Fats, lipids and blood coagulation

Saturated and n-3 fatty acids, postprandial lipaemia, and the combined effects of fatty acids and lipid-lowering drugs have been of principal interest in recent studies in the field of dietary fats, lipids and haemostasis. The sex-specific effect of individual saturated fatty acids on coagulation factor VII activity has been discovered, and the significant effect of factor VII R353Q polymorphism on the postprandial response has also been found. An increased intake of n-3 fatty acids or fat reduction when combined with intensive lifestyle interventions may lead to reduced thrombotic potential in type 2 diabetic patients and obese individuals. Furthermore, positive effects on haemostasis by combined treatment with long-chain n-3 fatty acids and simvastatin indicate that n-3 fatty acids may be of some relevance with lipid-lowering drugs. The possible unfavourable effect of n-3 fatty acids on plasminogen activator inhibitor-1 activity is still a matter of dispute, but recent studies suggest that n-3 fatty acids, including alpha-linolenic acid, may have antithrombotic effects by enhancing protein C activity.     

Relationship between lipids and aflatoxin biosynthesis

This paper describes the key role of lipids on fungal growth and of lipoperoxidation on the output of aflatoxin biosynthesis both in vitro and in vivo. In vitro BHA, BHT and cysteamine, depending their concentration, are capable of reducing or blocking aflatoxin output induced by lipoperoxides or halomethanes in cultures ofAspergillus flavus orA. parasiticus without affecting fungal growth. In vivo BHA and BHT significantly reduced aflatoxin production on wheat, maize and sunflower inoculated with aflatoxigenicAspergilli essentially by preventing fungal growth. In vivo the seeds surface lipids represent a very important carbon source for fungal growth.     

Relationship between the fatty acid composition of lipids and the viability of dried yeast Saccharomyces cerevisiae

Summary The fatty acid composition of the total lipids and phospholipids of intact and convectively dried yeast has been studied and an increase has been detected in the degree of unsaturation of fatty acids during dehydration. The validity of the inverse relation between the viability of the yeast population during dehydration and rehydration and the degree of unsaturation of the fatty acids has been demonstrated by regression analysis. The results may help to predict the viability of dried yeast.     

Phytoplankton carbon incorporation patterns and biochemical composition of particulate matter in the eastern North Atlantic subtropical region

Photosynthetic carbon metabolism and the biochemical compositionof late-winter phytoplankton assemblages in the eastern NorthAtlantic Ocean were studied during an oceanographic cruise carriedout in March 1992. Enhanced levels of phytoplankton biomassand primary production were linked to the subtropical front-AzoresCurrent (STF/AC) region. High values of the relative incorporationof carbon into proteins indicated that the phytoplankton wasgrowing at a rate close to the maximum growth rate. A very highpercentage of carbon was incorporated into the lipid fractionat southern latitudes, whereas incorporation into polysaccharidespeaked at the AC. In general, the biochemical composition ofparticulate matter reflected the observed patterns of photosynthatepartitioning. Latitudinal changes in phytoplankton species compositionaccounted for the geographical variability in ¹⁴C labellingpatterns. Turnover times of particulate matter estimated forthe STF/AC region were relatively low (3–5 days) and suggestedbalancedgrowth of the microplankton community over daily timescales.     

Investigation of associations between ABO blood groups and coagulation,fibrinolysis, total lipids,cholesterol, and triglycerides

Summary To investigate possible associations between ABO blood system and coagulability levels, fibrinolysis, total lipids, cholesterol, and triglycerides, the plasma and serum of 300 Rh-positive male blood donors were tested. The tests performed were: RT, PTT, K-PTT, PT, F.V, F.II, F.VIII, Complex II, VII, and X, TGT, fibrinogen, HAE 0.2, HAE 0.5, ELT, LIP, Col. 1, Col. 2 and TRI.Analysis of the laboratory data shows a lower coagulability in O blood group individuals. This result was obtained in coagulation tests (RT, PTT, and K-PTT) specific for factor VIII level. In addition, a higher sensitivity to the in vitro heparin anticoagulant effect in O group individuals was confirmed. Nevertheless, these conclusions are specific for Negroids, the same effects not being observed in Caucasians. None of the other laboratory tests revealed any differences related to either blood group or race.     

Relationship between chain collapse and secondary structure formation in a partially folded protein

Chain collapse and secondary structure formation are frequently observed during the early stages of protein folding. Is the chain collapse brought about by interactions between secondary structure units or is it due to polymer behavior in a poor solvent (coil‐globule transition)? To answer this question, we measured small‐angle X‐ray scattering for a series of β‐lactoglobulin mutants under conditions in which they assume a partially folded state analogous to the folding intermediates. Mutants that were designed to disrupt the secondary structure units showed the gyration radii similar to that of the wild type protein, indicating that chain collapse is due to coil‐globule transitions. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 651–658, 2014.     

Activated protein C, a protein at the crossroads between coagulation and inflammation

Sepsis is defined as a systemic response to infection, characterized by an intense inflammatory response linked to coagulation activation and fibrinolysis inhibition, two processes which are intimately associated. In a field where mortality remains very high, administration of activated protein C, a physiological coagulation inhibitor with cytoprotective properties, has demonstrated its effectiveness and was able to reduce mortality. Protein C belongs to a system that involves plasma proteins and endothelial cell receptors. In addition to well documented effects on coagulation and fibrinolysis, activated protein C exhibits anti-inflammatory, anti-apoptotic but also anti-histone activities. Indeed, a recent study focusing on the cytoprotective effects of activated protein C showed that extracellular histones are released during severe sepsis and may participate in the pathophysiology of severe sepsis. These histones appear to be new targets of activated protein C.     

Relationship between serum lipids, lipoproteins and pseudocholinesterase during organophosphate poisoning in rabbits

The activities of serum pseudocholinesterase, lecithin: cholesterol acyltransferase (LCAT) and gamma-glutamyltransferase in rabbits were investigated before and after dichlorvos administration in vivo. The effects of this organophosphate on some serum lipids and lipoprotein fractions were also determined. LCAT activity remained almost unaffected after organophosphate administration. However, serum gamma-glutamyltransferase and pseudocholinesterase activities markedly decreased. Dichlorvos markedly lowered both serum low density lipoprotein (LDL) and cholesterol contents, whereas high density lipoprotein (HDL) concentration increased and very low density lipoprotein (VLDL) remained unaffected. Triglycerides as well as esterified fatty acids increased significantly but the statistical changes in free fatty acid concentrations were not significant, because individual variations in fatty acid concentrations were high.     

Dietary protein and fatty acid composition of liver lipids in the rat

In order to compare the effects of different sources of dietary protein on the fatty acid composition of phosphatidylcholines (PC), phosphatidylethanolamines (PE), phosphatidylinositols (PI), cholesteryl esters and triacylglycerols, male rats were fed for a 4-week period on cholesterol-free, or cholesterol-containing, diets based on casein, or soybean protein and olive oil. The most conspicuous difference observed was the occurrence of significantly higher levels of 5,8,11-eicosatrienoic acid, 20:3 (n - 9), in the different lipid classes of casein-fed, compared with soybean protein-fed, animals. In the PI fraction of livers from the groups of rats fed casein diet, this fatty acid amounted to between 9.9 and 13.3% by weight of the total fatty acids. Phospholipids from livers of casein-fed rats contained increased levels of oleic acid, 18:1 (n - 9) (in PC and PE) and reduced levels of stearic acid (18:0). Moreover, in this group of rats PI contained a reduced level of arachidonic acid, 20:4 (n - 6). A casein-related decrease in the linoleic acid, 18:2 (n - 6), content of PC and PE was observed only in the rats fed on cholesterol-free diet. Effects on the fatty acid composition were also observed in the triacyglycerol and cholesteryl ester fractions, in which the rats fed casein diet showed higher levels of palmitoleic acid, 16:1 (n - 7) (cholesterol-supplemented diet) and lower values for linoleic acid, than the soybean protein-fed rats.     

The effects of inhibitors of protein synthesis on incorporation of lipids into myelin

Abstract— Following intracranial injections of puromycin, the incorporation of [³H]leucine into brain protein was inhibited by 80 per cent. Conversely, incorporation of [³⁵S]sulphate into sulphatide or [2-³H]glycerol into phosphatidyl choline was not inhibited. Under these conditions, appearance of labelled protein in myelin was inhibited by 90 per cent, while the appearance of newly labelled sulphatide and phosphatidyl choline in myelin membrane was not greatly affected. Experiments with cycloheximide gave similar results with phosphatidyl choline, but incorporation of [³⁵S]sulphate into total sulphatide was decreased by about 30 per cent in animals given cycloheximide. Neither puromycin nor cycloheximide had any inhibitory effect on galactocerebroside sulphotransferase.     

Relationship between base composition in non-coding DNA of genes and codon composition

The C + G percentage in third position of codons is linearly dependent on the C + G composition of flanking regions and introns. A similar relationship is shown for the first and second position which significantly influence the nature of amino acid sequence. If mutations would be oriented according to the local base composition, this will imply that genes of the same multigenic family would evolve at different rate.     

α2-Macroglobulin-protease reactions: Relationship of covalent bond formation,methylamine reactivity,and specific proteolysis

Experiments were performed to define the relation between covalent binding of enzymes to β₂-macroglobulin (α₂M), the specific proteolysis of α₂M subunits to 85K fragments, and the reactivity of the methylamine site on α₂M. We studied the reaction of α₂M with native trypsin, anhydrotrypsin, and two active lysyl-blocked derivatives, methyl-trypsin and dimethylmaleyl-trypsin, the last with reversibly modified amino groups that can be regenerated at low pH. The results were: (1) All enzymes tested reacted with α₂M but only native trypsin formed covalent complexes (not dissociable by sodium dodecyl sulfate). Trypsin and the lysyl-blocked enzymes caused complete proteolysis of the α₂M subunits, in agreement with previous studies. (2) The dimethyl-maleyl-trypsin became covalently bound to α₂M only after removing the blocking groups of the bound enzyme, indicating that sequential proteolysis and covalent bond formation is possible. Under the conditions used for deblocking, there was no change in the covalent/noncovalent binding ratio of native trypsin, anhydrotrypsin, or the other lysyl-blocked derivative, methyl-trypsin. (3) Native trypsin or anhydrotrypsin displaced methyl- or dimethylmaleyl-trypsin from their α₂M complexes but the newly bound enzymes with free amino groups did not form covalent bonds indicating that enzymes must remain in association with the inhibitor for the bond to form. (4) Methylamine reacts with noncovalent α₂M complexes but not with covalent complexes. (5) Methylamine-treated α₂M can still form complexes with trypsin but at a drastically reduced rate and only noncovalent complexes are formed. In summary, sequential proteolysis and covalent bond formation is possible under certain conditions, and there is a strong correlation between covalent binding and loss of methylamine reactivity. The latter observation is suggestive evidence for the identity of the covalent binding site of α₂M and the putative thiol ester of the methylamine site. The enzyme lysyl amino groups, are likewise possible candidates for attacking nucleophile at that site.     

A J-like protein influences fatty acid composition of chloroplast lipids in Arabidopsis

A comprehensive understanding of the lipid and fatty acid metabolic machinery is needed for optimizing production of oils and fatty acids for fuel, industrial feedstocks and nutritional improvement in plants. T-DNA mutants in the poorly annotated Arabidopsis thaliana gene At1g08640 were identified as containing moderately high levels (50-100%) of 16∶1Δ7 and 18∶1Δ9 leaf fatty acids and subtle decreases (5-30%) of 16∶3 and 18∶3 (http://www.plastid.msu.edu/). TLC separation of fatty acids in the leaf polar lipids revealed that the chloroplastic galactolipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) were the main lipid types affected by this mutation. Analysis of the inferred amino acid sequence of At1g08640 predicted the presence of a transit peptide, three transmembrane domains and an N-terminal J-like domain, and the gene was named CJD1 for Chloroplast J-like Domain 1. GFP reporter experiments and in vitro chloroplast import assays demonstrated CJD1 is a chloroplast membrane protein. Screening of an Arabidopsis cDNA library by yeast-2-hybrid (Y2H) using the J-like domain of CJD1 as bait identified a plastidial inner envelope protein (Accumulation and Replication of Chloroplasts 6, ARC6) as the primary interacting partner in the Y2H assay. ARC6 plays a central role in chloroplast division and binds CJD1 via its own J-like domain along with an adjacent conserved region whose function is not fully known. These results provide a starting point for future investigations of how mutations in CJD1 affect lipid composition.     

Interaction between biotin lipids and streptavidin in monolayers: formation of oriented two-dimensional protein domains induced by surface recognition

Highly specific ligand-receptor interactions generally characterize surface recognition reactions. Such processes can be simulated by streptavidin-biotin-specific binding. Biotin lipids have thus been synthesized, and their interaction with streptavidin (or avidin) at the air-water interface was directly shown by measurement of surface pressure isotherms and fluorescence microscopy. These proteins interact with the biotin lipid monolayer via specific binding or nonspecific adsorption. Both phenomena were clearly distinguished by use of the inactivated form of streptavidin. The binding of fluorescein-labeled streptavidin to monolayers was also directly observed by fluorescence microscopy. The fluorescence of the protein domains is directly related to the state of polarization of the exciting light. This anisotropy can only be explained by the formation of oriented two-dimensional biotin lipid-streptavidin domains.     

Age and gender differences in the relationship between heart rate reactivity and elevated serum lipids

Data on serum lipids and heart rate reactivity to a standardized mental arithmetic stressor were collected on 54 healthy men and 73 healthy women. Subjects' ages ranged from 18 to 61 years, allowing analysis of data from both young and older subgroups. MANOVA revealed that age was directly associated with elevated lipids in men and women. There was no significant relationship between reactivity and lipids for either older or young men. Conversely, young women showed a significant direct relationship between reactivity and Total Cholesterol and LDL, but this was not present for older women. Implications of these data for the previously reported hypothetical relationship between reactivity and lipids are discussed. Limitations of the study are examined, and future directions for research suggested.I wish to thank Dianne Vella for her assistance in aspects of data-collection. This research was supported by grant No. 900946 from the National Health & Medical Research Council.     

Relationship between polyreactive properties of immunoglobulins and level of lipids in them

Presented paper deals with the relationship between immunoglobulin polyreactive properties and its lipid composition. Serum blood immunoglobulin fraction of an intact rabbit as an experimental model was used. Immunoglobulins (Ig) obtained by this way were transformed into polyreactive immunoglobulins (PRIg) by treatment with chaotropic agent KSCN or reactive oxygen species (ROS) with usage of Fe2+, EDTA and ascorbic acid. It was demonstrated that native Ig were able to bind with immobilized antigen (ovalbumin) and this ability dramatically increased after transformation of Ig into PRIg. The high immunoreactivity of PRIg was associated with marked fall (by 2-3 fold) of total phospholipids as well as individual ones--sphingomyelin and phosphatidylcholine. The main fatty acids of the Ig and PRIg phospholipid fractions in the sequence to decreasing decrease were stearic, palmitic, oleic and linoleic acids. The treatment of Ig by chaotropic agent and ROS led to decrease of stearic acid and enhancement of oleic, linoleic and arachidonic acids. The level of free cholesterol of Ig did not differ from that of PRIg. At the same time the content of cholesterol esters of PRIg was substantially diminished if compare with Ig. The main fatty acids of the Ig and PRIg cholesterol ester fraction in the sequence to decreasing were arachidonic, stearic, oleic, linoleic and palmitoleic acids. Transformation of Ig into PRIg was accompanied by enhancement of stearic acid level and loss of docosapentaenoic, arachidonic and palmitoleic acids. The results presented here support the idea about non peroxidative manner of the phospholipid and cholesterol ester extrusion from Ig molecule under its transformation into PRIg. Rather the last process could be explained by the term of concurrent physico-chemical interaction of Ig molecule with chaotropic agent or ROS leading to fall of lipid content. The presented data for the first time provide us an opportunity to conclude that transformation of Ig into PRIG is associated with the marked loss of essential phospholipids and cholesterol esters by the Ig molecule. The probable implication of this process in development of immune imbalance under certain diseases associated with oxidative stress have been discussed.     

Relationship between milk composition and pup's growth in mice

The effect of lactation numbers (first, second, and third) on milk composition and the relationship between milk composition and pup's growth were examined in four strains of mice (SHN, SLN, C3H/He, and GR/A). Although the percentage of milk fat and moisture increased and decreased, respectively, through the first to third lactations in C3H/He mice, no differences in milk composition among lactation numbers were observed in the other mouse strains. In pooled data derived from the four strains of mice, there was no significant correlation between milk composition and body weight or growth rate of pups on Day 12 or 20 of the first lactation. In contrast, the amount of milk fat and moisture was the positively and negatively, respectively, correlated with the pup's growth parameters during the second and third lactations. This was reflected by a lower fat content and a higher moisture content in the milk of GR/A mice, especially evident during the second and third lactations and reduced pup's growth, when compared with the other mouse strains. Milk lactose and ash content was not correlated with pup's growth parameters. The results of our study stress the importance of milk fat for the growth of mouse pups.     

Relationship between antioxidant potential and oxidative damage to lipids, proteins and DNA in aged rats

Oxidative stress has been implicated to play a major role in aging and age-related diseases. In the present study, we investigated the effects of aging on the total antioxidant capacity, uric acid, lipid peroxidation, total sulfhydryl group content and damage to DNA in adult (6 months), old (15 months) and senescent (26 months) male Wistar rats. The antioxidant capacity, determined by phycoerythrin-based TRAP method (total peroxyl radical-trapping potential) was significantly decreased in the plasma and myocardium of old and senescent rats, whereas plasma level of uric acid was elevated in 26-month-old rats. Age-related decline in plasma and heart antioxidant capacity was accompanied by a significant loss in total sulfhydryl group content, increased lipid peroxidation and higher DNA damage in lymphocytes. Correlations between TRAP and oxidative damage to lipids, proteins and DNA suggest that the decline in antioxidant status may play an important role in age-related accumulation of cell damage caused by reactive oxygen species.     

Structural distinction between soluble and particulate protein kinase C species

A number of peripheral membrane proteins functioning as regulatory enzymes are distributed between soluble and particulate fractions upon homogenization and subcellular fractionation. One such enzyme, the Ca²⁺/phospholipid-dependent protein kinase, protein kinase C, was analyzed in order to examine this characteristic of differential localization. The soluble and particulate forms of this enzyme were purified to relative homogeneity, and their biochemical and biophysical properties were analyzed and compared. Based on biochemical activities, the particulate form required lower phospholipid concentrations for maximal activation than for the soluble species. The particulate species had a more hydrophobic structure as demonstrated by a hydrophobic fluorescence probe, and had almost 50% more -helical structures according to secondary structure estimation, determined from far ultra-violet-circular dichroism spectra (200–250 nm). Using Fourier transform infrared spectroscopy, specific lipid spectra were detected associated with the soluble protein kinase C species. Further analyses with a fluorescent neutral membrane probe suggested that there was more lipid associated with the purified particulate form, which was of a less mobile nature than those associated with the soluble species. These structural differences provide an explanation for the preferential localization of the enzyme and may prove to be the basis for distribution of other membrane-active peripheral membrane regulatory enzymes.