Palmitic Acid Esters Research Articles

Personalization of lipid-based oral dosage forms via filament-based 3D-printing

While filament-based 3D-printing (3DP) is the most utilized 3DP technology in the pharmaceutical field, it has not been demonstrated for processing of drug-loaded lipid-based formulations. This work exploits hexa-glycerol ester of palmitic acid (Pg6-C16-P) as an advanced lipid material, loaded with felodipine as a poorly soluble model drug, for fabricating novel oral solid dosage forms (OSDFs) via filament-based 3D-printing. After material melt-blending, the formulation was extruded using the liquid feeding approach to obtain a mechanically manageable, and hence 3D-printable, drug-loaded lipid filament. The fabrication of geometries with variable infill densities was demonstrated. The extent of infill density was found to significantly impact the optimal printing parameters required to achieve the desired shape. The solid-state analysis confirmed the amorphous state of felodipine after 3DP. The release rate of the drug was studied via in vitro dissolution test and showed to be tunable based on the tablet geometry. It was also possible to tailor the design of the dosage form to perform similarly to a commercial product. The formulation was evidenced as safe via in vitro toxicity studies with improved felodipine solubility. This study establishes filament-based 3DP as an alternative platform viable for fabricating advanced lipid-based OSDFs, and concurrently, promotes Pg6-C16-P as a promising and high performing 3DP lipid material for drug delivery.

Phytochemical Screening and Antidiabetic Efficacy of Balanites aegyptiaca Seed Extract and Their Silver Nanoparticles on Muscle and Pancreatic Cell Lines.

Balanites aegyptiaca (L.) Delile, a member of the Zygophyllaceae family, is commonly known as the desert date. This tree is famous for yielding edible fruits and is esteemed for its nutritional richness and diverse health advantages. The primary aim of this research was to assess the potential antidiabetic and cytotoxic effects of seed extracts from B. aegyptiaca and its AgNPs for the first time on C2C12 and MIN6 cells, focusing on glucose uptake and insulin secretion, respectively. Additionally, the seed extracts underwent column chromatography through different solvent systems, resulting in the isolation of five distinct fractions with a mixture of methanol and water as an eluting solvent in different ratios. Comprehensive characterization of the aqueous seed extract was carried out using GC-MS and UPLC-MS. The study determined that the aqueous seed extract exhibited no toxicity at any tested concentration (6.25-100 μg/mL) on both cell types. The calculated IC50 values were 206.00 and 140.44 μg/mL for C2C12 and MIN6 cells, respectively, for seeds of AgNPs. Additionally, the aqueous seed extract and their AgNPs significantly increased glucose uptake by 150.45% and 156.00% of the control in C2C12 cells at a concentration of 100 μg/mL. Insulin secretion was also notably enhanced by 3.47- and 3.92-fold of the control after administering seed extracts and AgNPs, respectively, at 100 μg/mL. GC-MS and UPLC-MS analyses identified various compounds across different categories. Notably, the F2 fraction (methanol and water in ratio of 80:20 as eluting solvent) exhibited the highest glucose uptake activity (156.81% of control), while the F3 fraction (methanol and water in ratio of 70:30 as eluting solvent) fraction demonstrated the highest insulin secretion activity (3.70 folds of the control) among all fractions at 100 μg/mL. GC-MS analysis was employed to characterize both fractions, aiming to identify the compounds contributing to their antidiabetic potential. The study's findings concluded that both seed extracts and their AgNPs possess significant antidiabetic properties, with elevated activity observed in the case of AgNPs in both assays. Various compounds, including diosgenin, oleic acid, linoleic acid and palmitic acid esters were detected in the seed extracts, known for their reported antidiabetic and hypoglycemic effects.

Development of a premix based on micellar casein for fortification of meat systems with vitamin A

A novel approach to the protection of unstable fat-soluble vitamins, using retinol as an example, is presented in this work. This method is based on introducing vitamin A molecules into casein micelles. Protective properties of micellar casein towards different forms of retinol (native vitamin and palmitic acid ester) in vitro and in emulsion-type meat products are investigated. A technology of the introduction using micellar casein concentrate (MCC) has been developed. Conditions similar to those in which vitamin molecules can be in meat emulsions during heat treatment are simulated in vitro. The optimal time of “encapsulation” (2 hours) and the need for additional surfactant (tween-80) are identified. The use of the casein micelles protection made it possible to increase the number of retinol molecules that did not undergo decomposition under model conditions (in vitro) from ~30% to ~80%. Using the vitamin premix the degree of degradation of vitamin molecules does not exceed 4% after heat treatment. Data received allowed us to determine the efficiency of the protection properties of casein micelles for unstable vitamin A molecules.

The chemical and biological characteristics of fatty acid esters of hydroxyl fatty acids.

With the continuous advancements in detection methods and the exploration of unknown substances, an increasing number of bioactive compounds are being discovered. Fatty acid esters of hydroxyl fatty acids (FAHFAs), a class of endogenous lipids found in 2014, exhibit various physiological activities, such as improving glucose tolerance and insulin sensitivity, stimulating insulin secretion, and demonstrating broad anti-inflammatory effects. Moreover, some FAHFAs are closely linked to intestinal health and can serve as potential biomarkers for gut health. Various FAHFAs have been observed in food, including palmitic acid esters of hydroxy stearic acids (PAHSA), oleic acid esters of hydroxy stearic acids (OAHSA), linoleic acid esters of hydroxy linoleic acid (LAHLA). As a type of lipid regularly consumed in the daily diet, it is highly important to ascertain the types and quantities of FAHFAs present in the diet. This article, based on existing research, provides a review of the analysis methods for FAHFAs, particularly focusing on the separation of chiral isomers. It also summarizes the sources and contents of dietary FAHFAs, emphasizing their bioavailability and impact on the gut. Understanding the beneficial effects of these lipids in the diet can serve as a valuable reference for the development of specific functional foods.

Utilizing Manilkara zapota seed oil for biodiesel production and conducting an investigation into its properties and characteristics

<p>Biodiesel can be derived from the oils of fruit seeds for powering diesel engines, and this study focuses on extracting biodiesel from Manilkara Zapota seeds, which are readily available and not intended for human consumption. The primary aim is to underscore the use of non-edible oils as a cost-effective raw material for biodiesel production. The research indicates a successful production yield of 92.45% using a 0.76% catalyst, an excess of 6 methanol equivalents in comparison to the oil, a temperature of 62°C, and a reaction time of 90 minutes. The biodiesel obtained from Manilkara Zapota seed oil predominantly comprises methyl esters of oleic, stearic, and palmitic acids, presenting a viable alternative to fossil diesel for unmodified diesel engines. Optimal performance variables for maximum conversion efficiency were determined as a 40°C reaction temperature, 6:1 alcohol-to-oil ratio, 120 minutes of experimental duration, and a 0.5wt.% NaOH catalyst. Among these variables, the alcohol-to-oil ratio was identified as the most influential, contributing 84.34% to the overall performance. The thermal profile of Manilkara Zapota Seeds biomass exhibited multistage decomposition behavior. The synthesized Manilkara Zapota Methyl Ester (MZME) derived from seed oil, using the optimized performance variables, complies with the ASME D 6751 and EN14214 Biodiesel Standards.</p>

Development and efficacy evaluation of a novel water-in-oil-in-water adjuvant for an inactivated foot-and-mouth disease vaccine

ABSTRACT To develop a novel water-in-oil-in-water (W/O/W) adjuvant and evaluate the effect on foot-and-mouth disease (FMD) inactivated vaccine, in this study, we prepared the novel nano-emulsion adjuvant based on QS-21 (BEA) which is composed of the mixture of mineral oil Marcol52, surfactant Tween80, oleate polyoxyethylene ether ester, polyoxyethylene palmitic acid ester and span80, cosurfactant polyethylene glycol and QS-21. The two-step emulsification method formed the W/O/W nano-emulsion with two films and three-phase structures. The effective particle diameter of the BEA was about 184 nm, and it has good thermal stability. Then, BEA was emulsified as an adjuvant to prepare for the inactivated FMDV vaccine, and BALB/c mice and pigs were immunized to evaluate its safety and immunization effect. The results showed that the inactivated BEA-FMDV vaccine significantly increased BALB/c mice and pigs' antibodies and cytokine IFN-γ in serum. Meanwhile, the pig-neutralizing antibodies were higher than control group. Safety tests found no symptoms of FMD or significant toxic reactions. After 28 days of immunization, the protection rate can reach 93.3%. The BEA vaccine had good stability at 4 °C, no stratification after 180 days, and the content of 146S in the vaccine did not decrease. In conclusion, the BEA prepared in this study is suitable for FMDV inactivated vaccine and is an effective adjuvant.

A membrane targeted multifunctional cationic nanoparticle conjugated fusogenic nanoemulsion (CFusoN): induced membrane depolarization and lipid solubilization to accelerate the killing of Staphylococcus aureus.

Bacterial infections caused by Staphylococcus aureus are one of the growing concerns for human health care management globally. Antibiotic-associated adverse effects and the emergence of bacterial resistant strains necessitate the development of an alternative yet effective approach. Nanoemulsion-based therapy has emerged as a potential therapeutic strategy to combat bacterial infestation. Herein, we designed a cationic metal nanoparticle-conjugated fusogenic nanoemulsion (CFusoN) as a lipid solubilizing nanovesicle for the effective treatment of S. aureus infection with a killing efficiency of 99.999%. The cationic nanoparticle-conjugated nanoemulsion (viz. NECNP) (24.4 ± 2.9 mV) electrostatically bound with the negatively charged bacterial cell membrane (-10.2 ± 3.7 mV) causing alteration of the bacterial surface charge. The fluorometric and flow cytometry studies confirmed the bacterial membrane depolarization and altered cell membrane permeability leading to cell death. The atomic force microscopic studies further demonstrated the damage of the cellular ultrastructure, while the transmission electron microscopic image and membrane lipid solubilization analysis depicted the solubilization of the bacterial membrane lipid bilayer along with the leakage of the intracellular contents. The cell membrane fatty acid analysis revealed that the methyl esters of palmitic acid, stearic acid and octadecadienoic acid isomers were solubilized after the treatment of S. aureus with CFusoN. The bactericidal killing efficiency of CFusoN is proposed to occur through the synergistic efficacy of the targeted attachment of CNP to the bacterial cells along with the lipid solubilization property of NE. Interestingly, NECNP didn't elicit any in vitro hemolytic activity or cytotoxicity against red blood cells (RBCs) and L929 fibroblast cells, respectively, at its bactericidal concentration. Furthermore, a porcine skin wound infection model exhibited the enhanced wound cleansing potency of CFusoN in comparison to the commercially available wound cleansers. The obtained antibacterial activity, biocompatibility and skin wound disinfection efficacy of the NECNP demonstrated the formulation of a cell targeted CFusoN as a promising translatable strategy to combat bacterial infection.

Enhanced performance of eco‐friendly PVC by utilizing halo‐benzothiazole derivatives and glucose/fatty acids esters

AbstractTwo halo‐benzothiazoles, namely, 2‐amino‐6‐bromo‐benzo[d]thiazole and 2‐amino‐6‐fluoro‐benzo[d]thiazole were utilized to modify poly(vinyl chloride) (PVC) and gave two modified PVC samples for biomedical applications. Prepared glucose‐fatty acid esters of palmitic, oleic, and octanoic acids were exploited as plasticizers to produce phthalate‐free eco‐friendly antimicrobial PVC. The chemical structures of all prepared compounds were confirmed via Fourier‐transform infrared spectroscopy. The thermal properties of modified PVC was determined using thermo gravimetric analysis and differential scanning calorimetry while weight loss and degree of discoloration were used to assess the photostability after UV irradiation whereas references heat and UV absorbers were applied for comparison. The plasticizing effect of each glucose‐fatty acid ester at 20% by weight on the mechanical properties of modified PVC was determined and referenced to diethylhexylphthalate. The results revealed that the mechanical properties as well as the thermal stability of plasticized modified PVCs were enhanced. Plasticizers migration from the polymeric matrix was also determined and the results showed low extent of migration of the prepared plasticizers. So, modified plasticized PVC samples exhibited improved physicochemical properties. Antimicrobial properties of all samples, using disc diffusion agar method, were evaluated against some Gram +ve bacteria like Staphylococcus aureus and Streptococcus mutans and Gram −ve bacteria like Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa and fungi strains used were Candida albicans, Aspergillus niger, and Aspergillus ochraceous. All investigated compounds showed improved antibacterial and antifungal activities.Highlights PVC was chemically modified with two halo‐benzothiazoles to obtain antimicrobial PVC. Glucose‐fatty acids esters were prepared as eco‐friendly plasticizers for PVC. Halo‐benzothiazoles improved both the thermal‐ and photo‐stability of PVC. Glucose‐fatty acid plasticizers enhanced the mechanical properties of PVC. The extent of migration of prepared plasticizers is lower than that of reference plasticizer.

Obtaining Biodiesel from Fat Extracted from Solid Waste Produced in the Fleshing Stage of Leather Manufacturing

Tallow is produced in leather manufacturing, particularly in the fleshing stage. It contains large amounts of fat, which, in most industrial plants, pollutes water and soil due to poor disposal. In order to take advantage of this solid waste, chemical processes have been applied to transform the fats extracted from tallow into biodiesel. In this work, an extraction technique involving non-polar solvents pressurized at 15 psi. With gasoline, a 51,0% fat recovery was obtained. The quality of the extracted fat was determined by analyzing density, viscosity, melting point, moisture, acidity, and acidity, peroxides, saponification, and iodine indices. Biodiesel was obtained via transesterification reactions with methanol and sodium hydroxide. The content of methyl esters produced in the reaction was characterized by means of gas chromatography coupled with mass spectrometry, where the methyl esters of palmitic and oleic fatty acids were identified as major components.

Phytochemical Analysis and Biological Activity of Salvia compressa Vent.

Salvia extracts have various biological activities and are rich sources of bioactive metabolites. We identified five phytochemicals from S. compressa extract and assessed its biological properties. The plant's shoots were extracted using dichloromethane, and the constituents were isolated using column chromatography. High-resolution NMR spectroscopy characterized the chemical structures of the compounds (1 - 5). The cytotoxic effect of the extract was examined against MCF-7 cells by MTT reduction assay, while cisplatin was tested as a reference cytotoxic compound. The antibacterial activity was assessed using nutrient broth micro-dilution (NBMD), and chloramphenicol was used as the positive control. Citrostadienol (1), β-sitosterol (2), two glyceride esters of linolenic, linoleic, and palmitic acids (3, 4), and geraniol (5) were isolated from S. compressa for the first time. The extract showed activity against MCF-7 breast cancer cells and reduced cell viability to 68.2 ± 13.1% compared to the control drug at the concentration of 50 µg/mL, while it was not active against seven test bacteria. The anti-complementary activity of the isolated steroids suggests S. compressa for future anti-inflammatory research.

Filament-based 3D-printing of placebo dosage forms using brittle lipid-based excipients

In order to expand the limited portfolio of available polymer-based excipients for fabricating three-dimensional (3D) printed pharmaceutical products, Lipid-based excipients (LBEs) have yet to be thoroughly investigated. The technical obstacle of LBEs application is, however their crystalline nature that renders them very brittle and challenging for processing via 3D-printing. In this work, we evaluated the functionality of LBEs for filament-based 3D-printing of oral dosage forms. Polyglycerol partial ester of palmitic acid and polyethylene glycols monostearate were selected as LBEs, based on their chemical structure, possessing polar groups for providing hydrogen-bonding sites. A fundamental understanding of structure–function relationship was built to screen the critical material attributes relevant for both extrusion and 3D-printing processes. The thermal behavior of lipids, including the degree of their supercooling, was the critical attribute for their processing. The extrudability of materials was improved through different feeding approaches, including the common powder feeding and a devised liquid feeding setup. Liquid feeding was found to be more efficient, allowing the production of filaments with high flexibility and improved printability. Filaments with superior performance were produced using polyglycerol ester of palmitic acid. In-house designed modifications of the utilized 3D-printer were essential for a flawless processing of the filaments.

Liposomal Form of 2,4-Dinitrophenol Lipophilic Derivatives as a Promising Therapeutic Agent for ATP Synthesis Inhibition.

Mitochondrial uncoupler 2,4-dinitrophenol (2,4-DNP) is a promising antidiabetic and antiobesity agent. Its clinical use is limited by a narrow dynamic range and accumulation in non-target sensitive organs, which results in whole-body toxicity. A liposomal formulation could enable the mentioned drawbacks to be overcome and simplify the liver-targeted delivery and sustained release of 2,4-DNP. We synthesized 2,4-DNP esters with carboxylic acids of various lipophilic degrees using carboxylic acid chloride and then loaded them into liposomes. We demonstrated the effective increase in the entrapment of 2,4-DNP into liposomes when esters were used. Here, we examined the dependence of the sustained release of 2,4-DNP from liposomes on the lipid composition and LogPoct of the ester. We posit that the optimal chain length of the ester should be close to the palmitic acid and the lipid membrane should be composed of phospholipids with a certain phase transition point depending on the desired release rate. The increased effect of the ATP synthesis inhibition of the liposomal forms of caproic and palmitic acid esters compared to free molecules in liver hepatocytes was demonstrated. The liposomes’ stability could well be responsible for this result. This work demonstrates promising possibilities for the liver-targeted delivery of the 2,4-DNP esters with carboxylic acids loaded into liposomes for ATP synthesis inhibition.

ATGL is a biosynthetic enzyme for fatty acid esters of hydroxy fatty acids

Branched fatty acid (FA) esters of hydroxy FAs (HFAs; FAHFAs) are recently discovered lipids that are conserved from yeast to mammals1,2. A subfamily, palmitic acid esters of hydroxy stearic acids (PAHSAs), are anti-inflammatory and anti-diabetic1,3. Humans and mice with insulin resistance have lower PAHSA levels in subcutaneous adipose tissue and serum1. PAHSA administration improves glucose tolerance and insulin sensitivity and reduces inflammation in obesity, diabetes and immune-mediated diseases1,4–7. The enzyme(s) responsible for FAHFA biosynthesis in vivo remains unknown. Here we identified adipose triglyceride lipase (ATGL, also known as patatin-like phospholipase domain containing 2 (PNPLA2)) as a candidate biosynthetic enzyme for FAHFAs using chemical biology and proteomics. We discovered that recombinant ATGL uses a transacylation reaction that esterifies an HFA with a FA from triglyceride (TG) or diglyceride to produce FAHFAs. Overexpression of wild-type, but not catalytically dead, ATGL increases FAHFA biosynthesis. Chemical inhibition of ATGL or genetic deletion of Atgl inhibits FAHFA biosynthesis and reduces the levels of FAHFA and FAHFA-TG. Levels of endogenous and nascent FAHFAs and FAHFA-TGs are 80–90 per cent lower in adipose tissue of mice in which Atgl is knocked out specifically in the adipose tissue. Increasing TG levels by upregulating diacylglycerol acyltransferase (DGAT) activity promotes FAHFA biosynthesis, and decreasing DGAT activity inhibits it, reinforcing TGs as FAHFA precursors. ATGL biosynthetic transacylase activity is present in human adipose tissue underscoring its potential clinical relevance. In summary, we discovered the first, to our knowledge, biosynthetic enzyme that catalyses the formation of the FAHFA ester bond in mammals. Whereas ATGL lipase activity is well known, our data establish a paradigm shift demonstrating that ATGL transacylase activity is biologically important.

Triacylglycerols containing branched palmitic acid ester of hydroxystearic acid (PAHSA) are present in the breast milk and hydrolyzed by carboxyl ester lipase

Breast milk is a complex mixture containing underexplored bioactive lipids. We performed an observational case-control study to compare the impact of delivery mode: caesarean section (CS) and vaginal birth (VB); and term (preterm and term delivery) on the levels of lipokines in human milk at different stages of lactation. Metabolomic analysis of the milk identified triacylglycerol estolides as a metabolic reservoir of the anti-inflammatory lipid mediator 5-palmitic acid ester of hydroxystearic acid (5-PAHSA). We found that triacylglycerol estolides were substrates of carboxyl ester lipase and 5-PAHSA-containing lipids were the least preferred substrates among tested triacylglycerol estolide isomers. This explained exceptionally high colostrum levels of 5-PAHSA in the VB group. CS and preterm birth negatively affected colostrum lipidome, including 5-PAHSA levels, but the lipidomic profiles normalized in mature milk. Mothers delivering term babies vaginally produce colostrum rich in 5-PAHSA, which could contribute to the prevention of intestinal inflammation in newborns.

Overexpression of Two New Acyl-CoA:Diacylglycerol Acyltransferase 2-Like Acyl-CoA:Sterol Acyltransferases Enhanced Squalene Accumulation in Aurantiochytrium limacinum.

Thraustochytrids are heterotrophic marine eukaryotes known to accumulate large amounts of triacylglycerols, and they also synthesize terpenoids like carotenoids and squalene, which all have an increasing market demand. However, a more extensive knowledge of the lipid metabolism is needed to develop thraustochytrids for profitable biomanufacturing. In this study, two putative type-2 Acyl-CoA:diacylglycerol acyltransferases (DGAT2) genes of Aurantiochytrium sp. T66, T66ASATa, and T66ASATb, and their homologs in Aurantiochytrium limacinum SR21, AlASATa and AlASATb, were characterized. In A. limacinum SR21, genomic knockout of AlASATb reduced the amount of the steryl esters of palmitic acid, SE (16:0), and docosahexaenoic acid, SE (22:6). The double mutant of AlASATa and AlASATb produced even less of these steryl esters. The expression and overexpression of T66ASATb and AlASATb, respectively, enhanced SE (16:0) and SE (22:6) production more significantly than those of T66ASATa and AlASATa. In contrast, these mutations did not significantly change the level of triacylglycerols or other lipid classes. The results suggest that the four genes encoded proteins possessing acyl-CoA:sterol acyltransferase (ASAT) activity synthesizing both SE (16:0) and SE (22:6), but with the contribution from AlASATb and T66ASATb being more important than that of AlASATa and T66ASATa. Furthermore, the expression and overexpression of T66ASATb and AlASATb enhanced squalene accumulation in SR21 by up to 88%. The discovery highlights the functional diversity of DGAT2-like proteins and provides valuable information on steryl ester and squalene synthesis in thraustochytrids, paving the way to enhance squalene production through metabolic engineering.

Potent and selective anticancer activity of half-sandwich ruthenium and osmium complexes with modified curcuminoid ligands.

We have recently reported a series of half-sandwich ruthenium(II) complexes with curcuminoid ligands showing excellent cytotoxic activities (particularly ionic derivatives containing PTA (PTA = 1,3,5-triaza-7-phosphaadamantane). In the present study, new members of this family of compounds have been prepared with the objective to investigate the effect of a long hydrophobic chain obtained by replacing the OH-groups, present in curcumin and bisdemethoxycurcumin, with the palmitic acid ester. We report the synthesis of ruthenium(II) and osmium(II) p-cymene derivatives containing palmitic acid curcumin ester ligands ((1E,3Z,6E)-3-hydroxy-5-oxohepta-1,3,6-triene-1,7-diyl)bis(2-methoxy-4,1-phenylene)dipalmitate (p-curcH) and ((1E,3Z,6E)-3-hydroxy-5-oxohepta-1,3,6-triene-1,7-diyl)bis(4,1-phenylene)dipalmitate (p-bdcurcH). Complexes [M(II)(cym)(p-curc)/(p-bdcurc)(Cl)] 1-4 (M = Ru or Os) are neutral, whereas [M(II)(cym)(p-curc)/(p-bdcurc)(PTA)][SO3CF3] 5-8 are salts obtained when the chloride ligand is replaced by the PTA ligand. Stability studies performed on 1-8 in DMSO-PBS under physiological conditions (pH = 7.4) indicate that the complexes remain intact. The complexes exhibit potent and selective cytotoxic activity against an ovarian carcinoma cell line and its cisplatin-resistant form (A2780 and A2780cis), and non-cancerous human embryonic kidney (HEK293T) cells. To define the structure-activity relationships (SAR), the compounds have been compared with other Ru(II) and Os(II) complexes with curcuminoid ligands previously reported. SAR data reveal that the bisdemethoxycurcumin complexes are generally more active and selective than analogous curcumin-containing complexes.

Chemical profiling and anti-breast cancer potential of hexane fraction of Sphaeranthus indicus flowers

Purpose: The current study aimed to determine the phytochemicals and anti-breast cancer potential of Sphaeranthus indicus.Methods: S. indicus flowers were extracted with methanol followed by fractionation using n-hexane. For the chemical composition of n-hexane fraction, qualitative phytochemical and GC-MS analysis were performed. The anti-proliferative activity was measured by MTT assay, whereas, cytotoxic and proapoptotic effects in MCF-7 (breast cancer) cells were determined using propidium iodide, 4′,6-diamidino- 2-phenylindole, dichlorofluorescin diacetate, and JC-1 staining through fluorescent microscopy.Results: The phytochemical analysis indicated presence of phytosterols, oils and resins in the nhexane fraction. GC-MS analysis showed that n-hexane fraction comprises of 11 compounds including methyl esters of caprylic acid, myristic acid, pentadecanoic acid, palmitic acid, margaric acid, stearic acid, oleic acid, elaidic acid, linoleic acid, linolenic acid and behanic acid. The tested fraction showed remarkable cytotoxic activity against breast cancer (MCF-7) cells while it was found less toxic towards non-cancerous (BHK-21) cells. Furthermore, morphological assessment through fluorescent microscopy revealed cytotoxic and apoptotic effects by improved cell membrane permeability, increased reactive oxygen species level, compromised mitochondrial activity and condensation of chromatin network.Conclusion: The n-hexane fraction of S. indicus contains phytosterols, oils and fatty acid methyl esters and produced apoptotic effect against breast cancer cells.

Fatty acids and their derivatives from Chlorella vulgaris extracts exhibit in vitro antimicrobial activity against the honey bee pathogen Paenibacillus larvae

The green microalga Chlorella vulgaris Beijerinck (Chlorellaceae) is widely used as a food supplement for humans and animals. In beekeeping practice, Chlorella vulgaris has potential as a pollen supplement. We studied whether Chlorella extracts display antimicrobial properties against Paenibacillus larvae, the causative agent of the honey bee bacterial disease American foulbrood. We identified components responsible for antimicrobial activity and evaluated the added values of Chlorella as a food supplement for honey bees. Different extracts (water, acetone, methanol) were prepared from Chlorella biomass (phototrophically and heterotrophically cultivated) and screened for antimicrobial activity against ERIC I and ERIC II genotypes of P. larvae. Active acetone extracts of phototrophically cultivated Chlorella vulgaris biomass were fractioned via preparative reverse-phase chromatography. Antimicrobial activity was detected for 9 of the resulting 33 fractions. Further analysis revealed the chemical composition of the active fractions. C. vulgaris extracts showed a significant antimicrobial effect against vegetative cells and spores of P. larvae strains of ERIC I and ERIC II genotypes. The lowest MIC of the most active acetone extract was 6.3 µg/mL for both tested genotypes. In the majority of the active fractions, monolinolenin, fatty acid linoleic acid, and methyl esters of linoleic and/or palmitic acid were identified via high-performance liquid chromatography coupled with high-resolution mass spectrometry analysis. Based on our results, we concluded that algal C. vulgaris food supplements not only contain nutritional but also potential prophylactic properties for honey bee health.

GC-MS ANALYSIS OF THREE PLANTS ESSENTIAL OILS AND THEIR EFFECT ON BACTERIAL SPOT DISEASE OF TOMATO

The aim of this present study is to evaluate the antimicrobial activities of three plants essential oils, Nigella (Nigella sativa), Eucalyptus (Cinnamomum camphora) and clove (Syzygium aromaticum) against plant pathogenic bacteria Xanthomonas vesicatoria and their influences on controlling the bacterial spot disease of tomato plants. The chemical composition of the oils was calculated by gas chromatograph/mass spectrometer (GC/MS) analysis. The results revealed that the major constituents of Nigella oil were fatty acid i.e. oleic acid (4.09%%), Fumaric acid (0.13%), Palmitic acid (4.95-1.65 %), Octadec-9-enoic acid (0.53%), myristic acid (0.78%), Cyclohexadecane (0.01%) and Hexanedioic acid bis (2-ethylhexyl) ester (75.02%). In addition, the main components of Eucalyptus oil were D-camphor (40.01%), linalool (20.57%), and cineole (10.86%). The chemical analysis showed also that clove oil contained eugenol (22.94%), eugenyl acetate (16.65%), caryophyllene (10.37 %) and 2-Pyridineethanol (6.08 %). In in vitro bioassay against X.anthomonas vesicatoria, the results demonstrated that among the three tested plant oils, Eucalyptus oil (10 %) recorded the maximum value was (1.73 cm) bacterial inhibition zone followed by Clove oil (1.50 cm). Results also showed spraying plants with the tested essential oils two days before bacterial inoculation recorded the lowest disease index comparing to the treatment where plants were treating with essential oils two days after bacterial inoculation. The obtained results also clearly illustrated that, Peroxidase (PO), polyphenoloxidase PPO and chitinase activities increased as a result of spraying tomato plants with the tested oils. Moreover, protein analysis confirmed that new protein bands with low molecular weight had a progressive relationship with reduction of bacterial spot disease severity on plants treated with the tested oils.

Formation kinetics of radiolytic lipid products in model food–lipid systems with gamma irradiation

AbstractIrradiation processes can change the lipid composition of foods. The major lipid radiolysis products are known to be 2‐alkylcyclobutanones and hydrocarbons, the formation kinetics of which is determined by the chemical nature of the inherent lipids. This information can be used to evaluate the irradiation history of lipid‐rich foods. A solid‐phase microextraction (SPME)‐assisted gas chromatography–mass spectrometry (GC–MS) technique was used to study the formation of volatile hydrocarbons and 2‐dodecylcyclobutanone (2‐DCB), a cyclobutanone formed from the esters of palmitic acid, in irradiated (0–50 kGy) tripalmitin and chicken fat. Thirty‐three volatile compounds (hydrocarbons, aldehydes, ketones) increased with irradiation dose to varying extents. Six major lipid radiolysis products were selected to analyze their formation kinetics: tridecane, 1‐tetradecene, tetradecane, 1‐pentadecene, pentadecane, and 2‐DCB. The concentration of the selected compounds increased linearly with irradiation dose following zeroth‐order kinetics, except for tridecane and tetradecane in tripalmitin, formation of which followed first‐order kinetics. The formation rate of hydrocarbons was higher in tripalmitin than in chicken fat (5× higher for pentadecane), while 2‐DCB formed faster (3.5×) in chicken fat than in tripalmitin. This difference was related to the diversity in the natural fatty acid composition of chicken fat as compared to the homogeneous structure of tripalmitin. The results of this study indicate that the concentrations of palmitic acid radiolysis products linearly increase with irradiation dose in general to offer potential to back‐trace irradiation dose in lipid‐rich foods.