Characterization of CDC group DF-3 by cellular fatty acid analysis.

Fourteen strains of Centers for Disease Control group DF-3 bacteria were examined for cellular fatty acid composition to evaluate their chemical relatedness to known bacterial species and groups. The fatty acids were liberated from whole cells by base hydrolysis, methylated, and analyzed by capillary gas-liquid chromatography. All group DF-3 strains possessed a distinct fatty acid profile which was characterized by large amounts (24%) of 12-methyltetradecanoate (a-C15:0), moderate amounts of saturated iso-branched-chain acids (i-C14:0 and i-C15:0), and small to moderate amounts of both branched- and straight-chain hydroxy acids (3-OH C15:0, i-3-OH C16:0, 3-OH C16:0, and i-3-OH C17:0). This fatty acid profile was unique as compared with the profiles of other bacteria we have previously tested but was most similar to the profiles of Capnocytophaga species.     

Chemical characterization of clinical isolates which are similar to CDC group DF-3 bacteria.

Six clinical isolates, taken from blood or wounds, that had biochemical characteristics most similar to Centers for Disease Control group DF-3 bacteria were examined for cellular fatty acid composition and isoprenoid quinone content to evaluate their chemical relatedness to known bacterial species and groups. The fatty acids were liberated from whole cells by base hydrolysis, methylated, and analyzed by capillary gas-liquid chromatography. The isoprenoid quinones were extracted from lyophilized whole cells and analyzed by reverse-phase high-performance liquid chromatography. All six strains, which were designated group DF-3-like, possessed a distinct fatty acid profile that was characterized by large amounts (greater than 20%) of 13-methyltetradecanoate (i-C15:0) and 12-methyltetradecanoate (a-C15:0), moderate amounts of saturated branched-chain 13-carbon acids (i-C13:0 and a-C13:0) and hexadecanoate (n-C16:0), and small to moderate amounts of both branched- and straight-chain hydroxy acids (i-3-OH-C15:0, 3-OH-C16:0, i-3-OH-C17:0, and 2-OH-C17:0). This fatty acid profile was unique compared with the profiles of group DF-3 and other bacteria we have previously tested and is useful for the rapid identification of group DF-3-like isolates. The isoprenoid quinone content of four group DF-3-like strains was similar, with ubiquinone-9 (Q-9) and Q-10 as their major quinones, while the other two group DF-3-like strains contained Q-7 as their major quinones, with smaller amounts of Q-8 and Q-9.     

Cellular fatty acids of Alcaligenes and Pseudomonas species isolated from clinical specimens.

The cellular fatty acid composition of 25 clinical isolates of Alcaligenes and Pseudomonas was determined by gas-liquid chromatography (GLC). The GLC fatty acid profiles of three species of Pseudomonas were markedly different from those of Alcaligenes. The most significant differences were the presence and relative amounts of hydroxy, branched-chain, and cyclopropane fatty acids. One of the major fatty acids in A. faecalis was a 17-carbon cyclopropane (17 delta) acid, whereas a 15-carbon branched-chain acid (13-methyl tetradecanoate) characterized isolates of P. putrefaciens. The determination of these fatty acids by GLC provides a rapid and specific means of distinguishing clinical isolates of Pseudomonas and Alcaligenes.     

Gas-liquid chromatographic analysis of cellular fatty acid methyl esters in Aeromonas species

The cellular fatty acids of 39 strains belonging to the genus Aeromonas (Aeromonas hydrophila, Aeromonas caviae, Aeromonas sobria, Aeromonas media, Aeromonas schubertii, Aeromonas veronii) were determined by high resolution gas-liquid chromatography. The fatty acid profiles were characterized by major amounts (60% or more) of one saturated (hexadecanoic acid = 16:0) and two unsaturated (hexadecenoic acid = 16:1 and octadecenoic acid = 18:1) acids. While the majority of the strains of the six species exhibited, qualitatively, very similar fatty acid compositions, only minor and inconsistent differences could be observed which would be useful for a distinction of the different taxons. The following fatty acids were qualitatively identified: 12:0, i-13:0, 14:0, 3-OH 13:0, i-15:0, 15:0, 2-OH 14:0, 3-OH 14:0, i-16:0, 16:1, 16:0, i-17:1, i-17:0, a-17:0, 17:0 cyclopropane, 17:1, 17:0, 18:1 (3 isomers), 18:0 and i-20:0. Excellent congruence was found in reproducibility studies. Fatty acid analyses show a great homogeneity within the group and the technique does not appear to be the ideal method in distinguishing between Aeromonas species.     

Fatty acid composition of gliding bacteria: oral isolates of Capnocytophaga compared with Sporocytophaga.

The extractable and bound lipids and cellular fatty acids of the gram-negative gliding bacteria, Capnocytophaga sputigena, C. gingivalis, and C. ochracea were compared to the non-host-related gliding bacterium Sporocytophaga myxococcoides. The extractable lipids represented between 17 and 28% of the cell dry weight, whereas only 2 to 4% of the lipids were in the bound fraction. The methyl esters of the cellular fatty acids were mainly aC15:0, which accounted for 69 to 73% of the total extractable fatty acids; S. myxococcoides had a similar distribution of branched-chain fatty acids; however, aC17:0 was the predominant fatty acid in this free-living gliding organism.     

Cellular fatty acid composition as an adjunct to the identification of asporogenous, aerobic gram-positive rods

Cellular fatty acid (CFA) compositions of 561 asporogenous, aerobic gram-positive rods were analyzed by gas-liquid chromatography as an adjunct to their identification when grown on blood agar at 35 degrees C. The organisms could be divided into two groups. In the first group (branched-chain type), which included coryneform CDC groups A-3, A-4, and A-5; some strains of B-1 and B-3; "Corynebacterium aquaticum"; Brevibacterium liquefaciens; Rothia dentocariosa; and Listeria spp., the rods had sizable quantities of antiesopentadecanoic (Ca15:0) and anteisoheptadecanoic (Ca17:0) acids. Other species with these types of CFA included B. acetylicum, which contained large amounts of isotridecanoic (Ci13:0) and anteisotridecanoic (Ca13:0) acids. CFAs useful for distinguishing among Jonesia denitrificans, Oerskovia spp., some strains of CDC groups B-1 and B-3, Kurthia spp., and Propionibacterium avidum were hexadecanoic (C 16:0) acid, isopentadecanoic (Ci15:0) acid, and Ca15:0). The second group (straight-chained type), which included Actinomyces pyogenes; Arcanobacterium haemolyticum; C. bovis; C. cystitidis; C. diphtheriae; C. flavescens, "C. gentalium"; C. jeikeium; C. kutscheri; C. matruchotii; C .minutissimum; C. mycetoides; C. pilosum; C. pseudodiphtheriticum; "C. pseudogenitalium"; C. pseudotuberculosis; C. renale; CDC groups 1, 2, ANF-1, D-2, E, F-1, F-2, G-1, G-2, and I-2; C. striatum; "C. tuberculostearicum"; C. ulcerans; C. vitarumen; C. xerosis; and Erysipelothrix rhusiopathiae, was typified by significant quantities of hexadecanoic (C16:0) and oleic acids (C18:cis9), with differences in the amounts of linoleic acid (C18:2), stearic acid (C18:0), an unnamed peak (equivalent chain length, 14.966), and small quantities of other known saturated and unsaturated fatty acids. CFA composition of these organisms was sufficiently discriminatory to assist in classification but could not be used as the sole means of identification.     

Cellular fatty acids of Flavobacterium meningosepticum and Flavobacterium species group IIb.

The cellular fatty acid profiles of Flavobacterium meningosepticum and Flavobacterium species group IIb were markedly different from those of related bacteria. The profiles were characterized by the presence of 13-methyl-tetradecanoate and three uncommon acids: 2-hydroxy-13-methyl-tetradecanoate, 15-methyl-hexadecanoate, and 3-hydroxy-15-methyl-hexadecanoate.     

Relationship between lipofuscin granules and polyunsaturated fatty acids in the housefly, Musca domestica

The fractional volume occupied by lipofuscin granules in epithelial cells of the midgut or oenocytes of abdominal fat body of 3-day-old and 13-day-old male houseflies was determined in two groups of flies by electron microscopic morphometry. One group had developed from larvae reared on diets containing no added polyunsaturated fatty acids and the second from larvae reared on diets containing added linoleic acid. No polyunsaturated fatty acids could be detected in the lipids of the first group of flies using a method which would have detected their presence in amounts greater than 0.1% of the total esterified fatty acids. The second group contained at least two hundred times more than this minimal level. The volume of lipofuscin granules increased significantly (p less than 0.01) (about threefold for the fat body and twofold in midgut cells) between 3 days and 13 days of age but no statistically significant difference was seen between the two groups of flies at the same age. The results show that if lipofuscin formation depends on the oxidation of polyunsaturated fatty acids in the housefly, then extremely small amounts of the acids are involved which lie below the detection limit of the methods employed. The age-associated small increase of extractable fluorescence seen previously in the linoleic acid group of flies is not associated with an increase in the lipofuscin granules.     

Isoprenoid quinone content and cellular fatty acid composition of Campylobacter species

A total of 36 strains of Campylobacter species were examined for isoprenoid quinones and cellular fatty acids. The isoprenoid quinone content was determined by reverse-phase high-pressure liquid chromatography, and the fatty acids were determined by capillary gas-liquid chromatography. All Campylobacter species contained menaquinone-6 (2-methyl-3-farnesyl-farnesyl-1,4-naphthoquinone) and a methyl-substituted menaquinone-6 (2,[5 or 8]-dimethyl-3-farnesyl-farnesyl-1,4-napthoquinone) as the major isoprenoid quinones. The latter menaquinone has not been reported in other bacteria and may prove to be a useful chemical marker of Campylobacter species. Campylobacter jejuni and most Campylobacter coli were distinguished from other campylobacteria by the presence of a C19 cyclopropane acid, and Campylobacter sputorum subsp. mucosalis differed from other species by the presence of lauric acid.     

Comparison of the effects of acid and base hydrolyses on hydroxy and cyclopropane fatty acids in bacteria

The cellular fatty acid compositions of Legionella oakridgensis, Brucella suis, Pseudomonas aeruginosa, and Francisella tularensis were compared after base hydrolysis (saponification), acid hydrolysis, and acid methanolysis procedures were used to release the fatty acids. The branched-chain, unsaturated, saturated, and ester-linked hydroxy acids were released as effectively with saponification at 100 degrees C for 30 min as with acid hydrolysis or acid methanolysis at 85 degrees C for 16 h. Although the amide-linked hydroxy acids were released more effectively by acid hydrolysis or acid methanolysis, these methods degraded the cyclopropane fatty acids, producing a number of new peaks or artifacts in the chromatograms. Cyclopropane fatty acids were not degraded by saponification, and at least 50% of the hydroxy acids were released when the cells were saponified with 15% NaOH in 50% aqueous methanol. Thus, the results show that saponification for 30 min at 100 degrees C with 15% NaOH, followed by methylation is an excellent method for routine fatty acid analysis of bacteria and for screening cultures whose identity and fatty acid composition are unknown.     

De novo arachidonic acid synthesis in Perkinsus marinus, a protozoan parasite of the eastern oyster Crassostrea virginica

The capability of synthesizing fatty acids de novo in the meront stage of the oyster protozoan parasite, Perkinsus marinus, was investigated employing stable-isotope-labeled precursors (1,2 13C-acetate and palmitic-d(31) acid). Fatty acid methyl esters derived from 1,2 13C-acetate and palmitic-d(31) acid were analyzed using gas chromatography/mass spectrometry and gas chromatography/flame ionization detection. Results revealed that in vitro cultured P. marinus meronts utilized 13C-acetate to synthesize a range of saturated and unsaturated fatty acids. The saturated fatty acids 14:0, 16:0, 18:0, 20:0, 22:0, 24:0 and the unsaturated fatty acids, 18:1(n-9), 18:2(n-6), 20:1(n-9), 20:2(n-6), 20:2(n-9), 20:3(n-6), 20:4(n-6) were found to contain 13C, after 7, 14, and 21 days incubation with the precursor. This indicates that meronts can synthesize fatty acid de novo using acetate as a substrate. Meronts efficiently elongated 16:0-d(31) to 18:0, 20:0, 22:0, 24:0, but desaturation activity was limited, after 7 and 14 days cultivation. Only a small quantity of 18:1-d(29) was detected. This suggests that meronts cannot directly convert exogenous palmitic acid or its products of elongation to unsaturated counterparts. The ability to synthesize 20:4(n-6) from acetate is particularly interesting. No parasitic protozoan has been reported to be capable of synthesizing long chain essential fatty acids, such as 20:4(n-6) de novo. Future study will be directed to determine whether the observed in vitro activities indeed reflect the in vivo activities, when meronts are associated with the host.     

Changes in the incorporation of free fatty acids upon the stimulation of human polymorphonuclear leukocytes

We have investigated the incorporation of free fatty acids into the cellular lipids of human polymorphonuclear leukocytes (PMN). Resting PMN incorporated both saturated and unsaturated fatty acids into triacylglycerol with only small amounts incorporated into the phospholipids. In contrast, PMN stimulated with the calcium ionophore A23187 incorporated significantly higher amounts of fatty acids, predominantly those other than arachidonic acid, into phosphatidylcholine and phosphatidylinositol, with reduced incorporation into triacylglycerol. Stimulation of PMN with serum-treated zymosan or the chemotactic peptide f-met-leu-phe but not phorbol myristate acetate, also increased the incorporation of fatty acids into these phospholipids. This stimulation-induced incorporation of fatty acids into cellular phospholipids was directed exlusively into position 2 of the lipid and probably reflects the reacylation of lysophospholipids after the release of arachidonic acid by phospholipase A2.     

辛伐他汀对高糖高脂诱导人骨髓间充质干细胞凋亡的影响

背景：研究表明,他汀类药物能够促进骨髓间充质干细胞的增殖与黏附能力,抑制高糖高脂培养下骨髓间充质干细胞的凋亡。 目的：观察辛伐他汀对高糖高脂诱导条件下人骨髓间充质干细胞凋亡的影响。 方法：将0.001,0.01,0.1,1.0 μmol/L辛伐他汀分别与高糖高脂诱导条件下人骨髓间充质干细胞培养48 h,以正常培养骨髓间充质干细胞及高糖高脂诱导条件下培养的骨髓间充质干细胞为对照。倒置显微镜下观察细胞形态,MTT法比较不同浓度辛伐他汀对高糖高脂环境下骨髓间充质干细胞存活率的影响,应用流式细胞术检测细胞凋亡,加入PI3K/Akt信号转导通路抑制剂LY294002后辛伐他汀对骨髓间充质干细胞凋亡的影响。 结果与结论：与高糖高脂诱导组比较,辛伐他汀0.01,0.1,1.0 µmol/L组骨髓间充质干细胞存活率均升高(P < 0.01),其中辛伐他汀浓度在0.1 μmol/L时骨髓间充质干细胞存活率升高最显著(P < 0.01);同时流式细胞仪检测结果显示,辛伐他汀0.01,0.1,1.0 µmol/L组细胞凋亡率下降(P< 0.01),其中0.1 µmol/L组凋亡率下降最显著(P < 0.01)。0.1 µmol/L辛伐他汀对骨髓间充质干细胞的影响可被LY294002阻断。说明辛伐他汀能抑制高糖高脂诱导条件下骨髓间充质干细胞的凋亡,其机制可能与PI3K/Akt信号途径有关。     

Omega-6/omega-3 essential fatty acid ratio in cultured beluga sturgeon

The balance of omega-6/omega-3 fatty acids is an important factor in human health. High omega-6 fatty acid content in the diet along with a high omega-6/omega-3 ratio can lead to the development of many chronic diseases. Conversely, higher levels of omega-3 fatty acids such as eicosapentaenoic acid (C20:5, EPA) and docosahexaenoic acid (C22:6, DHA) and lower omega-6/omega-3 ratio are desirable for human health. We determined the lipid content in beluga flesh which was 6.5?%. Monounsaturated fatty acids were the main class of fatty acids followed by polyunsaturated and saturated fatty acids. The EPA and DHA contents of cultured beluga flesh were lower than those of other cultured sturgeon species, which may be due to the contents of these fatty acids in the diet. The content of omega-6 fatty acids (25.7?%) was higher than omega-3 fatty acids (8.58?%), giving an omega-6/omega-3 ratio of 3.00.     

Cellular fatty acid composition of Campylobacter fetus.

The cellular fatty acid composition of 29 human and animal isolates of Campylobacter fetus was determined with gas-liquid chromatography. All strains contained small amounts of 3-hydroxytetradecanoic acid, suggesting the presence of lipid A. Each of 13 different blood or stool isolates of C. fetus subsp. jejuni obtained from humans or fowl contained a 19-carbon cyclopropane acid which was not present in C. fetus subsp. fetus (6 strains from cattle) or C. fetus subsp. intestinalis (10 strains from humans and cattle). These findings indicate that C. fetus subsp. jejuni can be rapidly differentiated from other C. fetus by gas-liquid chromatography analysis of cellular fatty acids.     

Unusual fatty acid substitution in lipids and lipopolysaccharides of Helicobacter pylori.

Cellular fatty acids, phospholipid fatty acids, and lipopolysaccharide fatty acids of four strains of Helicobacter pylori were analyzed by gas-liquid chromatography. The presence of myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, 19-carbon cyclopropane fatty acid, beta-hydroxypalmitic acid, and beta-hydroxystearic acid was confirmed. In phospholipids, myristic acid and 19-carbon cyclopropane fatty acid were the major fatty acids. Hydroxy fatty acids and unsaturated fatty acids were not detected or occurred only in small amounts. The major fatty acids of lipopolysaccharides were stearic acid, beta-hydroxypalmitic acid, and beta-hydroxystearic acid. Unsaturated fatty acids and 19-carbon cyclopropane fatty acid were not found. The unusual compositions of H. pylori phospholipid and lipopolysaccharide fatty acids may have important implications for the taxonomy, physicochemical membrane properties, and biological activity of lipopolysaccharides.     

Effects of fatty acids on motility retention by Treponema pallidum in vitro.

Treponema pallidum (Nichols virulent strain) was incubated under 75% N2 + 20% H2 + 5% CO2 in prereduced serum-free modified Eagle-Richter medium supplemented with different concentrations of various long-chain fatty acids complexed with fatty acid-free bovine serum albumin. Motility retention was greater in medium with oleic acid containing 15 rather than 2 mg of albumin per ml. Palmitic, stearic, oleic, or linoleic acid alone caused rapid loss of motility at concentrations as low as 5 microgram/ml. Elaidic acid (92 microgram/ml) alone had no effect on motility. Various combinations of saturated plus unsaturated fatty acids did not inhibit motility retention or were less inhibitory than either of the individual fatty acid components. The combination of palmitic plus oleic acids was least toxic. Rapid loss of motility occurred with pairs of unsaturated or saturated fatty acids, or with Tween 40, 60, or 80, alone or combined. Autoxidation of oleic acid resulted in decreased toxicity for T. pallidum but increased toxicity for baby hamster kidney cells.     

Fatty Acid Requirements of the Kazan 5 and Reiter Strains of Treponema pallidum

The fatty acid requirements of two avirulent treponemes were investigated by using a “lipid-poor” albumin-thioglycolate medium. The Kazan 5 and the Reiter stains of Treponema pallidum required a pair of fatty acids for growth. One member of the pair was saturated and the other was unsaturated. The saturated fatty acids could contain either an odd or even number of carbon atoms, but a chain length of at least 14 carbon atoms was necessary. Unsaturated fatty acids with one, two, or three double bonds were satisfactory if they contained 15 or more carbon atoms. The pair of fatty acids could be replaced with a single 18-carbon monounsaturated fatty acid if it was in the trans configuration rather than the naturally occurring cis form.     

Magnetic resonance spectroscopy and chromatographic methods identify altered lipid composition in human renal neoplasms

We report on the characterization of the lipid obtained from cortical and medullary normal human kidney tissue, benign renal neoplasms (oncocytoma) and 2 different types of malignant renal neoplasms (chromophobic cell carcinoma and clear cell carcinoma). The total lipid fractions were analyzed by 13C magnetic resonance spectroscopy and thin-layer chromatography, whereas the composition of the total fatty acids and the content of total cholesterol were determined by gas chromatography. alpha-Tocopherol was detected and quantified by high-performance liquid chromatography. The spectroscopic and chromatographic analysis revealed significant differences in the renal tissues examined. It was confirmed that cholesteryl esters (mainly oleate) are typical of clear cell renal carcinomas. Their potential role as prognostic and diagnostic factors is discussed, with particular emphasis on its capability to indicate the tumor diffusion in healthy renal parenchyma. alpha-Tocopherol is prevalent in clear cell carcinoma and it is present in nearly the same low amounts in cortex, medulla and chromophobic cell renal carcinoma. Q10 coenzyme and dolichols were detected by thin-layer chromatography and they are present in significant amounts in the cortex and the benign oncocytoma. Great variations were found in the distribution of saturated and unsaturated fatty acids, especially in the docosapentaenoic, docosahexaenoic and arachidonic acids and the corresponding omega-6/omega-3 fatty acids ratio.     

Effects of prostaglandin E2, analogs, fatty acids, and indomethacin on fibrinogen level.

Dose-response relations in rabbits for 3-h intravenous infusion of prostaglandin E2 (PGE2) and (15S)-15-methyl-1-prostaglandin E2 methyl ester (MePGE2) on plasma fibrinogen and systolic blood pressure were determined and described by regression equations. MePGE2 was 20 times more active than PGE2. Fibrinogen synthetic rate responses to PGE2 and MePGE2 were estimated. Infusion of the PGE2 precursor, arachidonic acid, elevated plasma fibrinogen, but fibrinogen response to 0.5-9 mg/kg arachidonic acid was unrelated to dose and half that given by 3 mg/kg PGE2. Slow infusion of several other fatty acids raised plasma fibrinogen as effectively as arachidonic acid, but prostaglandins D2 and F2alpha had only a slight effect. Infusion of 30 times the indomethacin dose that blocks platelet prostaglandin synthetase did not alter the plasma fibrinogen response to arachidonic acid. Indomethacin did not inhibit plasma fibrinogen elevations following ACTH or endotoxin infusion, or subcutaneous turpentine injection. Intravenous infusion of two cyclic ether prostaglandin endoperoxide analogs, (15S)-hydroxy-9alpha, 11alpha-(epoxymethano) prosta-5Z, 13E-dienoic acid, and (15S)-hydroxy-11alpha, 9alpha(epoxymethano) prosta-5Z, 13E-dienoic acid, failed to increase plasma fibrinogen.