJECTIVE: To identify in vitro effects of hydrochloric acid, valeric acid, and other volatile fatty acids (VFAs) on the pathogenesis of ulcers in the nonglandular portion of the equine stomach. SAMPLE POPULATION: Gastric tissues from 13 adult horses. PROCEDURE: Nonglandular gastric mucosa was studied by use of Ussing chambers. Short-circuit current (Isc) and potential difference were measured and electrical resistance and conductance calculated after tissues were bathed in normal Ringer's solution (NRS) or NRS and hydrochloric, valeric, acetic, propionic, and butyric acids. Treated tissues were examined histologically. RESULTS: Incubation in 60mM valeric acid at pH < or = 7.0 abruptly and irreversibly abolished Isc, which was followed by a slower decrease in resistance and an increase in conductance. Incubation in 60mM acetic, propionic, and butyric acids and, to a lesser extent, hydrochloric acid at pH < or = 7.0 significantly decreased Isc, which was followed by an increase in resistance and a decrease in conductance. CONCLUSIONS AND CLINICAL RELEVANCE: Incubation in valeric acid at pH < or = 7.0 caused a dramatic decrease in mucosal barrier function in the nonglandular portion of the stomach. Changes in barrier function attributable to exposure to valeric acid were associated with histopathologic evidence of cellular swelling in all layers of the nonglandular mucosa. Because of its high lipid solubility, valeric acid penetrates the nonglandular gastric mucosa, resulting in inhibition of sodium transport and cellular swelling. Valeric acid and other VFAs in gastric contents may contribute to the pathogenesis of ulcers in the nonglandular portion of the stomach of horses.
OBJECTIVE: To investigate the volatile fatty acids in gingival crevicular fluid (GCF) and to analyze the relationship between the levels of the volatile fatty acids and chronic periodontitis. METHODS: GCF samples taken from 37 patients with chronic periodontitis and 16 volunteers with healthy periodontal status were analyzed by capillary electrophoresis. RESULTS: The detection frequencies and concentrations of succinic acid, butyric acid and valeric acid were significantly higher in GCF of chronic periodontitis than in that of healthy group. The detection frequencies of propionic acid had no statistic difference between the two groups, but the concentrations of it was significantly higher in inflammation group. We also found that the concentrations of succinic acid, propionic acid and butyric acid were significantly lower in shallow pockets than that in deep pockets. CONCLUSIONS: The volatile fatty acids, especially succinic acid, propionic acid, butyric acid and valeric acid were associated significantly with the severity and inflammation of periodontal disease. The levels of succinic acid, propionic acid and butyric acid in GCF were related to pocket depth.
The effectiveness of wood vinegars was studied for controlling offensive odors from piggery wastes. Odorant chemicals and the sense of odors from piggery farm, covered with polyvinylchloride film to prevent the volatilization to ambient air, were measured in both laboratory and in situ in order to evaluate the degree of deodorization. The results obtained were as follows: 1. The odorant chemicals from piggery wastes were identified as ammonia, methyl sulfide, hydrogen sulfide, butyric acid and valeric acid. Among them, the major chemicals being responsible for the sense of odor were butyric acid and valeric acid. 2. In a laboratory experiment using an air tight vessel, the amount of wood vinegars needed for deodorization of piggery wastes was 6.6%; The removal efficiency of the odorant chemicals volatilized from the wastes was from 70 to 90% of control levels. 3. In situ test at a piggery farm showed that removal efficiency of odorant chemicals was similar to that of the laboratory experiment. In sensory tests, authorized by the Environmental Protection Agency of Japan, minimal offensive odors were observed with the addition of 6.6% wood vinegars. Furthermore, flies were rarely observed, indicating that wood vinegars may play an important role as a repellent.
The contents of volatile fatty acids in the rumen fluid were studied in calves at an early (seven weeks of age) and traditional weaning term (nine weeks of age). Weaning at the age of seven weeks enabled to save 16.2 kg of milk replacer per calf; there was also a lower content of concentrates in the starter feed with a supplement of Amylastim. The health condition of the calves was good in both groups. The average daily weight gains for the period from the second to the ninth week of age were 0.550 kg in the early weaned calves and 0.690 kg in the calves weaned at a normal time. Early weaning had a positive influence on the development of rumen metabolism. Calves weaned at the age of seven weeks, compared with those weaned at the age of nine weeks, had much higher concentrations of VFA (p less than 0.05) in rumen fluid (at the age of seven weeks: 130.49 mmol per litre vs. 111.53 mmol per litre; at the age of eight weeks: 119.74 mmol per litre vs. 96.98 mmol per litre). Early weaned calves had the statistically significantly (p less than 0.05) higher proportions of propionic acid, butyric acid, i-butyric acid, and valeric acid. Later-weaned calves had the significantly higher (p less than 0.05) contents of acetic acid and i-valeric acid.
Copolyesters of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) were produced by Burkholderia cepacia D1 at 30 degrees C in nitrogen-free culture solutions containing n-butyric acid and/or n-valeric acid. When n-valeric acid was used as the sole carbon source, the 3HV fraction in copolyester increased from 36 to 90 mol% as the concentration of n-valeric acid in the culture solution increased from 1 to 20 g/l. The addition of n-butyric acid to the culture solution resulted in a decrease in the 3HV fraction in copolyester. The copolymers biosynthesized by this method were mixtures of random copolymers having a wide variety of composition of the 3HV component. The melting points of the fractionated copolymers show a concave curve with the minimum at the 3HV content of approximately 40 mol%. The alpha-parameter of lattice indices of the P(3HB) crystal for the fractionated copolymers largely increased as the 3HV composition increased. Biodegradability of the copolymer increased with the lower content of 3HV composition and/or the lower crystallinity.