Lipolysis of natural long chain and synthetic medium chain galactolipids by pancreatic lipase-related protein 2

Monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) are the most abundant lipids in nature, mainly as important components of plant leaves and chloroplast membranes. Pancreatic lipase-related protein 2 (PLRP2) was previously found to express galactolipase activity, and it is assumed to be the main enzyme involved in the digestion of these common vegetable lipids in the gastrointestinal tract. Most of the previous in vitro studies were however performed with medium chain synthetic galactolipids as substrates. It was shown here that recombinant guinea pig (Cavia porcellus) as well as human PLRP2 hydrolyzed at high rates natural DGDG and MGDG extracted from spinach leaves. Their specific activities were estimated by combining the pH-stat technique, thin layer chromatography coupled to scanning densitometry and gas chromatography. The optimum assay conditions for hydrolysis of these natural long chain galactolipids were investigated and the optimum bile salt to substrate ratio was found to be different from that established with synthetic medium chains MGDG and DGDG. Nevertheless the length of acyl chains and the nature of the galactosyl polar head of the galactolipid did not have major effects on the specific activities of PLRP2, which were found to be very high on both medium chain [1786 ± 100 to 5420 ± 85 U/mg] and long chain [1756 ± 208 to 4167 ± 167 U/mg] galactolipids. Fatty acid composition analysis of natural MGDG, DGDG and their lipolysis products revealed that PLRP2 only hydrolyzed one ester bond at the sn-1 position of galactolipids. PLRP2 might be used to produce lipid and free fatty acid fractions enriched in either 16:3 n − 3 or 18:3 n − 3 fatty acids, both found at high levels in galactolipids.     

Impact of feeding and starvation on the lipid metabolism of the Arctic pteropod Clione limacina

Feeding and starvation experiments were carried out with Clione limacina sampled in Kongsfjorden (Svalbard, Arctic) during summer 2002. Dry mass and lipid mass, lipid class and fatty acid compositions were analysed. Specimens of C. limacina used for the feeding study had a mean length of 25 mm, a dry mass (DM) of 13.7 mg, and a moderate lipid content of 12.1%DM. Animals were allowed to ingest only one individual of its exclusive prey, Limacina helicina which had 8.0 mm in diameter, 21.4 mg DM and 8.7% lipid of ash-free DM. Five days after feeding, the dry mass of C. limacina had increased from 13.7 to 25.3 mg which corresponds to an uptake of about 80% of the ash-free DM (14.3 mg) of L. helicina. Lipid mass increased from 1.5 to 3.9 mg which is almost two times more the ingested lipid from L. helicina (1.2 mg lipid). Thus, the major portion of lipids was synthesised de novo by C. limacina from non-lipid compounds. These lipids were triacylglycerols (TAG) and 1-O-alkyldiacylglycerol ethers (DAGE), increasing from low proportions of 6.1% and 5.7% to 42.3% and 25.8%, respectively. Considerable de novo synthesis was observed for the monounsaturated fatty acids 16:1(n − 7), 17:1(n − 8), 18:1(n − 9), and 18:1(n − 7) and the alkyl moiety 16:0. The increase in the polyunsaturated fatty acids 22:6(n − 3), 20:5(n − 3), and 18:4(n − 3) corresponded with the amount available by ingestion of L. helicina, supporting that C. limacina is not able to synthesise polyunsaturates. After 15 days of digestion, dry mass and lipids dropped almost back to the initial values.During the 100-day starvation experiment, two groups of animals were separately considered as storage lipid-rich and lipid-poor animals because of their large differences in the amount and proportion of TAG and DAGE. Storage lipid-rich C. limacina were only found until day 50, whereas lipid-poor animals were present throughout the experiment. In the lipid-rich specimens, the levels of TAG were about twice that of DAGE. The proportions of TAG decreased considerably during the 50 days of starvation (from 48.3% to 25.1% of total lipid). DAGE, varying between 16.5% and 20.5%, showed only a small decrease. The lipid-poor animals survived 100 days of starvation, exhibiting low initial amounts and proportions of storage lipids which were nearly exhausted at the end. In all C. limacina specimens, the total lipid content remained almost constant showing that lipid and non-lipid components were simultaneously utilised. This implies that body shrinkage may be an important adaptation to long-term starvation. Based on these results, it is possible to estimate the potential survival period of lipid-rich C. limacina under food limitation. A model, which considers maturity and reproduction (egg production), reveals that lipid-rich specimens might be able to survive up to 260 days without food.     

Differential lipid and fatty acid profiles of photoautotrophic and heterotrophic Chlorella zofingiensis: Assessment of algal oils for biodiesel production

The objective of this study was to document and compare the lipid class and fatty acid composition of the green microalga Chlorella zofingiensis cultivated under photoautotrophic and heterotrophic conditions. Compared with photoautotrophic cells, a 900% increase in lipid yield was achieved in heterotrophic cells fed with 30 g L⁻¹ of glucose. Furthermore heterotrophic cells accumulated predominantly neutral lipids (NL) that accounted for 79.5% of total lipids with 88.7% being triacylglycerol (TAG); whereas photoautotrophic cells contained mainly the membrane lipids glycolipids (GL) and phospholipids (PL). Together with the much higher content of oleic acid (C18:1) (35.2% of total fatty acids), oils from heterotrophic C. zofingiensis appear to be more feasible for biodiesel production. Our study highlights the possibility of using heterotrophic algae for producing high quality biodiesel.     

Dietary n-6 PUFA deprivation for 15 weeks reduces arachidonic acid concentrations while increasing n-3 PUFA concentrations in organs of post-weaning male rats

Few studies have examined effects of feeding animals a diet deficient in n-6 polyunsaturated fatty acids (PUFAs) but with an adequate amount of n-3 PUFAs. To do this, we fed post-weaning male rats a control n-6 and n-3 PUFA adequate diet and an n-6 deficient diet for 15 weeks, and measured stable lipid and fatty acid concentrations in different organs. The deficient diet contained nutritionally essential linoleic acid (LA,18:2n-6) as 2.3% of total fatty acids (10% of the recommended minimum LA requirement for rodents) but no arachidonic acid (AA, 20:4n-6), and an adequate amount (4.8% of total fatty acids) of α-linolenic acid (18:3n-3). The deficient compared with adequate diet did not significantly affect body weight, but decreased testis weight by 10%. AA concentration was decreased significantly in serum (− 86%), brain (− 27%), liver (− 68%), heart (− 39%), testis (− 25%), and epididymal adipose tissue (− 77%). Eicosapentaenoic (20:5n-3) and docosahexaenoic acid (22:6n-3) concentrations were increased in all but adipose tissue, and the total monounsaturated fatty acid concentration was increased in all organs. The concentration of 20:3n-9, a marker of LA deficiency, was increased by the deficient diet, and serum concentrations of triacylglycerol, total cholesterol and total phospholipid were reduced. In summary, 15 weeks of dietary n-6 PUFA deficiency with n-3 PUFA adequacy significantly reduced n-6 PUFA concentrations in different organs of male rats, while increasing n-3 PUFA and monounsaturated fatty acid concentrations. This rat model could be used to study metabolic, functional and behavioral effects of dietary n-6 PUFA deficiency.     

Production of algae-based biodiesel using the continuous catalytic Mcgyan® process

This study demonstrates the production of algal biodiesel from Dunaliella tertiolecta, Nannochloropsis oculata, wild freshwater microalgae, and macroalgae lipids using a highly efficient continuous catalytic process. The heterogeneous catalytic process uses supercritical methanol and porous titania microspheres in a fixed bed reactor to catalyze the simultaneous transesterification and esterification of triacylglycerides and free fatty acids, respectively, to fatty acid methyl esters (biodiesel). Triacylglycerides and free fatty acids were converted to alkyl esters with up to 85% efficiency as measured by 300 MHz ¹H NMR spectroscopy. The lipid composition of the different algae was studied gravimetrically and by gas chromatography. The analysis showed that even though total lipids comprised upwards of 19% of algal dry weight the saponifiable lipids, and resulting biodiesel, comprised only 1% of dry weight. Thus highlighting the need to determine the triacylglyceride and free fatty acid content when considering microalgae for biodiesel production.     

Astaxanthin from the red crab langostilla (Pleuroncodes planipes): optical R/S isomers and fatty acid moieties of astaxanthin esters

The composition of the fatty acids of astaxanthin esters and the distribution of astaxanthin optical RS isomers in the esterified and unesterified astaxanthin fractions extracted from the meal of the pelagic red crab langostilla (Pleuroncodes planipes; Decapoda, Anomura) were determined. Astaxanthin diesters comprised approximately 70%, monoesterified astaxanthin approximately 12%, and unesterified astaxanthin approximately 10% of total carotenoids, respectively. Unidentified carotenes and minor yellow xanthophylls represented approximately 8% of the total carotenoids. Three astaxanthin diester fractions (ratio 5:4:1) and one monoester fraction were clearly distinguished by thin-layer chromatography, and fatty acid moieties were determined in all of them. Saturated fatty acids accumulated in astaxanthin diesters, but were reduced in the monoester fraction when compared to langostilla crude oil extract (CE). Astaxanthin diesters, but not monoesters were enriched in C16:0 and C18:1n-9, when compared to the CE. Astaxanthin monoesters were rich in polyunsaturated fatty acids (approximately 70% of total fatty acids), in particular C20:5n-3 and C22:6n-3. Acylation of astaxanthin in langostilla seems to be selective rather than specific. The three diesterified astaxanthin fractions of langostilla had a ratio of approximately 3:1:3 between the (3R,3'R)-, (3R,3'S)-, and (3S,3'S)-astaxanthin isomers, whereas in the monoesterified and unesterified fractions the ratio was approximately 4:1:4. The astaxanthin optical RS isomer composition indicates that langostilla is unable to racemize astaxanthin.     

Functional screening of a novel Δ15 fatty acid desaturase from the coccolithophorid Emiliania huxleyi

The coccolithophorid Emiliania huxleyi is a bloom-forming marine phytoplankton thought to play a key role as a biological pump that transfers carbon from the surface to the bottom of the ocean, thus contributing to the global carbon cycle. This alga is also known to accumulate a variety of polyunsaturated fatty acids. At 25 °C, E. huxleyi produces mainly 14:0, 18:4n − 3, 18:5n − 3 and 22:6n − 3. When the cells were transferred from 25 °C to 15 °C, the amount of unsaturated fatty acids, i.e. 18:1n − 9, 18:3n − 3 and 18:5n − 3, gradually increased. Among the predicted desaturase genes whose expression levels were up-regulated at low temperature, we identified a gene encoding novel ?15 fatty acid desaturase, EhDES15, involved in the production of n − 3 polyunsaturated fatty acids in E. huxleyi. This desaturase contains a putative transit sequence for localization in chloroplasts and a ?6 desaturase-like domain, but it does not contain a cytochrome b₅ domain nor typical His-boxes found in ?15 desaturases. Heterologous expression of EhDES15 cDNA in cyanobacterium Synechocystis sp. PCC 6803 cells increased the level of n − 3 fatty acid species, which are produced at low levels in wild-type cells grown at 30 °C. The orthologous genes are only conserved in the genomes of prasinophytes and cryptophytes. The His-boxes conserved in orthologues varied from that of the canonical ?15 desaturases. These results suggested the gene encodes a novel ?15 desaturase responsible for the synthesis of 18:3n − 3 from 18:2n − 6 in E. huxleyi.     

A moderate change in temperature induces changes in fatty acid composition of storage and membrane lipids in a soil arthropod

A moderate change in ambient temperature can lead to vital physiological and biochemical adjustments in ectotherms, one of which is a change in fatty acid composition. When temperature decreases, the composition of membrane lipids (phospholipid fatty acids) is expected to become more unsaturated to be able to maintain homeoviscosity. Although different in function, storage lipids (triacylglycerol fatty acids) are expected to respond to temperature changes in a similar way. Age-specific differences, however, could influence this temperature response between different life stages. Here, we investigate if fatty acid composition of membrane and storage lipids responds similarly to temperature changes for two different life stages of Orchesella cincta. Juveniles and adults were cold acclimated (15 °C → 5 °C) for 28 days and then re-acclimated (5 °C → 15 °C) for another 28 days. We found adult membranes had a more unsaturated fatty acid composition than juveniles. Membrane lipids became more unsaturated during cold acclimation, and a reversed response occurred during warm acclimation. Membrane lipids, however, showed no warm acclimation, possibly due to the moderate temperature change. The ability to adjust storage lipid composition to moderate changes in ambient temperature may be an underestimated fitness component of temperature adaptation because fluidity of storage lipids permits accessibility of enzymes to energy reserves.     

Deep-water shrimp (Pandalus borealis, Krøyer 1838) as indicator organism for fish-farm waste

A feeding experiment was carried out with deep-water shrimp (Pandalus borealis), a common benthic inhabitant of Norwegian fjords. Shrimp were reared in tanks for three months, fed either salmon feed or cod, and the fatty acid profile of their muscle tissue was monitored. The salmon feed pellets and cod had completely different fatty acid compositions, with significant differences in most of the analysed fatty acids, eight of them, 14:0, iso16:0, 16:1n7, 16:2n11, 18:3n3, 20:0, 22:0 and 22:1n11, differing between the two diets by a factor of 10 or more. The levels of the fatty acids in the diets differed also substantially from the levels in the shrimp tissue at the start of the experiment. The shrimp were fed ad libitum and both groups increased significantly in total fatty acid content in the course of the experiment. Nevertheless, the fatty acids made up no more than 6.5 mg g^− 1 muscle tissue, with 88 ± 1% of them bound in polar, membrane lipids. Their composition in the tissue did also change to a certain extent, but the resulting tissue composition did not reflect the fatty acid composition of the diets. Only two fatty acids, 18:2n6 and 18:3n3, clearly followed the dietary level. They are abundant in commercial salmon feed since they are rich in plant oils used as additives to the commercial feed. Determination of fatty acid levels in shrimp tissue may enable shrimp to be used as indicator organisms for the influence of organic fish-farm waste on the food web in adjacent areas.     

Aliphatic Chains of Esterified Lipids in Isolated Eyespots of Euglena gracilis var. bacillaris

Isolated eyespot granules of Euglena gracilis Klebs var. bacillaris Pringsheim contained approximately 6% lipids (based on protein). Separation of the lipid extracts by thin layer chromatography revealed four major fractions: wax esters, triacylglycerols, free fatty acids, and phospholipids. Methanolysis of each fraction yielded between 27 and 29 different fatty acids ranging from 12:0 to 22:6. Acetates of the fatty alcohols of the wax fraction consisted of 11:0 to 18:0 carbon chains, with 14:0 being the major component; unsaturated alcohols were not detected.     

Lipid, fatty acid and protein utilization during lecithotrophic larval development of Lithodes santolla (Molina) and Paralomis granulosa (Jacquinot)

During the larval development of the subantarctic king crab, Lithodes santolla, and stone crab, Paralomis granulosa, we compared changes in the carbon, fatty acid and protein contents of larvae reared under constant conditions from hatching to metamorphosis, either in presence or absence of food (Artemia spp. nauplii). In both species the feeding condition had no influence on any of the chemical parameters studied, indicating a fully lecithotrophic (i.e. non-feeding) mode of development from hatching of the first zoea to metamorphosis of the late megalopa. Dry mass and carbon contents at hatching were similar in the larvae of both species, but L. santolla contained initially higher total amounts of fatty acids and protein than P. granulosa. Both species utilized considerable portions of their total fatty acid pool which decreased logarithmically throughout the time of development. At metamorphosis, it was almost exhausted in P. granulosa, while L. santolla had consumed only about 60%. Protein utilization, in contrast, was higher in L. santolla (40%) than in P. granulosa (20%). Triacylglycerol was the principal storage lipid in both species, accounting initially for about 75% of the lipid fraction; it was strongly utilized during larval development. Phospholipid constituted the second largest lipid class; it also decreased in P. granulosa, but to a lesser extent in L. santolla. The major fatty acids of both species were 18:1(n−9), 20:5(n−3) and 16:0 as well as, in lower proportions, 18:1(n−7), 22:6(n−3), 16:1(n−7) and 18:0. Monounsaturated fatty acids represented the dominant group in L. santolla, whereas P. granulosa contained similar amounts of mono- and polyunsaturated fatty acids. In L. santolla, monounsaturated fatty acids, especially 16:1(n−7), were preferentially utilized as compared to polyunsaturates. Due to a particularly strong lipid utilization in P. granulosa, all individual fatty acids were largely depleted at metamorphosis, showing similar extents of consumption. L. santolla had higher initial lipid and protein stores that seem to be used more economically as compared to P. granulosa.     

Comparative characterization of aqueous dispersions and cast films of different chitin nanowhiskers/nanofibers

Water dispersions of TEMPO-oxidized α-chitin nanowhisker (TOChN), partially deacetylated α-chitin nanowhisker/nanofiber mixture (DEChN), HCl-hydrolyzed chitin nanowhisker (HHChN) and squid-pen β-chitin nanofiber (SQChN) were prepared, and the properties of nano-dispersions and their cast films were characterized between the four chitin nano-samples. Because SQChN has the highest aspect ratio, its 0.1% dispersion had the highest shear stress and viscosity at the same shear rate in the four chitin nano-samples, and showed gel-like behavior in the whole shear rate range from 10⁻³ to 10³ s⁻¹. AFM images of the self-standing films showed that film surfaces consisted of characteristic chitin nano-elements with different morphologies and degrees of orientation between the four chitin samples, whereas all chitin nanowhisker/nanofiber films had similar thermal degradation points at ∼200 °C. The DEChN film had the highest tensile strength of ∼140 MPa, elongation at break of ∼10% and light-transmittance of 87% at 400 nm. In contrast, the SQChN film had the lowest tensile strength, Young's modulus and light-transmittance. All chitin nanowhisker/nanofiber films had similar oxygen permeabilities of ∼1 mL μm m⁻² day⁻¹ kPa⁻¹, which was clearly lower than that (184 mL μm m⁻² day⁻¹ kPa⁻¹) of a poly(lactic acid) film.     

Bioenergetics of early life-history stages of the brachyuran crab Cancer setosus in response to changes in temperature

In many marine invertebrates, a latitudinal cline in egg size is considered an adaptive response to a decrease in temperature, and enhances the energetic fitness of their larvae at hatching. However, the amount of energy carried over from the egg to the larval stage depends on the metabolic efficiency of egg development. In the present study, eggs of the brachyuran crab Cancer setosus were sampled for their dry mass (DM), carbon (C), nitrogen (N), and fatty acid (FA) content throughout development from blastula stage until hatching of zoea 1-larvae at Antofagasta (23°S) and Puerto Montt 41°S (Chile) under different temperature treatments (12, 16 and 19 °C). Hatching zoea 1 larvae contained 60 ± 3% of the initial blastula egg C content, regardless of site or temperature. However, the ontogenetic decrease in egg C content was to a significantly higher extend based on the utilization of energy-rich FA at 12 °C (− 1.16 µg/egg) compared to the 19 °C treatments in Antofagasta and Puerto Montt (− 0.63 to − 0.73 µg FA per egg). At 19 °C egg-metabolism was based to a substantial extend on protein, which allowed for the saving of energy-richer lipids. We conclude that the production of larger eggs with high FA content appears to be adaptive not only to fuel the larval development, but is also a response to the prolonged egg developmental times at lower temperatures.     

The paradox high zooplankton biomass-low vegetal particulate organic matter in high turbidity zones: What way for energy transfer?

Estuarine ecosystems are characterized by high zooplanktonic biomasses, essentially constituted by copepods and mysids whose nutritional requirements are mainly provided by phytoplankton, an easily available carbon form. The Gironde estuary is characterized by high turbidities which limit light penetration in the water column and therefore primary production. Consequently, primary production is low and its availability for higher trophic level is very limited. The main goal of this study was to characterize the total vegetal particulate organic matter (POM) in high turbidity zones of the Gironde estuary during summer (a critical period characterized by high heterotrophic bacterial degradation and high zooplanktonic biomasses) and to analyse its utilization by zooplankton, using prey/predator experiments and trophic biomarkers (fatty acids). The specific goals were to define (i) how vegetal POM was exploited by the different zooplanktonic groups (protozoa, copepods and mysids) and (ii) which alternative preys could be used when vegetal POM was not sufficient to ensure their nutritional requirements.Chlorophyll biomass was very low in the MTZ during summer 2002 (0.48 ± 0.03 mg m^− 3). Total zooplankton grazing was low (19% d^− 1) probably due to a large contribution of detritus originating from terrestrial plants in vegetal POM compared to phytoplankton. The highest grazing pressure was exercised by the mysid Mesopodopsis slabberi due to its high abundances and by its almost entirely herbivorous diet (phytoplankton and small terrestrial detritus). Grazing rates (19.7 ± 4.2 and 9.6 μgC cop^− 1 d^− 1 for juveniles and adults, respectively) seemed to be sufficient to satisfy their daily carbon requirement. Grazing rate of the copepod Eurytemora affinis (139 ngC cop^− 1 d^− 1) seemed to be insufficient to cover its nutritional requirements and the copepods probably needed to complete a great part of their diet from protozoa. Grazing rates of the mysid Neomysis integer (24.7 ± 0.01 and 20.89 ± 8.45 μgC cop^− 1 d^− 1 for juveniles and adults, respectively) were higher than those of M. slabberi when feeding only on phytoplankton. However, when other preys were introduced in its environment, N. integer only fed on the copepod E. affinis with a preference for nauplii. The study revealed the great importance of protozoa and bacteria in the trophic transfers between vegetal POM and zooplankton in the MTZ during summer, despite the low protozoa grazing pressure on vegetal POM (3.1%). The detritic food chain probably implies various trophic transfers with little direct relationships between vegetal POM and zooplankton.     

The composition of the free fatty acids from Dendrolimus pini exuviae

The pine moth Dendrolimus pini effectively resists many insecticides, but it can be controlled by the use of bioinsecticides such as entomopathogenic fungi. In the use of microbial agents for the biocontrol of D. pini, it is important to identify the cuticular lipids of this pest if we are to understand the factors responsible for the preferential adhesion or selective repulsion of entomopathogenic fungi that are potentially useful in biocontrol. In this work the qualitative and quantitative analyses of free fatty acids in two exuviae extracts (petroleum ether and dichloromethane) and two developmental stages (larval-larval and larval-pupal molts) were studied. The free fatty acid composition of the epicuticular lipids from exuviae of D. pini was characterized chemically using gas chromatography (GC) and gas chromatography-electron impact mass spectrometry (GC-MS). Structural analyses of the dichloromethane extracts from larval-larval exuviae (LLE) and larval-pupal exuviae (LPE) revealed that the carbon numbers for the major acid moieties ranged from C_8:0 to C_34:0. Only C_23:0 was not identified in the LPE extract. The relative contents of fatty acids in the extracts varied from trace amounts to 34%. The fatty acids extracted by dichloromethane were essentially the same as those in the petroleum ether extract. We also identified dehydroabietic acid in the exuviae of D. pini. The respective quantities of dehydroabietic acid obtained from D. pini LLE and LPE were 1763 ± 103 μg/g exuviae and 11521 ± 1198 μg/g of exuviae.     

Plasmodium falciparum: Chemically defined medium for continuous intraerythrocytic growth using lipids and recombinant albumin

Dioleoylphosphatidylcholine and other phosphatidylcholines containing different fatty acid moieties were found to increase the ability of nonesterified fatty acids (NEFA) to sustain continuous intraerythrocytic growth of Plasmodium falciparum in the presence of specific proteins. Other phospholipids, including phosphatidylethanolamine, phosphatidylserine, and phosphatidic acid, were beneficial to parasite growth. Different combinations and concentrations of NEFA tested in the presence of phospholipids and bovine albumin had variable effects on parasite growth. The most effective combination for promoting parasite growth consisted of 30 μg/ml cis-9-octadecenoic acid (oleic acid) plus 15 μg/ml hexadecanoic acid (palmitic acid). Recombinant human albumin could replace bovine or human albumin in culture media enriched with structurally defined lipids. This study therefore established a chemically defined culture medium suitable for sustaining the growth of P. falciparum.     

n − 3, but not n − 6 lipid particle uptake requires cell surface anchoring

Omega-3 (n − 3) fatty acids are emerging as bioactive agents protective against cardiovascular disease. However, their cellular delivery pathways are poorly defined. Here we questioned whether the uptake of n − 3 triglyceride-rich particles (TGRP) is mediated by cell surface proteoglycans (PG) using LDL receptor (LDLR)+/+ and LDLR−/− cell models. LDLR+/+ but not LDLR−/− cells showed higher n − 6 over n − 3 TGRP uptake. Removal of cell surface proteins and receptors by pronase markedly enhanced the uptake of n − 3 but not n − 6 TGRP. Lactoferrin blockage of apoE-mediated pathways decreased the uptake of n − 6 TGRP by up to 85% (p < 0.05) but had insignificant effect on n − 3 TGRP uptake. PG removal by sodium chlorate in LDLR+/+ cells substantially reduced n − 3 TGRP uptake but had little effect on n − 6 TGRP uptake. Thus, while n − 6 TGRP uptake is preferentially mediated by LDLR-dependent pathways, the uptake of n − 3 TGRP depends more on PG and non-LDLR cell surface anchoring.     

Preparation and characterization of molecular weight fractions of glycosaminoglycan from sea cucumber Thelenata ananas using free radical depolymerization

A glycosaminoglycan from sea cucumber Thelenata anana (THG) was isolated as a polymer of molecular weight of around 70 kDa. Its low molecular weight derivatives were first prepared by free radical depolymerization with hydrogen peroxide in the presence of copper(II) ion. The parameters of the process were investigated by a high-performance gel permeation chromatography. Analyses of chemical composition and molecular weight distribution indicated that the fragmentation of the main-chain of THG occurred randomly, obeyed pseudo first-order kinetics, and produced species with rather narrow and unimodal distribution of molar mass. The characterization of different molecular weight fractions was investigated by using viscometry and atomic force microscopy (AFM). Analysis of molecular weight and intrinsic viscosity in terms of the known theories for unperturbed wormlike cylinder yielded 1201 ± 110 nm⁻¹, 15.3 ± 1.5 nm, and 1.5 ± 0.3 nm for molar mass per unit contour length M_L, persistence length q, and diameter d, respectively. The M_L and d values were approximately consistent with those observed by AFM. The present data suggest that THG may dissolve in 0.1 M aqueous NaCl as single-stranded helical chains.     

Fatty Acids of Extractable and Bound Lipids of Rhodomicrobium vannielii

Cells of Rhodomicrobium vannielii grown at 29 C in a lactate-containing medium were extracted at room temperature with organic solvents. The extractable fraction contained the bulk of the simple lipid (1.87% of cell dry weight) and complex lipids (phospholipids, 4.2%; sulfolipid, 0.01%), coenzyme Q (0.09%), and pigments (carotenoids 1.2%; bacteriochlorophyll, 1.9%). The cell residue contained the bound lipids (nonpolar fatty acid fraction, 1.86%; polar hydroxy fatty acids, 0.49%). The residue also contained poly-β-hydroxybutyric acid (0.2%), which was extracted in boiling chloroform. In both the simple and complex lipids, vaccenic acid (11-octadecenoic acid) was the largest single component (approximately 90% in each fraction). The fatty acids of the bound lipid contained 35% vaccenic acid, even- and odd-numbered saturated and unsaturated straight-chain fatty acids, cyclopropane-, branched-, and α- and β-hydroxy fatty acids. The extractable lipids contained only straight-chain saturated and unsaturated even-numbered fatty acids. Nearly 60% of hydroxy fatty acid fraction was α-hydroxydodecanoic acid (24%) and β-hydroxydodecanoic acid (34.5%). Coenzyme Q was crystallized and identified as Q₉ on the basis of melting point and chromatographic properties. Q₁₀ had been previously reported.