Long Chain Fatty Acid

A new and simple method was developed for the extraction and derivatization of fatty acid (FA) in milk. Lipid extraction of milk was carried out in H 2 SO 4 /methanol. Methylation was performed for 2 h at 608C, and FA methyl esters were recovered for chromatographic analysis by the addition of hexane. The method parameters were optimized and the simple method was compared to the official reference procedure for the extraction and methylation of FAs in milk samples. For most of the 24 FAs determined, similar or significantly higher recoveries were obtained by the simple method than by the conventional method. The simple method allows processing a high number of samples, at the same time, minimizing the sample manipulation and, consequently, the sample loss and contamination. In conclusion, the proposed method is simple, rapid, low cost, and achieves good results.

Background: Protein distribution profiles along the human intestinal tract of transporters involved in the absorption of cholesterol and long-chain fatty acids (LCFA) have been scarcely evaluated. Methodology/Principal Findings: In post-mortem samples from 11 subjects, intestinal transporter distribution profiles were determined via Western Blot. Differences in transporter protein levels were statistically tested using ANOVA and Tukey's Post Hoc comparisons. Levels in all segments were expressed relative to those in duodenum. Except for ABCG5 and FATP4, levels (mean6SEM) were the highest in the ileum. For ABCA1, ileal levels (1.8060.26) differed significantly from those in duodenum (P = 0.049) and proximal colon (0.9260.14; P = 0.029). ABCG8 levels in ileum (1.9160.30) differed from those in duodenum (P = 0.041) and distal colon (0.8460.22; P = 0.010) and jejunum (1.6460.26) tended to be higher than distal colon (0.8460.22; P = 0.087). Ileal NPC1L1 levels (2.5660.51) differed from duodenum levels (P = 0.019) and from distal colon (1.0960.22; P = 0.030). There was also a trend (P = 0.098) for higher jejunal (2.2360.37) than duodenal NPC1L1 levels. The levels of ABCG5 did not correlate with those of ABCG8. FAT/CD36 levels in ileum (2.0360.42) differed from those in duodenum (P = 0.017), and proximal and distal colon (0.8960.13 and 0.9760.15 respectively; P = 0.011 and P = 0.014). FABPpm levels in ileum (1.0460.13) differed from proximal (0.6460.07; P = 0.026) and distal colon (0.6660.09; P = 0.037). Conclusions/Significance: The distribution profiles showed a bell-shape pattern along the GI-tract with the highest levels in ileum for ABCA1, ABCG8, NPC1L1, FATCD36 and FABPm, suggesting a prominent role for ileum in transporter-mediated uptake of cholesterol and LCFAs.

Circulating long-chain fatty acids (LCFA) and glucose are the main sources for energy production in the heart. In the healthy heart the ratio of glucose and LCFA oxidation is sensitively balanced and chronic alterations in this substrate mix are closely associated with cardiac dysfunction. While it has been accepted for several years that cardiac glucose uptake is mediated by facilitated transport, i.e. by means of the glucose transport proteins GLUT1 and GLUT4, only in the last few years it has become clear that proteins with high-affinity binding sites to LCFA, referred to as LCFA transporters, are responsible for bulk LCFA uptake. Similar to the GLUTs, the LCFA transporters CD36 and FABP pm can be recruited from an intracellular storage compartment to the sarcolemma to increase the rate of substrate uptake. Permanent relocation of LCFA transporters, mainly CD36, from intracellular stores to the sarcolemma is accompanied by accumulation of lipids and lipid metabolites in the heart. As a consequence, insulin signalling and glucose utilization are impaired, leading to decreased contractile activity of the heart. These observations underline the particular role and interplay of substrate carriers for glucose and LCFA in modulating cardiac metabolism, and the development of heart failure. The signalling and trafficking pathways and subcellular machinery regulating translocation of glucose and LCFA transporters are beginning to be unravelled. More knowledge on substrate transporter recycling, especially the similarities and differences between glucose and LCFA transporters, is expected to enable novel therapies aimed at changing the subcellular distribution of glucose and LCFA transporters, thereby manipulating the substrate preference of the diseased heart to help restore cardiac function.

Cellular long-chain fatty acid (LCFA) uptake constitutes a process that is not yet fully understood. LCFA uptake likely involves both passive diffusion and protein-mediated transport. Several lines of evidence support the involvement of a number of plasma membrane-associated proteins, including fatty acid translocase (FAT)/CD36, plasma membrane-bound fatty acid binding protein (FABPpm), and fatty acid transport protein (FATP). In heart and skeletal muscle primary attention has been given to unravel the mechanisms by which FAT/CD36 expression and function are regulated. It appears that both insulin and contractions induce the translocation of intracellular stored FAT/CD36 to the plasma membrane to increase cellular LCFA uptake. This review focuses on this novel mechanism of regulation of LCFA uptake in heart and skeletal muscle in health and disease. The distinct signaling pathways underlying insulin-induced and contraction-induced FAT/CD36 translocation will be discussed and a comparison will be made with the well-defined glucose transport system involving the glucose transporter GLUT4. Finally, it is hypothesized that malfunctioning of recycling of these transporters may lead to intracellular triacylglycerol (TAG) accumulation and cellular insulin resistance. Current data indicate a pivotal role for FAT/CD36 in the regulation of LCFA utilization in heart and skeletal muscle under normal conditions as well as during the altered LCFA utilization observed in obesity and insulin resistance. Hence, FAT/CD36 might provide a useful therapeutic target for the prevention or treatment of insulin resistance.

In this study, we evaluated serum asymmetric dimethyl arginine (ADMA) level, homocysteine level and its clinical significance in rheumatoid arthritis (RA). The study included 40 patients with RA and 30 controls. Serum ADMA and homocysteine levels were measured using High Performance Liquid Chromatography system. Patients with DAS 28 scores ≤ 3.2, and > 3.2 were allocated into lower and high/moderate disease activity groups, respectively. Additionally patients were divided into 2 groups as early RA (disease duration ≤ 2 years) and established RA (duration of the disease ≥ 2 years). Functional disability was evaluated using Health Assessment Questionnaire (HAQ). Radiographs were scored using the modified Larsen score. Serum ADMA level was significantly higher than controls (P=0.008) in RA patients, while homocysteine level was comparable (P=0.059). Serum ADMA and homocysteine levels did not correlate with clinical and laboratory parameters of disease activity such as erythrocyte se...

Raillietina (Fuhrmannetaa) echinobothrida, (Magnin, 1881) cestode parasite of Gallus gallus domesticus is redescribed on the basis of type material from Aurangabad, Marathawada, Maharashtra, India. The present worms resemble with R. echinobothrida, (Magnin, 1881) in having all essential morphological characters. having scolex oval, rostellum elongated/rounded, presence of four suckers, short neck, mature proglottids are broader than long, testes rounded and excretory canal long tube. But the same differ due to number of testes. Hence it is redescribed. We carried out the phylogenetic analyses to observe molecular resemble of related proteins.

The fatty acids distribution pattern in triacylglycerols of three medicinal seed oils viz. Ipomea biloba (Convolvulaceae), Argemone mexicana (Papaveraceae) and Holarrhena antidysentrica (Apocynaceae) were determined using pancreatic lipase hydrolysis method. All the three seeds contain a high percentage of unsaturated acids (I. biloba-52.94%; A. mexicana-82.68%; H. antidysentrica-93.15%). I. biloba contains relatively greater amount of saturated acids (47.06%) than other two seeds. The lipolytic data revealed that linoleic acid dominates at 2-position of triacylglycerols whereas the other two seeds i.e. A. mexicana and H. antidysentrica show almost similar pattern of distribution with oleic acid preferentially acylated at 2-position.

Chlamydiae are obligatory intracellular parasites which are responsible for various acute and chronic diseases in animals and humans. The outer membrane of the chlamydial cell wall contains a truncated lipopolysaccharide (LPS) antigen, which harbors a group-specific epitope being composed of a trisaccharide of 3-deoxy-D-manno-oct-2-ulosonic (Kdo) residues of the sequence K-Kdo-(2C8)-K-Kdo-(2C4)-K-Kdo. The chemical structure was established using LPS of recombinant Escherichia coli and Salmonella enterica strains after transformation with a plasmid carrying the gene encoding the multifunctional chlamydial Kdo transferase. Oligosaccharides containing the Kdo region attached to the glucosamine backbone of the lipid A domain have been isolated or prepared by chemical synthesis, converted into neoglycoproteins and their antigenic properties with respect to the definition of cross-reactive and chlamydia-specific epitopes have been determined. The low endotoxic activity of chlamydial LPS is related to the unique structural features of the lipid A, which is highly hydrophobic due to the presence of unusual, long-chain fatty acids.

The acute in vitro and in vivo effects of longchain fatty acids (LCFAs) on the regulation of adrenergic lipolysis were investigated in human adipose tissue. The effect of a 2 h incubation, without or with LCFA (200 mmol/l), on basal and hormonally induced lipolysis was tested in vitro on isolated fat cells. The lipolytic response to epinephrine was enhanced by suppression of the antilipolytic a 2adrenergic effect. Then, healthy lean and obese male subjects performed a 45 min exercise bout at 50% of their heart rate reserve either after an overnight fast or 3 h after a highfat meal (HFM: 95% fat, 5% carbohydrates). Subcutaneous adipose tissue lipolysis was measured by microdialysis in the presence or absence of an a-antagonist (phentolamine). In vivo, a HFM increased plasma levels of nonesterified fatty acids in lean and obese subjects. In both groups, the HFM did not alter hormonal responses to exercise. Under fasting conditions, the a 2 -adrenergic antilipolytic effect was more pronounced in obese than in lean subjects. The HFM totally suppressed the a 2 -adrenergic antilipolytic effect in lean and obese subjects during exercise. LCFAs per se, in vitro as well as in vivo, suppress a 2 -adrenergic-mediated antilipolysis in adipose tissue. LCFA-mediated suppression of antilipolytic pathways represents another mechanism whereby a high fat content in the diet might increase adipose tissue lipolysis.

The acute in vitro and in vivo effects of longchain fatty acids (LCFAs) on the regulation of adrenergic lipolysis were investigated in human adipose tissue. The effect of a 2 h incubation, without or with LCFA (200 mmol/l), on basal and hormonally induced lipolysis was tested in vitro on isolated fat cells. The lipolytic response to epinephrine was enhanced by suppression of the antilipolytic a 2adrenergic effect. Then, healthy lean and obese male subjects performed a 45 min exercise bout at 50% of their heart rate reserve either after an overnight fast or 3 h after a highfat meal (HFM: 95% fat, 5% carbohydrates). Subcutaneous adipose tissue lipolysis was measured by microdialysis in the presence or absence of an a-antagonist (phentolamine). In vivo, a HFM increased plasma levels of nonesterified fatty acids in lean and obese subjects. In both groups, the HFM did not alter hormonal responses to exercise. Under fasting conditions, the a 2 -adrenergic antilipolytic effect was more pronounced in obese than in lean subjects. The HFM totally suppressed the a 2 -adrenergic antilipolytic effect in lean and obese subjects during exercise. LCFAs per se, in vitro as well as in vivo, suppress a 2 -adrenergic-mediated antilipolysis in adipose tissue. LCFA-mediated suppression of antilipolytic pathways represents another mechanism whereby a high fat content in the diet might increase adipose tissue lipolysis.

Among factors promoting mucosal hyperplasia after bowel resection, long-chain fatty acids may have a special role. The purpose of the present study was to evaluate the effects of high-fat diet (HFD) on early intestinal adaptation in rats with short bowel syndrome (SBS). Male Sprague-Dawley rats underwent either a bowel transection with re-anastomosis (Sham rats) or 75% small bowel resection (SBS rats). Animals were randomly assigned to one of three groups: Sham rats fed normal chow (Sham-NC); SBS rats fed NC (SBS-NC); and SBS rats fed HFD (SBS-HFD). Rats were killed on days 3 or 14. Body weight and parameters of intestinal adaptation (overall bowel and mucosal weight, mucosal DNA and protein, villus height, and crypt depth) were determined at time of killing. By day 3, SBS-HFD rats demonstrated higher duodenal and jejunal bowel and mucosal weights and ileal villus height and jejunal crypt depth vs SBS-NC rats. By day 14 SBS-HFD rats continued to demonstrate increased duodenal and jejunal bowel weight and duodenal mucosal weight vs SBS-NC animals. We conclude that early exposure to HFD both augmented and accelerated structural bowel adaptation in a rat model of SBS.

order to change substrate specificity of NXRG, five point mutations were introduced by site-directed mutagenesis. Mutated NXRG was placed under the control of a glyceraldehyde 3-phosphate dehydrogenase promoter and integrated into the genome of C. tropicalis K1. The resulting recombinant yeast, C. tropicalis KNV, showed XR activity of 80 mU/mg of proteins toward D-xylose, whereas activity toward L-arabinose was not detected. In fed batch fermentation for xylitol production, KNV showed lower arabitol/xylitol ratio (23%) than that of the control strain (33%).

The aim of this study was to characterize the heat treatment-induced changes in electrophoretic patterns of myofibrillar proteins. The study was carried out on longissimus thoracis (LT) muscle from two pig lines (Galia and Redone). The following post mortem metabolic and meat quality parameters were determined: pH 24h , lactate, residual glycogen, glycolytic potential and drip loss. Heat treatment at 100°C was applied for either 10 min or 30 min. SDS-PAGE electrophoresis was performed before and after cooking. The results showed that Galia-breed LT muscles had lower pH 24h and higher residual glycogen, glycolytic potential and drip loss than Redone-breed LT muscle. We found different electrophoretic patterns between raw and cooked meat for each breed. Electrophoretic pattern profiles after heating showed a decrease in myosin (up to 50%) and actin (up to 40%) together with new bands of lower molecular weight (mainly at 80 kDa, 60 kDa and 25 kDa). The appearance of these new bands could be explained either by heating-induced myosin and actin band breakdown or by changes in protein solubility and extractability. Research is currently being led to identify these proteins and thus confirm these hypotheses.

We aimed to study the protein and gene expression of some hepatic enzymes of lipid metabolism along with plasma biomarkers in grazing dairy cattle during the transition period. Blood and liver biopsies from a group of eight multiparous cows were sampled at -28, -14, +4, +14, +28 and +56 days relative to parturition. Peak concentrations of NEFA and beta-hydroxybutyric acid with high triacylglycerol content in the liver were recorded on day 4 postpartum. Consistent with blood biomarkers, the gene expression of carnitine palmitoyltransferase 1A (CPT1A) and acyl-CoA oxidase 1 (ACOX1) increased, whereas that of diacylglycerol O-acyltransferase 1 (DGAT1) decreased. Nevertheless, CPT1A protein expression did not change during all the period evaluated and ACOX1 protein expression increased on day 56 postpartum. In addition, the protein expression of peroxisome proliferator-activated receptor alpha (PPAR-alpha) increased on day 28 postpartum. On the other hand, DGAT1 protein expression decreased on day 14 postpartum. As expected, the expression of genes associated with fatty acid oxidation increased on the first days postpartum but, notably, protein expression was highest after transition. Since most infectious diseases and metabolic disorders in dairy cattle occur particularly on the first days postpartum, it is not so clear whether an increase in the oxidation capacity of the liver at that time could help to prevent disease and improve dairy production. The valuable results about protein expression of enzymes involved in liver lipid metabolism could help to better characterize the metabolism of dairy cattle during the transition period.

Inhibition of acetyl-CoA carboxylases has the potential for modulating long chain fatty acid biosynthesis and mitochondrial fatty acid oxidation. Hybridization of weak inhibitors of ACC2 provided a novel, moderately potent but lipophilic series. Optimization led to compounds 33 and 37, which exhibit potent inhibition of human ACC2, 10-fold selectivity over inhibition of human ACC1, good physical and in vitro ADME properties and good bioavailability. X-ray crystallography has shown this series binding in the CT-domain of ACC2 and revealed two key hydrogen bonding interactions. Both 33 and 37 lower levels of hepatic malonyl-CoA in vivo in obese Zucker rats.

Four yellow-pigmented group D enterococci of uncertain taxonomic position were isolated from several humans with severe infections. The results of DNA composition, DNA-DNA hybridization, fatty acid content, and biochemical property studies demonstrated that these organisms were slightly related to other previously described yellow-pigmented enterococcal species and constitute a new species, for which we propose the name Enterococcusjlavescens. The type strain of E. jlavescens is strain CCM 439.

Low concentrations of the long-chain fatty acids oleate and stearate inhibited all steps of the anaerobic thermophilic biogas process during digestion of cattle manure. The lag phase increased when the concentrations of oleate and stearate were 0.2 g/1 and 0.5 g/l, respectively, and no growth was found at concentrations of 0.5 g/1 for oleate and 1.0 g/1 for stearate. The toxic effect of these acids was permanent as growth did not occur when inhibited cultures were diluted to a non-inhibitory concentration. No adaptation to the fatty acids toxicity was observed by pre-exposing the cultures to non-inhibitory concentrations and the inhibitory response was the same as for cultures not pre-exposed to the fatty acids. Oleate was less inhibitory when added as a neutral oil in the form of the glycerol ester. This indicates that it is the free fatty acid that influences the bacterial activity. rial growth and biogas production . LCFA can be inhibitory at low concentrations (Nie-

The typical biorefinery platforms are sugar, thermochemical (syngas), carbon-rich chains, and biogas platforms, each offering unique advantages and disadvantages. The sugar platform uses hexose and pentose sugars extracted or converted from plant body mass. The thermochemical (syngas) platform entails a chemical or biological conversion process using pyrolysis or gasification of plants to produce biofuels. The carbon-rich chains platform is used to produce biodiesel from long-chain fatty acids or glycerides. In the present work, we suggest a new platform using volatile fatty acids (VFAs) for the production of biofuels and biochemicals production. The VFAs are short-chain fatty acids composed mainly of acetate and butyrate in the anaerobic digestion (AD) process, which does not need sterilization, additional hydrolysis enzymes (cellulose or xylanase), or a high cost pretreatment step. VFAs are easily produced from almost all kinds of biomass with low lignin content (terrestrial, aquatic, and marine biomass) by the AD process. Considering that raw material alone constitutes 40~80% of biofuel production costs, biofuels made from VFAs derived from waste organic biomass potentially offer significant economical advantage. © KSBB † Both authors contributed equally.

The growing challenge of drug resistance has intensified the search for new therapeutic targets against the virulent pathogen Mycobacterium tuberculosis (Mtb). The complex cell envelope of Mtb contains unique lipids, such as mycolic acids, which contribute to its survival under hostile conditions. While modern drugs like isoniazid inhibit mycolic acid biosynthesis through the fatty acid synthase II (FAS II) complex, alternative bypass pathways may facilitate the emergence of drug resistance. HtdX, a putative β-hydroxyacyl dehydratase gene conserved in the mycobacterial species, is hypothesized to play a role in these alternative fatty acid metabolism pathways. Although HtdX is expressed under nutrient-deficient conditions, its structural and functional characterization remains largely unexplored. This study presents the crystal structures of HtdX, revealing a MaoC-like dehydratase with a double hot-dog fold. Site-directed mutagenesis, enzyme kinetics, and fluorescence spectroscopy highlight the critical roles of the α2-β2 loop and the proline rich PP-loop in substrate specificity. The α2-β2 loop determines fatty acyl chain length specificity, while the PP-loop regulates the interaction between HtdX and the acyl carrier protein (AcpM). Computational predictions, complemented by molecular dynamics simulations and principal component analyses, establish that the N-terminal region of HtdX is essential for membrane binding. Overall, these findings offer insights into HtdX substrate specificity and provide theoretical understanding of its interaction with the membrane.

The function of adipose differentiation-related protein (ADRP) is known to be the uptake of long-chain fatty acids and formation of lipid droplets in lipid-accumulating cells. We hypothesized that ADRP might stimulate activated hepatic stellate cells (HSCs) to accumulate lipids, resulting in their transition to the quiescent state. In this study, cultured HSCs in fifth passages isolated from rat were infected by adenovirus vector expressing ADRP (Ad.GFP-ADRP), and morphologic and functional changes were evaluated in comparison with control HSCs infected by recombinant adenovirus-expressing ␤-galactosidase (Ad.LacZ). In Ad.GFP-ADRP-infected cells only, many tiny lipid droplets were apparent in the cytoplasm, while the outline of the cells was not changed. The ADRP was detected around the lipid droplets. In HSCs with intracellular actin filaments, the staining pattern of the filaments before and after infection with Ad.GFP-ADRP or Ad.LacZ did not differ. The cell proliferation rate was not influenced by infection with Ad.LacZ or Ad.GFP-ADRP. Type I collagen secretion from cells overexpressing ADRP was not significantly different from that of Ad.LacZ-infected cells. In our in vitro study, ADRP overexpression induced the formation of cytoplasmic lipid droplets in activated HSCs but could not convert other characteristics of the activated form into those of the quiescent form.

Our objective was to determine the effects of calcium salts of long-chain fatty acids (CLFA) and rumen-protected methionine (RPM) on plasma concentrations of ghrelin, glucagon-like peptide-1 (7-36) amide (GLP-1) and pancreatic hormones in lactating cows. Four midlactation Holstein cows were used in a 4 x 4 Latin square experiment in each 2-wk periods. Cows were fed corn silage-based diets with supplements of CLFA (1.5% added on dry matter basis), RPM (20 g/d), CLFA plus RPM and without supplement. Jugular blood samples were taken from 1 h before to 2 h after morning feeding at 10 min intervals on day 12 of each period. CLFA decreased DMI, but RPM did not affect DMI. Both supplements of CLFA and RPM did not affect metabolizable energy intake and milk yield and composition. Plasma NEFA, triglyceride (TG) and total cholesterol (T-Cho) concentrations were increased with CLFA alone, but increases of plasma TG and T-Cho concentrations were moderated by CLFA plus RPM. CLFA increased plasma ghrelin concentration and the ghrelin concentration with CLFA plus RPM was the highest among the treatments. Plasma GLP-1, glucagon and insulin concentrations were decreased with CLFA, whereas adding RPM moderated the decrease of plasma glucagon concentration by CLFA. These results suggest that the addition of Met to cows given CLFA increases plasma ghrelin and glucagon concentrations associated with the decrease in plasma TG and T-Cho concentrations.

The free and protein-bound ceramides of dog stratum corneum (SC) were analyzed by thin-layer chromatography after tape stripping of the abdomen of five dogs. The sphingoid bases were identified by gas-liquid chromatography as sphingosine, phytosphingosine, and 6hydroxysphingosine. Electrospray ionization-ion trap mass spectrometry was used to characterize the protein-bound ceramides containing sphingosine and omega-hydroxy long-chain fatty acids. Although the molecular species were the same ones in all dogs, wide quantitative variations in the patterns of SC ceramides were observed in different breeds of dogs. The free ceramide concentration changed with the depth of SC, with a higher concentration in the deep layers, whereas the concentration of protein-bound ceramides remained constant. These results show that canine SC is close to that of humans with respect to ceramides.

Several long-chain fatty acids (LCFA) are natural ligands of nonruminant peroxisome proliferator-activated receptor-γ (PPARG), which, along with its lipogenic target genes, is upregulated in bovine mammary tissue during lactation. Thus, PPARG might represent an important control point of bovine milk fat synthesis. We tested lipogenic gene network expression via quantitative PCR of 19 genes in bovine mammary epithelial cells cultured with 16:0, 18:0, cis-9 18:1, trans-10 18:1, trans-10,cis-12 18:2 [t10c12 conjugated linoleic acid (CLA)], 20:5, ethanol (control), and the PPARG agonist rosiglitazone (ROSI). Triplicate cultures were maintained for 12 h with 50 μM ROSI or 100 μM LCFA. Responses common to 16:0 and 18:0 relative to the control included significantly greater expression of INSIG1 (+298%, +92%), AGPAT6 (+137%, +169%), FABP3 (+755%, +338%), and FABP4 (+171%, 157%). These were coupled with greater intracellular lipid droplet formation and mRNA of ACSS2, LPIN1, SCD, and SREBF2 in response to 16:0, and greater DGAT1 and THRSP with 18:0. Trans-10 18:1 and t10c12 CLA reduced expression of FASN (-60%, -31%), SCD (-100%, -357%), and SREBF1 (-49%, -189%). Furthermore, t10c12 CLA downregulated ACSS2, FABP3, INSIG1, SREBF2, and THRSP expression. Expression of SREBF1 was lower with cis-9 18:1 (-140%) and 20:5 (-125%) compared with the control. This latter LCFA also decreased SCD, SREBF2, and LPL expression. No effects of LCFA or ROSI on PPARG were observed, but ROSI upregulated (+39% to +269%) expression of ACACA, FASN, LPIN1, AGPAT6, DGAT1, SREBF1, SREBF2, and INSIG1. Thus, these genes are putative PPARG target genes in bovine mammary cells. This is the first report showing a direct effect of trans-10 18:1 on bovine mammary cell lipogenic gene expression. The coordinated upregulation of lipogenic gene networks in response to ROSI and saturated LCFA offers support for PPARG activation in regulating bovine milk fat synthesis.

Background: L-Carnitine is the essential endogenous factor for the transport of long-chain fatty acids from the cytoplasm to within the mitochondrion where the β-oxidation process takes place. L-Carnitine is a superoxide scavenger and an antioxidant that possesses an anti-ischemic action and a stabilizing effect on cell membranes. It may be of help in liver ischemia reperfusion injury. Results regarding the effects of L-carnitine on liver ischemia and reperfusion injury are few and conflicting. Objective: The aim of this study was to investigate the efficacy of exogenous L-carnitine on lipid peroxidation and protecting liver at different stages of experimental total warm hepatic ischemia-reperfusion (TWHIR) procedure in rats. Methods: This experimental study in healthy, weanling, male Wistar rats (weighing 180-200 g) was conducted at the Experimental Animal

An ethnopharmacological investigation was conducted among the Baka pygmies of Dja biosphere reserve (Cameroon) to collect information on the antimalarial plants used in their daily life. Holarrhena floribunda is one of those plants. Extracts of the stem barks of H. floribunda showed remarkable inhibitory activity against drug-resistant strains of Plasmodium falciparum at doses of 1.02-18.53 µg/mL when tested in vitro against two parasite clones designated as Indochina (W-2) and Sierra Leone (D-6). The aqueous extract was the most active against Indochina (W-2), with IC 50 values of 1.02 µg/mL, while the ethanolic extract appeared to be the most active against Sierra Leone (D-6), with an IC 50 of 4.33 µg/mL. The bioassay-guided fractionation of the neutral fraction of the crude extract led to the isolation of lupeol (1) and its three new long-chain fatty acid ester derivatives, namely, 3-O-(3′-hydroxyeicosanoyl)lupeol (2), 3-O-[(2′-(tetracosyloxy)acetyl]lupeol (3), and 3-O-[(1′′-hydroxyoctadecyloxy)-2′-hydroxypropanoyl]lupeol (4). These new compounds displayed some in vitro inhibition activity against the chloroquine-resistant strain FCR-3 isolated from Gambia and the chloroquine-sensitive standard strain 3D7. The hydroxy group of the fatty acid side chain appears to decrease the observed activity.

Several mouse models for mitochondrial fatty acid β-oxidation (FAO) defects have been developed. So far, these models have contributed little to our current understanding of the pathophysiology. The objective of this study was to explore differences between murine and human FAO. Using a combination of analytical, biochemical and molecular methods, we compared fibroblasts of long chain acyl-CoA dehydrogenase knockout (LCAD -/-), very long chain acyl-CoA dehydrogenase knockout (VLCAD -/-) and wild type mice with fibroblasts of VLCAD-deficient patients and human controls. We show that in mice, LCAD and VLCAD have overlapping and distinct roles in FAO. The absence of VLCAD is apparently fully compensated, whereas LCAD deficiency is not. LCAD plays an essential role in the oxidation of unsaturated fatty acids such as oleic acid, but seems redundant in the oxidation of saturated fatty acids. In strong contrast, LCAD is neither detectable at the mRNA level nor at the protein level in men, making VLCAD indispensable in FAO. Our findings open new avenues to employ the existing mouse models to study the pathophysiology of human FAO defects.

Two genes, accB and accE , that form part of the same operon, were cloned from Streptomyces coelicolor A3(2). AccB is homologous to the carboxyl transferase domain of several propionyl coezyme A (CoA) carboxylases and acyl-CoA carboxylases (ACCases) of actinomycete origin, while AccE shows no significant homology to any known protein. Expression of accB and accE in Escherichia coli and subsequent in vitro reconstitution of enzyme activity in the presence of the biotinylated protein AccA1 or AccA2 confirmed that AccB was the carboxyl transferase subunit of an ACCase. The additional presence of AccE considerably enhanced the activity of the enzyme complex, suggesting that this small polypeptide is a functional component of the ACCase. The impossibility of obtaining an accB null mutant and the thiostrepton growth dependency of a tipAp accB conditional mutant confirmed that AccB is essential for S. coelicolor viability. Normal growth phenotype in the absence of the inducer was restored ...

RESUME -En Asie Centrale, l'élevage du chameau de Bactriane, du dromadaire et de leurs hybrides est destiné en priorité à la production de lait dont la consommation est assurée sous forme de lait cru ou plus généralement fermenté (shubat au Kazakhstan). Ces produits considérés comme des "produits terroir" fortement associés à une identité culturelle locale ont la réputation de présenter des propriétés médicinales et probiotiques auxquelles l'industrie agro-alimentaire commence à s'intéresser. Le shubat, notamment, présente des propriétés antibiotiques et un certain nombre d'effets prophylactiques. Il est utilisé en particulier dans le traitement de la tuberculose, de la gastro-entérite, des ulcères gastriques et pour l'alimentation du nourrisson. Les facteurs "santé" attribués au lait de chamelle et ses produits transformés peuvent être liés à certains de ses composants : lactoferrine, immunoglobuline, lysozyme, lactoperoxydase, vitamine C dont les facteurs de variation liés à l'alimentation, la saison, le stade physiologique sont à l'étude. Ces composants généralement présents dans des laits d'autres espèces, auraient la particularité chez la chamelle d'être thermorésistants et parfois, comme c'est le cas pour la lactoferrine ou la vitamine C de s'y trouver en quantité 10 à 100 fois plus importante que dans le lait de vache.

The objective of present study was to investigate the effect of feed additive on base of L-carnitine (30%) on some parameters of egg quality. Totally 40 White Leghorn laying hens were divided in two groups: control group (n=20) received drinking water without any additives and experimental group (n=20) received feed additive in dose 1 ml per 1.2 l of drinking water for a period of 5 days in 2 week intervals from 26 to 56 week of age. Hens in both groups were feeding ad libitum with commercial layer diet. A total of 200 eggs (100 eggs from both group) were collected to study for external and internal quality of eggs. Albumen index and Haugh unit were improved (p<0.05), while yolk index and yolk color were not affected by administration of L-carnitine (p˃0.05). The albumen percentage increased (p<0.05) and that yolk percentage decreased (p<0.05) in response to supplementation of L-carnitine. Addition of L-carnitine did not influence egg weight, egg shape index and egg shell quality (shell percentage, shell thickness, shell strength).

The objective of present study was to investigate the effect of feed additive on base of L-carnitine (30%) on some parameters of egg quality. Totally 40 White Leghorn laying hens were divided in two groups: control group (n=20) received drinking water without any additives and experimental group (n=20) received feed additive in dose 1 ml per 1.2 l of drinking water for a period of 5 days in 2 week intervals from 26 to 56 week of age. Hens in both groups were feeding ad libitum with commercial layer diet. A total of 200 eggs (100 eggs from both group) were collected to study for external and internal quality of eggs. Albumen index and Haugh unit were improved (p&lt;0.05), while yolk index and yolk color were not affected by administration of L-carnitine (p˃0.05). The albumen percentage increased (p&lt;0.05) and that yolk percentage decreased (p&lt;0.05) in response to supplementation of L-carnitine. Addition of L-carnitine did not influence egg weight, egg shape index and egg shell q...

Objective:To use the technique of complementation analysis to help define genofype and classify patients with clinical manifestations consistent with those of the disorders of peroxisome assembly, namely the Zellweger syndrome (ZS), neonatal adrenoleukodystrophy (NALD), infantile Refsum disease (IRD), and rhizomelic chondrodysplasia punctata (RCDP). Study design: Clinical findings, peroxisomal function, and complementation groups were examined in 173 patients with the clinical manifestations of these disorders. Results: In 37 patients (21%), peroxisome assembly was intact and isolated deficiencies of one of five peroxisomal enzymes involved in the 13-oxidation of fatty acids or plasmalogen biosynthesis were demonstrated. Ten complementation groups were identified among 93 patients (54%) with impaired peroxisome assembly and one of three phenotypes (ZS, NALD, or IRD) without correlation between complementation group and phenotype. Forty-three patients (25%) had impaired peroxisome assembly associated with the RCDP phenofype and belonged to a single complementation group. Of the 173 patients, 10 had unusually mild clinical manifestations, including survival to the fifth decade or deficits limited to congenital cataracts. Conclusions: At least 16 complementation groups, and hence genofypes, are associated with clinical manifestations of disorders of peroxisome assembly. The range of phenotype is wide, and some patients have mild involvement. (J PEDIATR 1995;I 27:13-22)

Foxtail millet (Setaria itnlica), consumed by the rural and tribal populations in South India, is a rich source of nutrients. Fermentation by endogenous microflora increased the total soluble sugars and reducing sugars with a simultaneous decrease in the starch content. The protein extractability and albumin/globulin fractions were improved. The beneficial long-chain fatty acid profile of raw flour was retained. Acetic and butyric acid were the major short-chain fatty acids produced. Most of these changes occurred in the first 24 h of fermentation.

BACKGROUND & AIMS-Long-chain fatty acid receptors G-protein-coupled receptor 40 (GPR40) (FFAR1) and GPR120 have been implicated in the chemosensation of dietary fats. I cells in the intestine secrete cholecystokinin (CCK), a peptide hormone that stimulates digestion of fat and protein, but these cells are rare and hard to identify. We sought to determine whether dietary fat-induced secretion of CCK is directly mediated by GPR40 expressed on I cells. METHODS-We used fluorescence-activated cell sorting to isolate a pure population of I cells from duodenal mucosa in transgenic mice that expressed green fluorescent protein under the control of the CCK promoter (CCK-enhanced green fluorescent protein [eGFP] bacterial artificial chromosome mice). CCK-eGFP cells were evaluated for GPR40 expression by quantitative reverse transcription polymerase chain reaction and immunostaining. GPR40 -/-mice were bred with CCK-eGFP mice to evaluate functional relevance of GPR40 on long-chain fatty acidstimulated increases in [Ca 2+ ]i and CCK secretion in isolated CCK-eGFP cells. Plasma levels of CCK after olive oil gavage were compared between GPR40 +/+ and GPR40 -/-mice. RESULTS-Cells that expressed eGFP also expressed GPR40; expression of GPR40 was 100fold greater than that of cells that did not express eGFP. In vitro, linoleic, oleic, and linolenic acids increased [Ca 2+ ]i; linolenic acid increased CCK secretion by 53% in isolated GPR40 +/+ cells that expressed eGFP. In contrast, in GPR40 -/-that expressed eGFP, [Ca 2+ ]i response to linoleic acid was reduced by 50% and there was no significant CCK secretion in response to linolenic acid. In mice, olive oil gavage significantly increased plasma levels of CCK compared with pregavage levels: 5.7-fold in GPR40 +/+ mice and 3.1-fold in GPR40 -/-mice.

The antioxidant activities of TBHQ, BHT and tocopherol mixtures at different concentrations (0, 100, 200, 400 mg/kg) have been evaluated by the application of Differential Scanning Calorimetry (DSC) and Rancimat apparatus. Rancimat apparatus that measures the secondary oxidation products was set at 110 ⁰C with an air flow of 18-20 L/h to measure the induction period (IP) of the samples. The DSC method was concerned with the oxidation of the samples in an oxygen flow DSC cell, while the cell temperature was set at isothermal temperature of 150⁰C. Oxidative induction time (T0), and the maximum of the peak (Tmax) were improved with increasing antioxidant concentrations. According to the results, tocopherols mixture was the most efficient antioxidant, followed by TBHQ and BHT. Furthermore, it was indicated that there is a good agreement (P&lt;0.0001) between both methods and values obtained. In conclusion, the DSC method is a fast, convenient and reliable method to predict the antioxida...

Published data on treatment of fatty acid oxidation defects are scarce. Treatment recommendations have been developed on the basis of observations in 75 patients with long-chain fatty acid oxidation defects from 18 metabolic centres in Central Europe. Recommendations are based on expert practice and are suggested to be the basis for further multicentre prospective studies and the development of approved treatment guidelines. Considering that disease complications and prognosis differ between different disor-Communicating editor: Bridget Wilcken

Binding of 13C-enriched oleic acid to bovine serum albumin and to three large proteolytic fragments of albumin--two complementary fragments corresponding to the two halves of albumin and one fragment corresponding to the carboxyl-terminal domain--yielded unique patterns of NMR resonances (chemical shifts and relative intensities) that were used to identify the locations of binding of the first 5 mol of oleic acid to the multidomain albumin molecule. The first 3 mol of oleic acid added to intact albumin generated three distinct NMR resonances as a result of simultaneous binding of oleic acid to three heterogeneous sites (primary sites). Two of these resonances were seen upon addition of 1 or 2 mol of oleic acid to fragments representing either the carboxyl-terminal half (residues 307-582) or the carboxyl-terminal domain (residues 377-582); the third resonance was seen upon addition of 1 mol of oleic acid to the fragment representing the amino-terminal half (residues 1-306). The reson...

The remarkable binding properties of serum albumin have been investigated extensively, but little is known about an important class of fatty acids, the very longchain saturated fatty acids (VLCFA; Ͼ 18 carbons). Although VLCFA are metabolized efficiently in normal individuals, they are markers for and possibly causative agents of several peroxisomal disorders. We studied the binding of [ 13 C]carboxyl-enriched arachidic (C20:0), behenic (C22:0), lignoceric (C24:0), and hexacosanoic (C26:0) acids to bovine serum albumin (BSA) by 13 C-NMR spectroscopy. For each VLCFA, the NMR spectra showed multiple signals at chemical shifts previously identified for long-chain fatty acids (12-18 carbons), suggesting stabilization of binding by similar, if not identical, interactions of the fatty acid carboxyl anion with basic amino acid residues. The maximal binding (mol of VLCFA/mol of BSA) and the number of observed binding sites decreased with increasing chain length, from 4-5 for C20:0, 3-4 for C22:0, and 2 for C24:0; we validated our previous conclusion that BSA has only one site for C26:0 (Ho, J. K., H. Moser, Y. Kishimoto, and J. A. Hamilton. 1995. J. Clin. Invest. 96: 1455-1463). Analysis of chemical shifts suggested that the highest affinity sites for VLCFA are low affinity sites for long-chain fatty acids. In competition experiments with 13 C-labeled C22:0 (3 mol/ mol of BSA) and unlabeled oleic acid, C22:0 bound to BSA in the presence of up to 4 mol of oleic acid/mol of BSA, but 1 mol was shifted into a different site. Our studies suggest that albumin has adequate binding capacity for the low plasma levels of VLCFA with 20 to 26 carbons, but the protein may not be able to bind longer chain VLCFA. -

We determined the structural features necessary for fatty acids to exert their action on K+ channels of gastric smooth muscle cells. Examination of the effects of a variety of synthetic and naturally occurring lipid compounds on K+ channel activity in cell-attached and excised membrane patches revealed that negatively charged analogs of medium to long chain fatty acids (but not short chain analogs) as well as certain other negatively charged lipids activate the channels. In contrast, positively charged, medium to long chain analogs suppress activity, and neutral analogs are without effect. The key requirements for effective compounds seem to be a sufficiently hydrophobic domain and the presence of a charged group. Furthermore, those negatively charged compounds unable to &quot;flip&quot; across the bilayer are effective only when applied at the cytosolic surface of the membrane, suggesting that the site of fatty acid action is also located there. Finally, because some of the effecti...

Background: Chronic ethanol (EtOH) consumption impairs gut-liver axis by causing dysbiosis, colonic epithelial tight junction (TJ) disruption and barrier dysfunction, a mechanism involved in the pathogenesis of alcoholic liver disease (ALD). Alcoholic dysbiosis is associated with reduced probiotic populations. Probiotics are known to protect the intestinal mucosa from various insults. We investigated the protective and mitigating roles of Lactobacillus plantarum (LP) in EtOH-induced gut and liver injury. Methods: Mice were fed EtOH (0% 2d, 1% 2d, 2% 2d, 4% 1 wk, 5% 1 wk and 6% 1 wk) in Lieber-DeCarli liquid diet with or without LP; control mice were pair fed with isocaloric maltodextrin. Gut permeability was measured by vascular-to-luminal flux of FITC-inulin in vivo. Plasma endotoxin level was measured to assess endotoxemia. TJ (occludin and ZO-1) and adherens junction (AJ) (E-cadherin and b-catenin) integrity was analyzed by confocal microscopy. Reduced and oxidized protein thiols were evaluated by fluorescence staining using BODIPY FL-N-(2aminoethyl) maleimide. Liver triglyceride (TG) and plasma AST/ALT were measured to assess liver injury. Expression of antioxidants (SOD1, SOD2, Trx1, catalase, GPX1 & Nrf2), cytokine (IL-1β & TNFα) and chemokine (MCP-1, CCL5, CXCL1 & CXCL2) genes were evaluated in liver and distal colon by qRT-PCR. Results: LP feeding attenuated EtOH-induced disruption of colonic epithelial TJ, increase in inulin permeability, and elevation of plasma endotoxin. EtOH feeding induced protein thiol oxidation in the colon, which was associated with down regulation of antioxidant gene expression, and LP blocked these effects of EtOH. EtOH up regulated the expression of cytokine and chemokine genes, which was attenuated by LP. LP also prevented EtOH-induced elevation of plasma AST/ALT and liver TG, down regulation of antioxidant genes and up regulation of cytokine/chemokine genes in liver and colon. In Caco-2 cell monolayers, EtOH caused an increase in inulin permeability and disruption of TJ that was blocked by LP pretreatment. LP effect on EtOH-induced TJ disruption and inulin permeability was blocked by AG1478 (EGFR kinase inhibitor) and anti-EGFR antibody. To determine the effect of EGFR in LP effect on alcoholic gut and liver injury EtOH feeding experiment was conducted in rTA-EGFR*Tg mice that expressed dominant negative EGFR by doxycycline-inducible mechanism. LP effect on alcoholic gut and liver injury was blocked by doxycycline, demonstrating the role of EGFR in the protective effect of LP. Furthermore, LP administration following the EtOH-induced gut and liver injury accelerated the recovery process. Conclusion: These data demonstrate that LP prevents and mitigates EtOH-induced gut and liver injury by blocking the down regulation of antioxidants and up regulation of cytokine/chemokine genes by EGFR-dependent mechanism.

A recently discovered peroxygenase from the fungus Marasmius rotula (MroUPO) is able to catalyze the progressive one-carbon shortening of medium and long-chain mono- and dicarboxylic acids by itself alone, in the presence of H2O2. The mechanism, analyzed using H218O2, starts with an α-oxidation catalyzed by MroUPO generating an α-hydroxy acid, which is further oxidized by the enzyme to a reactive α-keto intermediate whose decarboxylation yields the one-carbon shorter fatty acid. Compared with the previously characterized peroxygenase of Agrocybe aegerita, a wider heme access channel, enabling fatty acid positioning with the carboxylic end near the heme cofactor (as seen in one of the crystal structures available) would be at the origin of the unique ability of MroUPO shortening carboxylic acid chains.

A recently discovered peroxygenase from the fungus Marasmius rotula (MroUPO) is able to catalyze the progressive one-carbon shortening of medium and long-chain mono- and dicarboxylic acids by itself alone, in the presence of H2O2. The mechanism, analyzed using H218O2, starts with an α-oxidation catalyzed by MroUPO generating an α-hydroxy acid, which is further oxidized by the enzyme to a reactive α-keto intermediate whose decarboxylation yields the one-carbon shorter fatty acid. Compared with the previously characterized peroxygenase of Agrocybe aegerita, a wider heme access channel, enabling fatty acid positioning with the carboxylic end near the heme cofactor (as seen in one of the crystal structures available) would be at the origin of the unique ability of MroUPO shortening carboxylic acid chains.

All eukaryotic and prokaryotic organisms are thought to synthesize fatty acids using a type I or type II synthase. In addition, eukaryotes extend pre-existing long chain fatty acids using microsomal elongases (ELOs). We have found that Trypanosoma brucei, a eukaryotic human parasite that causes sleeping sickness, uses three elongases instead of type I or type II synthases for the synthesis of nearly all its fatty acids. Trypanosomes encounter diverse environments during their life cycle with different fatty acid requirements. The tsetse vector form requires synthesis of stearate (C18), whereas the bloodstream form needs myristate (C14). We find that trypanosome fatty acid synthesis is modular, with ELO1 converting C4 to C10, ELO2 extending C10 to C14, and ELO3 elongating C14 to C18. In blood, ELO3 downregulation favors myristate synthesis, whereas low concentrations of exogenous fatty acids in cultured parasites cause upregulation of the entire pathway, allowing the parasite to adapt to different environments.

All eukaryotic and prokaryotic organisms are thought to synthesize fatty acids using a type I or type II synthase. In addition, eukaryotes extend pre-existing long chain fatty acids using microsomal elongases (ELOs). We have found that Trypanosoma brucei, a eukaryotic human parasite that causes sleeping sickness, uses three elongases instead of type I or type II synthases for the synthesis of nearly all its fatty acids. Trypanosomes encounter diverse environments during their life cycle with different fatty acid requirements. The tsetse vector form requires synthesis of stearate (C18), whereas the bloodstream form needs myristate (C14). We find that trypanosome fatty acid synthesis is modular, with ELO1 converting C4 to C10, ELO2 extending C10 to C14, and ELO3 elongating C14 to C18. In blood, ELO3 downregulation favors myristate synthesis, whereas low concentrations of exogenous fatty acids in cultured parasites cause upregulation of the entire pathway, allowing the parasite to adapt to different environments.

Leaves, stem, roots and inflorescence of Solanum torvum Sw. were extracted in two different organic solvents (chloroform & methanol). Antibacterial and antifungal effects of the extracts were tested on fifteen (six Gram positive & nine Gram negative) human pathogenic bacteria and on eight pathogenic fungi. Methanolic extracts of roots of S. torvum exhibited promising antibacterial and antifungal effects on all organisms tested in comparison with that observed in the leaves, stems and inflorescence extracts. The toxicity of the extracts was in the following order; root>stem>inflorescence>leaf. The minimum inhibitory concentration (MIC) values of methanolic extract of roots of S. torvum were in the range between 64-128 µg mL-1. Chloroform extracts of roots were more toxic (LC 50 35.4629 ppm) than other extracts analyzed in Brine shrimp test. In conclusion, S. torvum appears to be an attractive material for the development of antimicrobial drugs and environment friendly biopesticides.