HEPATOLOGY, Issue 6 2005
Alan F. Hofmann
Experiments were performed in 2 volunteers to define the biotransformation and physiological properties of norursodeoxycholic acid (norUDCA), the C23 (C24 -nor) homolog of UDCA. To complement the in vivo studies, the biotransformation of norUDCA ex vivo using precision-cut human liver slices was also characterized. In the human studies, both a tracer dose given intravenously and a physiological dose (7.9 mmol, 3.0 g) given orally were excreted equally in bile and urine. By chromatography and mass spectrometry, the dominant biotransformation product of norUDCA in bile and urine was the C-23 ester glucuronide. Little N -acyl amidation (with glycine or taurine) occurred. The oral dose induced a sustained bicarbonate-rich hypercholeresis, with total bile flow averaging 20 ,L/kg/min, a rate extrapolating to 2 L/d. The increased bile flow was attributed to cholehepatic shunting of norUDCA as well to the lack of micelles in bile. Phospholipid and cholesterol secretion relative to bile acid secretion decreased during secretion of norUDCA and its metabolites, presumably also because of the absence of micelles in canalicular bile. When incubated with human liver slices, norUDCA was glucuronidated, whereas UDCA was conjugated with glycine or taurine. In conclusion, in humans, norUDCA is glucuronidated rather than amidated. In humans, but not animals, there is considerable renal elimination of the C-23 ester glucuronide, the dominant metabolite. NorUDCA ingestion induces a bicarbonate-rich hypercholeresis and evokes less phospholipid and cholesterol secretion into bile than UDCA. Molecules that undergo cholehepatic shunting should be powerful choleretics in humans. (HEPATOLOGY 2005;42:1391,1398.) [source]

Phospholipid signalling through phospholipase D and phosphatidic acid

IUBMB LIFE, Issue 8 2006
Rosanna Cazzolli
Abstract Phospholipase D (PLD) hydrolyzes the phosphodiester bond of the predominant membrane phospholipid, phosphatidylcholine producing phosphatidic acid and free choline. This activity can participate in signal transduction pathways and impact on vesicle trafficking for secretion and endocytosis, as well as receptor signalling. Phospholipids can regulate PLD activity directly, through specific intermolecular interactions, or indirectly, through their effect on the localization or activity of PLD's protein effectors. This short review highlights these various phospholipid inputs into the regulation of PLD activity and also reviews potential roles for PLD-generated phosphatidic acid, particularly a mechanism by which the phospholipid may participate in the process of vesicular trafficking. iubmb Life, 58: 457 - 461, 2006 [source]

Evidence for surfactant contributing to the gastric mucosal barrier of the horse

EQUINE VETERINARY JOURNAL, Issue 6 2000
M. T. ETHELL
Summary This study was undertaken to determine the hydrophobicity of the luminal surface of the equine stomach and to elucidate the ultrastructure of the lining imparting that property. Gastric and duodenal mucosal samples from 5 horses were collected immediately after euthanasia and subjected to surface contact angle measurement using a goniometer. Gastric mucosal samples from 4 horses and a foal were examined by electron microscopy following a fixation procedure known to preserve phospholipids and oligolamellar structures. Contact angles for the equine gastric glandular mucosal surface (mean ± s.e. 78.0 ± 11.0°) were greater than for the duodenum (33.4 ± 8.7°), (P = 0.003). The contact angles for gastric squamous tissue (50.4 ± 4.5°) tended to be greater than for duodenum (P = 0.15). Electron microscopy revealed the existence of surfactant as abundant osmiophilic phospholipid material within both squamous and glandular gastric mucosae. These results indicate the hydrophobic nature of the equine gastric mucosae. We propose that the water-repellent nature of the stomach contributes to the ,gastric mucosal barrier' and is imparted by surface-active phospholipid adsorbed to the surface. Phospholipids may also be utilised as a physical barrier to back-diffusion of acid by lining intracellular canaliculi and oxyntic ducts where other defence mechanisms are absent. [source]

Identification of phospholipids as new components that assist in the in vitro trimerization of a bacterial pore protein

FEBS JOURNAL, Issue 3 2001
Hans De Cock
The in vitro trimerization of folded monomers of the bacterial pore protein PhoE, into its native-like, heat- and SDS-stable form requires incubations with isolated cell envelopes and Triton X-100. The possibility that membranes could be isolated that are enriched in assembly factors required for assembly of the pore protein was now investigated. Fractionation of total cell envelopes of Escherichia coli via various techniques indeed revealed the existence of membrane fractions with different capacities to support assembly in vitro. Fractions containing mainly inner membrane vesicles supported the formation of trimers that were associated with these membrane vesicles. However, only a proportion of these trimers were heat- and SDS-stable and these were formed with slow kinetics. In contrast, fractions containing mainly outer membrane vesicles supported formation of high amounts of heat-stable trimers with fast kinetics. We identified phospholipids as active assembly components in these membranes that support trimerization of folded monomers in a process with similar characteristics as observed with inner membrane vesicles. Furthermore, phospholipids strongly stimulate the kinetics of trimerization and increase the final yield of heat-stable trimers in the context of outer membranes. We propose that lipopolysaccharides stabilize the assembly competent state of folded monomers as a lipochaperone. Phospholipids are involved in converting the folded monomer into new assembly competent intermediate with a short half-life that will form heat-stable trimers most efficiently in the context of outer membrane vesicles. These results provide biochemical evidence for the involvement of different lipidic components at distinct stages of the porin assembly process. [source]

Dihomo-,-linolenic acid inhibits tumour necrosis factor-, production by human leucocytes independently of cyclooxygenase activity

IMMUNOLOGY, Issue 3 2003
Maaike M. B. W. Dooper
Summary Dietary oils (such as borage oil), which are rich in ,-linolenic acid (GLA), have been shown to be beneficial under inflammatory conditions. Dihomo-GLA (DGLA) is synthesized directly from GLA and forms a substrate for cyclooxygenase (COX) enzymes, resulting in the synthesis of lipid mediators (eicosanoids). In the present study, the immunomodulatory effects of DGLA were investigated and compared with those of other relevant fatty acids. Freshly isolated human peripheral blood mononuclear cells (PBMC) were cultured in fatty acid (100 µm)-enriched medium for 48 hr. Subsequently, cells were stimulated with lipopolysaccharide (LPS) for 20 hr and the cytokine levels were measured, in supernatants, by enzyme-linked immunosorbent assay (ELISA). Phospholipids were analysed by gas chromatography. Fatty acids were readily taken up, metabolized and incorporated into cellular phospholipids. Compared with the other fatty acids tested, DGLA exerted pronounced modulatory effects on cytokine production. Tumour necrosis factor-, (TNF-,) and interleukin (IL)-10 levels were reduced to 60% of control levels, whereas IL-6 levels were not affected by DGLA. Kinetic studies showed that peak levels of TNF-,, occurring early after LPS addition, were inhibited strongly, whereas IL-10 levels were not affected until 15 hr after stimulation. Both the reduction of cytokine levels and the decrease in arachidonic acid levels in these cells, induced by DGLA, were dose dependent, suggesting a shift in eicosanoid-subtype synthesis. However, although some DGLA-derived eicosanoids similarly reduced TNF-, levels, the effects of DGLA were probably not mediated by COX products, as the addition of indomethacin did not alter the effects of DGLA. In conclusion, these results suggest that DGLA affects cytokine production by human PBMC independently of COX activation. [source]

Phospholipid signalling through phospholipase D and phosphatidic acid

Relationship Between Fragmentation of Myofibrils and Liberation of Phospholipids from Z-Disks Induced by Calcium Ions at 0.1 mM: Mechanism of Tenderization of Pork and Beef during Postmortem Aging

JOURNAL OF FOOD SCIENCE, Issue 9 2003
K. SHIMADA
ABSTRACT We studied the myofibril fragmentation using porcine and bovine myofibrils to determine the mechanism by which myofibrils are fragmented during postmortem aging of meat. Both myofibril fragmentation and liberation of phospholipids from Z-disks were maximally induced by calcium ions at 0.1 mM. These phenomena showed identical dependencies on pH and temperature in the presence of calcium ions at 0.1 mM. All phospholipids in Z-disks, except for lysophosphatidylethanolamine, had affinity for calcium ions. Therefore, we concluded that liberation of calcium-phospholipid compounds from Z-disks causes weakening of the Z-disk structure, resulting in the myofibril fragmentation during postmortem aging of meat. [source]

Quantification of suppression of bitterness using an electronic tongue

JOURNAL OF PHARMACEUTICAL SCIENCES, Issue 12 2001
Sou Takagi
Abstract Phospholipids, such as phosphatidic acid, suppress bitter taste without affecting other taste qualities. In the present study, we detected and quantified this suppression effect with an electronic tongue whose transducer is composed of several kinds of lipid/polymer membranes with different characteristics. We measured a phospholipid cocktail and various kinds of taste substances with five basic taste qualities. The responses to quinine hydrochloride and L -tryptophan, which have a bitter taste, were reduced as the phospholipid concentration was increased, and the responses to the other taste substances were not affected by the phospholipids, as with the human sensation test. Furthermore, the change of bitter interisity caused by phospholipid was quantified by principal component analysis and the , scale, which expresses the relationship between taste intensity and taste substance concentration. The results are compared with those of the human sensory test and discussed. © 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 90:2042,2048, 2001 [source]

Prerequisites for recombinant factor VIIa-induced thrombin generation in plasmas deficient in factors VIII, IX or XI

JOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 1 2006
T. LIVNAT
Summary.,Background:,Recombinant factor VIIa (rFVIIa) used for the treatment of hemophilia A or B patients with an inhibitor is hemostatically effective because it induces thrombin generation (TG), despite grossly impaired FVIII- and FIX-dependent amplification of FX activation. Tissue factor (TF) and or activated platelets were shown to be essential for the rFVIIa activity. Objective: To evaluate the relative effects of TF and phospholipids on rFVIIa-induced TG in FVIII-, FIX- and FXI-deficient plasmas. Methods: Phospholipids had an independent effect that was augmented by TF. The contribution of blood-borne TF in FVIII-, FIX- and FXI-deficient plasma to rFVIIa-induced TG was demonstrated by removing microparticles and use of anti-TF antibodies. Results: At increasing concentrations of rFVIIa, the dependence of rFVIIa-induced TG on TF declined, but the presence of phospholipids was essential. rFVIIa was also shown to activate purified FIX and FX in the presence of phospholipids and absence of TF. rFVIIa-induced TG was dramatically augmented in FVIII- or FIX-deficient plasma in which the level of FVIII or FIX was increased to 1 or 2 U dL,1. Conclusions: The data indicate that rFVIIa-induced TG is affected by TF, phospholipids, rFVIIa concentration, and the presence of FVIII and FIX. [source]

Phospholipids in liquid chromatography/mass spectrometry bioanalysis: comparison of three tandem mass spectrometric techniques for monitoring plasma phospholipids, the effect of mobile phase composition on phospholipids elution and the association of phospholipids with matrix effects

RAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 14 2009
Yuan-Qing Xia
Because plasma phospholipids may cause matrix effects in bioanalytical liquid chromatography/tandem mass spectrometry (LC/MS/MS) methods, it is important to establish optimal mass spectrometric techniques to monitor the fate of phospholipids during method development and application. We evaluated three MS/MS techniques to monitor phospholipids using positive and negative electrospray ionization (ESI). The first technique is based on using positive precursor ion scan of m/z 184, positive neutral loss scan of 141 Da and negative precursor ion scan of m/z 153. The second technique is based on using class-specific positive and negative selected reaction monitoring (SRM) transitions to monitor class-representative phospholipids. The third technique, previously reported, utilizes in-source collision-induced dissociation (CID)-based positive SRM of m/z 184,,,184. We recommend the all-inclusive technique 1 for use in qualitative assessment of all classes of phospholipids and technique 2 for use in quantitative assessment of class-representative phospholipids. Secondly, we evaluated the elution behaviors of the plasma phospholipids under different reversed-phase mobile phase conditions. The phospholipid-eluting strength of a mobile phase was mainly dependent on the type and amount (%) of the organic eluent and the strength increased in the order of methanol, acetonitrile and isopropyl alcohol. Under the commonly used gradient and isocratic elution schemes in LC/MS/MS bioanalysis, not all the phospholipids are eluted off the column. Thirdly, we investigated the association between phospholipids and matrix effects in positive and negative ESI using basic, acidic and neutral analytes. While the phospholipids caused matrix effects in both positive and negative ESI, the extent of ionization suppression was analyte-dependent and was inversely related to the retention factor and broadness of the phospholipids peaks. The lysophospholipids which normally elute earlier in reversed-phase chromatography are more likely to cause matrix effects compared to the later-eluting phospholipids in spite of the larger concentrations of the latter in plasma. Copyright © 2009 John Wiley & Sons, Ltd. [source]

Modeling Lipid Uptake in Expanded Polytetrafluoroethylene Vascular Prostheses and Its Effects on Mechanical Properties

ARTIFICIAL ORGANS, Issue 5 2000
Diego Mantovani
Abstract: The radial transport across the wall of expanded polytetrafluoroethylene (ePTFE) arterial prostheses has a significant effect on lipid uptake observed in prostheses implanted in humans, which has been postulated to be one of the causes associated with implant failure. The goal of this study was to stimulate radial transport on a lipidic dispersion across the wall of an ePTFE prosthesis and investigate its effects on the circumferential mechanical properties of the prosthesis. An in vitro model was developed to simulate the lipidic radial transport across the wall. Lipids contained in a phosphatidylcholine dispersion were used as the transported molecules. Lipid concentration profiles were obtained after exposing commercial ePTFE prostheses to various transmural pressure and/or lipidic concentration gradients. Phospholipids gradually accumulated up to the external reinforcing wrap of the prosthesis, which clearly acted as a rigid barrier against lipid infiltration. Tensile tests performed on the virgin samples showed that the wrap was much more rigid than the microporous part of the prosthesis. After the lipid simulation, the rigidity of the wrap decreased with respect to what was observed for the virgin prosthesis. Finally, some clinical implications of this phenomena are discussed. [source]

Evidence for surfactant contributing to the gastric mucosal barrier of the horse

Supplemental dietary flaxseed oil affects both neutral and phospholipid fatty acids in cultured tilapia

EUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY, Issue 8 2008
Nilson E. de Souza
Abstract This work aimed to evaluate the neutral lipid (NL) and phospholipid (PL) classes in tilapia (Oreochromis niloticus) muscle tissue. Tilapias were raised in captivity for a period of 5,months with increasing levels (0, 1.25, 2.50, 3.75, and 5.00%) of flaxseed oil [source of ,-linolenic acid (LNA), 18:3n -3] in substitution for sunflower oil (control). The NL/PL ratio was 1.9, and 45,fatty acids were determined for both classes of lipid. The class totals of n -3 acids always increased in all treatments, while the totals for n -6 acids always decreased (p,<0.05). For a given level of flaxseed oil, the LNA contents were consistently higher, including EPA (20:5n -3) and DHA (22:6n -3). Arachidonic acid (20:4n -6) remained high in the PL but was reduced as levels of dietary flaxseed oil were increased. The n -6/n -3 ratios decreased significantly with the rise in flaxseed oil content in all treatments, and highly unsaturated fatty acid contents increased with the levels of flaxseed oil. Overall, the influence of flaxseed oil on the fatty acid composition in the contributing NL and PL classes was to increase n -3 PUFA, thus raising the nutritional value of this freshwater fish meat and, consequently, contributing to the health of consumers. [source]

Altered fatty acid pattern of phospholipids and triglycerides in the submandibular gland of ,3-depleted rats

EUROPEAN JOURNAL OF ORAL SCIENCES, Issue 2 2007
Christine Delporte
Alteration of the phospholipid (PL) and triglyceride (TG) fatty acid pattern was recently documented in several organs of rats depleted in long-chain polyunsaturated ,3 fatty acid (,3 rats). This study extends such a knowledge to the submandibular gland. The total PL and TG content of the salivary gland was not different in control and ,3 rats. The sole ,3 fatty acids found in ,3 rats (C22:5,3 and C22:6,3) were present at levels 3,12 times lower than in control rats. The C22:5,3/C22:6,3 ratio was increased threefold in ,3 rats. The PL and TG C16:0/C16:1,7 and C18:0/C18:1,9 ratios were decreased in ,3 rats. The conversion of C18:2,6 to C20:4,6 and C22:4,6 appeared facilitated in the ,3 rats. Some of these rats were injected intravenously, 60,120 min before killing, with either a medium-chain triglyceride:fish oil emulsion or a control medium-chain triglyceride:olive oil emulsion. The former emulsion increased the PL C22:5,3 and C22:6,3 content and prevented the age-related decrease in C16:0/C16:1,7 and C18:0/C18:1,9 ratios otherwise also recorded in PL. In conclusion, these findings document an increased activity of ,9-desaturase, a more efficient conversion of C18:2,6 to its metabolites, and an impaired generation of C22:6,3 from C22:5,3 in ,3 rats. [source]

Plant fructans stabilize phosphatidylcholine liposomes during freeze-drying

FEBS JOURNAL, Issue 2 2000
Dirk K. Hincha
Fructans have been implicated as protective agents in the drought and freezing tolerance of many plant species. A direct proof of their ability to stabilize biological structures under stress conditions, however, is still lacking. Here we show that inulins (linear fructose polymers) isolated from chicory roots and dahlia tubers stabilize egg phosphatidylcholine large unilamellar vesicles during freeze-drying, while another polysaccharide, hydroxyethyl starch, was completely ineffective. Liposome stability was assessed after rehydration by measuring retention of the soluble fluorescent dye carboxyfluorescein and bilayer fusion. Inulin was an especially effective stabilizer in combination with glucose. Analysis by HPLC showed that the commercial inulin preparations used in our study contained no low molecular mass sugars that could be responsible for the observed stabilizing effect of the fructans. Fourier transform infrared spectroscopy showed a reduction of the gel to liquid-crystalline phase transition temperature of dry egg PtdCho by more than 20 °C in the presence of inulin. A direct interaction of inulin with the phospholipid in the dry state was also indicated by dramatic differences in the phosphate asymmetric stretch region of the infrared spectrum between samples with and without the polysaccharide. [source]

Contrasts in cortical magnesium, phospholipid and energy metabolism between migraine syndromes.

HEADACHE, Issue 4 2003
MD Boska
Neurology. 2002;58:1227-1233. BACKGROUND: Previous single voxel (31)P MRS pilot studies of migraine patients have suggested that disordered energy metabolism or Mg(2+) deficiencies may be responsible for hyperexcitability of neuronal tissue in migraine patients. These studies were extended to include multiple brain regions and larger numbers of patients by multislice (31)P MR spectroscopic imaging. METHODS: Migraine with aura (MWA), migraine without aura (MwoA), and hemiplegic migraine patients were studied between attacks by (31)P MRS imaging using a 3-T scanner. RESULTS: Results were compared with those in healthy control subjects without headache. In MwoA, consistent increases in phosphodiester concentration [PDE] were measured in most brain regions, with a trend toward increase in [Mg(2+)] in posterior brain. In MWA, phosphocreatine concentration ([PCr]) was decreased to a minor degree in anterior brain regions and a trend toward decreased [Mg(2+)] was observed in posterior slice 1, but no consistent changes were found in phosphomonoester concentration [PME], [PDE], inorganic phosphate concentration ([Pi]), or pH. In hemiplegic migraine patients, [PCr] had a tendency to be lower, and [Mg(2+)] was significantly lower than in the posterior brain regions of control subjects. Trend analysis showed a significant decrease of brain [Mg(2+)] and [PDE] in posterior brain regions with increasing severity of neurologic symptoms. CONCLUSIONS: Overall, the results support no substantial or consistent abnormalities of energy metabolism, but it is hypothesized that disturbances in magnesium ion homeostasis may contribute to brain cortex hyperexcitability and the pathogenesis of migraine syndromes associated with neurologic symptoms. In contrast, migraine patients without a neurologic aura may exhibit compensatory changes in [Mg(2+)] and membrane phospholipids that counteract cortical excitability. Comment: If the theory of hyperexcitability of migraine brain is correct, basic scientists will need to find clear markers for the neuronal abnormalities that underlie this excitability. Using their techniques, these researchers could not find such markers. SJT [source]

Amphiphilic Homochiral Oligopeptides Generated via Phase Separation of Nonracemic , -Amino Acid Derivatives and Lattice-Controlled Polycondensation in a Phospholipid Environment

HELVETICA CHIMICA ACTA, Issue 11 2003
Irina Rubinstein
Racemic S -ethyl thioesters of N, -stearoyllysine (=,S -ethyl (R,S)-2-amino-6-(stearoylamino)hexanethioate) and S -ethyl thioesters of , -stearyl glutamic acid (=stearyl (R,S)-4-amino-5-(ethylsulfanyl)-5-oxopentanoate) self-assemble as separated two-dimensional crystalline monolayers within an achiral phospholipid environment of racemic 1,2-dipalmitoylglycerol (DPG) and 1,2-dipalmitoylglycero-3-phosphoethanolamine (DPPE), as demonstrated by grazing-incidence X-ray-diffraction (GIXD) measurements performed on the surface of H2O. Lattice-controlled polycondensation within these crystallites with deuterium-enantiolabeled monomers was initiated by injecting aqueous solutions of Ag+ or I2/KI beneath the monolayers, which yielded mixtures of diastereoisomeric oligopeptides containing up to six to eight repeating units, as analyzed by MALDI-TOF mass spectrometry. Analysis of the diastereoisomeric distribution showed an enhanced relative abundance of the oligopeptides with homochiral sequences containing three or more repeating units. Within the DPPE monolayers, the nucleophilic amino group of the phospholipid operates as an initiator of polymerization at the periphery of the monomer two-dimensional crystallites. Enhanced relative abundance of enantiomerically enriched homochiral oligopeptides was obtained by the polycondensation of nonracemic monomers. This enhancement indicated a phase separation into racemic and enantiomorphous monomer crystallites within the phospholipid environment, although this separation could not be observed directly by GIXD. A possible role that might have been played by crystalline assemblies for the abiotic generation and amplification of oligopeptides with homochiral sequences is discussed. [source]

Novel biotransformation and physiological properties of norursodeoxycholic acid in humans,,

Relevance between lipid metabolism-associated genes and rat liver regeneration

HEPATOLOGY RESEARCH, Issue 8 2008
Cunshuan Xu
Aim:, Lipids are important in constituting cell structure and participating in many biological processes, particularly in energy supplementation to cells. The aim of the present study is to elucidate the action of lipid metabolism-associated genes on rat liver regeneration (LR). Methods:, Lipid metabolism-associated genes were obtained by collecting website data and retrieving related articles, and their expression changes in the regenerating rat liver were checked by the Rat Genome 230 2.0 array. Results:, In total, 280 genes involved in lipid metabolism were proven to be LR-associated by comparing the gene expression discrepancy between the partial-hepatectomy and sham-operation groups. The initial and total expression numbers of these genes occurring in the initial phase, G0/G1 transition, cell proliferation, cell differentiation, and structure,functional rebuilding of LR were 128, 33, 135, 6, and 267, 147, 1026, 306, respectively, illustrating that these genes were initially expressed mainly in the initiation stage and functioned in different phases. Upregulation (850 times) and downregulation (749 times), as well as 25 types of expression patterns, showed that the physiological and biochemical activities were diverse and complicated in LR. Conclusion:, According to the results of the chip detection, it was presumed that fatty acid synthesis at 24,66 h, leukotriene and androgen synthesis at 16,168 h, prostaglandin synthesis at 2,96 h, triglyceride synthesis at 18,24 h, glycosphingolipid synthesis at 0.5,66 h, metabolism of phosphatidyl inositol and sphingomyelin at 2,16 h, and cholesterol catabolism at 30,168 h were enhanced. Throughout almost the whole LR, the genes participating in estrogen, glucocorticoid, and progesterone synthesis, and triglyceride catabolism were upregulated, while phospholipid and glycosphingolipid catabolism were downregulated. [source]

The Rubino test for leprosy is a ,2 -glycoprotein 1-dependent antiphospholipid reaction

IMMUNOLOGY, Issue 1 2000
A. Panunto-Castelo
Summary We describe the isolation and identification of three components required for the Rubino reaction (RR), which is the rapid sedimentation of formalinized sheep red-blood cells (SRBC) initiated by serum from leprosy patients with defective Mycobacterium leprae -specific cell immunity. The Rubino reaction factor (RRF) required for this phenomenon, previously identified as an immunoglobulin M (IgM), was purified from leprosy patient serum by adsorption to formalinized SRBC. Purified RRF IgM, when added to formalinized SRBC, did not produce a positive RR. However, when the contact was carried out in the presence of normal human serum (NHS), cells rapidly sedimented. The purified cofactor from NHS contained two components of 70 000 and 50 000 molecular weight (MW), as determined by sodium dodecyl sulphate,polyacrylamide gel electrophoresis (SDS,PAGE). The latter was recognized by the RRF IgM on immunoblot and its N-terminal sequence indicated that it was ,2 -glycoprotein 1 (,2 -GP1), an anionic phospholipid-binding protein. Methanol-treated formalinized SRBC did not support the RR. Thin-layer chromatography of an extract of membranes indicated that the SRBC ligand was a cell-surface phospholipid. Cardiolipin inhibited the RR. These data demonstrate that the RR involves a trimolecular interaction in which IgM, ,2 -GP1 and an SRBC phospholipid participate. By analogy with the antiphospholipid antibodies (anti-PL) that occur in autoimmune processes, serum samples from 29 systemic lupus erythematosus patients with high levels of anticardiolipin antibodies were submitted to the RR. A positive RR was obtained for 45% (13 of 29 patients). These results modify the paradigm of the absolute specificity of the RR for leprosy and demonstrate that RRF IgM is a ,2 -GP1-dependent anti-PL. [source]

Lysophosphatidic acid induces ovarian cancer cell dispersal by activating Fyn kinase associated with p120-catenin

INTERNATIONAL JOURNAL OF CANCER, Issue 4 2008
Ruby Yun-Ju Huang
Abstract Lysophosphatidic acid (LPA), known as the "ovarian cancer activating factor," is a natural phospholipid involved in important biological functions, such as cell proliferation, wound healing and neurite retraction. LPA causes colony dispersal in various carcinoma cell lines by inducing morphological changes, including membrane ruffling, lamellipodia formation, cell,cell dissociation and single cell migration. However, its effects on cell,cell dissociation and cell,cell adhesion of ovarian cancer cells have not been studied. In our study, we showed that LPA induced sequential events of intercellular junction dispersal and "half-junction" formation in ovarian cancer SKOV3 cells and that Src-family kinases were involved in both processes, since the effects were abolished by the selective tyrosine kinase inhibitor PP2. LPA induced rapid and transient activation of Src family kinases, which were recruited to cell,cell junctions by increasing the association with the adherens junction protein p120-catenin. We identified the Src family kinase, Fyn, as the key component associated with p120-catenin after LPA stimulation in SKOV3 cells. Our study provides evidence that LPA induces junction dispersal in ovarian cancer SKOV3 cells by activating the Src family kinase Fyn and increasing its association with p120-catenin at the cell,cell junction. © 2008 Wiley-Liss, Inc. [source]

Effect of cream treatment on phospholipids and protein recovery in butter-making process

INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 4 2008
Michel Britten
Summary A simple approach is proposed to recover native protein and phospholipid fractions from butter-making process using equipments available in dairy-processing plant. A washing treatment was used to remove protein from the cream and increase the phospholipids purity in buttermilk. Cream from a first separation was diluted with milk ultrafitration permeate and separated a second time. A 10X dilution factor reduced protein concentration in the cream from 1.6 ± 0.2 to 0.52 ± 0.03%. As a result, the phospholipids to protein ratio in buttermilk increased from 53 ± 10 to 172 ± 7 mg g,1. In butter-making process, an important portion of total phospholipids (,26%) is retained in butter. Butter serum made from washed cream could then be used to produce phospholipid concentrates with phospholipids to protein ratio of 473 ± 3 mg g,1. Interestingly, the extracts from butter serum are characterised by a higher proportion of sphingomyelin compared with those from buttermilk. [source]

Lipid composition of retailed organic, free-range and conventional chicken breasts

INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 3 2007
Kishowar Jahan
Summary Lipid fractions of 20 retailed chicken breasts were correlated with production system: organic, corn-fed, free-range and conventional. Neutral lipid (NL), phospholipid (PL) and free fatty acids (FFA) were examined separately. Influence of production systems was found more pronounced in PL composition than NLs. Corn-fed and free-range NLs had higher contents of nutritionally beneficial eicosapentanoic acid (C20:5 n-3) and docosahexanoic acid (C22:6 n-3) than organic and conventional. Lower polyunsaturated fatty acids in organic and free-range PLs could be beneficial for tissue stability. Principal component product space for PLs showed clear clustering related to product category. In contrast, this was not observed with FFA except in the partial least square regression product space suggesting influences on NLs and PLs and FFA. PLs had lower contents of arachidonic acid than in earlier studies. Advantages were observed in lipid fractionation using advanced sorbent extraction matrices. [source]

Phospholipid signalling through phospholipase D and phosphatidic acid

Effects of modification of membrane lipid composition on Bacillus subtilis sporulation and spore properties

JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2009
K.K. Griffiths
Abstract Aims:, To determine effects of inner membrane lipid composition on Bacillus subtilis sporulation and spore properties. Methods and Results:, The absence of genes encoding lipid biosynthetic enzymes had no effect on B. subtilis sporulation, although the expected lipids were absent from spores' inner membrane. The rate of spore germination with nutrients was decreased c. 50% with mutants that lacked the major cardiolipin (CL) synthase and another enzyme for synthesis of a major phospholipid. Spores lacking the minor CL synthase or an enzyme essential for glycolipid synthesis exhibited 50,150% increases in rates of dodecylamine germination, while spores lacking enzymes for phosphatidylethanolamine (PE), phosphatidylserine (PS) and lysylphosphatidylglycerol (l-PG) synthesis exhibited a 30,50% decrease. Spore sensitivity to H2O2 and tert-butylhydroperoxide was increased 30,60% in the absence of the major CL synthase, but these spores' sensitivity to NaOCl or OxoneÔ was unaffected. Spores of lipid synthesis mutants were less resistant to wet heat, with spores lacking enzymes for PE, PS or l-PG synthesis exhibiting a two to threefold decrease and spores of other strains exhibiting a four to 10-fold decrease. The decrease in spore wet heat resistance correlated with an increase in core water content. Conclusions:, Changing the lipid composition of the B. subtilis inner membrane did not affect sporulation, although modest effects on spore germination and wet heat and oxidizing agent sensitivity were observed, especially when multiple lipids were absent. The increases in rates of dodecylamine germination were likely due to increased ability of this compound to interact with the spore's inner membrane in the absence of some CL and glycolipids. The effects on spore wet heat sensitivity are likely indirect, because they were correlated with changes in core water content. Significance and Impact of the Study:, The results of this study provide insight into roles of inner membrane lipids in spore properties. [source]

The prion/lipid hypothesis , further evidence to support the molecular basis for transmissible spongiform encephalopathy risk assessment

JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2007
P. Gale
Summary Defining the molecular structure of the transmissible spongiform encephalopathy (TSE) agent is important both for underpinning risk assessments and for developing and understanding decontamination strategies. Recent studies have shown that oligomeric particles comprising 14,28 prion protein (PrP) molecules are much more infectious than larger fibrils (prion rods) or indeed smaller oligomers (trimers) and PrP monomers. Here, results from deactivation studies (with alkali, heat, hexane or formaldehyde) are interpreted in terms of the infectious nucleation seed comprising 14,28 PrP molecules held together by interactions with amphipathic phospholipid (PL) or more probably sphingolipid (SL) from the host. According to the PrP/lipid hypothesis, the strength of the protein/lipid interactions accounts for the high thermostability of TSE infectivity and for differences in thermostability between strains. The implications of the molecular biophysics data for environmental TSE risk assessments are discussed with respect to behaviour in soil. While formaldehyde appears to cause inactivation of scrapie infectivity by increasing the ID50, the dose,response is complicated by apparent heterogeneity between hamster subpopulations in susceptibility. The process of inactivation by formaldehyde may be due to cross-linking the highly infectious 14,28 PrP oligomers into larger, but less infectious aggregates. This process appears more reversible in some hamster subpopulations than others. [source]

The infectivity of transmissible spongiform encephalopathy agent at low doses: the importance of phospholipid

JOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2006
P. Gale
Abstract The issue of whether the mechanism of infection is independent or co-operative for low doses of transmissible spongiform encephalopathy (TSE) agent is critical for risk assessment. The susceptibility (and hence ID50) of individuals with the same prion protein (PrP) genotype may vary considerably with a small proportion being very susceptible. Assuming independent action, the incubation period (IP) would continue to increase until the dose is below the ID50 of the most susceptible individuals in the experiment, at which point it would become constant. This may explain the observed increase in IP with decreasing dose below the apparent ID50 in experiments with untreated TSE agent. In contrast, IPs for autoclaved or NaOH-treated TSE agent increase greatly at doses phospholipid (PL). PL would play a structural role through mediating protein/lipid interactions with PrP. Heating or alkali treatment destroys the PL breaking up the nucleation seeds, which require long IPs to reform at low doses. Replenishing those inactivated PLs with host PL would explain how the phenotypic effect of long IP at low dose is lost on subpassage. It is proposed here that strain thermostability is controlled by the nature and strength of the PrP/PL interactions, which are independent of the host PrP genotype. Although repeated oral exposure to low doses of scrapie is less harmful than a single large exposure, this effect may reflect interference by competition rather than diminished risks due to a co-operative effect, and is of little importance for ,one-off' low-dose environmental exposures. [source]

Characterization of Aeromonas and Vibrio species isolated from a drinking water reservoir

JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2001
E.P. Ivanova
E.P. IVANOVA, N.V. ZHUKOVA, N.M. GORSHKOVA AND E.L. CHAIKINA. 2001. Aims: To study the phenotypic and chemotaxonomic (i.e. phospholipid and cellular fatty acid composition) characteristics of environmental Aeromonas spp. and Vibrio spp. isolated from a drinking water reservoir near Vladivostok City, and the application of some chemotaxonomic markers for discrimination of the two genera and species. Methods and Results: Presumptive Aeromonas species were dominant in surface water samples (up to 25% of the total number of bacteria recovered). These strains were consistent with respect to the cultural and biochemical properties used to define the species Aeromonas sobria (seven strains) and Aer. popoffii (three strains). Vibrio mimicus (two strains) and Vibrio metschnikovii (one strain) were identified according to phenotypic features and cellular fatty acid composition. Conclusions: Environmental Aer. sobria isolates were atypical in their ability to grow at 42°C, and were haemolytic, proteolytic and cytotoxic. Although it was present in a high proportion in the water samples, atypical Aer. sobria is not an indicator of polluted water. Significance and Impact of the Study: The incidence of Aeromonas in the drinking water reservoirs in the Far East of Russia is reported for the first time. [source]

Antihyperlipidemic activity of 3-hydroxymethyl xylitol, a novel antidiabetic compound isolated from Casearia esculenta (Roxb.) root, in streptozotocin-diabetic rats

JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 2 2010
Govindasamy Chandramohan
Abstract Casearia esculenta root (Roxb.) is widely used in traditional system of medicine to treat diabetes in India. An active compound, 3-hydroxymethyl xylitol (3-HMX), has been isolated, and its optimum dose has been determined in a short duration study and patented. In addition, the long-term effect of 3-HMX in type 2 diabetic rats on carbohydrate metabolism was investigated, and its antihyperglycemic effect was shown previously (Chandramohan et al., Eur J Pharmacol 2008;590:437,443). In this study we investigated the effect of 3-HMX on plasma and tissue lipid profiles in streptozotocin-induced diabetic rats. Diabetes was induced in adult male albino rats of the Wistar strain, weighing 180,200 g, by administration of streptozotocin (40 mg/kg of body weight) intraperitoneally. The normal and diabetic rats were treated with 3-HMX (40 mg/kg BW/day) for 45 days. The levels of total cholesterol, triglycerides, free fatty acids, and phospholipids were assayed in the plasma besides lipoprotein-cholesterol (high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and very low density lipoprotein-cholesterol (VLDL-C)) and tissues (liver, kidney, heart, and brain). Total cholesterol, triglyceride, free fatty acid, and phospholipid (LDL-C and VLDL-C in plasma only) levels increased in plasma and tissues significantly, whereas plasma HDL-C significantly decreased in diabetic rats. Treatment with 3-HMX or glibenclamide reversed the above-mentioned changes and improved toward normalcy. Histological study of liver also confirmed the biochemical findings. Thus administration of 3-HMX is able to reduce hyperglycemia and hyperlipidemia related to the risk of diabetes mellitus. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:95,101, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20317 [source]

Regulation of Wnt/,-catenin pathway by cPLA2, and PPAR,

JOURNAL OF CELLULAR BIOCHEMISTRY, Issue 2 2008
Chang Han
Abstract Cytosolic phospholipase A2, (cPLA2,) is a rate-limiting key enzyme that releases arachidonic acid (AA) from membrane phospholipid for the production of biologically active lipid mediators including prostaglandins, leukotrienes and platelet-activating factor. cPLA2, is translocated to nuclear envelope in response to intracellular calcium increase and the enzyme is also present inside the cell nucleus; however, the biological function of cPLA2, in the nucleus remains unknown. Here we show a novel role of cPLA2, for activation of peroxisome proliferator-activated receptor-, (PPAR,) and ,-catenin in the nuclei. Overexpression of cPLA2, in human cholangiocarcinoma cells induced the binding of PPAR, to ,-catenin and increased their association with the TCF/LEF response element. These effects are inhibited by the cPLA2, siRNA and inhibitors as well as by siRNA knockdown of PPAR,. Overexpression of PPAR, or treatment with the selective PPAR, ligand, GW501516, also increased ,-catenin binding to TCF/LEF response element and increased its reporter activity. Addition of AA and GW501516 to nuclear extracts induced a comparable degree of ,-catenin binding to TCF/LEF response element. Furthermore, cPLA2, protein is present in the PPAR, and ,-catenin binding complex. Thus the close proximity between cPLA2, and PPAR, provides a unique advantage for their efficient functional coupling in the nucleus, where AA produced by cPLA2, becomes immediately available for PPAR, binding and subsequent ,-catenin activation. These results depict a novel interaction linking cPLA2,, PPAR, and Wnt/,-catenin signaling pathways and provide insight for further understanding the roles of these key molecules in human cells and diseases. J. Cell. Biochem. 105: 534,545, 2008. © 2008 Wiley-Liss, Inc. [source]