Branched-chain Saturated Fatty Acids Research Articles

Updating the fatty acid profiles of retail bovine milk in China based on an improved GC-MS method: implications for nutrition.

The importance of food components to potential benefits and risks to human health is gradually being consumer awareness. Milk is an important part of the lipid content of the human diet, and there are few detailed reports on the fatty acid (FA) profiles of retail milk. In the study, we developed a gas chromatography-mass spectrometry (GC-MS) method to simultaneously determine 82 FAs, including 11 even-chain saturated FAs, 10 odd-chain saturated FAs, 9 branched-chain saturated FAs, 30 monounsaturated FAs, and 22 polyunsaturated FAs; this was applied to analyze samples (186 samples) of commercially available milk from 22 provinces throughout China and to evaluate the nutritional value of these samples based on FA-related indices. The results showed that the overall composition of milk FAs among the different regions was numerically similar, and minor FAs showed few differences. When considering the retail milk FA composition and dairy fat intake in China, regional variations have a limited impact on FA consumption. Moreover, milk accounts for approximately one-third and <10% of the maximum recommended intake of saturated FAs and trans-FAs in consumer diets, respectively. This study provides an updated report on the composition of FAs and the nutritional value of retail milk across China, which can serve as a reference for producers for future research on regulating milk FAs, for consumers to select milk, and for nutrition departments to formulate relevant nutritional guidance recommendations.

Fatty acids composition of actinobacteria isolated from mussels Mytilus galloprovincialis of the Black Sea Odesa Bay

Analysis of the fatty acid composition of total cellular lipids is important chemotaxonomic characteristic that is used to identify microorganisms, in particular actinobacteria, with the help of fatty acid spectra libraries. Actinobacteria are well known as producers of secondary metabolites, which are of pharmacological and commercial interest. A significant number of actinomycetes is associated with various marine benthic communities. The aim of this study was determination of the fatty acid composition and preliminary identification of actinobacteria isolated from mussels Mytilus galloprovincialis of the Black Sea Odesa Bay. Actinobacteria of 12 isolated strains were grown in a liquid medium at 28° C for 72 h. Methyl esters of fatty acids were determined on a gas chromatograph Agilent. The MIDI Sherlock microorganisms identification system was used to identify the studied strains. It was found that the fatty acids methyl esters profiles of studied actinobacteria strains were characterized by the predominance of 12-17 saturated branched-chain fatty acids with high content of 12-methyltridecanoic, 12-methyltetradecanoic, 14-methylpentadecanoic and 14-methylhexadecanoic fatty acids. All 12 actinobacteria strains isolated from mussels (Mytilus galloprovincialis) of Odesa Bay were identified as members of the Streptomyces genus. Keywords: Black Sea, fatty acid compositions, marine actinobacteria, Mytilus galloprovincialis, Streptomyces

Valproic Acid Initiates Transdifferentiation of the Human Ductal Adenocarcinoma Cell-line Panc-1 Into α-Like Cells.

Non-mesenchymal pancreatic cells are a potential source for cell replacement. Their transdifferentiation can be achieved by triggering epigenetic remodeling through e. g. post-translational modification of histones. Valproic acid, a branched-chain saturated fatty acid with histone deacetylase inhibitor activity, was linked to the expression of key transcription factors of pancreatic lineage in epithelial cells and insulin transcription. However, the potential of valproic acid to cause cellular reprogramming is not fully understood. To shed further light on it we employed next-generation RNA sequencing, real-time PCR, and protein analyses by ELISA and western blot, to assess the impact of valproic acid on transcriptome and function of Panc-1-cells. Our results indicate that valproic acid has a significant impact on the cell cycle, cell adhesion, histone H3 acetylation, and metabolic pathways as well as the initiation of epithelial-mesenchymal transition through acetylation of histone H3 resulting in α-cell-like characteristics. We conclude that human epithelial pancreatic cells can be transdifferentiated into cells with endocrine properties through epigenetic regulation by valproic acid favoring an α-cell-like phenotype.

Arsenic and Protein Expression: It might help to know the mechanism of As toxicity

Arsenic and Protein Expression: It might help to know the mechanism of As toxicity

Lipidomics of facial sebum in the comparison between acne and non-acne adolescents with dark skin

Lipidomics is advantageous in the study of sebum perturbations occurring in acne. An extended evaluation of the sebum lipid profiles in acne-prone sebaceous areas is lacking in dark skin. Yet, there is a void space in understanding how the building blocks of sebum lipids, i.e. individual fatty acids (FAs), are intertwined with acne-prone skin. We aimed to determine the sebum lipidome in facial areas of adolescents with and without acne in Nigeria. A cross-sectional analytical study was conducted in 60 adolescents/young adults divided in 30 acne patients (15F, 15M) and 30 age and sex-matched controls. Sebum samples obtained from foreheads and cheeks were analysed separately by gas chromatography–mass spectrometry (GCMS) and thin layer chromatography (HPTLC). Distributions of sebum components were investigated with multivariate ANOVA-simultaneous component analysis (ASCA). Sebum incretion in acne was paralleled by significantly higher abundance of triglycerides, wax esters, and squalene together with monounsaturated FAs (MUFAs), and straight chain saturated FAs (SFAs), especially those with odd-carbon chain, i.e. C13:0, C15:0, and C17:0. Profiling weight/weight percentage of individual components revealed that, in acne, the free FAs (FFAs) array was shifted towards higher relative abundance of the SFAs C15:0, C16:0, and C17:0 and lower percentage of the anteiso-branched FFAs with 12, 14, 16, and 18 carbons. In acne patients, MUFAs and PUFAs were quantitatively increased and decreased on foreheads and cheeks, respectively. Relative abundance of fatty alcohols was decreased in acne independent on the site. The results indicated that acne associates with site-specific derangement of the pathways regulating the balance among odd straight-chain and branched-chain SFAs, MUFAs, which included sapienate (C16:1n-10), PUFAs, and squalene.

Regulation of transgelin and GST-pi proteins in the tissues of hamsters exposed to sodium arsenite

Regulation of transgelin and GST-pi proteins in the tissues of hamsters exposed to sodium arsenite

Effects of fatty acid from deep-sea microorganisms on lipid bilayer membrane fluidity under high pressure: comparison of branched-chain and polyunsaturated fatty acid

In this study, the effects of unsaturated and saturated branchedchain fatty acids in biomembranes of microorganisms living under high temperature and high pressure on the fluidity of biomembranes were investigated by time-resolved fluorescence anisotropy measurements. First, the relationship between the order parameter S and the rotational diffusion coefficient Dw, which can be calculated from the fluorescence anisotropy measurements, and the motion of lipid molecules was investigated using lipids with three different structures, and it was found that the former was related to the spacing of lipid molecules and the latter to the motion of lipid molecules. Next, we investigated the S and Dw values of lipid bilayer membrane containing the saturated branched-chain fatty acid 12-methyltridecanoic acid (12-MTA) and the polyunsaturated fatty acid (PUFA) cis-4,7,10,13,16,19-docosahexaenoic acid (DHA). The results showed that 12-MTA increased the S value and decreased the Dw value. On the other hand, DHA tended to reduce the S value and increase the Dw value, albeit slightly. These results mean that 12-MTA narrows the molecular spacing of lipids and inhibits lipid molecular movement, while DHA tends to widen the molecular spacing of lipids and promote lipid molecular movement.

Effect of Pork Meat Replacement by Fish Products on Fatty Acid Content, Physicochemical, and Sensory Properties of Pork Pâtés

The aim of the study was to assess the influence of the addition of fish raw materials (roe or fish meat) on the quality and nutritional value of pork pâtés. The control group (n = 4) consisted of pork pâtés, I experimental group (n = 6) of pâtés with 20% addition of roe (perch and pike), and II group of pâtés with 20% addition of fish (perch and pike meat) (n = 6). The pâtés’ pH, color, and profiled texture analysis were instrumentally measured and water, protein, fat, ash, and fatty acid content were determined by reference methods. To assess the oxidative stability of lipids the measurement of peroxide number, thiobarbituric acid reactive substances and content of conjugated dienes and trienes was used. The degree of fat hydrolysis was determined on the basis of acid value. Sensory analysis was carried out using the scaling method, taking into account 12 unit quality characteristics. Products with roe and meat contained less fat (accordingly 15.9% and 14.1%) and showed lower calorific value (accordingly 225.6 and 208.6 kcal/100 g) compared to pork pâtés (20.2% of lipids, 267 kcal/100 g). Moreover, the addition of fish raw materials improved the index of nutritional quality for protein (from 3.2 to 3.9) and beneficially reduced the nutritional index for fat (from 2.2 to 1.9). Fish constituents modified, to a certain extent, the color, texture, and sensory properties of pâtés, while maintaining full acceptability in consumer assessment. The addition of fish roe significantly increased the healthful quality by improving the fatty acid profile of pâtés, in which the significantly highest content of n-3 fatty acids, including eicosapentaenoic and docosahexaenoic acids (accordingly 252.21, 43.17, and 107.94 mg/100 g product), as well the highest concentration of saturated branched chain fatty acids were determined (18.75 mg/100 g product).

Hierarchically porous FER zeolite obtained via FAU transformation for fatty acid isomerization

Skeletal isomerization of linear unsaturated fatty acids is important in the production of branched-chain saturated fatty acids with diverse applications. This reaction can be efficiently catalyzed by ferrierite (FER) zeolite. The reaction, however, suffers from diffusion limitations in the 10-membered ring channels. Herein, we report a method for the synthesis of hierarchically porous FER zeolite via transformation of FAU precursor driven by N-methylpyrrolidine (NMP) and amphiphile 1,2-dimethyl-3-hexadecyl-1H-imidazol-3-ium bromide (C16dMImz) as the structure-directing agent (SDA) and a mesoporogen, respectively, under hydrothermal conditions. This dual-template approach allows tuning the morphological and textural properties of the mesoporous FER materials by varying the concentration of the mesoporogen in the initial gel. The optimized FER sample is characterized by a high mesoporous volume (0.19 cm3 g-1), large external surface area (∼120 m2 g-1) and reduced crystal size in the a- and c-dimensions. This implies shortened diffusional pathways in the 10-membered ring channels. These modifications led to a significantly enhanced catalytic performance of hierarchical FER zeolite in the isomerization of fatty acids in comparison with a bulk FER reference zeolite.

Structural Identification of Monounsaturated Branched Chain Fatty Acid Methyl Esters by Combination of Electron Ionization and Covalent Adduct Chemical Ionization Tandem Mass Spectrometry.

Monounsaturated normal fatty acids (n-MUFA) and saturated branched chain fatty acids (BCFA) are structurally characterized by separate tandem mass spectrometry methods for double bond localization and for chain branching in their respective fatty acid methyl ester (FAME) derivatives; however, these methods have never been applied to branched monounsaturated FAME. Here, we report application of electron ionization (EI)-MS/MS and solvent-mediated covalent adduct chemical ionization (CACI)-MS/MS of monounsaturated BCFA methyl esters (MUBCFAME) of a chain length of 15-20 carbons. A novel system was used to implement CI with low vapor pressure reagents in a tabletop triple quadrupole MS. Anteiso-MUBCFA EI-MS/MS of the molecular ion (M) yields a characteristic diagnostic ion [M-29]+. iso-MUBCFA can be distinguished from n-MUFA by an ion intensity ratio of [M-32]+/[M-43]+, with iso-MUBCFA yielding a ratio greater than 1.7, while n-MUFA yields a ratio less than 1.0. Chain branching at the iso and anteiso positions, terminal isopropyl and sec-butyl, respectively, do not alter CACI-MS/MS diagnostic ions compared to normal BCFA, enabling double bond positions of MUBCFA to be determined with the analogous α and ω diagnostic ions from cleavage on both sides of the erstwhile double bond. Taken together, this straightforward FAME-based technique via combination of EI-MS/MS and CACI-MS/MS enables fundamental structural identification of MUBCFA without standards.

Sea Lions Develop Human-like Vernix Caseosa Delivering Branched Fats and Squalene to the GI Tract

Vernix caseosa, the white waxy coating found on newborn human skin, is thought to be a uniquely human substance. Its signature characteristic is exceptional richness in saturated branched chain fatty acids (BCFA) and squalene. Vernix particles sloughed from the skin suspended in amniotic fluid are swallowed by the human fetus, depositing BCFA/squalene throughout the gastrointestinal (GI) tract, thereby establishing a unique microbial niche that influences development of nascent microbiota. Here we show that late-term California sea lion (Zalophus californianus) fetuses have true vernix caseosa, delivering BCFA and squalene to the fetal GI tract thereby recapitulating the human fetal gut microbial niche. These are the first data demonstrating the production of true vernix caseosa in a species other than Homo sapiens. Its presence in a marine mammal supports the hypothesis of an aquatic habituation period in the evolution of modern humans.

Phytanic acid induced neurological alterations in rat brain synaptosomes and its attenuation by melatonin

Phytanic acid (3,7,11,15-tetramethylhexadecanoic acid) (Phyt) is a saturated branched chain fatty acid which originates after the breakdown of chlorophyll molecule, phytol. It plays an important role in a variety of metabolic disorders with peroxisomal impairments. The aim of our investigation was to evaluate the adverse effects of Phyt on synaptic functions by using synaptosomal preparation of rat brain as an in vitro model and the possible protective role of melatonin against Phyt-induced neurotoxicity. Melatonin is an antioxidant, secreted by the pineal gland. Melatonin and its metabolites have neuroprotective effects on cellular stress, by reducing reactive oxygen species (ROS) and reactive nitrogen species (RNS). In the present investigation, synaptosomes prepared from rat brain were co-treated with melatonin (10μM) and Phyt (50μM) for 2h. Co-treatment of Phyt with melatonin significantly restored the altered levels of protein carbonyl (PC) contents and lipid peroxidation (LPO). It also replenished the Phyt-induced alterations on the levels of non-enzymatic antioxidant defence reduced glutathione (GSH), enzymatic antioxidants such as catalase (CAT) and superoxide dismutase (SOD) and synaptosomal integral enzymes such as AChE, Na+, K+-ATPase and MAO. We observed that Phyt induced oxidative stress in synaptosomes as indicated by an elevation in the generation of ROS and melatonin was able to inhibit the elevated ROS generation. Moreover, the neurotoxic effects elicited by Phyt on NO level and membrane potential were totally prevented by the treatment of melatonin. The results of our investigation emphasize the potential use of melatonin as a nutraceutical and mitigatory agent against Phyt-induced oxidative stress.

Phytanic Acid-Induced Neurotoxicological Manifestations and Apoptosis Ameliorated by Mitochondria-Mediated Actions of Melatonin.

Phytanic acid, a saturated branched chain fatty acid and a major constituent of human diet, is predominantly found in dairy products, meat, and fish. It is a degradation product from the phytol side chain of chlorophyll. Degradation of PA is known to occur mainly in peroxisomes via α-oxidation and in mitochondria via β-oxidation. Due to its β-methyl group present at the 3-position of the carbon atoms, PA cannot be β-oxidized. Although alteration in the metabolism of PA may play an important role in neurodegeneration, the exact mechanism behind it remains to be evaluated. In this study, we have described the potential of PA to induce neurotoxicity as an in vitro model (neuronal cell line, SH-SY5Y cells). Cells were pretreated with melatonin (10μM) for 1h followed by with and without PA (100μM) for 24h. In the present study, our data has confirmed that PA markedly increased both intracellular reactive oxygen species and reactive nitrogen species levels. Our results have shown that PA treatment did not induce cell death by cleavage of caspase-3/PARP-1 mediated by mitochondria through intrinsic pathways; however, PA induced nitric oxide-dependent apoptosis in SH-SY5Y cells. Additionally, melatonin pretreatment reduced the cell death in SH-SY5Y cells. Melatonin also effectively exerted an antiapoptotic and anti-inflammatory action by regulating Bax, Bcl-2, p-NFκB, and iNOS expressions in SH-SY5Y cells. These results suggested that melatonin acted as an antioxidative and antiapoptotic agent by modulating ROS, apoptotic proteins, and inflammatory responses under BCFA-induced neurotoxic conditions. The protective effects of melatonin depend on direct scavenging activity of free radicals and indirect antioxidant effects. Further deciphering of the cellular and molecular mechanism associated with neuroprotection by melatonin is warranted in BCFA-induced neurotoxicity.

Structural diversity and possible functional roles of free fatty acids of the novel soil isolate Streptomyces sp. NP10.

Herein, a novel soil bacterium Streptomyces sp. NP10 able to grow outside usual streptomycetes optimum conditions (e.g., at 4°C, pH 9 and high NaCl concentration), exhibiting atypical hemolytic, DNAse, and cellulolytic activities, is described. This strain produces and excretes into the growth medium large amounts of free long-chain fatty acids (FAs). A concurrent lipidomics study revealed a large structural diversity of FAs with over 50 different n- and branched-chain, (un)saturated, and cyclopropane FAs (C7-C30) produced by this strain. Two of these, i-17:0cy9-10 and a-18:0cy9-10, represent new natural products and the first ever identified branched cyclopropane FAs. Both free and bound lipid profiles of Streptomyces sp. NP10 were dominated by saturated branched chain FAs (i-14:0, a-15:0, and i-16:0). Although these free FAs showed only a moderate antimicrobial activity, our results suggest that they could have an ecophysiological role in interspecies signaling with another soil microorganism Pseudomonas aeruginosa. This work represents the first comprehensive report on the structural diversity and complexity of the free FA pool in Streptomyces. A naturally occurring streptomycete, such as Streptomyces sp. NP10, which secretes significant amounts of free long-chain FAs (non-cytotoxic) into the medium, could be useful in microbial biodiesel production.

The use of 2-dimensional gas chromatography to investigate the effect of rumen-protected conjugated linoleic acid, breed, and lactation stage on the fatty acid profile of sheep milk

In this study, 2-dimensional gas chromatography (GC × GC) was used to obtain a detailed fatty acid (FA) profile of sheep milk and to evaluate the effects of a rumen-protected conjugated linoleic acid (rpCLA) supply, breed, days in milk (DIM), sampling period, and number of lambs suckling on the FA profile. Twenty-four ewes, from 3 autochthonous breeds of the Veneto Alps (Brogna, Foza, and Lamon), were housed in 6 pens (2 pens/breed), according to DIM (38±23 d) and body weight (61±13kg). The ewes and their offspring of 3 pens (1 pen/breed) were fed ad libitum a total mixed ration (control), and the other animals received the same diet supplemented with 12 g/d per ewe, plus 4 g/d for each lamb older than 30 d, of an rpCLA mixture. The study lasted 63 d. Two composite milk samples for each ewe were prepared during the first and second months of the trial. The pooled milk samples were analyzed in duplicate for FA profile by 2-dimensional gas chromatography, which allowed us to obtain a detailed FA profile of sheep milk, with 170 different FA detected, including many that were present in small concentrations. The milk relative proportions of individual FA, groups of FA, or FA indices were analyzed by PROC MIXED of SAS (SAS Institute Inc., Cary, NC), considering diet, breed, DIM, and sampling period as sources of variation. The random effect of animal was used to test diet, breed, and DIM, whereas the effects of period were tested on the residual. Breed had a small influence on milk FA profile, mainly on branched- and odd-chain FA. Within breed, animal repeatability for the relative proportions of milk FA was notable for almost all monounsaturated FA and for saturated FA with 14 to 19 carbon atoms, except C16:0, and less so for polyunsaturated FA. The inclusion of rpCLA (CLA cis-9,trans-11 and CLA trans-10,cis-12) increased the presence of the same CLA isomers in the milk as well as that of CLA trans-9,trans-11, and decreased the proportions of de novo-synthesized short-chain FA. From a cluster analysis based on the matrix of correlation coefficients among all FA relative proportions, 3 main FA groups were observed: the first included mainly odd- or branched-chain saturated FA, C18:0, C16:0 and CLA trans-10,cis-12; the second included monounsaturated FA or polyunsaturated FA with 16 to 20 carbons, CLA cis-9,trans-11, and CLA trans-9,trans-11; and the third included short- to medium-chain saturated FA, polyunsaturated FA with 2 to 5 double bonds, and 3 CLA isomers not affected by rpCLA addition (CLA trans-11,cis-13, CLA cis-9,cis-11, and CLA cis-10,cis-12).

Ion-Mobility Mass Spectrometry Assay for Incorporation of Phytanic Acid into Muscle Phospholipids

Human dysferlinopathies (Limb-Girdle Muscular Dystrophy 2b, Myoshi Myopathy) are muscle-wasting syndromes caused by mutations in the dysferlin protein. Dysferlin loss impairs sarcolemmal repair that may contribute to disease progression. One therapeutic approach is to treat subjects with compounds that fortify the natural membrane tendency to reseal after damage, and minimize the inflammation that impedes regeneration and amplifies tissue destruction. Phytanic acid is a saturated branched chain fatty acid comprised of a 16 carbon aliphatic chain with 4 methyl groups (4ME 16:0); it is incorporated into phospholipids and triglycerides. Model phospholipid bilayers containing phytanic acid exhibit greater electrical stability than bilayers composed of straight-chain phospholipids. Phytanic acid-containing phospholipids may improve the resistance of the muscle fiber sarcolemma to stretch-induced damage. To measure phytanic acid incorporation into muscle phospholipids, dysferlin-deficient A/J mice were maintained on a defined diet supplemented with 2% phytol for three weeks; control animals received the defined diet without phytol. Muscle tissue lipids were analyzed by mass spectrometry. The muscle phosphatidylcholine species profiles were similar between phytol and control diet, except that some additional species were detected in the phytol diet muscle. The two most abundant novel species (m/z 790.7 and 862.7) are tentatively identified as PC 20:0-16:0 and PC 20:0-22:6. To confirm the presence of phytanic acid, the ion mobility of these species was compared with diphytanoyl PC standard and endogenous straight chain PC phospholipids. The ion mobility is intermediate between PC species containing either zero or two phytanic acid chains, consistent with having one phytanic acid chain. We estimate that these species represent 5% of PC. Using this methodology, we will identify phytanic acid containing species in the other classes, in order to determine the total amount of phytanic acid-containing phospholipid incorporated in the muscle.

Serum phytanic and pristanic acid levels and prostate cancer risk in Finnish smokers.

Phytanic acid is a saturated branched-chain fatty acid found predominantly in red meat and dairy products, and may contribute to the elevated risks of prostate cancer associated with higher consumption of these foods. Pristanic acid is formed during peroxisomal oxidation of phytanic acid, and is the direct substrate of α-Methyl-CoA-Racemase (AMACR)—an enzyme that is consistently overexpressed in prostate tumors relative to benign tissue. We measured phytanic and pristanic acids as percentages of total fatty acids by gas chromatography-mass spectrometry in prediagnostic blood samples from 300 prostate cancer cases and 300 matched controls, all of whom were participants in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study supplementation trial and follow-up cohort. In addition to providing a fasting blood sample at baseline, all men completed extensive diet, lifestyle, and medical history questionnaires. Among controls, the strongest dietary correlates of serum phytanic and pristanic acids were saturated fat, dairy fat, and butter (r = 0.50 and 0.40, 0.46 and 0.38, and 0.40 and 0.37, respectively; all P-values <0.001). There was no association between serum phytanic acid and risk of total or aggressive prostate cancer in multivariate logistic regression models (for increasing quartiles, odds ratios (OR) and 95% confidence intervals (CI) for aggressive cancer were 1.0 (referent), 1.62 (0.97–2.68), 1.12 (0.66–1.90), and 1.14 (0.67–1.94), Ptrend = 0.87). Pristanic acid was strongly correlated with phytanic acid levels (r = 0.73, P < 0.0001), and was similarly unrelated to prostate cancer risk. Significant interactions between phytanic and pristanic acids and baseline circulating β-carotene concentrations were noted in relation to total and aggressive disease among participants who did not receive β-carotene supplements as part of the original ATBC intervention trial. In summary, we observed no overall association between serum phytanic and pristanic acid levels and prostate cancer risk. Findings indicating that the direction and magnitude of these associations depended upon serum levels of the antioxidant β-carotene among men not taking β-carotene supplements should be interpreted cautiously, as they are likely due to chance.

Bacillus pervagus sp. nov. and Bacillus andreesenii sp. nov., isolated from a composting reactor

Two strains, 8-4-E12(T) and 8-4-E13(T), were isolated from a biowaste composting reactor. Based on 16S rRNA gene sequences, both strains belong to the genus Bacillus. Strain 8-4-E12(T) was most closely related to the type strains of Bacillus shackletonii, B. acidicola, B. sporothermodurans and B. oleronius (96.4, 96.3, 96.0 and 95.6 % 16S rRNA gene similarity, respectively), whereas strain 8-4-E13(T) was most closely related to the type strain of Bacillus humi (96.5 % sequence similarity). Strains 8-4-E12(T) and 8-4-E13(T) shared 94 % 16S rRNA gene sequence similarity. The fatty acid profile of strain 8-4-E12(T) was dominated by saturated iso- and anteiso-branched fatty acids (iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0), and also contained considerable amounts of C16 : 0. The fatty acid profile of strain 8-4-E13(T) showed a predominance of iso-C15 : 0 (65 %), with smaller amounts of other saturated branched-chain fatty acids along with an unsaturated alcohol. Both strains contained diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine as major polar lipids. Additionally, strain 8-4-E12(T) contained an unknown lipid and strain 8-4-E13(T) two unknown (amino-)phospholipids. The diagnostic diamino acid found in the cell-wall peptidoglycan of 8-4-E12(T) and 8-4-E13(T) was meso-diaminopimelic acid. The predominant menaquinone was MK-7. The results of physiological and biochemical tests also allowed phenotypic differentiation of the two strains from each other and from related Bacillus species. On the basis of their phylogenetic, phenotypic and chemotaxonomic properties, strains 8-4-E12(T) and 8-4-E13(T) represent novel species of the genus Bacillus, for which the names Bacillus pervagus sp. nov. (type strain 8-4-E12(T) = DSM 23947(T) = LMG 27601(T)) and Bacillus andreesenii sp. nov. (type strain 8-4-E13(T) = DSM 23948(T) = LMG 27602(T)) are proposed.

13-Methyltetradecanoic Acid Exhibits Anti-Tumor Activity on T-Cell Lymphomas In Vitro and In Vivo by Down-Regulating p-AKT and Activating Caspase-3

13-Methyltetradecanoic acid (13-MTD), a saturated branched-chain fatty acid purified from soy fermentation products, induces apoptosis in human cancer cells. We investigated the inhibitory effects and mechanism of action of 13-MTD on T-cell non-Hodgkin’s lymphoma (T-NHL) cell lines both in vitro and in vivo. Growth inhibition in response to 13-MTD was evaluated by the cell counting kit-8 (CCK-8) assay in three T-NHL cell lines (Jurkat, Hut78, EL4 cells). Flow cytometry analyses were used to monitor the cell cycle and apoptosis. Proteins involved in 13-MTD-induced apoptosis were examined in Jurkat cells by western blotting. We found that 13-MTD inhibited proliferation and induced the apoptosis of T-NHL cell lines. 13-MTD treatment also induced a concentration-dependent arrest of Jurkat cells in the G1-phase. During 13-MTD-induced apoptosis in Jurkat cells, the cleavage of caspase-3 and poly ADP-ribose polymerase (PARP, a caspase enzymolysis product) were detected after incubation for 2 h, and increased after extending the incubation time. However, there was no change in the expression of Bcl-2 or c-myc proteins. The appearance of apoptotic Jurkat cells was accompanied by the inhibition of AKT and nuclear factor-kappa B (NF-κB) phosphorylation. In addition, 13-MTD could also effectively inhibit the growth of T-NHL tumors in vivo in a xenograft model. The tumor inhibition rate in the experimental group was 40%. These data indicate that 13-MTD inhibits proliferation and induces apoptosis through the down-regulation of AKT phosphorylation followed by caspase activation, which may provide a new approach for treating T-cell lymphomas.

A proapoptotic effect of valproic acid on progenitors of embryonic stem cell-derived glutamatergic neurons

Valproic acid (VPA) is a branched-chain saturated fatty acid with a long history of clinical use as an antiepileptic drug (AED). VPA is also known to inhibit histone deacetylases (HDACs) and to cause diverse effects on neural progenitor cells (NPCs) and neurons. Although the neuroprotective or neurodestructive effects of VPA have been investigated in heterogeneous cell populations, in this study, we used homogeneous populations of NPCs and glutamatergic cortical pyramidal neurons, which were differentiated from embryonic stem (ES) cells. At therapeutic concentrations, VPA had a proapoptotic effect on ES cell-derived NPCs of glutamatergic neurons, but not on their progeny. This effect of VPA most likely occurred through the inhibition of HDACs, because similar phenotypes were observed following treatment with other HDAC inhibitors (HDACis) such as trichostatin A and sodium butyrate. The proapoptotic phenotype was not observed when cells were exposed to a structural analog of VPA, valpromide (VPM), which has the same antiepileptic effect as VPA, but does not inhibit HDACs. Western blotting confirmed that treatment with HDACis, but not VPM, significantly increased the levels of histone H3 acetylation in NPCs. HDACi treatments did not affect the survival of neurons, although the acetylation levels were increased to a limited extent. These results, which are based on a homogeneous culture system, suggest that VPA inhibits HDAC activity and induces the apoptosis of NPCs that are fated to differentiate into glutamatergic neurons. The dose-dependent effects of VPA both on apoptosis and hyperacetylation of histone H3 in NPCs supported this notion. These cell type- and differentiation stage-specific effects of VPA imply that dysfunction of HDACs during pregnancy significantly increase the risk of congenital malformations associated with VPA administration.