Contribution of mono- and co-culture of Pseudomonas paralactis, Acinetobacter MN21 and Stenotrophomonas maltophilia to the spoilage of chill-stored lamb.

Exploring the contribution of common microorganisms to spoilage is of great significance in inhibiting spoilage in lamb. This work investigated the extent of protein degradation and profile changes of free amino acids (FAAs), free fatty acids (FFAs) and volatile organic compounds (VOCs) in lamb caused by single- and co-culture of the common aerobic spoilage bacteria, P. paralactis, Ac. MN21 and S. maltophilia. Meanwhile, some key VOCs produced by the three bacteria during lamb spoilage were also screened by orthogonal partial least square discriminant analysis and difference value in VOCs content between inoculated groups and sterile group. Lamb inoculated with P. paralactis had the higher total viable counts, pH, total volatile base nitrogen and TCA-soluble peptides than those with the other two bacteria. Some FAAs and FFAs could be uniquely degraded by P. paralactis but not Ac. MN21 and S. maltophilia, such as Arg, Glu, C15:0, C18:0 and C18:1n9t. Co-culture of the three bacteria significantly promoted the overall spoilage, including bacterial growth, proteolysis and lipolysis. Key VOCs produced by P. paralactis were 2, 3-octanedione, those by Ac. MN21 were 1-octanol, octanal, hexanoic acid, 1-pentanol and hexanoic acid methyl ester, and that by S. maltophilia were hexanoic acid. The production of extensive key-VOCs was significantly and negatively correlated with C20:0, C23:0 and C18:ln9t degradation. This study can provide a basis for inhibiting common spoilage bacteria and promoting high-quality processing of fresh lamb.

Exploring lipid digestion discrepancies between preterm formula and human milk: Insights from in vitro gastrointestinal digestion and the impact of added milk fat.

Lipids play a pivotal role in the nutrition of preterm infants, acting as a primary energy source. Due to their underdeveloped gastrointestinal systems, lipid malabsorption is common, leading to insufficient energy intake and slowed growth. Therefore, it is critical to explore the reasons behind the low lipid absorption rate in formulas for preterm infants. This study utilized a simulated in intro gastrointestinal digestion model to assess the differences in lipid digestion between preterm human milk and various infant formulas. Results showed that the fatty acid release rates for formulas IF3, IF5, and IF7 were 58.90 %, 56.58 %, and 66.71 %, respectively, lower than human milk's 72.31 %. The primary free fatty acids (FFA) and 2-monoacylglycerol (2-MAG) released during digestion were C14:0, C16:0, C18:0, C18:1n-9, and C18:2n-6, in both human milk and formulas. Notably, the higher release of C16:0 in formulas may disrupt fatty acid balance, impacting lipid absorption. Further investigations are necessary to elucidate lipid absorption differences, which will inform the optimization of lipid content in preterm infant formulas.

Revealing the intrinsic relationship between microbial communities and physicochemical properties during ripening of Xuanwei ham.

To clarify the relationship between microorganisms and physicochemical indicators of Xuanwei ham. Six ham samples for the first, second and third year were selected, respectively. The changes of physicochemical properties, the free fatty acids and microbial communities of Xuanwei ham were investigated by GC-MS and high-throughput sequencing technology. Results showed that scores of colour, overall acceptability, texture, taste and aroma were the highest in the third year sample. With increasing ripening time, moisture content, water activity (Aw), lightness (L*), springiness, and resilience decreased continuously, and yellowness (b*) was the highest in the second year sample. 31 free fatty acids were detected, and unsaturated fatty acids such as palmitoleic acid, oleic acid, and linoleic acid were the major fatty acids. The content of palmitoleic acid, oleic acid and eicosenoic acid increased significantly during processing. At the phylum level, the dominant bacteria were Proteobacteria and Firmicutes, and fungi were Ascomycota. At the genus level, the dominant bacteria were Staphylococcus and Psychrobacter, and fungi were Aspergillus. Correlation analysis showed that water content and Aw were closely related to microorganisms, and most unsaturated fatty acids were significantly correlated with microorganisms. These findings showed that microorganisms played an important role in the quality of Xuanwei ham, and provided a scientific basis for the quality control of Xuanwei ham.

Characterization of Palm Fatty Acid Distillate, Diacylglycerol Regioisomers, and Esterification Products Using High-Performance Size Exclusion Chromatography.

High-performance size exclusion chromatography (HPSEC) equipped with an evaporative light scattering detector (ELSD) was utilized for characterization of palm fatty acid distillate (PFAD) and its esterified products, with a particular focus on lipid profiles and diacylglycerol (DAG) regioisomers. The separation of triacylglycerol (TAG), DAG, monoacylglycerol (MAG), and free fatty acid (FFA) was achieved through a single 100-Å Phenogel column, coupled with a 2-cm C18 guard, utilizing toluene/acetic acid (100:0.25, v/v) as the mobile phase. This separation was based on size sieving principles and the interactions between the hydroxyl group(s) and the Phenogel matrix. The limit of detection (LOD) and limit of quantification (LOQ) for the esterified PFAD products analyzed by this method fell within the range of 4.8-5.5 µg/mL and 14.7-16.7 µg/mL, respectively. Additionally, the same column, paired with a 2-cm silica guard and a mobile phase comprised of toluene/isooctane/acetic acid (35:65:0.15, v/v/v), was used for the characterization of DAG regioisomers within the esterified PFAD. LODs and LOQs for sn-1,3-DAG and sn- 1,2-DAG were determined to be 39.2 and 118.7 µg/mL, and 32.8 and 99.5 µg/mL, respectively. Investigation of esterified PFAD products prepared using 4% H2SO4 at 120°C. After 2 h, the analysis revealed the highest MAG content at 31.85%, accompanied by 51.54% DAG, 2.35% TAG, and a residual 14.27% FFA. Notably, as the reaction time extended, the MAG content decreased, while both DAG and TAG levels exhibited an increasing trend. Further examination of DAG regioisomers during PFAD esterification, under varying catalyst concentrations (2-10%) and reaction temperatures (80-140°C), demonstrated a significant increase in the percentage of sn-1,3-DAG, inversely correlated with the reduction in FFA from 2% H 2 SO 4 and 80°C onwards. Remarkably, the percentage of sn-1,2-DAG remained relatively stable regardless of changes in catalyst concentrations or temperatures, confirming its susceptibility to isomerization into the thermodynamically more stable sn-1,3-DAG form. This study provides valuable insights into the composition and behavior of esterified PFAD products.

Analysis of changes in volatile compounds and evolution in free fatty acids, free amino acids, nucleotides, and microbial diversity in tilapia (Oreochromis mossambicus) fillets during cold storage.

BACKGROUND: Aquatic products are rich in nutrients and unique in flavor, and are popular among the public. However, aquatic products are extremely susceptible to quality degradation during storage, of which odor deterioration is the most obvious and influential aspect. Odor deterioration in aquatic products is widespread and severely affects overall flavor and quality. In this study, odor deterioration and flavor-related quality degradation of tilapia during cold storage are discussed, focusing on the changes in volatile compounds and the evolution of free fatty acids (FFAs), free amino acids (FAAs), nucleotides, and microbial diversity. RESULTS: A total of 63 volatile compounds were detected by gas chromatography-mass spectrometry, including 11 hydrocarbons, 10 alcohols, 6 aldehydes, 8 ketones, 6 esters, 9 aromatics, 3 phenols, and 10 other compounds. Microbial diversity analysis revealed that Acinetobacter, Psychrobacter, Vagococcus, and Myroides were the main dominant species of tilapia at the end of cold storage and predicted that microorganisms could influence the flavor of tilapia by participating in important metabolic pathways. Meanwhile, the evolution of FFAs, FAAs, and nucleotides also had a significant impact on odor deterioration, as evidenced by the contribution of unsaturated fatty acids (such as oleic acid and linoleic acid), Lys, and off-flavor nucleotides (HxR and Hx) to the undesirable flavor. Oxidation of oleic acid and linoleic acid resulted in changes in aldehydes, with Lys, HxR, and Hx being key flavor precursors and off-flavor contributors. CONCLUSION: This study contributes to a comprehensive overview of odor deterioration and the evolution of flavor-related quality in tilapia during cold storage, providing new insights into the regulation of overall flavor and quality. © 2023 Society of Chemical Industry.

High levels of short-chain fatty acids secreted by Candida albicans hyphae induce neutrophil chemotaxis via free fatty acid receptor 2.

Candida albicans belongs to our commensal mucosal flora and in immune-competent individuals in the absence of epithelial damage, this fungus is well tolerated and controlled by our immune defense. However, C. albicans is an opportunistic microorganism that can cause different forms of infections, ranging from superficial to life-threatening systemic infections. C. albicans is polymorphic and switches between different phenotypes (e.g. from yeast form to hyphal form). C. albicans hyphae are invasive and can grow into tissues to eventually reach circulation. During fungal infections, neutrophils in particular play a critical role for the defense, but how neutrophils are directed toward the invasive forms of fungi is less well understood. We set out to investigate possible neutrophil chemoattractants released by C. albicans into culture supernatants. We found that cell-free culture supernatants from the hyphal form of C. albicans induced both neutrophil chemotaxis and concomitant intracellular calcium transients. Size separation and hydrophobic sorting of supernatants indicated small hydrophilic factors as responsible for the activity. Further analysis showed that the culture supernatants contained high levels of short-chain fatty acids with higher levels from hyphae as compared to yeast. Short-chain fatty acids are known neutrophil chemoattractants acting via the neutrophil free fatty acid receptor 2. In line with this, the calcium signaling in neutrophils induced by hyphae culture supernatants was blocked by a free fatty acid receptor 2 antagonist and potently increased in the presence of a positive allosteric modulator. Our data imply that short-chain fatty acids may act as a recruitment signal whereby neutrophils can detect C. albicans hyphae.

Trajectory of stratum corneum lipid subclasses in the first year of life and associations with atopic dermatitis: A prospective cohort study.

BACKGROUND: Trajectories of stratum corneum (SC) lipid subclasses and their associations with infant atopic dermatitis (AD) are unclear. This study aimed to quantify the trajectories of 15 SC subclasses and carbon chain lengths and their associations with AD within 12 months. METHODS: In total, 213 newborns were enrolled at birth with nonlesional skin samples collected from the inner forearm at birth, 42 days, 3, 6, and 12 months, respectively. Lesional skin samples were collected from 120 AD patients at clinic with the disease onset within the first year of life. Mass spectrometry was applied to assess relative contents of 12 ceramide (CER), three free fatty acid (FFA) subclasses, and average carbon chain length (CCL). AD incident within 1 year old was diagnosed by dermatologists according to UK criteria. RESULTS: Sixty-four (30.0%) cases of ADs occurred in the cohort. All SC lipid subclasses and CCLs, but EOP varied significantly during the first year. AD infants showed lower NP but higher NS, NH, AP, hydroxy FFA, and CCL of FFAs compared with nonaffected infants. After normalization by age, the differences remained and were more pronounced in lesional skin of clinical AD infants compared with non-ADs. NS, NH, and CCL of FFAs in lesional skin of AD infants showed positive and significant correlations with the levels of transepidermal water loss at 3 month; some evidence supports a negative correlation for NP. CONCLUSIONS: We provide an overview of developmental trajectories of 15 CER and FFA subclasses across the first year of healthy infants and a link between the imbalance of some subclasses with the development of AD.

Effects of wildfire smoke PM2.5 on indicators of inflammation, health, and metabolism of preweaned Holstein heifers.

Wildfires are a growing concern as large, catastrophic fires are becoming more commonplace. Wildfire smoke consists of fine particulate matter (PM2.5), which can cause immune responses and disease in humans. However, the present knowledge of the effects of wildfire PM2.5 on dairy cattle is sparse. The present study aimed to elucidate the effects of wildfire-PM2.5 exposure on dairy calf health and performance. Preweaned Holstein heifers (N = 15) were assessed from birth through weaning, coinciding with the 2021 wildfire season. Respiratory rate, heart rate, rectal temperatures, and health scores were recorded and blood samples were collected weekly or twice a week for analysis of hematology, blood metabolites, and acute phase proteins. Hourly PM2.5 concentrations and meteorological data were obtained, and temperature-humidity index (THI) was calculated. Contribution of wildfires to PM2.5 fluxes were determined utilizing AirNowTech Navigator and HYSPLIT modeling. Mixed models were used for data analysis, with separate models for lags of up to 7 d, and fixed effects of daily average PM2.5, THI, and PM2.5 × THI, and calf as a random effect. THI ranged from 48 to 73, while PM2.5 reached concentrations up to 118.8 µg/m3 during active wildfires. PM2.5 and THI positively interacted to elevate respiratory rate, heart rate, rectal temperature, and eosinophils on lag day 0 (day of exposure; all P < 0.05). There was a negative interactive effect of PM2.5 and THI on lymphocytes after a 2-d lag (P = 0.03), and total white blood cells, neutrophils, hemoglobin, and hematocrit after a 3-d lag (all P < 0.02), whereas there was a positive interactive effect on cough scores and eye scores on lag day 3 (all P < 0.02). Glucose and NEFA were increased as a result of combined elevated PM2.5 and THI on lag day 1, whereas BHB was decreased (all P < 0.05). Contrarily, on lag day 3 and 6, there was a negative interactive effect of PM2.5 and THI on glucose and NEFA, but a positive interactive effect on BHB (all P < 0.03). Serum amyloid A was decreased whereas haptoglobin was increased with elevated PM2.5 and THI together on lag days 0 to 4 (all P < 0.05). These findings indicate that exposure to wildfire-derived PM2.5, along with increased THI during the summer months, elicits negative effects on preweaned calf health and performance both during and following exposure.

Wildfires contribute to fine particulate matter (PM2.5) pollution throughout the United States. Wildfire-PM2.5 exposure negatively affects human health and dairy cow production; however, the effects on calves are not known. We monitored preweaned calves exposed to natural wildfires to understand how wildfire-PM2.5 exposure affects calf health and performance. Calves exposed to wildfire PM2.5 and elevated temperature­humidity index (THI) experienced respiratory symptoms, alterations in blood cell composition and metabolism, and changes in circulating inflammatory proteins. These results suggest that PM2.5 along with increased THI induced an inflammatory response and alterations in energy metabolism that may contribute to calf health and performance deficits.

Influences of glyphosate residues and different concentrate feed proportions in dairy cow rations during early gestation on performance, blood parameters, functional properties and DNA damage of blood cells in cows and their offspring.

Maternal exposure to various stimuli can influence pre- and postnatal development of the offspring. This potential has been discussed for glyphosate (GLY), active substance in some non-selective herbicides. Accordingly, present study investigated putative effects of GLY residues in rations on cows and their offspring. Dams received either GLY-contaminated (GLY groups) or control (CON groups) rations combined with low (LC groups) or high (HC groups) concentrate feed proportions (CFP) for 16 weeks during mid- and late lactation and early gestation (59±4 days at beginning of GLY exposure; mean±SE). During this feeding trial, average daily GLY exposures of dams were 1.2 (CONLC), 1.1 (CONHC), 112.5 (GLYLC) and 130.3 (GLYHC) µg/kg body weight/d. After a depletion period (107±4 days; mean±SE) and calving, blood samples of dams and their calves were collected (5-345 min after birth) before calves were fed colostrum and analyzed for hematological and clinical-chemical traits, redox parameters, functional properties of leukocytes and DNA damage in leukocytes. No evidence for malformations of newborn calves could be collected. At parturition, most analyzed blood parameters were not affected by dietary treatment of dams during gestation. Significant GLY effects were observed for some traits, e.g. blood non-esterified fatty acids (NEFA) in calves. These deviations of GLY groups from CON groups likely resulted from strong time-dependent responses of NEFA levels within the first 105 minutes after birth and before colostrum intake (Spearman´s rank correlation R = 0.76, p<0.001). Additionally, significant GLY effects did not result in differences in measures that were beyond normally observed ranges questioning a pathological relevance. In summary, no evidence for teratogenic or other clear effects of GLY or CFP on analyzed parameters of dams and their newborn calves could be collected under applied conditions. However, detailed studies including GLY exposure during late and complete gestation period would be needed to rule out teratogenic effects.

Comparative analysis of intercellular lipid organization and composition between psoriatic and healthy stratum corneum.

The lipid composition and organization of the stratum corneum (SC) in patients with psoriasis and healthy subjects were compared using X-ray diffraction, Fourier-transform infrared spectroscopy (FT-IR), and ultraperformance liquid chromatography, combined with time-of-flight mass spectrometry (UPLC-TOFMS). In healthy SC (HSC), SC lipids formed two lamellar phases (long and short periodicity phases). Hexagonal and orthorhombic hydrocarbon-chain packing were observed in the lateral lipid organization at 30 °C via X-ray diffraction. In HSC, the lamellar phases and the hydrocarbon-chain packing organizations changed with elevated temperatures and finally disappeared. In these behaviors, the high-temperature hexagonal hydrocarbon-chain packing organization, which appeared above the orthorhombic hydrocarbon-chain packing organization, transformed to the liquid phase at about 90 °C in HSC. In psoriatic SC (PSC), hexagonal hydrocarbon-chain packing organization disappeared at about 65 °C with elevated temperatures. No high-temperature hexagonal hydrocarbon-chain packing organization were observed in PSC during heating process. Disorder of the hydrocarbon-chain packing of SC lipids was observed in PSC via FT-IR. In UPLC-TOFMS, free fatty acid (FFA) and ceramide (CER) compositions differed between patients with PSC and HSC. Specifically, the levels of ultra-long chain fatty acids containing CER and phytosphingosine-containing CER were decreased, while those of sphingosine and dihydrosphingosine-containing CER and unsaturated FFA were increased in PSC. Furthermore, FFA and CER carbon chain lengths decreased in patients with PSC. These results suggest that the alteration of SC lipid composition and the reduction of carbon chain lengths in PSC lowered the structural transformation temperature, thereby reducing barrier function.

Effect of shearing-induced lipolysis on foaming properties of milk.

BACKGROUND: The attraction of cappuccino-style beverages is attributed to the foam layer, as it greatly improves the texture, appearance, and taste of these products. Typical milk has a low concentration of free fatty acids (FFAs), but their concentration can increase due to lipolysis during processing and storage, which is detrimental to the foamability and foam stability of milk. There are contradictory results in reported studies concerning the effects of FFAs on the foaming properties of milk due to differences in milk sources, methods inducing lipolysis, and methods of creating foam. In this study, the foaming properties and foam structure of milk samples whose lipolysis was induced by ultra-turraxing, homogenisation, and microfluidisation (1.5-3.5 µ-equiv. mL-1 FFAs) were investigated. RESULTS: Compared with others, microfluidised milk samples had the smallest particle size, lowest absolute zeta potential, and highest surface tension; thus exhibited high foamability and foam stability, and very small and homogeneous air bubbles in foam structure. For all shearing methods, increasing FFA content from 1.5 to 3.5 µ-equiv. mL-1 markedly decreased the surface tension, foamability, and foam stability of milk samples. The FFA level that led to undesirable foam structure was 1.5 µ-equiv. mL-1 for ultra-turraxed milk samples and 2.5 µ-equiv. mL-1 for homogenised and microfluidised ones. CONCLUSION: Shearing-induced lipolysis greatly affected the physical properties of milk samples and subsequently their foaming properties and foam structure. At the same FFA level, lipolysis induced by microfluidisation was much less detrimental to the foaming properties of milk than lipolysis induced by ultra-turraxing and homogenisation. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Metabolic pathways altered by air pollutant exposure in association with lipid profiles in young adults.

Mounting evidence suggests that air pollution influences lipid metabolism and dyslipidemia. However, the metabolic mechanisms linking air pollutant exposure and altered lipid metabolism is not established. In year 2014-2018, we conducted a cross-sectional study on 136 young adults in southern California, and assessed lipid profiles (triglycerides, total cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol, very-low-density lipoprotein (VLDL)-cholesterol), and untargeted serum metabolomics using liquid chromatography-high-resolution mass spectrometry, and one-month and one-year averaged exposures to NO2, O3, PM2.5 and PM10 air pollutants at residential addresses. A metabolome-wide association analysis was conducted to identify metabolomic features associated with each air pollutant. Mummichog pathway enrichment analysis was used to assess altered metabolic pathways. Principal component analysis (PCA) was further conducted to summarize 35 metabolites with confirmed chemical identity. Lastly, linear regression models were used to analyze the associations of metabolomic PC scores with each air pollutant exposure and lipid profile outcome. In total, 9309 metabolomic features were extracted, with 3275 features significantly associated with exposure to one-month or one-year averaged NO2, O3, PM2.5 and PM10 (p < 0.05). Metabolic pathways associated with air pollutants included fatty acid, steroid hormone biosynthesis, tryptophan, and tyrosine metabolism. PCA of 35 metabolites identified three main PCs which together explained 44.4% of the variance, representing free fatty acids and oxidative byproducts, amino acids and organic acids. Linear regression indicated that the free fatty acids and oxidative byproducts-related PC score was associated with air pollutant exposure and outcomes of total cholesterol and LDL-cholesterol (p < 0.05). This study suggests that exposure to NO2, O3, PM2.5 and PM10 contributes to increased level of circulating free fatty acids, likely through increased adipose lipolysis, stress hormone and response to oxidative stress pathways. These alterations were associated with dysregulation of lipid profiles and potentially could contribute to dyslipidemia and other cardiometabolic disorders.

Anti-fatigue properties of the ethanol extract of Moringa oleifera leaves in mice.

BACKGROUND: Moringa oleifera (M. oleifera) leaves are rich in nutrients and bioactive ingredients. This study was aimed at evaluating the anti-fatigue effect of the ethanol extract of M. oleifera leaves (MLEE) on mice and its primary mechanism of action using a weight-loaded forced swimming test. In the present study, MLEE was prepared by ultrasound-assisted extraction, and its anti-fatigue effect and antioxidant capacity were evaluated in mice. Mice were administrated MLEE (320 mg kg-1 body weight) for 15 days. RESULTS: MLEE supplementation significantly increased levels of glucose and non-esterified fatty acids (NEFA), while decreasing levels of lactate and blood urea nitrogen in serum (P < 0.05); the levels of glycogen in the liver and muscle were also increased, as was the activity of glycogen synthase and the level of NEFA in muscle (P < 0.05). According to a Western blot analysis, MLEE increased the expression of AMPKα1, JNK, AKT and STAT3 in the muscle of mice. CONCLUSION: Our findings indicate that MLEE has an anti-fatigue effect via the AMPK-linked route, which enables it to control energy metabolism and enhance antioxidant enzyme activity. © 2023 Society of Chemical Industry.

Lipid metabolites, interleukin-6 and oxidative stress markers in follicular fluid and their association with serum concentrations in mares.

The application of trans-vaginal ovum pick up (OPU) and intracytoplasmic sperm injection (ICSI) is well established for commercial in vitro embryo production in horses. These assisted reproductive techniques are especially applied during the non-breeding season of the mare. However, little is known about how the health of the oocyte donor may affect the biochemical composition of the follicular fluid (FF) in small and medium-sized follicles routinely aspirated during OPU. This study aimed to investigate associations between systemic and FF concentrations of interleukin-6 (IL-6), total cholesterol, triglycerides, non-esterified fatty acids (NEFA), reactive oxygen metabolites (d-ROMs), biological antioxidant potential (BAP), and oxidative stress index (OSI) during the non-breeding season in mares. At the slaughterhouse, serum and FF of small (5-10 mm in diameter), medium (> 10-20 mm in diameter), and large (> 20-30 mm in diameter) follicles were sampled from 12 healthy mares. There was a strong positive association (P < 0.01) between the concentration of IL-6 in serum and those measured in small (r = 0.846), medium (r = 0.999), and large (r = 0.996) follicles. Serum concentrations of NEFA were positively correlated (P < 0.05) with those measured in small (r = 0.726), medium (r = 0.720), and large (r = 0.974) follicles. Values of total cholesterol and OSI in serum and medium follicles were significantly associated (r = 0.736 and r = 0.696, respectively). The serum concentrations of all lipid metabolites were markedly higher than those measured in FF of small- and medium-sized follicles. Values of IL-6 and OSI did not change significantly between serum and all follicle classes (P ≥ 0.05). To conclude, changes in the blood composition associated with inflammation, oxidative stress, and disturbed lipid metabolism of mares may lead to an inadequate oocyte microenvironment, which could affect oocyte quality and the success rate of OPU/ICSI programs. Further research should indicate whether these changes may ultimately affect in vitro oocyte developmental capacity and subsequent embryo quality.

Microphase transitions of Langmuir-Blodgett lipid-assembled monolayers with new types of ceramides, ultra-long-chain ceramide and 1-O-acylceramide.

HYPOTHESIS: Intercellular lipid lamellae, consisting of ceramide, cholesterol, and free fatty acids, are the primary pathways for substances in the stratum corneum (SC). The microphase transition of lipid-assembled monolayers (LAMs), mimicking an initial layer of the SC, would be affected by new types of ceramides: ceramide with ultra-long chain (CULC) and 1-O-acylceramide (CENP) with three chains in different direction. EXPERIMENTS: The LAMs were fabricated with varying the mixing ratio of CULC (or CENP) against base ceramide via Langmuir-Blodgett assembly. Surface pressure-area isotherms and elastic modulus-surface pressure plots were obtained to characterize π-dependent microphase transitions. The surface morphology of LAMs was observed by atomic force microscopy. FINDINGS: The CULCs favored lateral lipid packing, and the CENPs hindered the lateral lipid packing by lying alignment, which was due to their different molecular structures and conformations. The sporadic clusters and empty spaces in the LAMs with CULC were presumably due to the short-range interactions and self-entanglements of ultra-long alkyl chains following the freely jointed chain model, respectively, which was not noticeably observed in the neat LAM films and the LAM films with CENP. The addition of surfactants disrupted the lateral packing of lipids, thus weakening the LAM elasticity. These findings allowed us to understand the role of CULC and CENP in the lipid assemblies and microphase transition behaviors in an initial layer of SC.

Effects of milk types used in Antep cheese production on some cheese organoleptic quality parameters and brine composition during 5-month ripening.

Antep cheese is a local Turkish cheese characterized by scalding during production and ripened in brine. In this study, Antep cheeses were produced using mixtures of different milk types (cow, sheep, and goat milk) and ripened for 5 months. The composition, proteolytic ripening extension index (REI), free fatty acid (FFA) content, and volatile compounds of the cheeses and the variation of the brines were analyzed for the 5-month ripening period. Low proteolytic activity in cheese during ripening caused the cheeses to have low REI values (3.92%-7.57%), although it was observed that some parts of the water-soluble nitrogen fractions diffused into the brine, causing a lower REI. As a result of lipolysis during ripening, total FFA (TFFA) concentrations in all cheeses were increased, whereas the highest increases were detected in the concentrations of short-chain FFAs. The highest FFA concentrations were determined in cheese produced using goat milk, and the volatile FFA ratio in TFFA exceeded 10% in the third month of ripening. Although it was observed that the milk types used in the production had significant effects on the change of volatile compounds of the produced cheeses and their brines, the impact of the ripening time was more important. PRACTICAL APPLICATION: This study investigated Antep cheese made with different milk types. Volatile compounds and soluble nitrogen fractions were transferred to the brine by diffusion during ripening. The volatile profile of the cheese varied with milk type, but ripening time was the primary factor influencing volatile compounds. This suggests that the targeted organoleptic properties of the cheese are determined by ripening time and conditions. Additionally, changes in the brine's composition during the ripening process provide insight into how to manage the brine as waste.

High throughput FAMS - A fatty acid mass spectrometry method for monitoring polysorbate hydrolysis in QC.

Surfactant degradation in biopharmaceuticals has recently gained significant attention in the pharmaceutical industry. Specifically, hydrolytic degradation of polysorbates, leading to the release of free fatty acids potentially forming visible particles, is a key theme in technical development. To address this emerging topic, we present the development of a fully automated liquid-chromatography single quad mass detector method for the quantification of free fatty acids in biopharmaceuticals. For the first time, we have quantified the longer chain fatty acid degradation products of polysorbate, palmitic and stearic acid, allowing reliable detection and early critical insights for process improvements. This high-throughput method was validated underlining its robust performance in an interlaboratory trial as well as high flexibility allowing different robotic platforms and preparation techniques. The combination of automated sample preparation, separation by liquid chromatography and single quad mass detection makes the validated fatty acid mass spectrometry assay ready for routine use in a regulated environment.

Physicochemical Properties and Biological Activities of Koseret (Lippia adoensis Hochst. Var. Koseret) Seed and Leaf Oil Extracts.

BACKGROUND: Lippia adoensis Hoechst var. adoensis (wild variety) and variety koseret (cultivated variety) have been used as traditional medicine, condiments, and endemic to Ethiopia. OBJECTIVE: This study aimed to assess the physicochemical properties and biological activities of oil extracts from seed and leaves of koseret (L. adoensis var. koseret). METHODOLOGY: Soxhlet apparatus was used for oil extraction using hexane as a solvent. The oil quality assessment was based on oil yield, acid value, percent free fatty acid, and peroxide value, while the biological activities were investigated based on antioxidant and antimicrobial activities. The antimicrobial experiment was arranged as 2 x1x4 in a completely randomized factorial design with three replications. RESULTS: The result indicated that significantly higher oil yield (2.25%), acid value (2.66%) and free fatty acid (1.34%) were recorded for seed oil using the solvent extraction method. Leaf oil was recorded to have significantly higher values of DPPH (2,2- diphenyl-1- picrylhydrazyl), ascorbic acid and total carotenoid contents, but a lower value of hydrogen peroxide scavenging activity indicated that leaf oil presented higher antioxidant activity than seed oil in koseret. The koseret leaf oil demonstrated stronger antibacterial activity with a maximum zone of inhibition (14.50±0.21 mm), minimum inhibitory concentration (MIC, 0.25 µg/ml) and corresponding minimum bactericidal concentration (MBC, 0.25 µg/ml) against S. aureus. Furthermore, leaf oil has also presented stronger antifungal activity with a maximum zone of inhibition (14.83 mm), MIC (0.25 µg/ml), and minimum fungicidal concentration (MFC, 0.50 µg/ml) against Aspergillus versicolor. CONCLUSION: It can be concluded from the results of this study that leaf oil extract has demonstrated better biological activities, including both antioxidant and antimicrobial potentials, than seed oil in koseret.

Development and Validation of a New GC-FID Method for the Determination of Short and Medium Chain Free Fatty Acids in Wine.

A new analytical method for the determination of six volatile short and medium-chain fatty acids (acetic, propionic, isobutyric, isovaleric, hexanoic, and octanoic acids) through liquid-liquid extraction with diethyl ether, followed by GC-FID analysis, was developed and validated. The extraction conditions were optimized by evaluating the effect of the number of extractions (1 to 3) and the effect of the addition of salts (NaH2PO4, (NH4)2SO4, NaCl, (NH4)2SO4/NaH2PO4) to increase the concentration of the analytes in the ethyl ether phase. Results showed that a single extraction allows obtaining the highest sensitivity (due to the impossibility of evaporating the solvent to avoid losses of the analytes). The use of salting out agents, in particular, NaH2PO4, showed an important increase in the extraction extent, on average, 1.5 times higher as compared to the extraction performed without salt. The proposed method is rapid, requiring a total of 30 min for preparation and analysis, and it makes use of small amounts of sample (500 µL) and solvent (400 µL). The method was then applied to quantify the analytes in 5 white wines and 5 red wines, allowing to highlight some clear differences between red and white wines, with the red ones having a significantly higher amount of acetic acid (715.7 ± 142.3 mg/L in red wines and 351.5 ± 21.2 mg/L in white wines) and the white wines having a significantly higher amount of hexanoic and octanoic acid (6.1 ± 3.0 mg/L and 2.6 ± 0.8 mg/L, respectively, are the mean concentrations in white wines, and 4.7 ± 0.8 and 2.4 ± 0.4 mg/L, respectively, are the mean concentrations in red wines).

Free fatty acid profiling of Greek yogurt by liquid chromatography-high resolution mass spectrometry (LC-HRMS) analysis.

Yogurt is a fermented dairy product of high nutritional value, very popular in many parts of the world. Free fatty acids (FFAs), which are formed during fermentation, may cause changes in organoleptic properties of yogurt, and thus, the determination of FFAs is of importance. We present a liquid chromatography-high resolution mass spectrometry (LC-HRMS) method, which allows the simultaneous determination of a large set of common and uncommon FFAs in yogurt samples, avoiding any derivatization step. Twenty-five common saturated and unsaturated FAs, together with 21 saturated hydroxy fatty acids (SHFAs) and 17 saturated oxo fatty acids (SOFAs), were analyzed in 26 cow and 7 sheep Greek yogurt samples. A detailed analysis of bioactive SHFAs and SOFAs was carried out in yogurt samples for the first time. Differences at the concentrations of six common FAs and five oxidized FAs between the cow and sheep samples were observed. Based on these FAs, Principal Component Analysis (PCA) allows the discrimination of cow from sheep yogurt samples.