Synthesis of PEGylated amphiphilic block copolymers with pendant linoleic moieties by combining ring-opening polymerization and click chemistry.

This study focused on synthesizing and characterizing PEGylated amphiphilic block copolymers with pendant linoleic acid (Lin) moieties as an alternative to enhance their potential in drug delivery applications. The synthesis involved a two-step process, starting with ring-opening polymerization of ε-caprolactone (CL) and propargylated cyclic carbonate (MCP) to obtain PEG-b-P(CL-co-MCP) copolymers, which were subsequently modified via click chemistry. Various reaction conditions were explored to improve the yield and efficiency of the click chemistry step. The use of anisole as a solvent, N-(3-azidopropyl)linoleamide as a substrate, and a reaction temperature of 60°C proved to be highly efficient, achieving nearly 100% conversion at a low catalyst concentration. The resulting copolymers exhibited controlled molecular weights and low polydispersity, confirming the successful synthesis. Furthermore, click chemistry allows for the attachment of Lin moieties to the copolymer, enhancing its hydrophobic character, as deduced from their significantly lower critical micelle concentration than that of traditional PEG-b-PCL systems, which is indicative of enhanced stability against dilution. The modified copolymers exhibited improved thermal stability, making them suitable for applications that require high processing temperatures. Dynamic light scattering and transmission electron microscopy confirmed the formation of micellar structures with sizes below 100 nm and minimal aggregate formation. Additionally, 1H NMR spectroscopy in deuterated water revealed the presence of core-shell micelles, which provided higher kinetic stability against dilution.

Young honeybees show learned preferences after experiencing adulterated pollen.

Pollen selection affects honeybee colony development and productivity. Considering that pollen is consumed by young in-hive bees, and not by foragers, we hypothesized that young bees learn pollen cues and adjust their preferences to the most suitable pollens. To assess whether young bees show preferences based on learning for highly or poorly suitable pollens, we measured consumption preferences for two pure monofloral pollens after the bees had experienced one of them adulterated with a deterrent (amygdalin or quinine) or a phagostimulant (linoleic acid). Preferences were obtained from nurse-aged bees confined in cages and from nurse bees in open colonies. Furthermore, we tested the bees' orientation in a Y-maze using a neutral odour (Linalool or Nonanal) that had been previously associated with an amygdalin-adulterated pollen. Consumption preferences of bees, both in cages and in colonies, were reduced for pollens that had been adulterated with deterrents and increased for pollens that had been supplemented with linoleic acid. In the Y-maze, individuals consistently avoided the odours that they had previously experienced paired with the deterrent-adulterated pollen. Results show that nurse-aged bees associate pollen-based or pollen-related cues with either a distasteful/malaise experience or a tasty/nutritious event, leading to memories that bias their pollen-mediated response.

Green Extraction by Ultrasound, Microencapsulation by Spray Drying and Antioxidant Activity of the Tucuma Coproduct (Astrocaryum vulgare Mart.) Almonds.

The industrial processing amazon fruits, like tucuma, generates a large amount of coproducts with large nutritional potential. Thus, this work obtained the oily extract of the tucuma almonds coproducts by green extraction using palm oil by the ultrasound method and then microencapsulated by atomization and verification of its antioxidant activity. Thermogravimetric techniques, infrared spectroscopy, scanning electron microscopy, moisture content, water activity were applied to characterize the microparticles. Total carotenoids were determined by UV spectroscopy and antioxidant activity was measured by 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid and co-oxidation in the system ß-carotene/linoleic acid. The oily extract and microparticle had total carotenoid contents of 3.305 mg/100 g ± 0.01 and 2.559 mg/100 g ± 0.01, respectively. The antioxidant activity assessed through the 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid value was 584.75 µM/trolox ± 0.01 (oily extract) and 537.12 µM/trolox ± 0.01 (microparticle) were determined. In the system ß-carotene/linoleic acid showed oxidation of 49.9% ± 1.8 lipophilic extract and 43.3% ± 2.3 microparticle. The results showed that the oily extract of the tucuma almond coproduct can be used as a carotenoid-rich source and microencapsuled with possible application for functional foods production.

Structural Studies of the Cutin from Two Apple Varieties: Golden Delicious and Red Delicious (Malus domestica).

The cuticle, a protective cuticular barrier present in almost all primary aerial plant organs, has a composition that varies between plant species. As a part of the apple peel, cuticle and epicuticular waxes have an important role in the skin appearance and quality characteristic in fresh fruits destined for human consumption. The specific composition and structural characteristics of cutin from two apple varieties, "golden delicious" and "red delicious", were obtained by enzymatic protocols and studied by means of cross polarization magic angle spinning nuclear magnetic resonance (CP-MAS 13C NMR), attenuated total reflection infrared spectroscopy (ATR-FTIR), and mass spectrometry, and were morphologically characterized by specialized microscopy techniques (atomic force microscopy (AFM), confocal laser scanning microscopy (CLMS), and scanning electron microscopy (SEM)). According to CP-MAS 13C NMR and ATR-FTIR analysis, cutins from both varieties are mainly composed of aliphatics and a small difference is shown between them. This was corroborated from the hydrolyzed cutins analysis by mass spectrometry, where 9,10,18-trihydroxy-octadecanoic acid; 10,20-Dihydroxy-icosanoic acid; 10,16-dihydroxy hexadecenoic acid (10,16-DHPA); 9,10-epoxy-12-octadecenoic acid; and 9,10-epoxy-18-hydroxy-12-octadecenoic acid were the main monomers isolated. The low presence of polysaccharides and phenolics in the cutins obtained could be related to the low elastic behavior of this biocomposite and the presence of cracks in the apple cutin's surface. These cracks have an average depth of 1.57 µm ± 0.57 in the golden apple, and 1.77 µm ± 0.64 in those found in the red apple. The results obtained in this work may facilitate a better understanding that mechanical properties of the apple fruit skin are mainly related to the specific aliphatic composition of cutin and help to much better investigate the formation of microcracks, an important symptom of russet formation.

Characterization of a new GmFAD3A allele in Brazilian CS303TNKCA soybean cultivar.

KEY MESSAGE: We molecularly characterized a new mutation in the GmFAD3A gene associated with low linolenic content in the Brazilian soybean cultivar CS303TNKCA and developed a molecular marker to select this mutation. Soybean is one of the most important crops cultivated worldwide. Soybean oil has 13% palmitic acid, 4% stearic acid, 20% oleic acid, 55% linoleic acid and 8% linolenic acid. Breeding programs are developing varieties with high oleic and low polyunsaturated fatty acids (linoleic and linolenic) to improve the oil oxidative stability and make the varieties more attractive for the soy industry. The main goal of this study was to characterize the low linoleic acid trait in CS303TNKCA cultivar. We sequenced CS303TNKCA GmFAD3A, GmFAD3B and GmFAD3C genes and identified an adenine point deletion in the GmFAD3A exon 5 (delA). This alteration creates a premature stop codon, leading to a truncated protein with just 207 residues that result in a non-functional enzyme. Analysis of enzymatic activity by heterologous expression in yeast support delA as the cause of low linolenic acid content in CS303TNKCA. Thus, we developed a TaqMan genotyping assay to associate delA with low linolenic acid content in segregating populations. Lines homozygous for delA had a linolenic acid content of 3.3 to 4.4%, and the variation at this locus accounted for 50.83 to 73.70% of the phenotypic variation. This molecular marker is a new tool to introgress the low linolenic acid trait into elite soybean cultivars and can be used to combine with high oleic trait markers to produce soybean with enhanced economic value. The advantage of using CS303TNKCA compared to other lines available in the literature is that this cultivar has good agronomic characteristics and is adapted to Brazilian conditions.

Effect of Supercritical Carbon Dioxide Extraction Parameters on the Biological Activities and Metabolites Present in Extracts from Arthrospira platensis.

Arthrospira platensis was used to obtain functional extracts through supercritical carbon dioxide extraction (SFE-CO2). Pressure (P), temperature (T), co-solvent (CX), static extraction (SX), dispersant (Di) and dynamic extraction (DX) were evaluated as process parameters through a Plackett-Burman design. The maximum extract yield obtained was 7.48 ± 0.15% w/w. The maximum contents of bioactive metabolites in extracts were 0.69 ± 0.09 µg/g of riboflavin, 5.49 ± 0.10 µg/g of α-tocopherol, 524.46 ± 0.10 µg/g of ß-carotene, 1.44 ± 0.10 µg/g of lutein and 32.11 ± 0.12 mg/g of fatty acids with 39.38% of palmitic acid, 20.63% of linoleic acid and 30.27% of γ-linolenic acid. A. platensis extracts had an antioxidant activity of 76.47 ± 0.71 µg GAE/g by Folin-Ciocalteu assay, 0.52 ± 0.02, 0.40 ± 0.01 and 1.47 ± 0.02 µmol TE/g by DPPH, FRAP and TEAC assays, respectively. These extracts showed antimicrobial activity against Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922 and Candida albicans ATCC 10231. Overall, co-solvent was the most significant factor for all measured effects (p < 0.05). Arthrospira platensis represents a sustainable source of bioactive compounds through SFE using the following extraction parameters P: 450 bar, CX: 11 g/min, SX: 15 min, DX: 25 min, T: 60 °C and Di: 35 g.

A Fatty Acids Mixture Reduces Anxiety-Like Behaviors in Infant Rats Mediated by GABAA Receptors.

Fatty acids (C6-C18) found in human amniotic fluid, colostrum, and maternal milk reduce behavioral indicators of experimental anxiety in adult Wistar rats. Unknown, however, is whether the anxiolytic-like effects of fatty acids provide a natural mechanism against anxiety in young offspring. The present study assessed the anxiolytic-like effect of a mixture of lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, elaidic acid, and linoleic acid in Wistar rats on postnatal day 28. Infant rats were subjected to the elevated plus maze, defensive burying test, and locomotor activity test. Diazepam was used as a reference anxiolytic drug. A group that was pretreated with picrotoxin was used to explore the participation of γ-aminobutyric acid-A (GABAA) receptors in the anxiolytic-like effects. Similar to diazepam, the fatty acid mixture significantly increased the frequency of entries into and time spent on the open arms of the elevated plus maze and decreased burying behavior in the defensive burying test, without producing significant changes in spontaneous locomotor activity. These anxiolytic-like effects were blocked by picrotoxin. Results suggest that these fatty acids that are contained in maternal fluid may reduce anxiety-like behavior by modulating GABAergic neurotransmission in infant 28-day-old rats.

Complexes between ovalbumin nanoparticles and linoleic acid: Stoichiometric, kinetic and thermodynamic aspects.

Stoichiometric, kinetic and thermodynamic aspects of complex formation between heat-induced aggregates of ovalbumin (ovalbumin nanoparticles, OVAn) and linoleic acid (LA) were evaluated. Extrinsic fluorescence data were fitted to modified Scatchard model yielding the following results: n: 49±2 LA molecules bound per OVA monomer unit and Ka: 9.80±2.53×10(5)M. Kinetic and thermodynamic properties were analyzed by turbidity measurements at different LA/OVA monomer molar ratios (21.5-172) and temperatures (20-40°C). An adsorption approach was used and a pseudo-second-order kinetics was found for LA-OVAn complex formation. This adsorption process took place within 1h. Thermodynamic parameters indicated that LA adsorption on OVAn was a spontaneous, endothermic and entropically-driven process, highlighting the hydrophobic nature of the LA and OVAn interaction. Finally, Atomic Force Microscopy imaging revealed that both OVAn and LA-OVAn complexes have a roughly rounded form with size lower than 100nm.

Cholesterol induces surface localization of polyphenols in model membranes thus enhancing vesicle stability against lysozyme, but reduces protection of distant double bonds from reactive-oxygen species.

The main scope of the present study was to analyze the membrane interaction of members of different classes of polyphenols, i.e. resveratrol, naringenin, epigallocatechin gallate and enterodiol, in model systems of different compositions and phase states. In addition, the possible association between membrane affinity and membrane protection against both lipid oxidation and bilayer-disruptive compounds was studied. Gibbs monolayer experiments indicated that even though polyphenols showed poor surface activity, it readily interacted with lipid films. Actually, a preferential interaction with expanded monolayers was observed, while condensed and cholesterol-containing monolayers decreased the affinity of these phenolic compounds. On the other hand, fluorescence anisotropy studies showed that polyphenols were able to modulate membrane order degree, but again this effect was dependent on the cholesterol concentration and membrane phase state. In fact, cholesterol induced a surface rather than deep into the hydrophobic core localization of phenolic compounds in the membranes. In general, the polyphenolic molecules tested had a better antioxidant activity when they were allowed to get inserted into the bilayers, i.e. in cholesterol-free membranes. On the other hand, a membrane-protective effect against bilayer permeabilizing activity of lysozyme, particularly in the presence of cholesterol, could be assessed. It can be hypothesized that phenolic compounds may protect membrane integrity by loosely covering the surface of lipid vesicles, once cholesterol push them off from the membrane hydrophobic core. However, this cholesterol-driven distribution may lead to a reduced antioxidant activity of linoleic acid double bonds.

Antioxidant activity of the aqueous and methanolic extracts of coffee beans (Coffea arabica L.).

BACKGROUND: Coffee is important source of natural antioxidants in the diet, such as phenolic compounds, alkaloids, mainly caffeine, diterpenes (cafestol and kahweol) and Maillard reaction products formed during roasting. METHODS: In aqueous and methanolic extracts of coffee (Coffea arabica L.) roasted using traditional techniques from Brazil (B), Colombia (C), Ethiopia (E), Kenya (K) and coffee roasted using an industrial technique from Brazil (T), the phenolic and caffeine content as well as antioxidant properties were determined. RESULTS: Comparing the results from water and methanolic extracts it should be noted that the highest amount of phenolics was determined for a methanolic extract of coffee roasted using the industrial technique (650.96 mg GAE/g DW) and a water extract of Kenya coffee (461.63 mg GAE/g DW). Caffeine content was on average two times higher in all methanolic extracts than in water extracts. The radical scavenging activity of aqueous extracts was found to be higher than methanolic extracts. The highest antioxidant scavenging activity was determined for C (EC50 = 1.16 mg DW/ml) and E (EC50 = 1.3 mg DW/ml) water extracts. Compared to water extracts methanolic extracts showed significantly higher reducing power, ability to chelate Fe2+, inhibition of linoleic acid peroxidation and inhibition of lipoxygenase. CONCLUSIONS: This study demonstrated that the methanolic extracts obtained from different types of coffee exhibit potential anti-inflammatory and antioxidant properties. The highest antioxidant activity was shown by traditionally roasted coffees from Colombia and Ethiopia.

Linoleic acid binding properties of ovalbumin nanoparticles.

In the present work, ovalbumin (OVA) solutions (10 g/L, 50 mM NaCl, pH 7.5) were heat-treated at 75, 80 and 85°C (namely, OVA-75, OVA-80 and OVA-85, respectively), from 0 to 25 min. OVA nanoparticles (OVAn) around 100 nm were obtained. For 3 min of heat treatment, OVAn sizes increased with temperature, but for a heating time longer than 10 min, OVA-75 showed the highest size values. OVAn surface hydrophobicity increased 6-8 folds in comparison with native OVA and wavelength blue shifts of 25-30 nm in maximum fluorescence intensity were registered. These results suggest that buried hydrophobic residues were exposed to the aqueous medium. Binding experiments with linoleic acid (LA) as polyunsaturated fatty acid (PUFA) model were carried out. Firstly, binding ability of OVAn was determined from LA titration curves of intrinsic fluorescence measurements. OVA-85 at 5 min presented the highest binding ability and it was used for further binding properties studies (turbidity, particle size distribution--PSD--analysis and ζ-potential measurements). Turbidity measurement and PSD analysis showed that OVAn-LA nanocomplexes were formed, avoiding LA supramolecular self-assembly formation. The union of LA to OVAn surface confers them significant lower ζ-potential and larger size. Hence, fluorescence and ζ-potential results showed that LA would bind to OVAn by mean of hydrophobic interactions. Information derived from this work could be important to potentially use OVAn as PUFA vehiculization with applications in several industrial sectors (food, pharmaceutical, cosmetics, etc.).

Antioxidant and rheological properties of guava jam with added concentrated grape juice.

BACKGROUND: Guava jam enriched by the addition of concentrated grape juice in the proportion of 30% (w/w) (ENR) was studied for its antioxidant potential and rheological behavior. Total phenolics content, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH(•)) scavenging activity and ß-carotene-linoleic acid coupled oxidation assays were used to evaluate the antioxidant properties of the samples during storage for 90 days at room temperature (25°C). RESULTS: The ENR showed a total phenolic content of 11.09 g GAE kg(-1) of jam at production, more than two-fold the phenolic content presented by the standard formulation (STA). For the ENR formulation the antioxidant capacity increased in almost 20% (P ≤ 0.05) with time on the ß-carotene assay and decreased with time on the DPPH assay. The enriched guava jam exhibited a non-Newtonian shear-thinning behavior at temperatures ranging from 25 to 55°C, and, moreover, presented higher stability than the standard formulation when exposed to temperature variation. CONCLUSION: ENR constitutes an original food product with a notable antioxidant potential, even greater than the potential presented by the standard guava jam, worldwide appreciated delicacy.

Effect of limited enzymatic hydrolysis on linoleic acid binding properties of ß-lactoglobulin.

ß-Lactoglobulin (BLG) is a member of lipocalin family, proteins with ability to bind small hydrophobic ligands, such as retinol, vitamins and fatty acids. Moreover, BLG is susceptible to protease action producing a wide range of polypeptides depending on the hydrolysis degree (HD). In the present work, the effect of limited enzymatic hydrolysis on fatty acid binding properties of BLG was studied. Linoleic acid (LA) was used as a model fatty acid. Limited enzymatic hydrolysis was performed using α-chymotrypsin immobilised on agarose microparticles. BLG hydrolysates were produced at HD: 1%, 3% and 5%. In order to determine the influence of HD on BLG molecular weight SDS-PAGE was used. BLG structural modification and LA binding properties were monitored by means of fluorescence spectroscopic techniques. The increase in HD produced: (i) a BLG degradation and a molecular weight distribution of BLG hydrolysates and (ii) an increased exposition of buried hydrophobic residues, however it was observed a decrease in surface hydrophobicity possibly due to a deterioration of hydrophobic protein domains. It was observed that enzymatic hydrolysis treatment produced a decrease in BLG ability for binding LA. It was concluded that limited enzymatic hydrolysis could deteriorate the specific site on BLG structure necessary for binding LA.

A thin-layer chromatography autographic method for the detection of inhibitors of the Salmonella PhoP-PhoQ regulatory system.

INTRODUCTION: The PhoP-PhoQ system from Salmonella enterica serovar Typhimurium controls the expression of factors that are critical for the bacterial entry into host cells and the bacterial intramacrophage survival. Therefore it constitutes an interesting target to search for compounds that would control Salmonella virulence. Localisation of such compounds in complex matrixes could be facilitated by thin-layer chromatography (TLC) bioautography. OBJECTIVE: To develop a TLC bioautography to detect inhibitors of the PhoP-PhoQ regulatory system in complex matrixes. METHODS: The TLC plates were covered by a staining solution containing agar, Luria-Bertani medium, 5-bromo-4-chloro-3-indolyl-ß-D-galactopyranoside (X-gal), kanamycin and a S. typhimurium strain that harbours a reporter transcriptional lacZ-fusion to an archetypal PhoP-activated gene virK. After solidification, the plate was incubated at 37°C for 16 h. RESULTS: A bioautographic assay suitable for the localisation of inhibitors of the PhoP-PhoQ system activity in S. enterica serovar Typhimurium present in a complex matrix is described. The assay was used to analyse a series of hydrolysed extracts prepared by alkaline treatment of crude plant extracts. Bioassay-guided analysis of the fractions by NMR spectroscopy and MS led to the identification of linolenic and linoleic acids as inhibitory input signals of the PhoP-PhoQ system. CONCLUSION: A practical tool is introduced that facilitates detection of inhibitors of the Salmonella PhoP-PhoQ regulatory system. The assay convenience is illustrated with the identification of the first naturally occurring organic compounds that down-regulate a PhoP-PhoQ regulatory system from a hydrolysed extract.

Linoleic acid induces opening of connexin26 hemichannels through a PI3K/Akt/Ca(2+)-dependent pathway.

Connexin hemichannel (Cx HC) opening is involved in physiological and pathological processes, allowing the cellular release of autocrine/paracrine signaling molecules. Linoleic acid (LA) is known to modulate the functional state of connexin46 (Cx46) HCs. However, the molecular mechanism involved in this effect, or whether LA affects HCs constituted of other connexins, remains unknown. Here, we report the effects of LA on HCs in HeLa cells that express Cx26, one of the main Cxs in the cochlear sensory epithelium. Cx26 HC activity (dye uptake) was increased in a concentration-dependent manner by bath application of LA and inhibited by HC blockers. Moreover, intracellular BAPTA, a Ca(2+) chelator, and PI3K/AKT inhibitors were found to reduce the LA-induced Cx26 HC opening, suggesting that the LA effect is mediated by an increase of free intracellular Ca(2+) concentration and activation of the PI3K/Akt-dependent pathway. The LA-induced increase in free intracellular Ca(2+) concentration was mainly due to Ca(2+) influx through Cx26 HCs. In addition, the involvement of SH groups was ruled out, because dithiothreitol (DTT) did not block the LA-induced dye uptake. LA also increased the membrane current mediated by Cx26 HCs expressed in Xenopus oocytes and the dye uptake in HeLa cells expressing Cxs 32, 43 or 45. Since LA is an essential polyunsaturated fatty acid, its effect on HCs might be relevant to cell growth as well as to cellular functions of differentiated cells such as audition.

Spectroscopic and thermooxidative analysis of organic okra oil and seeds from Abelmoschus esculentus.

With changes in human consumption from animal fats to vegetable oils, the search for seed types, often from unconventional vegetable sources has grown. Research on the chemical composition of both seed and oil for Brazilian Okra in South America is still incipient. In this study, flour and oil from organic Okra seeds (Abelmoschus esculentus L Moench), grown in northeastern Brazil were analyzed. Similar to Okra varieties from the Middle East and Central America, Brazilian Okra has significant amounts of protein (22.14%), lipids (14.01%), and high amounts of unsaturated lipids (66.32%), especially the oleic (20.38%) and linoleic acids (44.48%). Oil analysis through PDSC revealed an oxidation temperature of 175.2 °C, which in combination with low amounts of peroxide, demonstrates its resistance to oxidation and favors its use for human consumption.

Biflavonoids from the unripe fruits of Clusia paralicola and their antioxidant activity.

Investigation of the green fruits of Clusia paralicola (Clusiaceae) led to the isolation and characterization of two 3,8"-biflavonoids, 2R, 3S, 2"R, 3"R-GB1-7"-O-beta-glucoside (1) and 2R, 3S, 2"R, 3,8"-binaringenin-7"-O-beta-glucoside (2), together with four known compounds: beta-sitosterol, stigmasterol, beta-amyrin, and epicatechin. The structures were established from the IR, LC-ESI-MS and NMR spectral data, including 2D-NMR experiments. The absolute configurations of 1 and 2 were determined by CD spectra. The total extract and the biflavonoids demonstrated significant antioxidant activity in DPPH, ABTS, and betacarotene/linoleic acid tests.

Genetic and chemical evaluation of Trypanosoma brucei oleate desaturase as a candidate drug target.

BACKGROUND: Trypanosomes can synthesize polyunsaturated fatty acids. Previously, we have shown that they possess stearoyl-CoA desaturase (SCD) and oleate desaturase (OD) to convert stearate (C18) into oleate (C18:1) and linoleate (C18:2), respectively. Here we examine if OD is essential to these parasites. METHODOLOGY: Cultured procyclic (insect-stage) form (PCF) and bloodstream-form (BSF) Trypanosoma brucei cells were treated with 12- and 13-thiastearic acid (12-TS and 13-TS), inhibitors of OD, and the expression of the enzyme was knocked down by RNA interference. The phenotype of these cells was studied. PRINCIPAL FINDINGS: Growth of PCF T. brucei was totally inhibited by 100 µM of 12-TS and 13-TS, with EC(50) values of 40±2 and 30±2 µM, respectively. The BSF was more sensitive, with EC(50) values of 7±3 and 2±1 µM, respectively. This growth phenotype was due to the inhibitory effect of thiastearates on OD and, to a lesser extent, on SCD. The enzyme inhibition caused a drop in total unsaturated fatty-acid level of the cells, with a slight increase in oleate but a drastic decrease in linoleate level, most probably affecting membrane fluidity. After knocking down OD expression in PCF, the linoleate content was notably reduced, whereas that of oleate drastically increased, maintaining the total unsaturated fatty-acid level unchanged. Interestingly, the growth phenotype of the RNAi-induced cells was similar to that found for thiastearate-treated trypanosomes, with the former cells growing twofold slower than the latter ones, indicating that the linoleate content itself and not only fluidity could be essential for normal membrane functionality. A similar deleterious effect was found after RNAi in BSF, even with a mere 8% reduction of OD activity, indicating that its full activity is essential. CONCLUSIONS/SIGNIFICANCE: As OD is essential for trypanosomes and is not present in mammalian cells, it is a promising target for chemotherapy of African trypanosomiasis.

Stingless bees: chemical differences and potential functions in Nannotrigona testaceicornis and Plebeia droryana males and workers.

Cuticular wax, abdominal and cephalic extracts of foraging workers and males of Nannotrigona testaceicornis and Plebeia droryana, from the "Aretuzina" farm in São Simão, SP, Brazil, were analyzed by GC-MS. The principal constituents were hydrocarbons, terpenes, aldehydes, esters, steroids, alcohols, and fatty acids. Interspecific differences for both cuticular wax and cephalic extracts were found. The composition of cuticular wax and cephalic extracts was similar at the intraspecific level, with minor component differences between males and workers. Abdominal extracts differentiated sexes (male and worker) at the intraspecific and interspecific levels. The main chemical components in abdominal extracts of N. testaceicornis workers and males were geranylgeranyl acetate and (Z)-9-nonacosene, respectively. The principal components of abdominal extracts from P. droryana workers and males were tetradecanal and unsaturated fatty acids (linoleic and linolenic acids), respectively. A secondary alcohol, (S)-2-nonanol, was detected in Plebeia droryana males only, but not in workers. Preliminary field experiments showed that (S)-(+)-2-heptanol and (S)-(+)-2-heptanol/(S)-(+)-2-nonanol (1:1) attracted workers of P. droryana, N. testaceicornis,and Frieseomelitta silvestrii. However, males did not respond suggesting that these compounds do not function as alarm or recruitment pheromones. In addition, racemic mixtures were inactive.

Potential oilseed crops from the semiarid region of northeastern Brazil.

The caatinga semi-arid ecosystem of northeastern Brazil is characterized by a dry, spiny and predominantly deciduous shrub/forest vegetation, and many species there are potential sources of renewable resources for the oleochemical industry. The present research determined the oil content and fatty acid profiles of seeds from eight caatinga species. Seed oils were extracted in a Soxhlet system, and their fatty acid content identified by GC-MS. Oil content varied between 20.2% in Tabebuia impetiginosa (Mart.) Standl. (Bignoniaceae) and 46.4% in Barnebya harleyi (W.R. Anderson & B. Gates) Malpighiaceae. Anemopaegma laeve DC. (Bignoniaceae) had the highest oleic acid content (63.4%), while high levels of linoleic acid were found in Banisteriopsis pubipetala (Juss.) Cuatrec. (42.8%) and B. harleyi (31.9%) (both Malpighiaceae). Palmitic acid was the major fatty acid (50%) in Hippocratea volubilis (L.) (Celastraceae). High levels of linoleic and linolenic acids were found in Croton adamantinus Mull. Arg. (Euphorbiaceae), averaging 44.2% and 45.2% respectively. Gadoleic acid in was the most abundant fatty acid in the oil produced by Serjania lethalis A. St. Hill. (Sapindaceae), averaging 69.6%. B. pubipetala, B. harleyi, C. adamantinus, and H. volubilis were identified as promising species for cultivation.