16-hydroxyhexadecanoic Acid Research Articles

Effects on the development of Spodoptera frugiperda feeding on diets spiked with Solanum sisymbriifolium extracts

Aiming to evaluate the effects of Solanum sisymbriifolium extracts on the development of Spodoptera frugiperda, a leaf dichloromethane extract was obtained and subjected to further purification following a bioguided methodology. The structure of the two main compounds isolated from S. sisymbriifolium Type IV glandular trichomes have been completely elucidated by a combination of chemical and spectroscopic methods. They are glycosides of 3-(R)-hydroxypalmitic acid and xylo (β-1-5) furanoarabinoside (Compound 1) and its 3-O-palmitoyl derivative (Compound 2). These compounds greatly influenced the survival and the development of S. frugiperda, when compared to an in vitro test control. The extracts delayed the development, decreased survival and promote abnormalities in the immatures (larvae and pupae). They also showed increasing toxicity towards S. frugiperda in the purification following a bioguided fractionation. The pure compounds had the most deleterious activity, increasing the larval and pupal toxicity (100% mortality for Compound 2 at 2.50 mg mL-1 and 90% mortality for Compound 1 at 1.00 mg mL-1).

(S)-1-(1-Methylpyridin-2-yl)-3-aminopiperidine as a novel derivatization reagent capable of enantiomeric separation and enhanced ESI-MS/MS detection for chiral carboxylic acids

A novel derivatization reagent, (S)-1-(1-methylpyridin-2-yl)-3-aminopiperidine (MPAPp), was designed and developed for the enantiomeric separation by conventional reversed-phase liquid chromatography (LC) and the sensitive electrospray ionization-tandem mass spectrometry (ESI-MS/MS) detection of chiral carboxylic acids. MPAPp reacted with the carboxylic acids at 60 °C within 5 min in the presence of a condensation agent to produce the positively-charged and diastereomeric derivatives. The MPAPp-derivatization enabled the satisfactory enantiomeric separation not only of the α- and β-chiral carboxylic acids [ketoprofen (KET), etodolac and 3-hydroxypalmitic acid], but also of the γ-chiral carboxylic acid [2-(γ-carboxyethyl)-6-hydroxy-2,7,8-trimethylchroman (γ-CEHC)] with the resolution (Rs) values of 1.35–1.82. The derivatization also worked for the enantiomeric separation of the ε-chiral carboxylic acid [α-lipoic acid, separation factor (α) = 1.04]. The detection responses of the MPAPp-derivatives were 25–500-times greater than the intact carboxylic acids, therefore, the low femtomolar derivatives were fully detected. The trace detection of a γ-CEHC enantiomer (limit of detection, 1.9 fmol on column) in saliva was achieved by the MPAPp-derivatization followed by LC/ESI-MS/MS. The pharmacokinetic profiles of the KET enantiomers during/after the MOHRUS® patch application were also obtained from the saliva analysis using the MPAPp-derivatization based LC/ESI-MS/MS method, which was precise (intra- and inter-assay relative standard deviations, <6.6% and <4.6%, respectively) and accurate (96.2–105.3%) within the range of 35–280 pg/mL for each enantiomer.

A metabolomics approach to characterize raw, pasteurized, and ultra-high temperature milk using ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry and multivariate data analysis

We developed a metabolomics workflow using ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry to determine the effect of thermal treatment on milk composition and metabolites based on multivariate data analysis. We analyzed raw, pasteurized, and UHT milk samples. The samples were first centrifuged to remove the fat layer and mixed with methanol to precipitate proteins. Subsequently, the supernatant was analyzed by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry in electrospray negative mode. Mass spectral data were acquired in MSE mode, a technique whereby both precursor and fragment mass spectral are simultaneously acquired by alternating between low and high collision energy (CE) during a single analytical run, to enable metabolite identification. Based on multivariate data analysis, these markers were significantly affected by thermal treatment. Among the 8 potential markers, we identified 7 oxylipids (9-hydroxydecanoic acid, 12-hydroxydodecanoic acid, 2-hydroxymyristic acid, 3-hydroxytetradecanoic acid, 5-hydroxyeicosatetraenoic acid, 3-hydroxyhexadecanoic acid, and 10-hydroxyoctadecanoic acid) and 1 phospholipid (LysoPE, hexadecanoyl-lysophosphatidylethanolamine). The oxylipids seemed to be adequate for distinguishing UHT milk from raw and pasteurized milk. The structures of the 8 potential markers were identified and characterized using informatics software. Our metabolomics workflow provides a fast approach for the identification of various types of milk.

High vulnerability of the heart and liver to 3-hydroxypalmitic acid-induced disruption of mitochondrial functions in intact cell systems.

Patients affected by long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency predominantly present severe liver and cardiac dysfunction, as well as neurological symptoms during metabolic crises, whose pathogenesis is still poorly known. In this study, we demonstrate for the first time that pathological concentrations of 3-hydroxypalmitic acid (3HPA), the long-chain hydroxyl fatty acid (LCHFA) that most accumulates in LCHAD deficiency, significantly decreased adenosine triphosphate-linked and uncoupled mitochondrial respiration in intact cell systems consisting of heart fibers, cardiomyocytes, and hepatocytes, but less intense in diced forebrain. 3HPA also significantly reduced mitochondrial Ca2+ retention capacity and membrane potential in Ca2+ -loaded mitochondria more markedly in the heart and the liver, with mild or no effects in the brain, supporting a higher susceptibility of the heart and the liver to the toxic effects of this fatty acid. It is postulated that disruption of mitochondrial energy and Ca2+ homeostasis caused by the accumulation of LCHFA may contribute toward the severe cardiac and hepatic clinical manifestations observed in the affected patients.

Biocatalytic Route for the Synthesis of Oligoesters of Hydroxy-Fatty acids and ϵ-Caprolactone.

Developments of past years placed the bio-based polyesters as competitive substitutes for fossil-based polymers. Moreover, enzymatic polymerization using lipase catalysts has become an important green alternative to chemical polymerization for the synthesis of polyesters with biomedical applications, as several drawbacks related to the presence of traces of metal catalysts, toxicity and higher temperatures could be avoided. Copolymerization of ϵ-caprolactone (CL) with four hydroxy-fatty acids (HFA) from renewable sources, 10-hydroxystearic acid, 12-hydroxystearic acid, ricinoleic acid, and 16-hydroxyhexadecanoic acid, was carried out using commercially available immobilized lipases from Candida antarctica B, Thermomyces lanuginosus, and Pseudomonas stutzeri, as well as a native lipase. MALDI-TOF-MS and 2D-NMR analysis confirmed the formation of linear/branched and cyclic oligomers with average molecular weight around 1200 and polymerization degree up to 15. The appropriate selection of the biocatalyst and reaction temperature allowed the tailoring of the non-cyclic/cyclic copolymer ratio and increase of the total copolymer content in the reaction product above 80%. The catalytic efficiency of the best performing biocatalyst (Lipozyme TL) is evaluated during four reaction cycles, showing excellent operational stability. The thermal stability of the reaction products is assessed based on TG and DSC analysis. This new synthetic route for biobased oligomers with novel functionalities and properties could have promising biomedical applications.

Soil metagenome-derived 3-hydroxypalmitic acid methyl ester hydrolases suppress extracellular polysaccharide production in Ralstonia solanacearum

Autoinducers are indispensable for bacterial cell–cell communication. However, due to the reliance on culture-based techniques, few autoinducer-hydrolyzing enzymes are known. In this study, we characterized soil metagenome-derived unique enzymes capable of hydrolyzing 3-hydroxypalmitic acid methyl ester (3-OH PAME), an autoinducer of the plant pathogenic bacterium Ralstonia solanacearum. Among 146 candidate lipolytic clones from a soil metagenome library, 4 unique enzymes capable of hydrolyzing the autoinducer 3-OH PAME, termed ELP86, ELP96, ELP104, and EstDL33, were selected and characterized. Phylogenetic analysis revealed that metagenomic enzymes were novel esterase/lipase candidates as they clustered as novel subfamilies of family I, V, X, and family XI. The purified enzymes displayed various levels of hydrolytic activities towards 3-OH PAME with optimum activity at 40–50 °C and pH 7–10. Interestingly, ELP104 also displayed N-(3-oxohexanoyl)-L-homoserine lactone hydrolysis activity. Heterologous expression of the gene encoding 3-OH PAME hydrolase in R. solanacearum significantly decreased exopolysaccharide production without affecting bacterial growth. mRNA transcription analysis revealed that genes regulated by quorum-sensing, such as phcA and xpsR, were significantly down-regulated in the stationary growth phase of R. solanacearum. Therefore, metagenomic enzymes are capable of quorum-quenching by hydrolyzing the autoinducer 3-OH PAME, which could be used as a biocontrol strategy against bacterial wilt.

Role of 3-Hydroxy Fatty Acid-Induced Hepatic Lipotoxicity in Acute Fatty Liver of Pregnancy.

Acute fatty liver of pregnancy (AFLP), a catastrophic illness for both the mother and the unborn offspring, develops in the last trimester of pregnancy with significant maternal and perinatal mortality. AFLP is also recognized as an obstetric and medical emergency. Maternal AFLP is highly associated with a fetal homozygous mutation (1528G>C) in the gene that encodes for mitochondrial long-chain hydroxy acyl-CoA dehydrogenase (LCHAD). The mutation in LCHAD results in the accumulation of 3-hydroxy fatty acids, such as 3-hydroxy myristic acid, 3-hydroxy palmitic acid and 3-hydroxy dicarboxylic acid in the placenta, which are then shunted to the maternal circulation leading to the development of acute liver injury observed in patients with AFLP. In this review, we will discuss the mechanistic role of increased 3-hydroxy fatty acid in causing lipotoxicity to the liver and in inducing oxidative stress, mitochondrial dysfunction and hepatocyte lipoapoptosis. Further, we also review the role of 3-hydroxy fatty acids in causing placental damage, pancreatic islet β-cell glucolipotoxicity, brain damage, and retinal epithelial cells lipoapoptosis in patients with LCHAD deficiency.

Microbial degradation of poly(ε-caprolactone) in a coastal environment

Biodegradable plastics may be a solution for eliminating large marine plastic debris, such as fishing gear, which causes environmental damage. The biodegradable polyester poly(ε-caprolactone) (PCL) is degraded in various environments; however, no reports have examined PCL-degrading microbes inhabiting coastal environments, despite the fact that marine plastic debris accumulates in this environment. Therefore, in this study, we isolated a PCL-degrading bacterium (strain TKCM 64) from marine plastic debris in coastal seawater (Okinoshima Park, Chiba, Japan) for the first time. Genetic and biochemical analyses showed that strain TKCM 64 was closely related to Pseudomonas pachastrellae JCM12285T. The maximum specific growth rate (0.4 h−1) of the isolate was observed in the absence of NaCl at 30 °C, and the strain was able to grow, even at 1.20 M NaCl, suggesting that the strain could belong to a marine species showing halotolerance. Strain TKCM 64 degraded PCL film at a rate of 1.39 ± 0.09 mg cm−2·day−1. The strain showed PCL hydrolytic activity, which was induced in the presence of PCL and its hydrolysate: 6-hydroxyhexanoic acid (6HH), in addition to a major component of cutin, 16-hydroxyhexadecanoic acid. Thus, we concluded that the PCL hydrolysate 6HH, as an analog of cutin hydrolysate, functioned as an inducer of a PCL hydrolase that is a kind of cutinase in strain TKCM 64. In addition, Pseudomonas pachastrellae was found to be a key bacterial species involved in PCL degradation in coastal environments.

Potential application of oat-derived ceramides in improving skin barrier function: Part 1. Isolation and structural characterization.

The impaired epidermal barrier and skin dryness in chronic skin conditions such as atopic dermatitis, psoriasis and aged skin are associated with the depletion of ceramides (CERs) in the stratum corneum. Previously, the beneficial effects of phyto-CERs, mainly from wheat and rice, in replenishing the depleted epidermal CERs and restoring the skin barrier have been shown. However, very few efforts have been made to exploit CERs from other plants for dermal applications. In an attempt to explore alternative plant source of CERs, glucosylceramides (GlcCERs) were isolated from the lipid extract of Ethiopian oat grain (Avena abyssinica). The GlcCER species were separated on a reversed phase HPLC and the structure of individual GlcCERs were identified by tandem MS with atmospheric pressure chemical ionization interface. The glycosidic linkage of the GlcCERs was cleaved by acid treatment and the predominant CERs species were isolated using column chromatography and preparative LC-MS. Further structural characterization of the CERs was made by HR/ESI-MS and NMR analyses. All the detected oat-derived GlcCER species consisted of C18 dihydroxy sphingoid bases amide-linked with α-hydroxylated saturated fatty acids (C16-C24). The two predominant GlcCER species consisted of sphingenine (d18:1) amide-linked to hydroxypalmitic acid (h16:0) and hydroxyarachidic acid (h20:0). The molecular formulae of the two major CERs assigned by HR/ESI-MS were identical to the ones identified by LC/APCI-MS/MS. The structural information was also supported by 1H, 13C, 1H COSY NMR and HMBC spectral analyses. The amount of GlcCERs in oat grain, quantified by HPTLC, was found to be 193.5mg/kg. The results indicated the similarity of oat CERs with commercial plant CERs (with comparable GlcCER content) suggesting its potential as source of CERs for oral (as dietary supplements) as well as topical applications.

Structural profiling of wax biopolymer from Pinus roxburghii Sarg. needles using spectroscopic methods

Pinus roxburghii Sarg. is the most abundant species in Himalayan region. The needles of the species largely contribute to the forest biomass and remain the major cause of forest fires leading to climate change, biodiversity loss, etc. Intriguingly, the layer of needles contains wax, a biomacromolecule with potential chemical functionalities for value addition. In the present study, a distinctive approach towards complete structural analysis of the isolated wax in its native state has been done using 1H, 13C, HSQC, HMBC, COSY, TOCSY along with GC–MS of the methyl esters of constituent fatty acids. The wax was isolated in a quantitative yield of 1.64% and analyses suggest that it is a polymer of linearly attached fatty acid esters which on hydrolysis yielded three types of ω-hydroxy fatty acids viz. 12-hydroxydodecanoic acid, 14-hydroxytetradecanoic acid and 16-hydroxyhexadecanoic acid in a ratio of 1:1:2 respectively. Complete assignments for a carbonyl group, α-, β- and other methylenes present in wax were achieved; corroborating the presence of polyester. In particular, identification of wax structure was accomplished through NMR; thereby providing a lead towards future structural analysis of waxes in their native form. The study would also be helpful to generate commercially important compounds derived from pine needle wax. This will offer an opportunity for utilisation of pine needle biomass: a root cause of Himalayan forest fires.

Aquidulcibacter paucihalophilus gen. nov., sp. nov., a novel member of family Caulobacteraceae isolated from cyanobacterial aggregates in a eutrophic lake.

A bacterial strain, TH1-2T, was isolated from cyanobacterial aggregates in the eutrophic Lake Taihu, Jiangsu Province, China. Cells were observed to be Gram-negative, slightly curved and rod-shaped. Optimal growth was obtained at pH 7.0 (range 5.5-8.5) and 30°C (range 20-37°C) in trypticase soy broth (TSB) without NaCl. Growth was not observed in TSB with 1.0% (w/v) NaCl added. The cells were found to be positive for oxidase and catalase activities. The major fatty acids were identified as 3-hydroxy hexadecanoic acid (C16:0 3-OH), C16:1 ω5c and summed feature 8 (consisting of C18:1 ω6c and cis-11-Octadecenoic acid (C18:1 ω7c). The major polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The 16S rRNA gene sequence of strain TH1-2T was found to be phylogenetically related to those of Glycocaulis abyssi MCS 33T and Brevundimonas naejangsanensis BIO-TAS2-2T (90.5 and 90.0% similarity, respectively). The genomic G+C content of strain TH1-2T was 55.6mol% based on whole genome calculations. Average nucleotide identities (ANI) and the digital DNA-DNA hybridizations (DDH) for complete genomes ranged from 66.84 to 67.32 and 21.3 to 31.8% between strain TH1-2T and type strains within the family Caulobacteraceae, higher than those between strain TH1-2T and the strains within the family Hyphomonadaceae. The phenotypic, chemotaxonomic and phylogenetic properties, and genome analysis, indicate that strain TH1-2T (=CGMCC 1.12979T=LMG 28362T) represents a novel species in a new genus within the family Caulobacteraceae; thus, the name Aquidulcibacter paucihalophilus gen. nov., sp. nov., is proposed.

Isolation and structural characterization of glucosylceramides from Ethiopian plants by LC/APCI-MS/MS

Chronic skin conditions such as atopic dermatitis, psoriasis and aged skin are characterized by defective skin barrier and dryness which are associated with reduced levels of skin ceramides (CERs). The beneficial effects of plant-derived CERs for skin hydration and skin barrier recovery have been shown in several studies. Although plenty of glucosylceramide (GlcCER)-based dietary supplements meant for skin barrier improvement have been marketed, there are limited commercial sources of plant GlcCERs. In an attempt to explore alternative GlcCER sources, a reversed phase LC–MS/MS method with atmospheric pressure chemical ionization (APCI) interface was developed for separation and structural identification of GlcCERs isolated from three plants. The GlcCERs were extracted from the seeds of grass pea (Lathyrus sativus L.), Ethiopian mustard (Brassica carinata) and haricot bean (Phaseolus vulgaris) and purified by column chromatography and preparative LC–MS. The individual GlcCER species were further separated and qualitatively analyzed by LC/APCI-MS/MS. The amount of GlcCERs in each plant was quantified by HPTLC. All GlcCER species detected in the three plants consisted of C18 di/trihydroxy sphingoid bases amide linked with hydroxy fatty acids (C14-C24). The trihydroxy SBs were acylated with very long chain FAs (C22-C24). The major GlcCERs derived from grass pea, Ethiopian mustard and haricot bean are composed of sphingenine (d18:1) linked to hydroxypalmitic acid (h16:0), 4-hydroxy-8-sphingenine (t18:1) coupled with hydroxynervonic acid (h24:1) and sphingadienine (d18:2) joined with h16:0, respectively. The GlcCERs contents in haricot bean (161.2mg/kg) and grass pea (130.0mg/kg) were found to be higher compared to Ethiopian mustard (71.8mg/kg). This qualitative and quantitative information suggests that the two plants of the Fabaceae family (haricot bean and grass pea) are potential alternative sources of GlcCERs for their use in products meant for the recovery of skin barrier function. The LC/APCI-MS/MS method described here has proven to be reliable for the screening of other potential plants containing GlcCERs.

Synthesis, characterization and enzymatic degradation of copolymers of ε-caprolactone and hydroxy-fatty acids

Abstract New copolymers of ε-caprolactone with three hydroxy-fatty acids, 12-hydroxy stearic acid, 16-hydroxyhexadecanoic acid and ricinoleic acid, were synthesized by catalytic polyesterification. The reactions were carried out in solvent-free systems and in organic solvents as well, using tin(II) 2-ethylhexanoate as catalyst, at different temperatures and molar ratios of the comonomers. Cyclic and linear polymeric products with medium molar weight of about 2000 Da have been synthesized and their chemical structures were confirmed by FT-IR, NMR and MALDI-TOF MS analysis. The synthesis parameters were optimized and the ε-caprolactone/hydroxy acid molar ratio was set as 5:1, according to mass spectrometry results. The biodegradability of the newly synthesized polymers was studied in the presence of Candida antarctica B lipase in phosphate buffer solutions (pH=7.4), at 37°C. The weight-loss profile emphasized the degradation of the 16-hydroxyhexadecanoic acid based polymer samples at more than 50% of their initial weight in 18 days of incubation in the presence of the lipase. The composition of the degradation products was assessed using the GC-MS technique and displayed residues of the comonomers moieties.

A comparison of the immunological potency of Burkholderia lipopolysaccharides in endotoxemic BALB/c mice.

Lipopolysaccharide is one of the virulence factors of the soil-borne pathogens Burkholderia pseudomallei, B. thailandensis, B. cenocepacia and B. multivorans, which cause septic melioidosis (often in B. pseudomallei infections but rarely in B. thailandensis infections) or cepacia syndromes (commonly in B. cenocepacia infections but rarely in B. multivorans infections). The inflammatory responses in Burkholderia LPS-induced endotoxemia were evaluated in this study. Prior to induction, the conserved structures and functions of each purified LPS were determined using electrophoretic phenotypes, the ratios of 3-hydroxytetradecanoic to 3-hydroxyhexadecanoic acid and endotoxin units. In an in vitro assay, cytokine expression of myeloid differentiation primary response gene 88 and Toll/IL-1 receptor domain containing adapter-inducing INF-β-dependent signaling-dependent signaling differed when stimulated by different LPS. Endotoxemia was induced in mice by s.c. injection as evidenced by increasing serum concentrations of 3-hydroxytetradecanoic acid and the septic prognostic markers CD62E and ICAM-1. During endotoxemia, splenic CD11b+ I-A+ , CD11b+ CD80+ , CD11b+ CD86+ and CD11b+ CD11c+ subpopulations increased. After induction with B. pseudomallei LPS, there were significant increases in splenic CD49b NK cells and CD14 macrophages. The inflamed CD11b+ CCR2+ , CD11b+ CD31+ , CD11b+ CD14+ , resident CD11b+ CX3 CR1+ and progenitor CD11b+ CD34+ cells showed delayed increases in bone marrow. B. multivorans LPS was the most potent inducer of serum cytokines and chemokines, whereas B. cenocepacia LPS induced relatively low concentrations of the chemokines MIP-1α and MIP-1β. Endotoxin activities did not correlate with the virulence of Burkholderia strains. Thus factors other than LPS and/or other mechanisms of low activity LPS must mediate the pathogenicity of highly virulent Burkholderia strains.

Disturbance of mitochondrial functions provoked by the major long-chain 3-hydroxylated fatty acids accumulating in MTP and LCHAD deficiencies in skeletal muscle

The pathogenesis of the muscular symptoms and recurrent rhabdomyolysis that are commonly manifested in patients with mitochondrial trifunctional protein (MTP) and long-chain 3-hydroxy-acyl-CoA dehydrogenase (LCHAD) deficiencies is still unknown. In this study we investigated the effects of the major long-chain monocarboxylic 3-hydroxylated fatty acids (LCHFA) accumulating in these disorders, namely 3-hydroxytetradecanoic (3HTA) and 3-hydroxypalmitic (3HPA) acids, on important mitochondrial functions in rat skeletal muscle mitochondria. 3HTA and 3HPA markedly increased resting (state 4) and decreased ADP-stimulated (state 3) and CCCP-stimulated (uncoupled) respiration. 3HPA provoked similar effects in permeabilized skeletal muscle fibers, validating the results obtained in purified mitochondria. Furthermore, 3HTA and 3HPA markedly diminished mitochondrial membrane potential, NAD(P)H content and Ca2+ retention capacity in Ca2+-loaded mitochondria. Mitochondrial permeability transition (mPT) induction probably underlie these effects since they were totally prevented by cyclosporin A and ADP. In contrast, the dicarboxylic analogue of 3HTA did not alter the tested parameters. Our data strongly indicate that 3HTA and 3HPA behave as metabolic inhibitors, uncouplers of oxidative phosphorylation and mPT inducers in skeletal muscle. It is proposed that these pathomechanisms disrupting mitochondrial homeostasis may be involved in the muscle alterations characteristic of MTP and LCHAD deficiencies.

Changes in cell surface properties and biofilm formation efficiency in Azospirillum brasilense Sp245 mutants in the putative genes of lipid metabolism mmsB1 and fabG1

The previously obtained insertion mutants ofAzospirillum brasilense Sp245 in the genes mmsBl and fabG1 (strains SK039 and Sp245.1610, respectively) were characterized by impaired flagellation and motility. The putative products of expression of these genes are 3-hydroxyisobutyrate dehydrogenase and 3-oxoacyl-[acyl-carrier protein] reductase, respectively. In the present work, A. brasilense- Sp245 strains SK039 and Sp245.1610 were found to have differences in the content of 3-hydroxyhexadecanoic, hexadecanoic, 3-hydroxytetradecanoic, hexadecenoic, octadecenoic, and nonadecanoic acids in their lipopolysaccharide prepa- rations, as well as in cell hydrophobicity and hemagglutination activity and dynamics of cell aggregation, in biomass amount, and in the relative content of lipopolysaccharide antigens in mature biofilms formed on hydrophilic or hydrophobic surfaces.

Deregulation of mitochondrial functions provoked by long-chain fatty acid accumulating in long-chain 3-hydroxyacyl-CoA dehydrogenase and mitochondrial permeability transition deficiencies in rat heart--mitochondrial permeability transition pore opening as a potential contributing pathomechanism of cardiac alterations in these disorders.

Mitochondrial trifunctional protein and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiencies are fatty acid oxidation disorders biochemically characterized by tissue accumulation of long-chain fatty acids and derivatives, including the monocarboxylic long-chain 3-hydroxy fatty acids (LCHFAs) 3-hydroxytetradecanoic acid (3HTA) and 3-hydroxypalmitic acid (3HPA). Patients commonly present severe cardiomyopathy for which the pathogenesis is still poorly established. We investigated the effects of 3HTA and 3HPA, the major metabolites accumulating in these disorders, on important parameters of mitochondrial homeostasis in Ca(2+) -loaded heart mitochondria. 3HTA and 3HPA significantly decreased mitochondrial membrane potential, the matrix NAD(P)H pool and Ca(2+) retention capacity, and also induced mitochondrial swelling. These fatty acids also provoked a marked decrease of ATP production reflecting severe energy dysfunction. Furthermore, 3HTA-induced mitochondrial alterations were completely prevented by the classical mitochondrial permeability transition (mPT) inhibitors cyclosporin A and ADP, as well as by ruthenium red, a Ca(2+) uptake blocker, indicating that LCHFAs induced Ca(2+)-dependent mPT pore opening. Milder effects only achieved at higher doses of LCHFAs were observed in brain mitochondria, implying a higher vulnerability of heart to these fatty acids. By contrast, 3HTA and docosanoic acids did not change mitochondrial homeostasis, indicating selective effects for monocarboxylic LCHFAs. The present data indicate that the major LCHFAs accumulating in mitochondrial trifunctional protein and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiencies induce mPT pore opening, compromising Ca(2+) homeostasis and oxidative phosphorylation more intensely in the heart. It is proposed that these pathomechanisms may contribute at least in part to the severe cardiac alterations characteristic of patients affected by these diseases.

Isolation and characterization of Maclura (Maclura pomifera) extracts obtained by supercritical fluid extraction

The chemical composition of the CO2 extract of Maclura fruit (Osage orange) was analyzed. The major constituents of the CO2 extracts were triterpene alcohol esters, along with the triglycerides of C16 and C18 fatty acids. Column chromatography of the CO2 extract yielded lupeol ester of hexadecanoic acid (2), lupeol ester of 3-hydroxyhexadecanoic acid (β-hydroxy palmitic acid) (3) trace of lupeol (1), butyrospermol ester of hexadecanoic acid (5), butyrospermol acetate (6) and trace of butyrospermol (4) as the least polar components. By consequent extraction with acetone of the lipid free Maclura material an extract was obtained, in which isoflavonoids osajin (7) and pomiferin (8) (natural antioxidant) were found in high yields. The structure of isolated compounds was established by spectroscopic data (IR, 1H and 13C NMR and MS) and some chemical reactions. The 3 is new natural product, not yet described in literature.

&lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; MTCC 7925 as a Biofactory for Production of the Novel SCL-LCL-PHA Thermoplastic from Non-Edible Oils

Novel short-chain-length-long-chain-length polyhydroxyalkanoate (SCL-LCL-PHA) copolymer production was examined with Pseudomonas aeruginosa MTCC 7925 under supplementation of non-edible oils such as karanja, jatropha, mahua, and castor oils, and their respective cakes for cost reduction. Polymer yield reached up to 4.66 g/l (63.7% dry cell wt., dcw) with a mol fraction of 89.7:4.2:2.7:3.4 of 3- hydroxybutyric acid (3HB): 3-hydroxyvaleric acid (3HV): 3-hydroxyhexadecanoic acid (3HHD): 3-hydroxyoctadecanoic acid (3HOD) units under the interactive condition of low nitrogen concentration with 0.5% (v/v) jatropha oil in combination with its cake extract, followed by 3.94 g/l (59.6% dcw) with a mol fraction of 91.6:3.3:2.5:2.6 of 3HB: 3HV: 3HHD: 3HOD with castor oil and its cake extracts. The novel co-polymer not only depicted material properties analogous to the common plastics but also better melting temperature (Tm), glass-transition temperature (Tg), elongation-to-break value and Young’s modulus than the homopolymer of poly-3-hydroxybutyrate (PHB). As compared to our previous report where palm oil and its cakes were used, a cost reduction of 54% was observed with the non-edible jatropha oil with its cakes. This opens up possibility for further study at pilot-scale level for low-cost production and future recommendations. Keywords: Low-cost carbon sources, NH4NO3, KH2PO4, Polyhydroxyalkanoates, Pseudomonas aeruginosa MTCC 7925, Non-edible oils, SCL-LCL-PHA co-polymer

Pectin-lipid self-assembly: influence on the formation of polyhydroxy fatty acids nanoparticles.

Nanoparticles, named cutinsomes, have been prepared from aleuritic (9,10,16-trihidroxipalmitic) acid and tomato fruit cutin monomers (a mixture of mainly 9(10),16-dihydroxypalmitic acid (85%, w/w) and 16-hydroxyhexadecanoic acid (7.5%, w/w)) with pectin in aqueous solution. The process of formation of the nanoparticles of aleuritic acid plus pectin has been monitored by UV-Vis spectrophotometry, while their chemical and morphological characterization was analyzed by ATR-FTIR, TEM, and non-contact AFM. The structure of these nanoparticles can be described as a lipid core with a pectin shell. Pectin facilitated the formation of nanoparticles, by inducing their aggregation in branched chains and favoring the condensation between lipid monomers. Also, pectin determined the self-assembly of cutinsomes on highly ordered pyrolytic graphite (HOPG) surfaces, causing their opening and forming interconnected structures. In the case of cutin monomers, the nanoparticles are fused, and the condensation of the hydroxy fatty acids is strongly affected by the presence of the polysaccharide. The interaction of pectin with polyhydroxylated fatty acids could be related to an initial step in the formation of the plant biopolyester cutin.