Oleic Research Articles

Effect of Oleic Acid Supplementation and Microoxygenation on Volatilome of Wine Prepared from Grape Hybrid H27 Using S. cerevisiae MK680910

Effect of Oleic Acid Supplementation and Microoxygenation on Volatilome of Wine Prepared from Grape Hybrid H27 Using S. cerevisiae MK680910

Dietary nicotinamide improves growth performance and ameliorates liver lipid profiles by alleviating glycerophospholipid and sphingolipids metabolism in swamp eel (Monopterus albus)

The effect of dietary niacinamide (NAM) on the growth performance and liver lipid metabolism of swamp eel have been poorly investigated. Therefore, a 120-day feeding trial was conducted to provide different levels of NAM in diet and an untargeted lipidome analysis was performed to address this subject. We found that dietary NAM improved the growth performance of swamp eel. It also decreased the contents of glucose and lipids, including glucose, glycogen, TG, and NEFA, as well as fat accumulation in liver, but increased HDL-C content. We speculated that NAM inhibited lipid accumulation in liver by decreasing the key enzyme activity of LPL, HL and LPS. Fatty acid profile analysis showed that the contents of MUFA and PUFA, such as palmitoleic acid, oleic acid, and eicosapentaenoic acid were significantly increased, while the contents of SFA, such as myristic acid, stearic acid, and palmitic acid were decreased in liver by dietary NAM. Lipidome analysis also indicated that the majority of differential lipid metabolites belonged to glycerophospholipids and sphingolipids, which were enriched in the pathways of insulin resistance, adipocytokine signaling pathway, cholesterol metabolism, fat digestion and absorption, lipid and atherosclerosis, and lipid cell lipolysis regulation. These findings gave scientific evidences and novel insights into the metabolic responses to dietary NAM in swamp eel, and provided basic knowledge for using NAM as a feed additive in swamp eel production.

Continuous Flow Microreactors for the High-efficiency Enzymatic Synthesis of 10-Hydroxystearic Acid from Oleic Acid.

Converting fatty acids into specialty chemicals is sustainable but hindered by the low efficiency and thermal instability of current oleic acid hydratases, along with mass transfer limitations in emulsion reactions. This study introduces an optimized continuous flow micro-reactor (CFMR) that efficiently transforms oleic acid at low (15 g·L-1) and high (50 g·L-1) concentrations, improving reaction efficiency and overcoming key conversion barriers. The first CFMR model showed reaction speeds surpassing traditional batch stirred tank reactors (BSTR). Optimizations were performed on three key components: liquid storage, mixer, and reaction section of the CFMR, with each round's best conditions carried into the next. This achieved a space-time yield of 597 g·L-1·d-1 at a 15 g·L-1 oleic acid load. To further enhance the yield, we optimized the emulsifier system to solve incomplete emulsification and developed a two-component feed microreactor (TCFMR) that addressed substrate and product inhibition at high loads, reaching a 91% conversion of 50 g·L-1 oleic acid in 30 minutes, with a space-time yield of 2312 g·L-1·d-1. These advancements represent significant progress in utilizing fatty acids and advancing sustainable chemical synthesis.

Computational Designed and Optimized Liposomal Curcumin-Embedded Bifunctional Cross-Linked Hydrogels for Wound Healing

Curcumin (CUR) bifunctional cross-linked nanocomposite hydrogels are presented as an efficient method for CUR delivery in wound healing. CUR-loaded liposomes (CUR-Ls) were optimized using the Box–Behnken design to augment particle size, size distribution, zeta potential, and CUR concentration. The antioxidant activity and cytotoxicity of CUR-Ls were assessed. Hyaluronic acid (HA)/poly(vinyl alcohol) (PVA) hydrogels were optimized with a central composite design; then, poly(N-vinylpyrrolidone-co-itaconic acid) (PNVP-ITA) was synthesized to enrich the properties of the hydrogels. The drug release kinetics of the CUR-L@HA/PVA/PNVP-ITA hydrogels were studied. Skin recovery was investigated in vivo on rat dorsal skin. The optimized CUR-Ls were constructed from 2.7% Tween® 20, 0.04% oleic acid, and 8.1% CUR, yielding nano-CUR-L with a narrow size distribution, negative surface charge, and CUR content of 19.92 ± 0.54 µg/mg. CUR-Ls improved the antioxidant effects of CUR. The optimized hydrogel contained 5% HA and 10% PVA. PNVP-ITA improved the properties of the hydrogels via enhanced cross-linking. CUR-Ls exhibited a more rapid release than CUR, whereas the hydrogels enhanced CUR release via a diffusion-controlled mechanism. CUR-L@HA/PVA/PNVP-ITA hydrogels improved the skin recovery rate compared to the commercial patch after 5 days. Therefore, the optimized CUR-L@HA/PVA/PNVP-ITA hydrogels facilitated skin recovery and could be a promising nanocomposite for wound dressings.

Tracking the storage stability in sesame (Sesamum indicum L.): impact of accelerated storage on storability characteristics, seed quality, phytochemical content, and fatty acids

Storage stability under non-optimal conditions is an essential characteristic of Sudanese sesame. To understand opportunities to improve storage stability in sesame, seed quality, storability characteristics, content of fatty acids and phytochemicals, and antioxidant capacity were evaluated in ten Sudanese genotypes subjected to high temperature (55 °C) and humidity (60% RH) for 16 and 32 days. The accelerated storage increased seed color, linoleic acid, fungal growth, and peroxide value, while oil content, oleic acid, water activity, phytochemicals, and antioxidant capacity decreased (P < 0.05). The germination rate and content of saturated fatty acids were retained despite the storage (P > 0.05). The landrace Abusundoug showed better storage stability than the other genotypes due to generally low fatty acids and high phytochemical contents. The differences in storage stability in the Sudanese genotypes underscore the need for their further evaluation and use in breeding programs to improve sesame shelf life and quality.

Synthesis of Nanoscale ZSM-5 Zeolites for the Catalytic Cracking of Oleic Acid into Light Olefins and Aromatics

Synthesis of Nanoscale ZSM-5 Zeolites for the Catalytic Cracking of Oleic Acid into Light Olefins and Aromatics

Biochemical profile of the native seaweed fly Fucellia maritima (Haliday, 1838) (Diptera: Anthomyiidae) reared on five different substrates

Abstract The sustainability of aquaculture production is crucial for the industry’s long-term viability. Diversification of feed ingredients is a key component in achieving this sustainability. Considered a promising alternative to traditional feed ingredients, terrestrial insect meals often lack sufficient n-3 polyunsaturated fatty acids (PUFA). This study explores the potential of marine insect species, specifically evaluating the influence of five different substrates on the growth and composition of the European native seaweed fly, Fucellia maritima. The substrates tested include a brown seaweed (Fucus sp.), a green seaweed (Ulva sp.), a red seaweed (the invasive Agarophyton vermiculophyllum), the invasive freshwater hyacinth (Eichhornia crassipes), and fish processing waste (codfish Gadus morhua frames). Results show no significant difference in the number of individuals per substrate, although Fucus sp. had a higher total number of individuals completing their life cycle. In contrast, feeding on codfish frames resulted in a lower number of completed life cycles, while E. crassipes led to no survival. Protein content in adult flies ranged from 55.2% to 56.7%, and in pupae, it ranged from 46.1% to 48.7% across different feeding substrates. Notably, pupae fed with Ulva sp., exhibited the highest protein content at 48.7%. Lipid content in adult flies ranged from 10.0% to 13.0%, while in pupae, it ranged from 8.8% to 11.4%. Codfish frames had the highest lipid content in both pupae (11.4%) and adults (13%). The most abundant fatty acid in Fucus sp. – fed pupae and adult flies was oleic acid (18:1 n-9), while palmitoleic acid (16:1 n-7) dominated in other treatments. It is worth highlighting that docosahexaenoic acid (22:6 n-3) was reported for the first time in a seaweed fly, namely when supplied with fish processing waste. These findings suggest that Fucellia maritima may be a promising complementary ingredient for formulating marine aquafeeds.

Anti-inflammatory activity and identification of two ‘Sarpagan’ indole alkaloids from the leaf methanolic extract of Rauvolfia densiflora (Wall.) Benth. ex Hook. f.

The Apocynaceae plant group contains many bioactive compounds that are pharmacologically relevant and are used as potential medicine for many diseases. Rauvolfia densiflora Benth. ex Hook.f. (syn- Rauvolfia verticillata (Lour.) Baill.) coming under the same family known for its diverse medicinal properties. The current study was carried out to evaluate the bioactive compounds present in the extract using GC-MS analysis and to find out the anti-inflammatory potential of the extract using in-vitro methods. GC-MS analysis revealed the presence of 16 bioactive compounds with diverse pharmacological activities. Among them, Oleic Acid, 17-hydroxy-, methyl ester, trans-decahydroquinoline, and Spiro-(1,3-dioxolane-2, 3’-indolin]-2’-one possesses more important properties. Two important sarpagan indole alkaloids were also identified namely Sarpagan-16-carboxylic acid or (Z) Akuammidine and sarpagan-17-ol or Vellosiminol, which are pharmacologically important compounds with anti-inflammatory, anti-asthmatic activities. The in-vitro anti-inflammatory activity was carried out using the COX assay. COX assay showed 55.27% inhibition at a concentration of 200 μg/mL with an IC50 value of 155.38 μg/mL, this is an indication that the methanolic leaf extract R. densiflora has the potential to be developed as a non-steroidal anti-inflammatory drug (NSAID).

In vitro versus cryo-induced capacitation of bovine spermatozoa, part 3: Compositional and molecular changes to the plasma membrane

The aim of this study was to assess the level of membrane cryodamage through the levels of selected capacitation and apoptosis-associated proteins, together with compositional membrane changes in capacitated (CAP), cryopreserved (CRYO) and non-capacitated bovine spermatozoa (CRTL). Sperm kinetic parameters were analyzed by the computer assisted sperm analysis (CASA) while the capacitation patterns were examined with the chlortetracycline (CTC) assay. In the case of DNA integrity, sperm chromatin structure assay and aniline blue staining were used. For the quantification of fatty acid content gas chromatography was performed. Using Western blotting the expression of capacitation (protein kinase C – PKC; phospholipases A2 and Cζ – PLA2, PLCζ; soluble adenylyl cyclase 10 – sAC10) and apoptosis-associated (apoptosis regulator Bax; B-cell lymphoma 2 – Bcl-2; caspase 3) proteins were evaluated. Data indicate a significant decline (p < 0.0001) of sperm kinetic parameters and higher occurrence (p < 0.0001) of DNA fragmentation in the CRYO group. CTC assay revealed a significant increase of acrosome-reacted spermatozoa in the CRYO group when compared to others. Compositional changes in the sperm membrane were visible as a notable decline of docosahexaenoic acid (p < 0.0001) associated with a significant decrease of membrane cholesterol (p < 0.05) and proteins (p < 0.0001) in the CRYO group while the amount of palmitic, stearic, oleic, and linoleic acid increased (p < 0.0001) significantly. Protein expression of all capacitation-associated proteins (PKC, PLA2, PLCζ, sAC10) was significantly down-regulated (p < 0.001; p < 0.0001) in the CRYO group. Relative quantification of apoptosis-associated proteins revealed increased Bax and decreased Bcl-2 levels in the CRYO group, except for caspase-3, which remained without significant changes.

Drug Binding and Delivery with Supramolecular System Based on Sodium Carboxymethylcellulose and Viologen Calix[4]resorcinol.

In the present work, a simple supramolecular approach was used to spontaneously form nanoparticles from sodium carboxymethylcellulose and viologen calix[4]resorcinol as a result of joint self-assembly in water at room temperature. Supramolecular interactions between them led to the formation of nanoparticles, the morphology and properties of which depend on polymer/macrocycle ratio. When there is excess of macrocycle, the nanoparticles bind lipophilic oleic acid, and when there is excess of polymer, they bind hydrophilic doxorubicin. Interestingly, the solubilization of lipophilic quercetin into these nanoparticles was significantly increased compared to the pure polymer and macrocycle, regardless of their ratio. The nanoparticles of triple composition (polymer/macrocycle/drug) have a more effective penetration ability into these cells than those of double composition (polymer/drug). The presented results are the first step towards the creation of new nanotechnologies to improve the bioavailability and delivery of bioactive components.

Utility of Arabidopsis KASII Promoter in Development of an Effective CRISPR/Cas9 System for Soybean Genome Editing and Its Application in Engineering of Soybean Seeds Producing Super-High Oleic and Low Saturated Oils.

This study reports the use of the Arabidopsis KASII promoter (AtKASII) to develop an efficient CRISPR/Cas9 system for soybean genome editing. When this promoter was paired with Arabidopsis U6 promoters to drive Cas9 and single guide RNA expression, respectively, simultaneous editing of the three fatty acid desaturase genes GmFAD2-1A, GmFAD2-1B, and GmFAD3A occurred in more than 60% of transgenic soybean lines at T2 generation, and all the triple mutants possessed desirable high-oleic traits. In sharp contrast, not a single line underwent simultaneous editing of the three target genes when AtKASII was replaced by the widely used AtEC1.2 promoter. Furthermore, our study showed that the stable and inheritable mutations in the high-oleic lines did not alter the overall contents of oil and protein or amino acid composition while increasing the oleic acid content up to 87.6% from approximately 23.8% for wild-type seeds, concomitant with 34.4- and 3.7-fold reductions in linoleic and linolenic acid, respectively. Collectively, this study demonstrates that the AtKASII promoter is highly promising for optimization of the CRISPR/Cas9 system for genome editing in soybean and possibly beyond.

Phytocompound profiling and GC-MS analysis of Lantana camara leaf extract

The primary objective of this investigation was to conduct an initial analysis of the chemical composition of Lantana camara leaves, with a specific focus on identifying potential pharmacologically active compounds. Gas chromatography-mass spectrometry (GC-MS) was employed to perform a preliminary phytochemical screening on various leaf extracts. The results identified multiple classes of phytoconstituents in Lantana camara leaves, including steroids, terpenoids, flavonoids, quinones, carbohydrates, alkaloids, and phenols. A detailed analysis of the crude aqueous extract using GC-MS identified several biologically active components, such as 13-docosenamide, alpha-hydroxyisocaproic acid, cyclo(L-prolyl-L-valine), and 2,5-piperazinedione etc. These compounds were present in varying proportions, indicating a complex chemical composition within the leaves of Lantana camara. The study's findings suggest that these identified chemicals may contribute to the plant's pharmacological properties and hold significant potential for innovative medical applications. The plant's bioactive compounds, including pyrrolo[1,2-a]pyrazine-1,4-dione, hexadecanoic acid, hexahydro and oleic acid, point to a wide range of possible therapeutic benefits. This investigation provides valuable insights into the chemical constituents of Lantana camara and underscores the potential significance of these compounds in medicinal research. Furthermore, it opens up promising avenues for future studies and the exploration of new pharmaceutical opportunities.

Abstract PR-06: Dietary fats dictate pancreatic cancer fate via phospholipid saturation

Abstract Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer death in the US and is anticipated to become the second within the next decade. Epidemiologic studies support a strong link between high fat diet (HFD) consumption and increased PDAC risk, however, the translational relevance of diet research has been limited by inconsistencies in fat source and consumption across human populations and mouse studies. Therefore, whether and how specific dietary fatty acids drive cancer development is poorly understood. To address this gap in knowledge, we performed a diet screen, comprised of 12 isocaloric HFDs differing solely in fat source and representing the diversity of modern human fat consumption, in an oncogenic Kras-driven mouse model (KC: Kras LSL-G12D /+;Pdx1-Cre) that closely mimics the genetic and histologic features of human PDAC progression. Unexpectedly, we discovered that diets rich in oleic acid – a monounsaturated fatty acid (MUFA) typically associated with good health – enhanced pancreatic tumorigenesis. Furthermore, we observed a strong positive correlation between increased tumor burden and diets exhibiting a high ratio of MUFAs to polyunsaturated fatty acids (PUFAs), concordant with high MUFA to PUFA levels in the plasma and in pancreatic phospholipids. Membrane phospholipid incorporation of MUFAs (relative to PUFAs) suppresses lipid peroxidation and cell death via ferroptosis. Consistent with this, pancreata from mice fed a MUFA-rich diet showed decreased expression of genes involved in lipid peroxidation and reduced histologic evidence of lipid peroxidation end products compared to a PUFA-rich diets. Furthermore, primary acinar cells from mice fed high MUFA diets were resistant to cell death upon treatment with ferroptosis inhibitors. Conversely, diets rich in PUFAs increased PUFA incorporation into pancreatic phospholipids, activated transcriptional programs of lipid peroxidation, and intercepted Kras-driven tumor progression in vivo. Collectively, these data support a role for dietary fatty acids in molding the pancreatic phospholipid landscape, altering lipid peroxidation and ferroptosis sensitivity to dictate tumor fate. Citation Format: Christian F Ruiz, Xiangyu Ge, Rylee McDonnell, Daniel McQuaid, Jennifer Kaplan, Guangtao Li, Michael C Rudolph, Fred F Gorelick, John Wysolmerski, Daniel Canals, John D Haley, Matthew Rodeheffer, Mandar D Muzumdar. Dietary fats dictate pancreatic cancer fate via phospholipid saturation [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr PR-06.

Preparation and In vitro/Ex vivo Evaluation of Nanoemulsion-Based in Situ Gel for Intranasal Delivery of Lasmiditan

Lasmiditan (LAS) was formulated as a nanoemulsion based in situ gel (NEIG)with the aim of improving its oral bioavailability via application intranasally. The solubility of LAS in oils, emulsifiers, and co-emulsifiers was determined to identify nanoemulsion (NE)components. Phase diagrams were constructed to identify the area of nanoemulsification. LAS NE was formulated using the spontaneous nanoemulsification method. Four NEs (F19, F24, F31, and F34) containing 7-15 % oleic acid (OA) as an oily phase, 40-55% labrasol (LR), and transcutol (TC) as emulsifier mixture at (1:1), (2:1), (3:1), and (1:2) ratio with 30-53 % (w/w) aqueous phase, having suitable optical transparency of 95–98%, globule size of 104-140 nm and polydispersity of 0.253–0.382 were selected for ex vivo permeation study. F31 with the highest flux value (2.32 ± 0.01 mg/cm2.min) relative to the other NEs. It achieves an enhancement ratio of 3.3 as compared to LAS aqueous suspension (8% LAS) also it achieves a significantly higher value of permeability coefficient. F31 was selected for the incorporation of different percentages of pH-sensitive in situ gelling polymer (Carbopol 934) to prepare NEIGs 4,5 and 6. The gel strength, pH, gelation time, and viscosity were predicted for the prepared NEIGs. In vitro release and ex vivo, nasal permeation were determined for NEIG5, which exerts comparable release and permeation values as F31 with more residence time in order to overcome the normal nasal physiological clearance.

Abstract C039: Investigating ER stress responses in pancreatic ductal adenocarcinoma under lipid imbalance

Abstract Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer-related death, with a five-year survival rate of 12%. Dense desmoplasia results in poor intratumoral perfusion and elevated interstitial fluid pressure, creating regions with severe oxygen and nutrient deprivation. One consequence is a deficiency in unsaturated fatty acids (uFAs), as the de novo synthesis of uFAs requires oxygen. Our results indicate that PDAC cells experience endoplasmic reticulum (ER) stress and activate subsequent ER stress responses under uFA- limited conditions in vitro. ER stress responses are governed by three major stress sensors: inositol-requiring enzyme 1α (IRE1α), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). When IRE1α is activated, XBP1 mRNA is spliced, resulting in an activated transcription factor, XBP1s, which modulates lipid metabolism. XBP1s is upregulated in PDAC compared to PanIN and normal tissue in murine and human samples. PDAC cells have reduced viability under uFA-limited conditions. Moreover, ER stress is activated under uFA deficiency, and the stress response is diminished upon rescue by monounsaturated oleic acids. To study the role of ER stress responses, we genetically depleted PERK, ATF6, or XBP1 in PDAC cells. The depletion of these pathways rescued PDAC cell viability. Interestingly, B-I09, an inhibitor of XBP1 splicing, led to a mild tumor reduction in vivo. We have found that "basal-like" patient tumors are significantly enriched in the ER stress response pathways based on bulk patient RNA-seq datasets. Therefore, we hypothesize that the two PDAC subtypes “basal” and “classical” might have different responses or reliance on the ER stress pathways. We have created isogenic subtype models to represent basal and classical subtypes by modulating epithelial to mesenchymal (EMT) factors and deltaNP63. Signaling studies suggest that there are no differences between the subtypes’ response to uFA-limited conditions. We are currently assessing the role of XBP1s and IRE1α in vivo by implanting the gene-edited cells into subcutaneous and orthotopic tumor models. Furthermore, we are pursuing spatial mass spectrometry to study the lipid environment in PDAC tumors and investigate whether lipid imbalance is the main cause of ER stress activation in the tumor microenvironment (TME). Overall, our studies show that ER stress response is upregulated in PDAC tumors, and stress response promotes cell death under in vitro TME-like conditions. Subtype features such as EMT and deltaNP63 do not alter PDAC cells’ ER stress responses, and had minimal impact on their stress response reliance. Citation Format: Yanqing (Christine) Jiang, Xu Han, John Tobias, Carson Poltorack, Mai Wang, Pamela Burgess-Jones, Nathan Coffey, Brian Keith, Celeste Simon. Investigating ER stress responses in pancreatic ductal adenocarcinoma under lipid imbalance [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr C039.

PSLBI-22 Identifying plasma metabolites influencing body weight in Salmonella challenged growing pigs: A machine learning approach

Abstract Body weight (BW) is an important component of pig productivity. Understanding the metabolic factors affecting growth rate is key to guiding nutritional strategies and bringing economic benefits to pig farms. Machine learning (ML) algorithms have been used to capture patterns in large datasets and predict animal production traits based on integrated information of performance data and blood records. However, utilizing ML models and metabolomic profiling to predict BW of pigs in small datasets with challenged animals has yet to be reported. Therefore, this study investigated use of plasma metabolites in predicting BW of pigs at the growing stage. This simulation used data from gilts (n = 40; BW = 25.6 ± 2.3 kg) raised over a 28-d trial. Plasma samples and BW were collected on d 0, 14, and 28 after 8 h of fasting. Ninety plasma metabolites were identified and quantified from an untargeted metabolomics assay. The development dataset included 30, and the validation dataset contained 10 unique animals with repeated measures in time. The metabolites were log-transformed, Pareto-scaled and missing values were imputed using the minimum nonzero value method. The BW prediction model was performed in R with the caret package. The optimal hyper-parameter configuration for the random forest and neural network algorithms was evaluated based on repeated cross-validation. The final model was the random forest composed of 400 trees with 45 randomly selected predictors. Model accuracy was estimated on repeated cross-validation with 10-folds and 5 repeats and compared with the validation set based on the coefficient of determination (R2), the root mean squared error (RMSE), and Pearson correlation (r). The metabolites with greater contribution with variable importance score above 10 to the model development were extracted. Aiming to understand differences in BW categories, the model response was visualized with an alluvial diagram for the 3 variables with the greatest metabolite importance, and BW was classified as high, medium, and low. The main metabolites driving BW were lysine, oleic acid, aminomalonic acid, myo-inositol, and linoleic acid. The RMSE after cross-validation was 7.22 kg of BW (r = 0.70), and R2 = 0.39. For pigs with a high BW, the predicted BW was explained by generally less plasmatic concentrations of lysine, oleic acid, and aminomalonic acid. Lysine is the main amino acid for muscle protein accretion, whereas oleic acid is for energy substrate. Aminomalonic acid is a metabolite resulting from glycine metabolism and is important in reversing oxidative damage caused by pathogens. Therefore, decreased plasma lysine, amino malonic acid, and oleic acid may indicate faster growth and lean deposition, leading to heavier BW in pigs. Using random forests helped to connect the plasma dynamics to BW and is a promising avenue to understand the main drivers in BW of group-housed pigs.

PSLBI-7 Coating extruded kibble with increasing levels of free fatty acids does not decrease palatability or impact e-nose odor profiles

Abstract Complete uncoated extruded kibble was coated with 5% by weight premium poultry fat (5,000 ppm Naturox, 0.57% FFA; Tyson, Springdale, AR) spiked with 0, 5, 10, 15, or 20 percent free fatty acids (Oleic acid, Millipore Sigma. Burlington, MA), and 3% liquid chicken-based palatant (Tyson, Springdale, AR). Kibbles were then utilized in oral palatability (n = 20 dogs; 10 male, 10 female), aromatic palatability (n = 20 dogs; 10 male, 10 female), and apparent total tract digestibility (ATTG; n = 36 dogs; 18 male, 18 female). Kibble odors were by ultra-fast gas chromatography e-nose (AlphaMOS, Toulouse, France), and SPME GC-MS (MUMC, Columbia, MO). In the aromatic palatability trials, there were no significant differences in first approach (P ≥ 0.21), percent interaction time (P ≥ 0.16), or interaction ratio (P ≥ 0.94). In the oral palatability trials, there were no significant differences in first approach (P ≥ 0.26), first bite (P ≥ 0.50), percent consumed (P ≥ 0.15), or intake ratio (P ≥ 0.59). There was a significant increase in fat digestibility among diets coated with 20% FFA compared with the uncoated diet (P = 0.02), with intermediate diets showing a slight numerical increase but no significant difference. However, there was no significant difference in protein, carbohydrate, or dry matter digestibility (P ≥ 0.06). Principle component analysis of e-nose peaks showed minimal odor changes between samples (DI = -3). SPME GC-MS analysis identified 16 compounds correlated with FFA percentage (r2 ≥ 0.50). When used to coat kibble, poultry fat with up to 20% FFA did not appear to negatively impact palatability or overall odor profile, though it did modify specific volatile compounds.

Chloro-aluminum phthalocyanine encapsulation into liquid crystalline nanodispersions enhance skin penetration and phototoxicity in skin cancer cells

Non-melanoma skin cancer is one of the most common tumors in the world. Photodynamic therapy (PDT) is currently a non-invasive therapeutic option for the treatment of skin cancer and actinic keratosis. Chloro-aluminum phthalocyanine (AlClPc) is a second-generation photosensitizer that has been studied for application in PDT. However, it has the disadvantage of self-aggregating in a biological environment, which impairs its therapeutic action, making it a candidate for nanocarrier delivery. Among the nanocarriers, liquid crystalline nanodispersions (LCN) are biocompatible and present unique organizational characteristics that make them suitable for skin delivery of drugs. Herein, a new LCN based on oleic acid (OA), Phosal 75SA® containing or not D-α-Tocopherol polyethylene glycol 1000 succinate (TPGS) was developed and characterized aiming its application for skin cancer. The selected LCN with AlClPc showed nanometric size (152.3 ± 1.93 nm), adequate PDI (0.231 ± 0.009) and encapsulation efficiency (70.26 ± 3.87 %) and stability for 30 days. Also, LCN were characterized by polarized light microscopy and small angle X-ray diffraction (SAXS) resulting in hexagonal liquid crystalline nanodispersions. In the in vitro skin penetration test, LCN-AlClPc showed better penetration into the deeper skin layers than the control. LCN-AlClPc showed greater phototoxicity in the A431 tumor cell line than the control in the same cell line and also greater phototoxicity in A431 when compared to L929 fibroblast cells. Thus, LCN developed could improve the phototoxicity and skin penetration in vitro and are potential drug delivery systems for the treatment of non-melanoma skin cancer.

139 Metabolic regulation of intramuscular adipose tissue development

Abstract Acetate arguably is the most important substrate for de novo fatty acid biosynthesis in bovine adipose tissue. However, including glucose in incubation media in vitro strongly stimulates fatty synthesis from acetate; additionally, glucose carbon is essential for triacylglycerol synthesis from acetate. Our laboratory is unique in that we utilize bovine intramuscular (i.m.) and subcutaneous (s.c.) adipose tissue explants dissected from beef longissimus muscle immediately post-exsanguination to study bovine adipose tissue metabolism. Four decades ago, using this experimental technique, we established that glucose is a primary substrate for fatty acid biosynthesis in i.m. adipose tissue. Recent research in our laboratory has addressed the hypothesis that acetate and/or long-chain fatty acids (LCFA) differentially affect lipid filling of i.m. and s.c. adipose tissue. We had previously demonstrated that, whereas s.c. adipose tissue is highly responsive to isoproterenol (a non-selective ß-adrenergic agonist), i.m. adipose tissue is relatively refractory to isoproterenol. This subsequently was confirmed in our laboratory utilizing primary cultures of bovine s.c. and i.m. adipocytes. We currently are focusing on the G-coupled protein receptors-43 (GPR43) and GPR120. Several previous studies from our laboratory have documented GPR43 expression in i.m. and s.c. adipose tissue, and we more recently demonstrated GPR120 expression in bovine adipose tissues. Acetate is a ligand for GPR43, whereas LCFA are ligands for GPR120. The binding of ß-adrenergic agonists to ß-adrenergic receptors increases the production of cyclic adenosine monophosphate (cAMP), whereas binding of acetate and LCFA to GPR43 and GPR120, respectively, decreases cAMP production. Thus, binding of circulating acetate and LCFA to their respective GPR receptors putatively should lead to increase lipid filling of bovine adipose tissues. A current study from our laboratory demonstrated that acetate had no effect on forskolin-stimulated cAMP production in i.m. or s.c. adipose tissue explants unless the culture medium also included oleic acid (18:1n-9). A subsequent study confirmed that oleic acid and palmitic acid (16:0), but not acetate, strongly depressed forskolin-stimulated cAMP production in s.c. adipose tissue explants; oleic and palmitic acid were much less effective in depressing cAMP production in i.m. adipose tissue explants. Unusually, GPR43 expression was detected in neither s.c. adipose tissue nor i.m. adipose tissue in our most recent study, whereas GPR120 expression was readily detected in both adipose tissue types. We conclude the following from these recent studies: LCFA binding to GPR120 is more effective than acetate binding to GPR43 in promoting lipid filling in bovine adipose tissue; and LCFA binding to GPR120 putatively would promote greater lipid filling in s.c. adipose tissue than in i.m. adipose tissue.

Molecular profiling of triacylglycerols in bovine and goat milk powders via ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry

Triacylglycerols (TAG) play several nutritional, technological, and functional roles in the dairy industry, representing approximately 98 % of milk fat. In the present study, the total fatty acid composition and the sn-2 fatty acid positional distribution of bovine and goat milk powders fat were analyzed by using the gas chromatography-flame ionization detector. Furthermore, the identification of TAG molecular species from milk powders was conducted using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The separation of milk TAG was accomplished by using isopropanol/acetonitrile (90:10, v/v) as mobile phase A, and water/acetonitrile (60:40, v/v) as mobile phase B. Myristic, palmitic, stearic, and oleic acids were identified as the most abundant fatty acids making up 11.55 and 10.71 %, 33.23 and 30.64 %, 10.86 and 12.52 %, and 26.74 and 21.80 % in the bovine and goat milk powders, respectively. In bovine milk, 221 molecular species of TAG were identified, with the predominant molecular species (LaCyO, CaLaPa, CyPaM, OCaCa, CaGCy, GLaCo) detected at a retention time of 6.362 min, with a relative abundance of 8.28 %. In goat milk, 202 molecular species were identified, with the major molecular species (CaLaM, CaPCa, CaCyS, CaCoAra, CaBuBe, LaPCy, LaCoS, LaBuAra, MPCo, MCyM, MBuS, PBuP, CyCoBe) detected at a retention time of 6.106 min, making up 8.14 % of the total TAG detected. These results provide valuable insights into milk fat composition for the dairy sector, aiding in the development of specialized dairy products.