Normal Fatty Acids Research Articles

A-066 Does essential fatty acid deficiency alter ethanolamine plasmalogen concentrations in red blood cells? Implications for clinical testing

Abstract Background Plasmalogens are glycerophospholipids characterized by a fatty alcohol at the sn-1 position, a polyunsaturated fatty acid at the sn-2 position, most commonly docosahexaenoic acid (DHA) or arachidonic acid, and a polar head group at the sn-3 position, mainly phosphoethanolamine. Markedly reduced plasmalogens are seen in peroxisome biogenesis defects, including Zellweger spectrum disorders (ZSD), since the first two steps of plasmalogen synthesis occur in peroxisomes. Disruption of the peroxisomal β-oxidation pathway also causes DHA deficiency in ZSD patients. Since plasmalogens are composed of fatty acids, the deficiency of DHA or other essential fatty acids could further decrease the plasmalogen level in ZSD patients and lower the results of plasmalogen testing in patients without ZSD. To test this hypothesis, we evaluated the impact of essential fatty acid (EFA) deficiency on plasmalogens in patients with ZSD and individuals with EFA deficiency due to unrelated causes. Methods Red blood cell (RBC) samples were obtained from individuals without ZSD referred to ARUP laboratories for clinical EFA testing (n = 80; age range: birth - 89 years), and from patients with ZSD (n = 7; age range birth - 9 years). Three ZSD patients were older and with a less severe phenotype. A total of 18 intact ethanolamine plasmalogen species were extracted from packed RBCs and quantified using a XEVO TQ-XS Mass Spectrometer in conjunction with Ultra-High Performance Liquid Chromatography (Waters). Total plasmalogen values were also calculated; plasmalogen deficiency was defined as ≤60% of age-matched control median. Fatty acids (FAs) were quantified by gas chromatography negative chemical ionization-mass spectrometry (GC-NCI-MS). EFA deficiency was diagnosed by increased Triene/Tetraene ratio (Mead acid/Arachidonic acid). Data was analyzed using Prism v.8.3.0 software (La Jolla, CA). The study was approved by the Institutional Review Board of the University of Utah. Results Out of the 80 samples from individuals without ZSD referred for EFA testing, 18% had normal fatty acid profile, 57% were receiving dietary supplements and 25% had elevated Triene/Tetraene ratio suggesting EFA deficiency. EFA deficiency was accompanied by low DHA only in two individuals without ZSD, whereas it was present in most of the samples from ZSD patients evaluated (5/7). None of the individuals without ZSD were deficient in plasmalogens. Moreover, when compared to individuals without ZSD with a normal FA profile, there was no significant difference observed with EFA deficiency or FA supplementation. Although our ZSD cohort is small, no correlation between EFA deficiency and plasmalogen levels was also seen in ZSD: 2 patients with milder phenotype had EFA deficiency but normal plasmalogens, 2 patients had EFA deficiency and low plasmalogens, 2 patients with low plasmalogens had low DHA without EFA deficiency, and 1 patient had low plasmalogens but normal EFA. Conclusions Plasmalogen quantitation in RBCs by LC-MS/MS is not significantly affected by EFA deficiency. Although more data is warranted to elucidate the correlation between plasmalogen levels and ZSD phenotype, plasmalogens can be within normal reference intervals in ZSD regardless of EFA status.

#4990 OMEGA-3 FATTY ACID ACTIVATES MITOCHONDRIAL BIOGENESIS AND PINK1 DEPENDENT MITOPHAGY IN KIDNEY AND HEART OF ADENINE INDUCED UREMIC RATS

Abstract Background and Aims Mitochondrial homeostasis is controlled by biogenesis, dynamics, and mitophagy. Mitochondrial dysfunction plays a central role in cardiovascular and renal disease and omega-3 fatty acids (FA) are beneficial for cardiovascular disease. We aimed to investigate whether omega-3 FA regulates the expression of mediators of mitochondrial biogenesis, dynamics, and mitophagy in kidney and heart of adenine-induced uremic rats. Method Sprague-Dawley rats were divided into three groups: normal control, adenine control, and omega-3 FA. Uremia was induced by feeding diets containing 0.75% adenine and 2.5% protein for the first 3 weeks. During next 4 weeks, they were received 2.5% protein with or without omega-3 FA (300 mg/kg/day). The renal and cardiac expression of PGC-1α, SIRT1/3, Nrf2, DRP-1, OPA1, Mfn1/2, PINK1, BNIP3 and NIX were examined by western blot analysis. The qPCR was used to determine mitochondrial DNA (mtDNA). Results Compared to normal, serum creatinine and heart weight/body weight in adenine control was increased and improved in omega-3 FA group. Compared with normal, PGC-1α, SIRT1/3 and Nrf2 were down-regulated in kidney and heart of adenine control. PGC-1α expression of kidney and heart was recovered in omega-3 FA group. DRP-1 of kidney was up-regulated but DRP-1 of heart was down-regulated in adenine control. DRP-1 of heart was recovered in omega-3 FA group. PINK1, BNIP3 and NIX were down-regulated in heart of adenine control and recovered in omega-3 FA group. PINK1 was down-regulated but BNIP3 and NIX were up-regulated in kidney of adenine control and those were mitigated in omega-3 FA group. MtDNA was decreased in kidney and heart of adenine control group but mtDNA of heart was recovered in omega-3 FA group. Conclusion DRP-1 related with mitochondrial fission may oppositely work in uremic kidney and heart. Omega-3 FA is beneficial for mitochondrial homeostasis by activating mitochondrial biogenesis and PINK1 dependent mitophagy in kidney and heart of uremic rats.

Analysis of Circulating Fatty Acid Profiles in Free-Ranging and Managed Care Marine Toads (Rhinella marina) with a Comparison of Whole-Blood Vial and Whole-Blood Dried Blood Spot Card Analyses

This study provides novel data on circulating concentrations of whole-blood fatty acids (FAs) in marine toads (Rhinella marina) via dried blood spot (DBS) card technology as a potential amphibian model species. Free-ranging (n = 10) animals were compared to managed populations fed two diet strategies for 60 days (n = 6 per diet). Thirty-six individual FAs were analyzed, with 28 found in significant reportable quantities. Eight FA groupings were represented. Traditional whole-blood vial (WBV) FA percentages were also collected and compared to DBS after managed care for 60 days. Results showed eleven individual FAs and four FA groups were higher in free-ranging toads (n = 10; p ≤ 0.05), while three FAs and three groups were higher in managed care (n = 12; p ≤ 0.05). FA concentrations compared between DBS cards and WBV at day 60 generally agreed, although two individual FAs and one grouping were higher in DBS (p ≤ 0.05). When free-ranging FAs were analyzed by sex, four individual FAs and two groupings were higher in females, while four individual FAs and one grouping were higher in males. Understanding normal FA circulating levels and how husbandry changes them may impact amphibian health. Additionally, DBS cards may provide a convenient sampling tool for fieldwork.

Rational synthesis and lithium storage properties of hierarchical nanoporous TiO2(B) assemblies with tailored crystallites and architectures

In comparison to the common anatase, rutile and brookite phases, the bronze phase TiO2 (TiO2(B)) is rarely prepared, and obtaining unique TiO2(B) structures, especially those with complex configurations remains a great challenge. This work presents a completely new synthetic approach for fabricating hierarchical nanoporous TiO2(B) assemblies with tailored crystallites and architectures via the reaction between tetrabutyl titanate and normal fatty acids. Three different kinds of normal fatty acids, i.e., pentanoic acid, hexanoic acid, and nonanoic acid were utilized as the sole solvent. After a simple solvothermal treatment, nanoporous TiO2(B) microspheres constructed by [001]-elongated ultrathin nanorods, randomly aggregated ultrafine nanocrystals, and crystallographically oriented nanocrystals were successfully produced separately. Further investigation revealed that the morphology of the hierarchical assemblies could be modified by using foreign substrates to adjust the growth dynamics of TiO2(B) crystals. As a good illustration, by introducing graphene nanosheets into the tetrabutyl titanate-pentanoic acid system, nanosized [001]-elongated-ultrathin-nanorod-constructed nanoporous TiO2(B) assemblies were obtained, which exhibited superior performance as an anode in Li-ion batteries. This work can not only shed new light on TiO2(B) crystallization, but also provide an effective solution for the rational design of complex TiO2(B) micro-/nanoarchitectures for desired applications.

A mathematical method for calibrating the signal drift in liquid chromatography - mass spectrometry analysis

Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) has become the most versatile analytical tool for profiling small-molecule compounds and increasingly been applied in many fields. Nevertheless, LC-MS based quantification still face some challenges, such as signal drift in LC-MS, which may affect the validity of the obtained data and lead to misinterpretation of biological results. Here, we established a calibration method known as “RIM” to compensate the signal drift of LC-MS. To this end, a mixture of d4-2-dimethylaminoethylamine (d4-DMED)-coded normal fatty acids (C5–C23) was used as calibrants to construct RIM calibration. With the addition of calibrants, not only the MS signal drift, but also the mass accuracy and LC retention time can be calibrated, thereby improving the reliability of quantitative data. The effectiveness of RIM was carefully validated using a human serum extract spiked with 34 standards and then RIM was applied for rat brain untargeted metabolome research. In addition, to expand the functionality and flexibility of RIM for data handling, we generated a MATLAB-based RIM program, which implements the above concepts and allows automatic data process. Taken together, the proposed RIM method has potential application in large-scale quantitative study of complex samples.

Using deep eutectic solvents to improve the biocatalytic reduction of 2-hydroxyacetophenone to (R)-1-phenyl-1,2-ethanediol by Kurthia gibsonii SC0312

The effects of five deep eutectic solvents (DESs) on the production of (R)-1-phenyl-1,2-ethanediol from 2-hydroxyacetophenone catalyzed by Kurthia gibsonii SC0312 were investigated in this study. Of these, choline chloride/1,4-butanediol (ChCl/Bd) showed excellent biocompatibility and suitably increased the cell membrane permeability while having a little impact on the structure of DNA. Indeed, ChCl/Bd at the concentration of 2 % increased the catalytic rate of the cells by 22 %. The other DESs did not stimulate the catalytic capacity of the cells, despite some increases in the cell membrane permeability. Additionally, the conformation of DNA was visibly changed when adding the other examined DESs except for choline chloride/triethylene glycol. The DESs modified the fatty acid composition of cellular membrane, decreased the relative amount of iso-C14:0 and increased the relative amount of normal C15:0. Meanwhile, the DESs were able to improve the relative ratio of normal fatty acids to branched fatty acids. Finally, a highly efficient reduction of 80 mM 2-hydroxyacetophenone by K. gibsonii SC0312 in the ChCl/Bd-containing system was established, affording (R)-1-phenyl-1,2-ethanediol in 80 % yield and optical purity >99 % at 30 mg/mL wet cells. This work offers a promising approach for the preparation of (R)-1-phenyl-1,2-ethanediol from 2-hydroxyacetophenone using K. gibsonii SC0312.

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.

Functional Characterization of Three Novel Genes Encoding Diacylglycerol Acyltransferase (DGAT) from Oil-Rich Tubers of Cyperus esculentus.

Cyperus esculentus is probably the only plant that is known to accumulate large amounts of oil in its tubers. However, the underlying metabolic mechanism and regulatory factors involved in oil synthesis of tubers are still largely unclear. In this study, one gene encoding type I diacylglycerol acyltransferase (DGAT) (CeDGAT1) and two genes encoding type II DGAT (CeDGAT2a and CeDGAT2b) from C. esculentus were identified and functionally analyzed. All three DGAT genes were found to be expressed in tuber, root and leaf tissues. CeDGAT1 is highly expressed in roots and leaves, whereas CeDGAT2b is dominantly expressed in tubers. Furthermore, the temporal expression pattern of CeDGAT2b is well coordinated with the oil accumulation in developing tubers. When each CeDGAT was heterologously expressed in triacylglycerol (TAG)-deficient mutant of Saccharomyces cerevisiae, Arabidopsis thaliana wild type or its TAG1 mutant with AtDGAT1 disruption, only CeDGAT2b showed the ability to restore TAG biosynthesis with lipid body formation in yeast mutant, enhance seed oil production of Arabidopsis wild type and rescue multiple seed phenotypes of TAG1 mutant. In addition, CeDGAT2b was shown to have a substrate preference for unsaturated fatty acids toward TAG synthesis. Taken together, our results indicated that CeDGAT2b from C. esculentus is an actively functional protein and is most likely the major contributor to tuber oil biosynthesis containing common fatty acids, in contrast to oil-rich seeds and fruits where DGAT1 plays a more central role than DGAT2 in oil production accumulating normal fatty acids, whereas DGAT2 is a primary regulator for oil synthesis rich in unusual fatty acids.

ELOVL3, 6 and 7 Specifically Elongate Saturated Branched Chain Fatty Acids and Compete with Normal Fatty Acids

IntroductionBranched chain fatty acids (BCFA) are saturated fatty acids (SFA) terminating with either an isopropyl or isobutyl group. They are common components in human diets, and are synthesized de novo in human skin sebaceous and meibomian glands as well as vernix caseosa. They are known to have structure specific activities but little known of the pathways for their endogenous interconversion. Multiple human ELOVLx (elongase of very long chain fatty acids) genes are responsible for interconversion of saturated fatty acids. Our objectives are to perform the first investigation of the activity of ELOVL on BCFA using anteiso‐15:0 and iso‐18:0 as examples, to characterize the substrate specificity to BCFA, and to investigate competition between BCFA and normal (n‐) SFA (n‐16:0 and n‐18:0) for the ELOVL‐mediated elongation.MethodsHuman breast cancer cell line MCF7 was used as the experiment model. MCF7 cells were grown in MEM‐α media with 10% FBS in a humidified environment at 37°C with 5% CO2. The open reading frame of ELOVL transcripts (ELOVL1, ELOVL3, ELOVL6 and ELOVL7) were cloned into a pcDNA3.1 expression vector. MCF7 cells were transfected with specific ELOVL transgene vector along with empty vector (control) for 24h using Polyplus jetPRIME transfection reagent. Then albumin‐bound fatty acids, anteiso‐15:0, iso‐18:0, n‐16:0 and n‐18:0 were dosed into cells at 0–100 μM and allowed to incubate for additional 24h. Cells were washed twice with 1 × PBS, harvested by trypsinization, fatty acid methyl esters were analyzed by GC‐FID and GC‐MS.ResultsELOVL6 chiefly elongated anteiso‐15:0 à anteiso‐17:0; ELOVL3 predominantly elongated iso‐18:0 à iso‐20:0; ELOVL7 showed moderate elongase activity toward both anteiso‐15:0 and iso‐18:0. n‐16:0 competed with anteiso‐15:0 for ELOVL6 mediated elongation, and n‐18:0 competed with iso‐18:0 for ELOVL3 mediated elongation. These latter results verified the characterized elongation activities.ConclusionHere for the first time we showed the substrate specificity of human elongase to BCFA systematically, and verified it by establishing competition between BCFA and n‐SFA.Support or Funding InformationInternal Fund

Phytosphingosine degradation pathway includes fatty acid α-oxidation reactions in the endoplasmic reticulum

Although normal fatty acids (FAs) are degraded via β-oxidation, unusual FAs such as 2-hydroxy (2-OH) FAs and 3-methyl-branched FAs are degraded via α-oxidation. Phytosphingosine (PHS) is one of the long-chain bases (the sphingolipid components) and exists in specific tissues, including the epidermis and small intestine in mammals. In the degradation pathway, PHS is converted to 2-OH palmitic acid and then to pentadecanoic acid (C15:0-COOH) via FA α-oxidation. However, the detailed reactions and genes involved in the α-oxidation reactions of the PHS degradation pathway have yet to be determined. In the present study, we reveal the entire PHS degradation pathway: PHS is converted to C15:0-COOH via six reactions [phosphorylation, cleavage, oxidation, CoA addition, cleavage (C1 removal), and oxidation], in which the last three reactions correspond to the α-oxidation. The aldehyde dehydrogenase ALDH3A2 catalyzes both the first and second oxidation reactions (fatty aldehydes to FAs). In Aldh3a2-deficient cells, the unmetabolized fatty aldehydes are reduced to fatty alcohols and are incorporated into ether-linked glycerolipids. We also identify HACL2 (2-hydroxyacyl-CoA lyase 2) [previous name, ILVBL; ilvB (bacterial acetolactate synthase)-like] as the major 2-OH acyl-CoA lyase involved in the cleavage (C1 removal) reaction in the FA α-oxidation of the PHS degradation pathway. HACL2 is localized in the endoplasmic reticulum. Thus, in addition to the already-known FA α-oxidation in the peroxisomes, we have revealed the existence of FA α-oxidation in the endoplasmic reticulum in mammals.

Identification of molecular species of acylglycerols of Philippine wild edible mushroom, Ganoderma lucidum

We identified the lipid species of the solvent extract from liquid-cultured mycelium of G. lucidum NRRL66208. The molecular species of acylglycerols containing all normal fatty acids were identified. One hundred and three molecular species of acylglycerols containing all normal fatty acids were identified by the MS fragmentation involved their constituent fatty acids. The chain lengths of the constituent fatty acids were from 14 to 26 carbon atoms and the numbers of double bonds were from zero to three. The contents of the molecular species of triacylglycerols in the mushroom lipid extract in decreasing order were: OOP (2.45%), OOO (1.94%), LLP (1.92%), OLP (1.80%), LLO (1.57%), OPP (1.30%), OOL (1.09%), OOS (0.88%), and LLL (0.85%). Forty one molecular species of acylglycerols out of 103 contained fatty acids with odd numbered carbon atoms. The fatty acids with odd numbered carbon atoms were 15:0, 17:0, 17:1, 19:0, 19:1, 21:0, 23:0 and 25:0. The total content of fatty acids with odd numbered carbon atoms was about 6.6% of all of the fatty acids of the triacylglycerols containing three normal fatty acids, and contents of individual fatty acids with odd numbered carbon atoms were as: 15:0 (2.76%), 17:0 (2.13%), 17:1 (1.35%), 19:0 (0.20%), 19:1 (0.17%). The contents of fatty acids with odd numbered carbon atoms in mushroom were high compared to those in higher plants. This is the first report on the molecular species of acylglycerols in mushroom.

Polarization Observations of Adsorption Behavior of Fatty Acids Using Optical Anisotropy of Liquid Crystal

Chemical interactions and formation of adsorbed films of additives strongly affect the tribological behavior in boundary lubrication. In order to examine the relationship between the adsorption behavior and friction properties of fatty acids, polarization observations using a liquid crystal and friction measurements were performed. A nematic liquid crystal [4-pentyl-4′-cyanobiphenyl (5CB)] and normal fatty acids with 16–22 C chains were base fluid and additive, respectively. The adsorption behavior of the fatty acids was observed using polarization microscopy and analyzing the optical anisotropy of 5CB. The adsorbed film of fatty acids induced a change in the orientation of 5CB from planar to homeotropic. The expansion speed of the homeotropic domain increased as the chain length of the fatty acid decreased. The friction coefficient of the test lubricants was measured in a steel–steel sliding contact using a low-speed reciprocating tribometer. At the first cycle of reciprocating sliding motion, the lower friction coefficient was measured using fatty acids with the shorter alkyl chain length. The friction coefficient at the first cycle showed a strong correlation with the expansion speed of the homeotropic domain. These results show that the shorter-chain fatty acids form an adsorbed film highly oriented perpendicular to the solid surface, and the adsorbed film provides a lower friction coefficient at the first cycle.

Ratios of Regioisomers of the Molecular Species of Triacylglycerols in Lesquerella (Physaria fendleri) Oil Estimated by Mass Spectrometry

AbstractThe ratios of regioisomers of 72 molecular species of triacylglycerols (TAG) in lesquerella oil were estimated using the electrospray ionization mass spectrometry of the lithium adducts of TAG in the HPLC fractions of lesquerella oil. The ratios of ion signal intensities (or relative abundances) of the fragment ions from the neutral losses of fatty acids (FA) as α‐lactones at the sn‐2 position (MS3) of the molecular species of TAG were used as the ratios of the regioisomers. The order of the preference of FA incorporation at the sn‐2 position of the molecular species of TAG in lesquerella was as: normal FA > OH18 (monohydroxy FA with 18 carbon atoms) > diOH18 > OH20 > diOH20, while in castor was as: normal FA > OH18 > OH20 > diOH18 > triOH18. Elongation (from C18 to C20) was more effective than hydroxylation in lesquerella to incorporate hydroxy FA at the sn‐1/3 positions. The block of elongation in lesquerella may be used to increase the content of hydroxy FA, e.g., ricinoleate, at the sn‐2 position of TAG and to produce triricinolein (or castor oil) for industrial uses. The content of normal FA at the sn‐2 position was about 95 %, mainly oleate (38 %), linolenate (31 %) and linoleate (23 %). This high normal FA content (95 %) at the sn‐2 position was a big space for the replacement of ricinoleate to increase the hydroxy FA content in lesquerella oil. The content of hydroxy FA at the sn‐1/3 positions was 91 % mainly lesquerolic acid (85 %) and the content of normal FA was 6.7 % at the sn‐1/3 position in lesquerella oil.

An Empirical Equation for Estimation of Kinematic Viscosity of Fatty Acid Methyl Esters and Biodiesel

AbstractKinematic viscosity (µ) is an important physical property of fatty acid methyl esters (FAME) and biodiesel. In this work, the Martin's rule of free energy additivity is extended to cover the kinematic viscosity of saturated and unsaturated FAME commonly found in nature. The proposed model can also be extended to estimate kinematic viscosity of biodiesel. The kinematic viscosity of a FAME or a biodiesel can be easily estimated from its carbon number (z), number of double bonds (nd) at different temperatures (T) without a prior knowledge of the viscosity of individual FAME. Both zave and nd(ave) can be derived from its fatty acid composition. Thus, kinematic viscosity of biodiesel at temperatures between 20 and 100 °C and at atmospheric pressure can be estimated. The average absolute deviation (AAD) estimated at 20–100 °C for saturated, unsaturated FAME, biodiesels and biodiesel blends are 4.15, 3.25, 6.95 and 2.79 %, respectively. The biodiesels collected in this study (191 data points) have the zave and nd(ave) between 14.10 and 17.96 and 0.21–1.54, respectively. The standard deviation was 0.249. The proposed model would be good for estimation of viscosity of biodiesel containing normal fatty acids, generally found in biodiesel feed stocks.

PHYSICOCHEMICAL CHARACTERISTICS, FATTY ACID COMPOSITION AND NUTRITIONAL EVALUATION OF FOUR MINOR OILS

Four minor seed oils from Abroma augusta (AA), Annona squamosa (AS), Bombax ceiba (BC) and Ceiba pentandra (CP) plants were refined and refined seed oils were analyzed for their physicochemical properties. All the refined seed oils had normal physicochemical characteristics. The seed oils except CP also had usual fatty acid composition and contained a higher proportion of unsaturated fatty acids namely linoleic and oleic, a suitable criterion of edibility of vegetable oils. In addition to normal fatty acids, the refined CP seed oil also contained 10.6% cyclopropenoid fatty acids (CPFA), therefore, is not recommended as a dietary source of fat unless a suitable method is developed for its removal or detoxification. Nutritional evaluation of the refined seed oils were carried out in rats by feeding the respective oils and peanut oil as control at 10% level in a 20% protein diet with adequate vitamins and minerals. The refined CP seed oil showed poor growth performance, low FER, altered lipid metabolism and histopathological abnormalities in the liver which indicate the unsuitability of it for edible purpose. Growth performances, FER, serum and liver lipids, organ weights and histopathological findings of other three minor oils were satisfactory and comparable to that of peanut oil, therefore, they can be considered for edible use.

Botryococcus braunii as a Phycoremediation Tool for the Domestic Waste Water Recycling from Cooum River, Chennai, India

The micro algae are the green photosynthetic micro organisms, which are the primary producers of the aquatic ecosystems. The green micro algae have evolved long back approximately two and a half billion years ago and initiated the process of photosynthesis in the world. The Botryococcus braunii is a green photosynthetic micro alga fixes atmospheric CO2 for energy production and it is well known for high quantity of lipid accumulation. Along with normal fatty acids the long chain different types of hydrocarbon production exudates outside of the cells are the characteristic features of this specific micro alga. At the same time, this unique micro alga has been reported to grow well in the sewage waste water and can be used to remove toxic metals and unwanted nutrients from waste water. In this present study, a new and novel strain of Botryococcus braunii has been isolated from the temple ponds around the Chennai city, Tamil Nadu, India. The effective treatment of domestic waste water from different sites around Chennai city using Botryococcus braunii has been well evaluated in this current study. The physiochemical parameters like COD, BOD, electrical conductivity (EC) and ammonia have been removed more than 70% from domestic waste water especially in Adyar River (AR) domestic waste water when compared to other waste water. The growth kinetics of the Botryococcus braunii strain in the domestic waste water AR and NB (Napier’s bridge) have shown good results when compared to other samples. The micro alga B. braunii strain is well known for its high lipid production and which was achieved from the biomass harvested from the domestic waste water sample AR. The total carotenoid content was also found in the micro algal biomass from the sample AR. This overall research study suggested that the implication of some important micro algae for the treatment of waste water can remediate the water resource as well as the biomass yield can be economically useful for many applications.

Production of polyol oils from soybean oil by bioprocess: results of microbial screening and identification of positive cultures

Recently we reported methods for microbial screening and production of polyol oils from soybean oil through bioprocessing (Hou and Lin, 2013). Soy-polyol oils (oxygenated acylglycerols) are important starting materials for the manufacture of polymers such as polyurethane. Currently, they are produced by a two-step chemical process involving epoxidation and subsequent opening of the oxirane ring. A total of 650 cultures isolated from soil samples collected from a biodiesel plant in Iowa and from soil and water samples collected from Peoria, IL were screened for the production of polyol oil from soybean oil. Out of these 650 cultures screened, 50 cultures were positive for converting soybean oil to two product groups: diacylglycerol polyol oil (polyol DAGs) and diacylglycerol (DAD) with normal fatty acids. These two product groups can be separated by a two-solvent system TLC. In this study, we identified the 11 most active of these positive cultures using 16S rRNA gene analysis. The 11 strains investigated were found to be bacterial and no active yeast strains were found in the present study. Phylogenetic analysis identified two strains A01-35 and E03-25 as Acinetobacter haemolyticus, a single strain was identified as Pseudomonas aeruginosa, the other active strains were identified as Pseudomonas protegens.

Regulation of Hypothalamic Neuronal Sensing and Food Intake by Ketone Bodies and Fatty Acids

Metabolic sensing neurons in the ventromedial hypothalamus (VMH) alter their activity when ambient levels of metabolic substrates, such as glucose and fatty acids (FA), change. To assess the relationship between a high-fat diet (HFD; 60%) intake on feeding and serum and VMH FA levels, rats were trained to eat a low-fat diet (LFD; 13.5%) or an HFD in 3 h/day and were monitored with VMH FA microdialysis. Despite having higher serum levels, HFD rats had lower VMH FA levels but ate less from 3 to 6 h of refeeding than did LFD rats. However, VMH β-hydroxybutyrate (β-OHB) and VMH-to-serum β-OHB ratio levels were higher in HFD rats during the first 1 h of refeeding, suggesting that VMH astrocyte ketone production mediated their reduced intake. In fact, using calcium imaging in dissociated VMH neurons showed that ketone bodies overrode normal FA sensing, primarily by exciting neurons that were activated or inhibited by oleic acid. Importantly, bilateral inhibition of VMH ketone production with a 3-hydroxy-3-methylglutaryl-CoA synthase inhibitor reversed the 3- to 6-h HFD-induced inhibition of intake but had no effect in LFD-fed rats. These data suggest that a restricted HFD intake regimen inhibits caloric intake as a consequence of FA-induced VMH ketone body production by astrocytes.

Identification of Tetraacylglycerols in Lesquerella Oil by Electrospray Ionization Mass Spectrometry of the Lithium Adducts

AbstractTetraacylglycerol (an acylglycerol estolide) contains an acyl chain attached to the hydroxyl group of another acyl chain attached to the glycerol backbone. Lesquerolic acid (Ls, OH1420:111) is the main fatty acid in lesquerella oil and may be used industrially for the manufacture of biodegradable industrial products. Electrospray ionization mass spectrometry of the lithium adducts of acylglycerols in the high‐performance liquid chromatography fractions from the seed oil of lesquerella (Physaria fendleri) was used to identify thirteen tetraacylglycerols. They were LsLsLsLn, LsLsLsL, LsLs‐OH20:2‐O, LsLsLsO, LsLsLnLn, LsLsLLn, LsLsOLn, LsLsLL, LsLsOL, LsLsOP, LsLsOO, LsLsLS and LsLsOS. The OH20:2 was auricolic acid (OH1420:211,17). For the four tetraacylglycerols containing one normal fatty acid (non‐hydroxy fatty acid), LsLsLsLn, LsLsLsL, LsLs‐OH20:2‐O and LsLsLsO, the normal fatty acids were all directly attached to the glycerol backbone, not to the hydroxyl group of fatty acids. We propose that the biosynthetic precursors (triacylglycerol acyltransferase) of these four tetraacylglycerols were LsLsLn, LsLsL, LsLsO (Ls‐OH20:2‐O) and LsLsO individually. LsLsO and Ls‐OH20:2‐O were equally active as the biosynthetic precursors for LsLs‐OH20:2‐O. For LsLsLS, linoleate were all attached to the glycerol backbone and LsLsL was proposed to be the biosynthetic precursor. For LsLsOS, stearate were all attached to the glycerol backbone and LsLsS was proposed to be the biosynthetic precursor. For the other seven tetraacylglycerols containing two normal fatty acids, LsLsAB, the biosynthetic precursors could be both LsLsA and LsLsB.

Mass Spectrometry of the Lithium Adducts of Diacylglycerols Containing Hydroxy FA in Castor Oil and Two Normal FA

AbstractCastor oil can be used in industry. The molecular species of triacylglycerols containing hydroxy fatty acids (FA) in castor oil have been identified. We report here the identification of twelve diacylglycerols (DAG) containing hydroxy FA in castor oil using positive ion electrospray ionization mass spectrometry of the lithium adducts. They were RR (diricinolein, R is ricinoleate), RL, RS, R‐diOH18:0, R‐diOH18:1, R‐diOH18:2, R‐triOH18:0, R‐triOH18:1, R‐triOH18:2, diOH18:0‐diOH18:1, diOH18:1‐diOH18:1 and diOH18:1‐diOH18:2. The MS2 fragment ions, [M + Li − FA]+ and [FA + Li]+, from the lithium adducts of DAG containing hydroxy FA (one or two hydroxy FA), were used for the identification. The additional fragment ions from the neutral losses of FA lithium salts [M + Li − FALi]+ were used for the identification of eleven DAG containing two normal FA in a soybean oil bioconversion product. The MS2 fragment ions from the neutral losses of FA lithium salts [M + Li − FALi]+ were not detected from the DAG containing hydroxy FA. The DAG containing FA with more hydroxyl groups than the other FA on the same DAG molecule tended to have a prominent fragment ion [FA + Li]+ and an undetectable fragment ion [M + Li − FA]+ while the FA was the more hydroxylated FA. Also the less hydroxylated FA of a DAG tended to have a prominent fragment ion [M + Li − FA]+ and an undetectable fragment ion [FA + Li]+ while the FA was the less hydroxylated FA.