TAG Content Research Articles

Phospholipase C signal mediated the glucose-induced changes of glucose absorption and lipid accumulation in the intestinal epithelial cells of yellow catfish Pelteobagrus fulvidraco.

In present study, we explored the effects and the underlying mechanisms of phospholipase C (PLC) mediating glucose-induced changes in intestinal glucose transport and lipid metabolism by using U-73122 (a PLC inhibitor). We found that glucose incubation activated the PLC signal and U-73122 pre-incubation alleviated the glucose-induced increase in plcb2, plce1 and plcg1 mRNA expression. Meanwhile, U-73122 pre-treatment blunted the glucose-induced increase in sodium/glucose co-transporters 1/2 mRNA and protein expressions. U-73122 pre-treatment alleviated the glucose-induced increase in TAG content, BODIPY 493/503 fluorescence intensity, lipogenic enzymes (glucose 6-phospate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), malic enzyme and fatty acid synthase (FAS)) activity and the mRNA expressions of lipogenic genes and related transcription factors (6pgd, g6pd, fas, acca, srebp1 and carbohydrate response element-binding protein (chrebp)) in intestinal epithelial cells of yellow catfish. Further research found that U-73122 pre-incubation mitigated the glucose-induced increase in the ChREBP protein expression and the acetylation level of ChREBP in HEK293T cells. Taken together, these data demonstrated that the PLC played a major role in the glucose-induced changes of glucose transport and lipid metabolism and provide a new perspective for revealing the molecular mechanism of glucose-induced changes of intestinal glucose absorption, lipid deposition and metabolism.

Accumulation and deposition of triacylglycerols in the starchy endosperm of wheat grain

A combination of lipidomics, transcriptomics and bioimaging has been used to study triacylglycerol synthesis and deposition in the developing starchy endosperm of wheat. The content of TAG increased between 14 and 34 days after anthesis, from 50 to 115 mg/100 g dry wt and from about 35 to 175 mg/100 g dry wt in two experiments. The major fatty acids were C16 (palmitic C16:0 and palmitoleic C16:1) and C18 (stearic C18:0, oleic C18:1, linoleic C18:2 and linolenic C18:3), with unsaturated fatty acids accounting for about 75–80% of the total throughout development. Linoleic acid (C18:2) was the major component at all stages and the proportion increased during development. Transcript profiling indicated that predominant route to TAG synthesis and oil accumulation is via the Kennedy pathway and diacylglycerol acyltransferase (DGAT) activity. Confocal microscopy of stained tissue sections showed that TAG accumulated in droplets which are associated with protein and concentrated in the starchy endosperm cells below the sub-aleurone cells. Transcripts encoding 16kd oleosins were also expressed, indicating that the oil droplets are in part stabilised by oleosin proteins.

Effects of dietary curcumin on growth, antioxidant capacity, fatty acid composition and expression of lipid metabolism-related genes of large yellow croaker fed a high-fat diet.

A 10-week feeding trial was conducted to investigate the effect of dietary curcumin (CC) on growth antioxidant responses, fatty acid composition, and expression of lipid metabolism-related genes of large yellow croaker fed a high-fat diet (HFD). Four diets (lipid level at 18 %) were formulated with different levels of curcumin (0, 0·02, 0·04 and 0·06 %). The best growth performance was found in the 0·04 % curcumin group, with the body and hepatic lipid levels lower than the control group (0 % CC). The content of TAG, total cholesterol and LDL-cholesterol was the least in the 0·06 % curcumin group. The lowest malondialdehyde and the highest superoxide dismutase, catalase and total antioxidant capacity were observed in the 0·04 % curcumin group. The 0·04 % curcumin group had higher expression of Δ6fad, elovl5 and elovl4 and showed higher hepatic n-6 and n-3 PUFA. Expression of ppara, cpt1, and aco was significantly increased, while expression of srebp1 and fas was dramatically decreased in curcumin groups compared with the control group. Overall, 0·04 % curcumin supplementation could mitigate the negative effects caused by HFD and promote growth via reducing hepatic lipid deposition, improving antioxidant activity and increasing PUFA of large yellow croaker. To conclude, abnormal hepatic lipid deposition was probably due to increased fatty acid oxidation and reduced de novo synthesis of fatty acids.

Metabolically engineered rice biomass and grain using genes associated with lipid pathway show high level of oil content.

Increasing lipid content using metabolic engineering methods in different parts of plant, including, leaves and stem can be considered as an innovative platform for achieving more energy and biofuel in more green habits. Two key enzymes, including, diacylglycerol acyltransferase (DGAT) and phospholipid:diacylglycerol acyltransferase (PDAT) catalyze the final step of TAG assembly. WRINKLED1 (WRI1) is one of the important transcription factors which regulate the fatty acid biosynthesis network and TAG accumulation by balancing carbon flux between carbohydrates and lipids. In addition, oleosin encoding gene (OLE) can protect TAGs from degradation by packing into oil bodies.In the current study, four important genes involved in TAG assembly and protection (i.e., AtDGAT1 and AtPDAT, AtWRI1, and AtOle) were overexpressed under a constitutive promoter in rice crop. TAG content of transgenic seeds increased significantly (P ≤ 0.05) by 26% in compared with those of control plants. Oleic and palmitic acid contents were significantly increased by 28% (from 32 to 41) and 27% (11 to 14) in seeds of transgenic plants in compared with controls, respectively. Our results showed an increase in the total grain and leaf oil contents by 70% (from 1.1 to 1.87%) and 22.5% (from 1.88 to 2.3%) in the metabolically engineered lines, respectively. This is the first report of transformation in rice for enhancing oil content and energy density in its seeds and vegetative parts. Such metabolically engineered crops would be cultivated for production much more oils in seeds and straw for food and biodiesel consequently.

TAG pathway engineering via GPAT2 concurrently potentiates abiotic stress tolerance and oleaginicity in Phaeodactylum tricornutum

BackgroundDespite the great potential of marine diatoms in biofuel sector, commercially viable biofuel production from native diatom strain is impractical. Targeted engineering of TAG pathway represents a promising approach; however, recruitment of potential candidate has been regarded as critical. Here, we identified a glycerol-3-phosphate acyltransferase 2 (GPAT2) isoform and overexpressed in Phaeodactylum tricornutum.ResultsGPAT2 overexpression did not impair growth and photosynthesis. GPAT2 overexpression reduced carbohydrates and protein content, however, lipid content were significantly increased. Specifically, TAG content was notably increased by 2.9-fold than phospho- and glyco-lipids. GPAT2 overexpression elicited the push-and-pull strategy by increasing the abundance of substrates for the subsequent metabolic enzymes, thereby increased the expression of LPAAT and DGAT. Besides, GPAT2-mediated lipid overproduction coordinated the expression of NADPH biosynthetic genes. GPAT2 altered the fatty acid profile in TAGs with C16:0 as the predominant fatty acid moieties. We further investigated the impact of GPAT2 on conferring abiotic stress, which exhibited enhanced tolerance to hyposaline (70%) and chilling (10 ºC) conditions via altered fatty acid saturation level.ConclusionsCollectively, our results exemplified the critical role of GPAT2 in hyperaccumulating TAGs with altered fatty acid profile, which in turn uphold resistance to abiotic stress conditions.

Comparative study of graphic-based tag clouds: theory and experimental evaluation for information search

PurposeGraphic-based tag clouds aim to visually represent tag content and tag structure, and then to better represent tagged information for later search. However, few studies have clarified the features among varied visualization approaches involved in graphic-based tag clouds and compared them for the purpose of information search.Design/methodology/approachAfter reviewing four kinds of graphic-based tag clouds, an experimental demonstration was conducted in our study to verify how user performs in information search for a general seeking task by using them. Precision ratio, recall ratio, clicks on search and time for search were four variables tested in the experiment. Also, two supplementary tests were respectively carried out to manifest how graphic-based tag clouds contributed to the identification of target tags and tag clusters.FindingsThe experimental results showed that compared to tag content visual tag structure was more important to find related tags from tag clouds for information search. In addition, tag clouds that visually represented the semantic relationships within tags could make user more confident about their search result and carry out a shorter learning process during searching, which signified a tag-based information search path when visual elements were applied.Originality/valueThis research is one of the first to illustrate systematically the graphic-based tag clouds and their impacts on information search. The research findings could suggest on how to build up more effective and interactive tag clouds and make proposition for the design of search user interface by using graphic-based tag clouds.

SHR-Zbtb16 minimal congenic strain reveals nutrigenetic interaction between Zbtb16 and high-sucrose diet.

Both prenatal and postnatal excessive consumption of dietary sucrose or fructose was shown to be detrimental to health and contributing to pathogenesis of metabolic syndrome. Our knowledge of genetic determinants of individual sensitivity to sucrose-driven metabolic effects is limited. In this study, we have tested the hypothesis that a variation of metabolic syndrome-related gene, Zbtb16 (Zinc Finger and BTB Domain Containing 16 will affect the reaction to high-sucrose diet (HSD) content in "matched" nutritional exposition settings, i.e. maternal HSD with re-exposition to HSD in adulthood vs. standard diet. We compared metabolic profiles of adult males of spontaneously hypertensive rats (SHR) and a single-gene, minimal congenic strain SHR-Zbtb16 fed either standard diet or exposed to HSD prenatally throughout gestation and nursing and again at the age of 6 months for the period of 14 days. HSD exposition led to increased adiposity in both strains and decrease of glucose tolerance and cholesterol (Ch) concentrations in majority of low-density lipoprotein (LDL) particle classes and in very large and large high-density lipoprotein (HDL) in SHR-Zbtb16 male offspring. There was a similar pattern of HSD-induced increase of triacylglycerols in chylomicrons and very low-density lipoprotein (VLDL) of both strains, though the increase of (triacylglycerol) TAG content was clearly more pronounced in SHR. We observed significant STRAIN*DIET interactions for the smallest LDL particles as their TAG content decreased in SHR-Zbtb16 and did not change in SHR in response to HSD. In summary, we provide evidence of nutrigenetic interaction between Zbtb16 and HSD in context of pathogenesis of metabolic syndrome.

Comprehensive lipid and metabolite profiling in healthy adults with low and high consumption of fatty fish: a cross-sectional study.

Fish consumption is associated with reduced risk of CVD, which may be partly mediated by alterations in plasma lipids, such as HDL-cholesterol. However, comprehensive analyses of associations between fatty fish consumption and lipoprotein subclass profile are limited and show inconsistent results. Therefore, the aim of the present exploratory study was to investigate the association between fatty fish consumption and lipoprotein subclass particle concentrations and composition, with an emphasis on HDL. We performed a comprehensive plasma metabolite profiling in 517 healthy adults, using a targeted high-throughput NMR spectroscopy platform. The participants were divided into tertiles based on consumption of fatty fish, reported through a validated FFQ. We compared the concentration of metabolites between the participants in the lowest and highest tertiles of fatty fish consumption. We show that high consumers of fatty fish (>223 g/week, median intake 294 g/week) had higher particle concentrations and content of total lipids, free cholesterol and phospholipids in large and extra-large HDL particles and higher content of total cholesterol, cholesteryl esters and TAG in large HDL particles than low consumers (<107 g/week, median intake 58 g/week). Using fatty fish consumption as a continuous variable, we found that fatty fish consumption was associated with lower levels of the inflammation marker glycoprotein acetyls. In conclusion, high consumers of fatty fish seem to have a more favourable HDL-cholesterol-related lipoprotein profile and anti-inflammatory phenotype than low consumers of fatty fish. Thus, these data support the current Norwegian dietary recommendations for fish consumption regarding CVD risk.

2298-PUB: Estradiol Protects Female ApoE KO Mice against Nonalcoholic Steatohepatitis

Lifestyle and increased consumption of high fat diets contribute greatly to the development of obesity, insulin resistance, type 2 diabetes (DM2), and cardiovascular diseases, which are less prevalent in young women than in men of the same age or postmenopausal women. One of the consequences of the Western lifestyle and high fat diet is Nonalcoholic Fatty Liver Disease (NAFLD) and its aggressive form, Nonalcoholic Steatohepatitis (NASH), which can progress to cirrhosis and hepatocellular cancer (HCC) and is rapidly becoming the leading cause for end-stage liver disease or liver transplantation. Therefore, we evaluated if female mice were protected against Western Diet (WD)-induced NASH in apolipoprotein E (ApoE) KO animals. Female ApoE KO mice ovariectomized (OVX) or sham operated (SHAM) were fed a WD for 7 weeks. Whole-body fat was increased by ∼65% (P&amp;lt;0.001) in OVX mice associated with an increase of glucose intolerance of by ∼75% (P&amp;lt;0.01) and increased hepatic triglycerides (TAG) by 130% (P&amp;lt;0.01) compared with SHAM mice. OVX mice also displayed increased plasma ALT (∼130, P&amp;lt;0.001) and AST (∼65%, P&amp;lt;0.05) compared with SHAM mice. These data were associated with increased hepatic inflammation in OVX mice, demonstrated by an increased of F4/80 of by ∼70% (P&amp;lt;0.01). Hepatic fibrosis was also increased in OVX mice by ∼80% (P&amp;lt;0.001) compared with SHAM mice. Estradiol treatment of OVX mice reversed some of these effects, reducing whole-body fat (P&amp;lt;0.01), glucose intolerance (P&amp;lt;0.01), and hepatic TAG content (P&amp;lt;0.001). Taken together, these data support the hypothesis that estradiol protects OVX mice from WD-induced NASH and glucose intolerance, by protecting female mice against hepatic steatosis, inflammation and fibrosis. Disclosure G.V. Moreira: None. S.L. Matos: None. F.N. Camargo: None. L. Araujo: None. G.M. Murata: None. C.R.O. Carvalho: None. J. Camporez: None. Funding Fundação de Amparo a Pesquisa do Estado de São Paulo (2018/04956-5)

Mouse Model of Non-Alcoholic Fatty Liver Disease That Replicates the Human Disease

Abstract Objectives To develop a mouse model of non-alcoholic fatty liver disease (NAFLD) that replicates the characteristic of the disease in humans, including development of obesity, dysbiosis, fibrosis and liver tumors. Methods Agouti yellow (Ay) mice, which have hyperphagia, were fed a Western diet (WD) (42% kcal from fat, 341 g/Kg sucrose, and 0.2% cholesterol) and a drinking solution containing fructose and glucose equivalent to 42 g/L of high fructose corn syrup (HFCS). Wild-type mice fed a low-fat diet (LFD) (10% kcal from fat) served as lean controls. After 16 weeks of diet treatment, tissues were collected and analyzed. Obesity was evaluated via body weight and body composition. NAFLD was evaluated by histology and confirmed by liver lipid quantification, plasma ALT and AST levels, expression of inflammation-related genes, and fibrosis-specific staining. Gene expression profile was evaluated by RNAseq. Gut microbiota dysbiosis was evaluated by 16S rRNA metagenomics. Results Ay mice fed the Western diet and HFCS for 16 weeks developed obesity and NAFLD. Histological evaluation determined that the mice developed non-alcoholic steatohepatitis (NASH) with steatosis, liver injury, inflammation, and fibrosis. This was confirmed by the following analyses: In these mice, steatosis, quantified by liver TAG content, increased 4X when compared to lean controls. Liver injury, assessed by measuring plasma ALT and AST, increased 11X and 4X, respectively. Inflammation, evaluated by liver mRNA expression of Tnf and CCl2, increased 6X and 12X, respectively. Fibrosis, quantified morphometrically, increased 2.5X. Gene expression profiling showed increases in inflammation- and fibrosis-related pathways, similar to NASH in humans. In addition, these mice developed gut microbiota dysbiosis, with increased Bacteroidetes and Proteobacteria and decreased Firmicutes, as reported in NASH in humans. Finally, 60% of the Ay mice developed liver tumors when fed the WD + HFCS diet for one year. Conclusions Ay mice fed a Western diet and HFCS developed obesity, gut microbiota dysbiosis, and NASH, including fibrosis and tumors, replicating the characteristics of NASH in humans. We present this as a new model for studying NASH physiopathology and testing new therapies. Funding Sources NIH CUNY.

Evaluation of diurnal responses of Tetradesmus obliquus under nitrogen limitation

Tetradesmus obliquus is an oleaginous microalga with high potential for triacylglycerol production. We characterized the biochemical composition and the transcriptional landscape of T. obliquus wild-type and the starchless mutant (slm1), adapted to 16:8 h light dark (LD) cycles under nitrogen limitation. In comparison to the nitrogen replete conditions, the diurnal RNA samples from both strains also displayed a cyclic pattern, but with much less variation which could be related to a reduced transcription activity in at least the usually highly active processes. During nitrogen limitation, the wild-type continued to use starch as the preferred storage compound to store energy and carbon. Starch was accumulated to an average content of 0.25 g·gDW−1, which is higher than the maximum observed under nitrogen replete conditions. Small oscillations were observed, indicating that starch was being used as a diurnal energy storage compound, but to a lesser extent than under nitrogen replete conditions. For the slm1 mutant, TAG content was higher than for the wild-type (average steady state value was 0.26 g·gDW−1 for slm1 compared to 0.06 g·gDW−1 for the wild-type). Despite the higher TAG content in the slm1, the conversion efficiency of photons into biomass components for the slm1 was only half of the one obtained for the wild-type. This is related to the observed decrease in biomass productivity (from 1.29 gDW·L−1·day−1 for the wild-type to 0.52 gDW·L−1·day−1 for the slm1). While the transcriptome of slm1 displayed clear signs of energy generation by degrading TAG and amino-acids during the dark period, no significant variation of these metabolites could be measured. When looking through the diurnal cycle, the photosynthetic efficiency was lower for the slm1 mutant compared to the wild-type especially during the second half of the light period, where starch accumulation occurred in the wild-type.

Spruce Galactoglucomannan-Stabilized Emulsions Enhance Bioaccessibility of Bioactive Compounds

The increasing public awareness of health and sustainability has prompted the development of functional foods rich in health-promoting ingredients. Processing technologies and sustainable multifunctional ingredients are needed for structuring these formulations. Spruce galactoglucomannan (GGM), the main hemicelluloses in softwood cell walls, are an abundantly available, emerging sustainable food hydrocolloid that have the ability to efficiently emulsify and stabilize oil-in-water emulsions. In this study, we illustrate how this lignocellulosic stabilizer affects the digestion of polyunsaturated fatty acids (PUFAs) in vitro. A 100% decrease in the initial TAG content was observed during the in vitro digestion, suggesting that complete hydrolysis of the TAGs was achieved by the digestive enzymes. Besides, no release of mono-, di-, and oligosaccharides or phenolic compounds from GGM was detected. Our results demonstrate that the GGM-stabilized emulsion could potentially deliver lipophilic bioactive ingredients and enhance their bioaccessibility. In addition, this bio-stabilizer itself would remain stable in the upper gastrointestinal track and serve as a prebiotic for gut microbiota. We anticipate GGM to complement or even replace many of the conventional carriers of bioactive components in future health care products and functional foods.

CRISPR/Cas9 Based Knockout of miR-145 Affects Intracellular Fatty Acid Metabolism by Targeting INSIG1 in Goat Mammary Epithelial Cells.

MiR-145 modulates fatty acid metabolism by regulating the expression of fatty acid metabolism-related genes in goat mammary epithelial cells. Previous studies using RNAi methods have clarified the function of miR-145 in lipogenesis. However, there are limiting factors such as short-term and inconsistent inhibition efficiency in RNAi method. On the basis of previous miR-145 functional studies, this study aims to knock out miR-145 and validate the function using CRISPR/Cas9 technology. We successfully obtained the single cell clone which had single nucleotide deletion around the Drosha processing site. The expression of miR-145 was significantly decreased, and the mRNA and protein expression of target gene INSIG1 were both increased by RT-qPCR and Western blot. The expression of fatty acid metabolism-associated gene (DGAT1, AGPAT6, TIP47, ADFP, CD36, ACSL1, ATGL, ACOX, CPT1A, FADS2, ELOVL5, PPARA, SCD1, FASN, and ACACA) were decreased. The contents of triacylglycerol and cholesterol were significantly inhibited. The percentage of C17:0 and C18:0 saturated fatty acid increased. Taken together, these data suggested that knockout of miR-145 could inhibit TAG and cholesterol contents and affect fatty acid composition through regulating the expression of fatty acid metabolism-related genes. These findings provide a sufficient theoretical basis for improving goat milk quality by miR-145.

Application of N-carbamylglutamate in Rex rabbits to reduce body fat deposition and its possible mechanism.

N-Carbamylglutamate (NCG) has been shown to enhance arginine synthesis and improve growth performance in animals. However, the effect of NCG on body fat deposition remains unknown. This study examined the effects of NCG on body fat deposition and evaluated the potential mechanisms involved. Rex rabbits (3 months old) were assigned to one of four dietary groups and supplemented with NCG at the following different concentrations in a feeding trial that lasted 67 d: 0 (control), 0·04, 0·08, and 0·12 %. NCG supplementation increased serum concentrations of arginine and proline by activating intestinal carbamoylphosphate synthase-І at the posttranscriptional level. Final body weights and growth performance were not affected by dietary NCG levels. However, NCG-treated rabbits had lower perirenal and subcutaneous fat percentages, serum TAG content, and hepatic fatty acid synthase (FAS) activity and increased NO synthase activity and serum levels of NO, growth hormone (GH), and insulin-like growth factor 1 (IGF-1). There were significant positive correlations between TAG content and perirenal fat percentage, as well as FAS activity and perirenal fat percentage, but significant negative correlations between TAG and NO levels, and FAS activity and IGF-1 level in rabbits after NCG treatment. NCG supplementation did not affect hepatic health indicators, except for serum ammonia concentrations, which were decreased in NCG-treated rabbits. Our results suggest that NCG can serve as a dietary supplement to reduce unfavourable fat deposition through inhibiting hepatic lipogenesis in animals since it appears to have no negative effects on growth performance or hepatic health.

Modeling biomass production during progressive nitrogen starvation by North Swedish green microalgae

Five newly isolated green algal species from Northern Sweden and one culture collection strain were tested for their ability to remove nitrogen and accumulate carbohydrates and neutral lipids (TAGs) under progressive nitrogen starvation. All six microalgal species increased biomass during N starvation, the amount of proteins decreased, and species dependent either TAGs and/or carbohydrates accumulated. Biomass of the algal strains Coelastrella sp. 3-4, Scenedesmus sp. B2-2 and S. obliquus RISE (UTEX 417) had very low final TAG content (≤3.4%) and high carbohydrate content (>41%) at the end of the starvation period. C. astroideum RW10 accumulated 9.2% TAGs and 53.9% carbohydrates during N-starvation; due to its modest growth rate (1.60 g/L and 1.06 1/day) resulting in low final biomass concentration, its cumulativeTAG and carbohydrate productivity were poor (175 mgTAG/system and 1.03 gCARBS/system). C. vulgaris 13-1 preferentially accumulated TAGs (10.3%) over carbohydrates (35%), with low minimal and maximal N quotas (2.27 and 11.6 mM/gDW) in its biomass and a very high growth rate (1.86 1/day) and cumulative TAGs productivity (278 mgTAG/system). Desmodesmus sp. RUC2 had the highest final biomass concentration (3.48 g/L) as well as cumulative TAG and carbohydrate productivity (269 mgTAG/system and 1.79 gCARBS/system). This species had the lowest minimal and maximal N quotas (1.58 and 8.50 mM/gDW) of all tested species, it can produce high amounts of biomass even when the available nitrogen concentration is low.A Droop's mathematical model with four basic parameters was applied to interpret the experimental data on N assimilation and biomass production under N starvation. The model corresponded well to the experimental data and therefore can successfully be applied to predict biomass production and N assimilation in Nordic algal species.

Separation and purification of triglyceride from nyamplung (Calophyllum inophyllum) seed oil as biodiesel feedstock by using continuous countercurrent extraction

Calophyllum inophyllum or commonly called nyamplung in Indonesia has the potential to be used as a biodiesel feedstock due to its high oil content. The purpose of this study was to determine the effect of feed flowrate and solvent to oil mass ratio on triglycerides (TAG) and free fatty acid (FFA) contents in non-polar lipid fraction (NPLF) of C. inophyllum seed oil by using continuous counter-current extraction. The contents of TAG and FFA in NPLF of C. inophyllum seed oil each sample points in continuous countercurrent extraction equipment. It was expected that the TAG became purer and suitable for biodiesel purpose. Various factors applied in this study were the effect of solvent to oil mass ratio and the effect of n-hexane + oil to methanol feed flowrate in liter per hour. Crude C. inophyllum seed oil contained 63.91% TAG, 15.76% FFA, 12.25% monoglycerides (MAG), dan 4.66% diglycerides (DAG). Separation and purification of TAG were carried out by using a packed column with the principle of countercurrent flow using n-hexane and methanol technical grade as solvents. The product of extraction analyzed TAG content qualitatively by using TLC and quantitatively using HT-GC, while NPLF from each sample points were analyzed using HT-GC. The optimum results were obtained by using a solvent to oil mass ratio of 6 (w:w) and (n-hexane + oil) to methanol feed flowrate of 6:15 (L/h:L/h) with 92.85% content of TAG, 2.19% content of FFA and 74.79% yield of NPLF.

Silencing of RpATG8 impairs the biogenesis of maternal autophagosomes in vitellogenic oocytes, but does not interrupt follicular atresia in the insect vector Rhodnius prolixus.

Follicular atresia is the mechanism by which the oocyte contents are degraded during oogenesis in response to stress conditions, allowing the energetic resources stored in the developing oocytes to be reallocated to optimize female fitness. Autophagy is a conserved intracellular degradation pathway where double-membrane vesicles are formed around target organelles leading to their degradation after lysosome fusion. The autophagy-related protein 8 (ATG8) is conjugated to the autophagic membrane and has a key role in the elongation and closure of the autophagosome. Here we identified one single isoform of ATG8 in the genome of the insect vector of Chagas Disease Rhodnius prolixus (RpATG8) and found that it is highly expressed in the ovary during vitellogenesis. Accordingly, autophagosomes were detected in the vitellogenic oocytes, as seen by immunoblotting and electron microscopy. To test if autophagosomes were important for follicular atresia, we silenced RpATG8 and elicited atresia in vitellogenic females by Zymosan-A injections. We found that silenced females were still able to trigger the same levels of follicle atresia, and that their atretic oocytes presented a characteristic morphology, with accumulated brown aggregates. Regardless of the difference in morphology, RpATG8-silenced atretic oocytes presented the same levels of protein, TAG and PolyP, as detected in control atretic oocytes, as well as the same levels of acidification of the yolk organelles. Because follicular atresia has the ultimate goal of restoring female fitness, we tested if RpATG8-silenced atresia would result in female physiology and behavior changes. Under insectarium conditions, we found that atresia-induced control and RpATG8-silenced females present no changes in blood meal digestion, survival, oviposition, TAG content in the fat body, haemolymph amino acid levels and overall locomotor activity. Altogether, we found that autophagosomes are formed during oogenesis and that the silencing of RpATG8 impairs autophagosome biogenesis in the oocytes. Nevertheless, regarding major macromolecule degradation and adaptations to the fitness costs imposed by triggering an immune response, we found that autophagic organelles are not essential for follicle atresia in R. prolixus.

Physicochemical and biochemical characterizations of some Tunisian seed oils

Four Tunisian vegetable oils extracted from seeds (Nigella sativa,Opuntia ficus indica,Pistacia lentiscusandHibiscus sabdariffa) have been characterized in this study. The following parameters were determined: acidity, peroxide value, saponification value, specific extinction coefficients K232, K270, chlorophylls and carotenoids content. The triglyceride and tocopherol compositions of the oils were determined using reversed phase high performance liquid chromatography with diode array detection (HPLC-DAD) and the fatty acids (FA) and phytosterol compositions were determined using a gas chromatography (GC) with a flame ionization detector (FID). Polyunsaturated fatty acids were predominant in all tested samples except inPistacia lentiscusoil where monounsaturated fatty acids were predominant. Major FA were linoleic and oleic acids. β-sitosterol was the most abundant phytosterol. All samples had high content of TAGs with an equivalent carbon number of 44, 46 and 48.Nigella sativaoil had the highest content of tocopherols.

Unraveling enhanced membrane lipid biosynthesis in Chlamydomonas reinhardtii starchless mutant sta6 by using an electrospray ionization mass spectrometry-based lipidomics method

The unicellular green alga Chlamydomonas reinhardtii, a well-established model organism, has been widely used in dissecting glycerolipid metabolism in oxygenating photosynthetic organisms. In previous studies, it has been found that shunting carbon precursors from the starch synthesis pathway can lead to a 10-fold increase in TAG content as compared to the wild type, but it is unknown whether inactivation of AGPase may affect membrane lipids biosynthesis. The study aims to investigate global changes in lipid metabolism and homeostasis in the starchless mutant C. reinhardtii sta6. By utilizing an electrospray ionization/mass spectrometry (ESI/MS)-based lipidomics approach, a total of 105 membrane lipid molecules of C. reinhardtii were resolved, including 16 monogalactosyldiacylglycerol (MGDG), 16 digalactosyldiacylglycerol (DGDG), 11 phosphatidylglycerol (PG), 6 sulfoquinovosyldiacylglycerol (SQDG), 49 diacylglyceryl-N,N,N-trimethylhomoserine (DGTS), 2 phosphatidylethanolamine (PE), and 5 phosphatidylinositol (PI) molecules. The quantitative results indicated that the membrane lipid profiles were similar between the two C. reinhardtii strains grown under both low- and high-light conditions, but the cellular contents of a great number of lipids were altered in sta6 due to the defect in starch biosynthesis. Under low-light conditions, sta6 accumulated more PI, MGDG, DGDG but less amounts of DGTS as compared to WT. Under high light, sta6 cells contained higher content membrane lipids than cc-124, except for PG, which is more or less similar in both strains. Our results demonstrate that the cellular membrane lipid homeostasis underwent profound changes in the starchless mutant, and thereby its physiological impact remains to be explored.

Study of UV-B Mutation Effect on pH Resistance and Lipid Production of Microalgae Botryococcus braunii

Microalgae Botryococcus braunii is a potential biodiesel producer as an alternative for fossil fuels due to its high lipid content. UV-B mutations were carried out to see the effect in microalgae growth at various pHs (3-8). Reduction of nitrogen levels was carried out to see the effect on the growth and lipid production of microalgae. UV-B mutation increased the ability of growth and resistance of B. braunii against low pH. Under low nitrogen conditions, the growth of B. braunii cells would not continue for a longer time. B. braunii which grow in nitrogen depletion medium produced lipid content greater than normal nitrogen. UV-B light mutation also increased the lipid content of B. braunii. At 7 days of incubation, the mutation not only increased lipid content, but also significantly increased the TAG content of B. braunii lipids.