Thematic Review Series: Phospholipases: Central Role in Lipid Signaling and Disease: A new era of secreted phospholipase A2

Among more than 30 members of the phospholipase A₂ (PLA₂) superfamily, secreted PLA₂ (sPLA₂) enzymes represent the largest family, being Ca²⁺-dependent low-molecular-weight enzymes with a His-Asp catalytic dyad. Individual sPLA₂s exhibit unique tissue and cellular distributions and enzymatic properties, suggesting their distinct biological roles. Recent studies using transgenic and knockout mice for nearly a full set of sPLA₂ subtypes, in combination with sophisticated lipidomics as well as biochemical and cell biological studies, have revealed distinct contributions of individual sPLA₂s to various pathophysiological events, including production of pro- and anti-inflammatory lipid mediators, regulation of membrane remodeling, degradation of foreign phospholipids in microbes or food, or modification of extracellular noncellular lipid components. In this review, we highlight the current understanding of the in vivo functions of sPLA₂s and the underlying lipid pathways as revealed by a series of studies over the last decade.     

AMP-activated protein kinase: an emerging drug target to regulate imbalances in lipid and carbohydrate metabolism to treat cardio-metabolic diseases: Thematic Review Series: New Lipid and Lipoprotein Targets for the Treatment of Cardiometabolic Diseases

The adenosine monophosphate-activated protein kinase (AMPK) is a metabolic sensor of energy metabolism at the cellular as well as whole-body level. It is activated by low energy status that triggers a switch from ATP-consuming anabolic pathways to ATP-producing catabolic pathways. AMPK is involved in a wide range of biological activities that normalizes lipid, glucose, and energy imbalances. These pathways are dysregulated in patients with metabolic syndrome (MetS), which represents a clustering of major cardiovascular risk factors including diabetes, lipid abnormalities, and energy imbalances. Clearly, there is an unmet medical need to find a molecule to treat alarming number of patients with MetS. AMPK, with multifaceted activities in various tissues, has emerged as an attractive drug target to manage lipid and glucose abnormalities and maintain energy homeostasis. A number of AMPK activators have been tested in preclinical models, but many of them have yet to reach to the clinic. This review focuses on the structure-function and role of AMPK in lipid, carbohydrate, and energy metabolism. The mode of action of AMPK activators, mechanism of anti-inflammatory activities, and preclinical and clinical findings as well as future prospects of AMPK as a drug target in treating cardio-metabolic disease are discussed.     

Effects of linoleic and gamma-linolenic acids (efamol evening primrose oil) on fatty acid-binding proteins of rat liver

Summary We have studied the effects of Efamol evening primrose oil (EPO) on fatty acid-binding proteins (L-FABP) of rat liver. EPO contains 72% cis-linoleic acid and 9% cis-gamma linolenic acid. EPO has been clinically used for treatment of a number of diseases in humans and animals. EPO is also known to lower cholesterol level in humans and animals. Feeding of an EPO supplemented diet to rats (n = 9) for 2 months decreases the oleate binding capacity of purified L-FABP of rat liver whereas the palmitate binding activity was increased by 38%. However, EPO feeding did not alter the L-FABP concentrations significantly as measured by using the fluorescence fatty acid probe, dansylamino undecanoic acid. Endogenous fatty acid analysis of L-FABPs revealed significant qualititative and quantitative changes in fatty acid pattern after EPO feeding. EPO feeding decreased the endogenous palmitate level by 53% and oleate level by 64% in L-FABPs and also EPO feeding decreased the total endogenous fatty acid content from 62 nanomole per mg of protein to 42 nanomole per mg of L-FABP (n = 3).     

Comparison of fatty acid composition in total lipid of diapause and non-diapause larvae of Cydia pomonella （Lepidoptera： Tortricidae）

Seasonal changes in the fatty acid composition of the total lipid extracted from the whole body of Cydia pomonella L. larvae were determined by gas chromatography. The six most abundant fatty acids in both non-diapause and diapause larvae of codling moth were oleic （35%-39%）, palmitic （23%-33%）, linoleic （16%-30%）, palmitoleic （5%-10%）, stearic （1.5%-3.0%） and linolenic acids （1.0%-2.5%）. This represents a typical complement of Lepidopteran fatty acids. The fatty acid composition of total lipid of C. pomonella larvae was related to diapause. In similarity to most other reports, the proportion of unsaturated fatty acids increased in diapause initiation state. The total lipid of diapause larvae contained more linoleic acid （25.8% vs. 16.1%） and less palmitic acid （24.7% vs. 33.4%）, than that of non-diapause larvae. The weight percentage of linoleic acid （C 18：2） increased from 16% to 26% from early-August through early-September during transition to diapause, while palmitic acid （C16：0） decreased from 33% to 25% at the same time. These changes resulted in an increase in the ratio of unsaturated to saturated fatty acids （UFA/SFA） from 1.72 in non-diapause larvae to 2.63 in diapause larvae.     

Lipid composition of the pea aphid,Acyrthosiphon pisum (Harris) (homoptera: Aphididae), reared on host plant and on artificial media

Polar and neutral lipids and their constitutive fatty acids were quantified in the pea aphid, Acyrthosiphon pisum (Harris), grown on host plant or on a lipid free artificial diet. The results were compared to determine if lipids were involved in the suitability of the diet for continuous rearing of this A. pisum biotype. For apterous adults grown on plants, the lipids were characterized by a low amount of neutral lipids (2.5% weight/fresh weight) almost entirely (96.4%) composed of hexanoyl and sorboyl dimyristin. These storage lipids were higher in the alatae (3.8%), probably correlated with potential flight activity. The phospholipid amounts were identical in these two morphs (1.3–1.4% weight/fresh weight), comprised mainly of phosphatidylethanolamines (52%) and phosphatidylcholines (40.6%). These phospholipids contained a still unidentified fatty acid, with a retention time close to that of linolenic acid and synthesized by the aphid or its bacterial symbionts (not found in plants). The apterous adult aphids reared on an artificial diet showed an accumulation of neutral lipids (8.9% for the first generation); this increase was shown to be slightly greater for the hexanoyl and sorboyl triglycerides. In contrast, the phospholipids decreased in aphids reared on an artificial diet (1.1% and 0.9%, respectively, for first and second generation), correlated with a phospholipid fatty acid profile significantly deficient in C18:3 and in the unidentified peculiar fatty acid. These phospholipids are essential components of biological membranes and a diet-driven deficient synthesis in some of their components may result in the observed symptoms. © 1992 Wiley-Liss, Inc.     

Total lipid and fatty acid compositions of <Emphasis Type="Italic">Lertha sheppardi</Emphasis> (Neuroptera: Nemopteridae) during its main life stages

Total lipid and the fatty acid compositions of phospholipid and triacylglycerol fractions, prepared from eggs, 3^rd instars of larvae, pupae, male and female adults of Lertha sheppardi, were analyzed by gas chromatography and gas chromatography-mass spectrometry. The effect of diet (adults’ nutrition) on fatty acid composition of L. sheppardi adults was also investigated. Total lipid of L. sheppardi considerably increased in adults compared with immature stages. There was a significant decrease in total lipid level in larval stage in contrast with egg stage. Qualitative analysis revealed the presence of 14 fatty acids during all stages. The major components were C16 and C18 saturated and unsaturated components which are ubiquitous to most animal species. In addition to these components, one odd-chain (C17:0) and prostaglandin precursor fatty acids were found. The fatty acid profiles of phospholipids and triacylglycerols were substantially different. In phospholipid fraction, monounsaturated fatty acids were the major proportion of fatty acids in both sex of adults and pupae, whereas polyunsaturated fatty acids were the most dominant fatty acids in eggs and 3^rd instars. Results of triacylglycerol fraction revealed that fatty acid composition of eggs had higher level of C16:1, C18:0 and C18:3n-3 content than that of 3^rd instars and pupae, which suggests accumulation of energetic and structural reserve materials during embryonic development. At more advanced developmental stages, mainly in adult females, the amount of C16:1 increased once again, which may be related to the need for accumulation of sufficient energy and of carbon reservoir in the developing new vitellum. Percentages of C18:1 were significantly high in adult stages compared to other stages. These findings indicate that the accumulation and consumption of fatty acids fluctuate through different development stages. Diet did not effect the fatty acid composition of L. sheppardi adults.     

The fatty acid compositions of predator Piocoris luridus （Heteroptera： Lygaeidae） and its host Monosteria unicostata （Heteroptera： Tingidae） reared on almond

The changes in fatty acid compositions during nutritional interaction among almond Amygdalus communis Linnaeus （Rosales： Rosaceae） （host plant）, lacebug Monosteria unicostata （Mulsant and Rey） （Heteroptera： Tingidae） and its predator Piocoris luridus Fieber （Heteroptera： Lygaeidae） were determined by gas chromatography and gas chromatography-mass spectrometry analyses. The fatty acid profiles of phospholipids and triacylglycerols were substantially different. Unlike the general observations for virtually most terrestrial insects, arachidonic and eicosapentaenoic acids were detected in high proportions of phospholipid fractions in both insects, especially in P. luridus. Also the almond tissues provide very little oleic acid to the herbivore diet, yet both insect species developed high proportions of this component. Our data reveals instances of specific accumulation of fatty acid biosynthesis, elongation/desaturation, and not incorporating selected fatty acids into cellular lipids.     

The distribution and diversity of Chironomidae (Insecta: Diptera) in western Finnish Lapland, with special emphasis on shallow lakes

Aim Predictions of aquatic ecosystem change with global warming require basic data that accurately reflect the environmental conditions underlying species distributions. However, in remote arctic areas such baseline data are scarce. We assess the influence of environmental variables on chironomid distribution and taxon richness in shallow, isothermal lakes in a poorly studied arctic region. We pay particular attention to community variation along the treeline ecotonal zone where many environmental variables change abruptly in a relatively small area. Location Lake transect in Finnish Lapland spanning from boreal coniferous forest to arctic tundra. Methods Chironomid assemblages were determined from surface‐sediment samples of 50 shallow (< 10 m) natural lakes. Abundance and taxon richness data were related to 24 limnological variables using canonical ordination techniques (DCA, CCA, RDA). A Monte Carlo permutation procedure was used to assess the explanatory power of single variables. Between‐vegetation zone differences of richness were tested for statistical significance using one‐way anova . Results In total, 7771 chironomid head capsules were identified, consisting of 13 species, 10 species groups, four subgenera, 41 genera, four genus groups, five types and three with uncertain taxonomic affiliation. A hump‐shaped relationship between taxon richness and elevation was noted along the study transect with a peak in taxon richness occurring in mountain birch woodland lakes at middle elevations, decreasing then towards both warmer and colder ends of the elevation/temperature gradient. Of the individual parameters, sediment organic content, total organic carbon, pH, and lake‐specific air temperature accounted for the greatest amount of variation in the chironomid data. Main conclusions Maximum taxon richness occurred at mid‐elevations where aquatic algae also reached their maximum diversity. This area coincides with an ecotonal transitional zone, which seems more likely to account for the peak in species richness. Our study demonstrates that the factors most strongly affecting chironomids in Finnish Lapland (i.e. temperature, and ecosystem features) are those that with great probability will also change as a result of future climate change. This will likely have an effect on the distribution of chironomids in subarctic and arctic areas.     

特种五谷虫脂肪酸的体外抗肿瘤、抗HIV-1整合酶活性及组成分析

为研究特种五谷虫(大头金蝇Chrysomya megacephala蝇蛆的俗称)脂肪酸的体外抗肿瘤、抗HIV-1整合酶活性,并确定其组分构成,通过溶剂萃取和油脂酶解获得2种脂肪酸FA1和FA2,采用MTT法/SRB法测定其体外抗人白血病细胞HL-60/人肺癌细胞A-549活性,以及采用Biotin-ELISA法检测其对HIV-1整合酶的抑制作用;并由GC-MS分析确定其脂肪酸组成。结果表明：FA1和FA2对人白血病细胞/人肺癌细胞均有显著的抑制活性,其IC₅₀值在35～65 μg/mL;对HIV-1整合酶同样具有强烈的抑制活性,IC₅₀值分别为86.7 μg/mL和98.5 μg/mL。GC-MS分析表明,FA1和FA2化学组成相似,均含有15%～16%的多不饱和脂肪酸（PUFA),其中有2个组分为ω-6 PUFA。提示特定培养的五谷虫,其脂肪酸成分具有显著的体外抗肿瘤、抗HIV-1整合酶活性,其中含有的PUFA,尤其ω-6 PUFA,可能是主要活性组分;FA2在来源上和FA1存有相关性。     

Combined effect of vanadium(V) and chromium(III) on lipid peroxidation in liver and kidney of rats

Since chromium(III) was demonstrated to have antioxidative action, we have decided to study the effect of this element on V-induced LPO in liver and kidney of rats. Outbred 2-month-old, albino male Wistar rats received daily, for a period of 12 weeks: group I (control), deionized water to drink; group II, sodium metavanadate (SMV) solution at a concentration of 0.100mgV/mL; group III, chromium chloride (CC) solution at a concentration of 0.004mgCr/mL and group IV, SMV-CC solution at a concentration of 0.100mgV and 0.004mgCr/mL. The particular experimental groups took up with drinking water about 8.6mgV/kg b.w./24h (group II), 0.4mgCr/kg b.w./24h (group III), 9mgV and 0.36mgCr/kg b.w./24h (group IV). The V- or Cr-treated groups had higher concentrations of these two elements in liver and kidney compared to the controls. The administration of vanadium alone caused a significant decrease in fluid intake and in body weight gain compared to the controls. In liver supernatants obtained from all tested rats a statistically significant increase in MDA concentration was demonstrated in spontaneous LPO in comparison with the control rats. Moreover, in rats intoxicated with vanadium alone a statistically significant increase in liver MDA level was observed in the presence of 100microM NaVO(3). Instead, in supernatants of liver received from rats treated with chromium alone, a statistically significant increase in MDA concentration in comparison with the controls was found in the presence of 400microM NaVO(3). In kidney supernatants obtained from rats treated with chromium alone, a statistically significant increase in lipid peroxidation was shown in the presence of 30microM FeSO(4) and 400microM NaVO(3). These results show that the tested doses of vanadium(V) and chromium(III) ingested by rats with their drinking water caused significant alterations in internal organs, especially in liver. Under the conditions of our experiment, Cr(III) did not demonstrate antioxidant action, it rather had an oxidant effect.     

Historical and biomechanical analysis of integration and dissociation in molluscan feeding,with special emphasis on the true limpets (Patellogastropoda: Gastropoda)

Modifications of the molluscan feeding apparatus have long been recognized as a crucial feature in molluscan diversification, related to the important process of gathering energy from the environment. An ecologically and evolutionarily significant dichotomy in molluscan feeding kinematics is whether radular teeth flex laterally (flexoglossate) or do not (stereoglossate). In this study, we use a combination of phylogenetic inference and biomechanical modeling to understand the transformational and causal basis for flexure or lack thereof. We also determine whether structural subsystems making up the feeding system are structurally, functionally, and evolutionarily integrated or dissociated. Regarding evolutionary dissociation, statistical analysis of state changes revealed by the phylogenetic analysis shows that radular and cartilage subsystems evolved independently. Regarding kinematics, the phylogenetic analysis shows that flexure arose at the base of the Mollusca and lack of flexure is a derived condition in one gastropod clade, the Patellogastropoda. Significantly, radular morphology shows no change at the node where kinematics become stereoglossate. However, acquisition of stereoglossy in the Patellogastropoda is correlated with the structural dissociation of the subradular membrane and underlying cartilages. Correlation is not causality, so we present a biomechanical model explaining the structural conditions necessary for the plesiomorphic kinematic state (flexoglossy). Our model suggests that plesiomorphically the radular teeth must flex laterally as they pass over the bending plane as a result of the mechanical restrictions in the flexible but inelastic subradular membrane and close association between subradular membrane and cartilages. Relating this model to the specific character states of the clades, we conclude that lack of flexure in patellogastropods is caused by the dissociation of the subradular membrane and cartilage supports. J. Morphol. 241:175–195, 1999. © 1999 Wiley‐Liss, Inc.     

Isomerization of octadecapentaenoic acid (18:5n‐3) in algal lipid samples under derivatization for GC and GC‐MS analysis

During gas chromatography (GC) analysis of fatty acid (FA) composition of the dinoflagellate Gymnodinium kowalevskii, we found unex‐pectedly low and irreproducible content of all‐cis‐3,6,9,12,15‐octadecapentaenoic acid (18:5n‐3), which is an important chemotaxonomic marker of several classes of microalgae. We compared chromatographic behavior of 18:5n‐3 methyl ester and other GC derivatives obtained using different conventional methods of derivatization. The use of methods based on saponification or base‐catalyzed transesterification resulted in a mixture of double‐bond positional isomers of 18:5. On a SUPELCOWAX 10 column, the equivalent chain length (ECL) value for authentic 18:5n‐3 methyl ester was 20.22, whereas the main component after base‐catalyzed methylation had ECL 20.88. Attempts to prepare N‐acyl pyrrolidides or 4,4‐dimethyloxazoline (DMOX) derivatives of 18:5n‐3 also gave inadequate results. These derivatives also showed a main peak corresponding to isomerized 18:5. Mass spectra for both DMOX and pyrrolidide derivatives of this compound showed the base peak at m/z 139, probably corresponding to 2,6,9,12,15‐18:5 acid. Of all methods tested for methylation, only derivatization with 5% HCl or 1% sulphuric acid in methanol gave satisfactory results. Therefore, GC or GC‐mass spectrometry analyses of algal lipids containing 18:5n‐3 may be inaccurate when base‐catalyzed methods of FA derivatization are applied. The best and simplest way to avoid incorrect GC results is to use standard acid‐catalyzed methylation.     

Effects of phospholipase A(2) and its products on structural stability of human LDL: relevance to formation of LDL-derived lipid droplets

Hydrolysis and oxidation of LDL stimulate LDL entrapment in the arterial wall and promote inflammation and atherosclerosis via various mechanisms including lipoprotein fusion and lipid droplet formation. To determine the effects of FFA on these transitions, we hydrolyzed LDL by phospholipase A(2) (PLA(2)), removed FFA by albumin, and analyzed structural stability of the modified lipoproteins. Earlier, we showed that heating induces LDL remodeling, rupture, and coalescence into lipid droplets resembling those found in atherosclerotic lesions. Here, we report how FFA affect these transitions. Circular dichroism showed that mild LDL lipolysis induces partial β-sheet unfolding in apolipoprotein B. Electron microscopy, turbidity, and differential scanning calorimetry showed that mild lipolysis promotes LDL coalescence into lipid droplets. FFA removal by albumin restores LDL stability but not the protein conformation. Consequently, FFA enhance LDL coalescence into lipid droplets. Similar effects of FFA were observed in minimally oxidized LDL, in LDL enriched with exogenous FFA, and in HDL and VLDL. Our results imply that FFA promote lipoprotein coalescence into lipid droplets and explain why LDL oxidation enhances such coalescence in vivo but hampers it in vitro. Such lipid droplet formation potentially contributes to the pro-atherogenic effects of FFA.     

New records and updated checklist of mosquitoes (Diptera: Culicidae) from Lao People's Democratic Republic,with special emphasis on adult and larval surveillance in Khammuane Province

A list of mosquitoes from the Nakai Nam Theun National Protected Area along the Nam Theun, Nam Mon, Nam Noy, and Nam On rivers, Nakai District, Khammuane Province, Lao People's Democratic Republic (Lao PDR) is presented. Fifty‐four mosquito taxa were identified, including 15 new records in the Lao PDR. A fragment of the mtDNA cytochrome c oxidase subunit I (COI) gene, barcode region, was generated for 34 specimens, and together with four specimens already published, it represented 23 species in eight genera. In addition, an updated checklist of 170 mosquito taxa from Lao PDR is provided based on field collections from Khammuane Province, the literature, and specimens deposited in the Smithsonian Institution, National Museum of Natural History (SI‐NMNH), Washington, DC, U.S.A. This paper provides additional information about the biodiversity of mosquito fauna in Lao PDR.     

Evaluation of groundwater contamination for fluoride and nitrate in semi-arid region of Nirmal Province,South India: A special emphasis on human health risk assessment (HHRA)

The main aim of this study was to assess the groundwater quality and human health risks of fluoride and nitrate contamination in Nirmal Province, South India, where groundwater is the primary source for drinking water. Hazard quotient (HQ) and total hazard index (THI) were calculated to estimate the non-carcinogenic risk to men, women, and children using the most substantial method recommended by United States Environmental Protection Agency (USEPA). The results of the study reveal that 26% and 20.59% of groundwater samples have significantly high nitrate and fluoride concentrations, exceeding the maximum permissible limits set by Bureau of Indian Standards (45 mg/L and 1.5 mg/L, respectively). Therefore, ingestion of high fluoride and nitrate water could be the chief reason for health risk in the study region. The total non-carcinogenic health risks for men, women, and children ranged from 2.95E?01 to 4.07E+00, 3.49E?01 to 4.80E+00, and 3.99E?01 to 5.50E+00, respectively. Moreover, 67.65%, 79.41%, and 82.35% of the total collected groundwater samples exceeded the permissible limit for acceptable total health index (THI = 1) for men, women, and children, respectively. Therefore, the health risk assessment suggests that children face higher health risk than men and women in the study region.