Acid Profile Research Articles

Aroma volatiles and sensory quality of organically-grown apple cultivars after dynamic controlled atmosphere (DCA) storage and comparison with CA-stored fruit with and without 1-methylcyclopropene (1-MCP)

Long-term storage of many apple cultivars can be especially challenging for organic production where use of 1-methylcyclopropene (1-MCP), a crop protectant that controls ripening is disallowed. For many cultivars, controlled atmosphere (CA) storage alone, is inadequate for managing disorders and ripening. Dynamic controlled atmosphere (DCA), by determining the pO2 just above the lower oxygen limit (LOL), permits the use of lower pO2 setpoints without any negative impacts of anaerobiosis. Consequently, DCA can replace 1-MCP for maintenance of fruit quality. The goal of this study was to determine where this held true for a selection of old and new release organically-grown cultivars in the context of sensory perception. Cultivars included ‘Enterprise’ and ‘GoldRush’ harvested in 2018 (year 1), and ‘Honeycrisp’, ‘Golden Delicious’, ‘Jonagold’, and ‘Fuji’ harvested in 2020 (year 2). The LOL was monitored for DCA using chlorophyll fluorescence (CF) and compared with conventional static CA with or without 1-MCP treatment at harvest. DCA storage suppressed ripening to a similar level as 1-MCP plus CA in both years as indicated by reduced ester biosynthesis and, for some cultivars, maintenance of flesh firmness. However, ripening suppression did not impact the overall sensory preference of ‘Enterprise’, ‘Honeycrisp’, or ‘Fuji’, as all cultivars retained firmness and tartness regardless of storage regime and even negatively impacted it for ‘Goldrush’, an especially tart and firm cultivar. Sensory liking of ‘Golden Delicious’ and ‘Jonagold’ which have more typical diminishing firmness and acid profiles were most benefited by either 1-MCP treatment or DCA storage. The results highlight that use of storage technologies that suppress ripening beyond conventional CA storage are best employed on cultivars with more conventional softening and acid metabolism profiles as opposed to new cultivars that were selected primarily against these ripening phenotypes. The results also demonstrate the value of sensory evaluation compared with instrumental measurements to reveal the influence of different storage regimes on human perception.

Metabolomics of IgE-Mediated Food Allergy and Oral Immunotherapy Outcomes based on Metabolomic Profiling.

The immunometabolic mechanisms underlying variable responses to oral immunotherapy (OIT) in patients with IgE-mediated food allergy are unknown. To identify novel pathways associated with tolerance in food allergy, we used metabolomic profiling to find pathways important for food allergy in multi-ethnic cohorts and responses to OIT. Untargeted plasma metabolomics data were generated from the VDAART healthy infant cohort (N=384), a Costa Rican cohort of children with asthma (N=1040), and a peanut OIT trial (N=20) evaluating sustained unresponsiveness (SU, protection that lasts after therapy) versus transient desensitization (TD, protection that ends immediately afterwards). Generalized linear regression modeling and pathway enrichment analysis identified metabolites associated with food allergy and OIT outcomes. Compared with unaffected children, those with food allergy were more likely to have metabolomic profiles with altered histidines and increased bile acids. Eicosanoids (e.g., arachidonic acid derivatives) (q=2.4×10 -20 ) and linoleic acid derivatives (q=3.8×10 -5 ) pathways decreased over time on OIT. Comparing SU versus TD revealed differing concentrations of bile acids (q=4.1×10 -8 ), eicosanoids (q=7.9×10 -7 ), and histidine pathways (q=0.015). In particular, the bile acid lithocholate (4.97[1.93,16.14], p=0.0027), the eicosanoid leukotriene B4 (3.21[1.38,8.38], p=0.01), and the histidine metabolite urocanic acid (22.13[3.98,194.67], p=0.0015) were higher in SU. We observed distinct profiles of bile acids, histidines, and eicosanoids that vary among patients with food allergy, over time on OIT and between SU and TD. Participants with SU had higher levels of metabolites such as lithocholate and urocanic acid, which have immunomodulatory roles in key T-cell subsets, suggesting potential mechanisms of tolerance in immunotherapy. - Compared with unaffected controls, children with food allergy demonstrated higher levels of bile acids and distinct histidine/urocanic acid profiles, suggesting a potential role of these metabolites in food allergy. - In participants receiving oral immunotherapy for food allergy, those who were able to maintain tolerance-even after stopping therapyhad lower overall levels of bile acid and histidine metabolites, with the exception of lithocholic acid and urocanic acid, two metabolites that have roles in T cell differentiation that may increase the likelihood of remission in immunotherapy. This is the first study of plasma metabolomic profiles of responses to OIT in individuals with IgE-mediated food allergy. Identification of immunomodulatory metabolites in allergic tolerance may help identify mechanisms of tolerance and guide future therapeutic development.

Effects of verjus acidification on muscat canelli grape juice and wines

Our study aimed to investigate the use of verjus (green grape juice) as an acidifying agentfor improving the overall acid profile and balance of wine, lowering the pH and alcohol content and creating a sustainable winemaking practice that can be utilised in warm grape-growing regions, such as Texas, where pH levels range from 3.5 to over 4.0 in white juice and wine. Muscat Canelli wines were produced using two different treatments for pH and acidity balance. Verjus was produced by pressing under-ripe grapes resulting from crop thinning practices and added at two different rates of 2 % and 11 % to juice obtained from ripe grapes prior to fermentation. For all the wines produced, pH, titratable acidity (expressed as g/L Tartaric Acid), alcohol, glucose-fructose, l-malic acid, tartaric acid, potassium, polyphenols were analysed, and preference testing was performed via consumer sensory panels. Results obtained indicate that verjus can be used to balance pH and acidity as well as lower alcohol content. Consumer preference ranking indicated no significant differences between any of the treatments, while acidity rankings correctly distinguished the wines according to their acid levels. This work shows that the use of verjus as a pre-fermentation acidification agent could be a potential tool for winemaking to help mitigate the negative effects associated with climate change.

Probiotic Characteristics of Streptococcus thermophilus and Lactobacillus bulgaricus as Influenced by New Food Sources.

The current research aimed to evaluate the potential effects of Solanum mammosum, Dioon mejiae, and Amanita caesarea on Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus survival and performance after exposure to different harsh conditions such as bile, acid, gastric juice, and lysozyme to mimic the digestive system from mouth to the intestine. Probiotic protease activity was observed to evaluate the proteolytic system. Probiotics were cultured in a broth mixed with plant material, and after incubation, the results were compared to the control sample. Therefore, plant material's total phenolic compound, total carotenoid compound, antioxidant activity, sugar profile, and acid profile were obtained to discuss their impact on the survival of probiotics. The results indicate that Amanita caesarea negatively affected probiotic survival in the bile tolerance test and positively affected Lactobacillus bulgaricus in the protease activity test. Otherwise, the other plant material did not change the results significantly (p > 0.05) compared to the control in different tests. Consequently, Solanum mammosum and Dioon mejiae had no significant effects (p > 0.05) in increasing probiotic survival.

Insights into the organic matter composition of soda lakes in the Pantanal, Brazil, through fatty acids analysis in sediments.

We conducted a geochemical analysis on three sediment cores collected from soda and freshwater lakes in the Pantanal region, in Brazil. Our objective was to identify the primary sources of organic matter associated with the Pleistocene-Holocene transition, as previously documented in that area. The Nhecolandia region, located in the Pantanal, encompasses over 10,000 lakes, with approximately 10% of them exhibiting alkaline characteristics. Soda lakes became highly saline and alkaline after ~ 910calyr BP, which influences biogeochemistry and aquatic ecology. They have high electrical conductivity and pH can reach 10.5. In contrast to freshwater, soda lakes are absent of surrounding vegetation. Literature suggests a strong influence by the Last Glacial Maximum on the region. We hypothesized that periods of aridity and increased precipitation influenced the composition of organic matter present in sediments and preserved within these cores. Our analysis focused on examining the presence and distribution of fatty acids, organic carbon content, and total nitrogen. In general, the cores exhibited two distinct parts in terms of organic matter sources: the upper sections of the cores were primarily composed by terrestrial sources, identified by the presence of long-chain fatty acids, while the deeper sections were dominated by aquatic sources, therefore short-chain fatty acids. We did not find significant difference among fatty acid profile that could distinguish freshwater from soda lakes, the only remarkable difference was the occurrence of saturated fatty acids, which is lower in freshwater lake. These findings suggest the occurrence of humid and arid periods in the region. The C/N ratio displayed a similar trend to the fatty acid's profiles, exhibiting an abrupt change that was likely induced by climate variations. Although diagenesis can alter the composition of organic matter and, subsequently, the C/N values, it is noteworthy that the abrupt change observed in the Salina da Ponta (soda lake) core corresponds to 3,200years BP. This coincides with documented climate changes that occurred during the Holocene. Our study revealed the influence of past climatic conditions on the sources and variations of organic matter in sediment cores from the Pantanal's soda and freshwater lakes. Consideration of diagenesis and climate variations is crucial for interpreting sedimentary records.

A feasibility study of various joining techniques for three-dimensional printed polylactic acid and wood-reinforced polylactic acid biocomposite

The current research assesses various joining techniques such as an adhesive bond, direct three-dimensional printing, and ultrasonic welding for dissimilar three-dimensional printed thermoplastics such as polylactic acid and wood-reinforced polylactic acid biocomposite. This study is the first of its kind to determine an effective technique for joining three-dimensional printed polylactic acid/wood-polylactic acid profiles. Mechanical responses such as lap shear strength and shore D hardness of the various joints are investigated and compared experimentally. The results highlight that 15%–17% higher shear strength can be obtained for ultrasonically welded joints compared with direct three-dimensional printing of polylactic acid and wood polylactic acid lap joints. The macroscopic investigation of the ultrasonic welded polymeric joint exhibits a good level of melting of polymers and wetting in the interface. This results in an inter-molecular diffusion of polymeric chains and entanglement of polymers under respective conditions.

Effect of cereal bran phenolic profile on Maillard reaction products formation during hydrothermal treatment

Maillard reaction products (MRP) contribute to sensory quality of various foods. Whole grains (WG) are rich in phenols which may influence Maillard reaction pathways during thermal processing and impact WG product sensory attributes. This study investigated how WG phenolic profile affects MRP formation. Amylase-hydrolyzed wheat (white and red) and sorghum (white, red, tannin) brans were hydrothermally processed at 150 °C/6 min, and characterized for MRP using colorimetry, fluorescence spectroscopy, HPLC-MS/MS, and HS-SPME/GC–MS. Bran phenolic structure, and to a lesser extent content, had larger influence on MRP formation than protein/amino acid profile. Polymeric tannins (both in situ and when added to wheat brans) strongly inhibited volatile and non-volatile MRP intermediates and melanoidin formation, likely via their carbocation depolymerization intermediates trapping furans. Principle component analysis demonstrated clear segregation of volatiles formation based on bran phenolic profile. Phenolic composition should be considered in WG product formulation and processing to achieve desired MRP formation.

Microclimate of Grape Bunch and Sunburn of White Grape Berries: Effect on Wine Quality.

This research aimed to evaluate the composition of wines made with white grapes which are particularly susceptible to sunburn symptoms due to the absence of anthocyanin. Sunburn is a complex physiological dysfunction leading to browning or necrosis of berry tissues. In vintage 2021, the canopy of 'Verdeca' grapevines grown in Salento, South Italy, was differently managed by sun exposing or shading the bunches. Micrometeorological conditions were studied at different levels. Grapes were vinified, comparing the winemaking with and without skin maceration. The vegetative-productive balance of plants was not substantially modified. On the contrary, a significant effect was observed on the quality and quantity of grapes produced: smaller berries with sunburn symptoms were found on unshaded bunches. This influenced the percentage distribution among skin, pulp and seeds, causing a decrease in must yield of up to 30%. The pH was significantly higher in macerated wines made using shaded grapes, due to a lower titratable acidity and to significant impacts on the acid profile. Obviously, maceration produced a higher extraction of phenolics in wines, which reached their maximum in wines made with sunburned grapes. The absorbance at 420 nm, index of yellow color, was also significantly higher in sunburned grapes, indicating greater oxidation. Even though excessive grape sun-exposure could negatively affect the perception of white wines made without maceration (resulting in more oxidative character), the sensory quality of orange/amber wines was not significantly impacted by the presence of sunburned grapes. Thus, this winemaking technique could be particularly interesting to set up a production strategy adapted to viticultural regions strongly affected by climate change.

The Potential of Traditional ‘Gaja’ and New Breed Lines of Waxy, Blue and Purple Wheat in Wholemeal Flour Fermentation

The aim of this study was to analyse and compare the acidity, microbiological and colour characteristics, fatty (FA) and amino (AA) acid profiles, biogenic amine (BA) and gamma-aminobutyric acid (GABA) concentrations, and macro- and microelement contents in non-treated (non-fermented) and fermented wholemeal cereal flours of ‘Gaja’ (traditional wheat) and new breed lines DS8888-3-6 (waxy wheat), DS8548-7 (blue wheat) and DS8535-2 (purple wheat). Independent fermentations were undertaken with selected strains of Pediococcus acidilactici, Liquorilactobacillus uvarum and Lactiplantibacillus plantarum. The results revealed that all the wholemeal cereal flours of the analysed wheat varieties are suitable for fermentation with the selected strains because all the fermented samples showed lactic acid bacteria (LAB) viable counts higher than 8.00 log10 CFU/g and desirable low pH values. In most of the cases, fermentation increased the concentration of essential amino acids in the wholemeal cereal samples, and the LAB strain used for fermentation proved to be a significant factor in all the essential amino acid content of wholemeal wheat (p ≤ 0.0001). When comparing the non-fermented samples, the highest GABA content was found in ‘Gaja’ and waxy wheat samples (2.47 µmol/g, on average), and, in all the cases, fermentation significantly increased GABA concentration in the wholemeal cereals. On the other hand, total levels of biogenic amines in wholemeal samples ranged from 22.7 to 416 mg/kg. The wheat variety was a significant factor in all the analysed macro- and microelement contents (p ≤ 0.0001) in the wholemeal cereals. Furthermore, fermentation showed to be a significant factor in most of the FA content of the wholemeal cereal samples. Finally, fermentation can also contribute to improving the biological and functional value of wholemeal wheat flours (by increasing essential amino acids and GABA concentrations); however, safety parameters (e.g., biogenic amines) also should be taken into consideration when optimizing the most appropriate technological parameters.

Lipid metabolism and m6A RNA methylation are altered in lambs supplemented rumen-protected methionine and lysine in a low-protein diet

BackgroundMethionine or lysine has been reported to influence DNA methylation and fat metabolism, but their combined effects in N6-methyl-adenosine (m6A) RNA methylation remain unclarified. The combined effects of rumen-protected methionine and lysine (RML) in a low-protein (LP) diet on lipid metabolism, m6A RNA methylation, and fatty acid (FA) profiles in the liver and muscle of lambs were investigated. Sixty-three male lambs were divided into three treatment groups, three pens per group and seven lambs per pen. The lambs were fed a 14.5% crude protein (CP) diet (adequate protein [NP]), 12.5% CP diet (LP), and a LP diet plus RML (LP + RML) for 60 d.ResultsThe results showed that the addition of RML in a LP diet tended to lower the concentrations of plasma leptin (P = 0.07), triglyceride (P = 0.05), and non-esterified FA (P = 0.08). Feeding a LP diet increased the enzyme activity or mRNA expression of lipogenic enzymes and decreased lipolytic enzymes compared with the NP diet. This effect was reversed by supplementation of RML with a LP diet. The inclusion of RML in a LP diet affected the polyunsaturated fatty acids (PUFA), n-3 PUFA, and n-6 PUFA in the liver but not in the muscle, which might be linked with altered expression of FA desaturase-1 (FADS1) and acetyl-CoA carboxylase (ACC). A LP diet supplemented with RML increased (P < 0.05) total m6A levels in the liver and muscle and were accompanied by decreased expression of fat mass and obesity-associated protein (FTO) and alkB homologue 5 (ALKBH5). The mRNA expressions of methyltransferase-like 3 (METTL3) and methyltransferase-like 14 (METTL14) in the LP + RML diet group were lower than those in the other two groups. Supplementation of RML with a LP diet affected only liver YTH domain family (YTHDF2) proteins (P < 0.05) and muscle YTHDF3 (P = 0.09), which can be explained by limited m6A-binding proteins that were mediated in mRNA fate.ConclusionsOur findings showed that the inclusion of RML in a LP diet could alter fat deposition through modulations of lipogenesis and lipolysis in the liver and muscle. These changes in fat metabolism may be associated with the modification of m6A RNA methylation.Graphical abstractA systematic graph illustrates the mechanism of dietary methionine and lysine influence on lipid metabolism and M6A. The green arrow with triangular heads indicates as activation and brown-wine arrows with flat heads indicates as suppression.

Aromatic Amino Acids: Phenylalanine and Tyrosine in Patients with Hypertension and Coronary Artery Disease

Aim. To evaluate changes in the profile of aromatic amino acids (AAA) in patients with cardiovascular diseases (CVD): hypertension and coronary artery disease (CАD) in comparison with healthy study participants.Material and methods. One hundred and thirty-one participants were included in the study: 58 participants were included in the hypertension group, 46 in the CАD group, and 27 participants without signs of CVD in the control group. We used ultrahigh-performance liquid chromatography in combination with a triple quadrupole analyzer to measure plasma AAA: phenylalanine and tyrosine (Phe, Tyr) in all study participants. The association of AAA with biochemical blood test parameters, echocardiography (EchoCG) parameters, blood pressure level and clinical characteristics was analyzed.Results. A statistically significant difference in the level of concentration of Phe and Tyr was revealed (p=0,002 and p=0,024, respectively), comparing the three groups. Post-hoc analysis showed differences in the circulating level of both amino acids in patients with CAD vs the control group (Phe p=0,008 and Tyr p=0,020). Also a statistically significant difference in the level of Phe of the hypertension and CАD groups (p=0,017) was found. A negative correlation of low-density lipoproteins (LDL) with the level of Phe (r=-0,685, p&lt;0,05) and Tyr (r=-0,583, p&lt;0,05), as well as the level of Phe with total cholesterol (r=-0,461, p&lt;0,05) was found in the group without CVD. In the hypertension group, only a weak positive correlation was found between very low-density lipoproteins and AAA levels (Phe r=0,326 and Tyr r=0,365, p&lt;0,05), while in patients with CAD, the level of Phe and Tyr was negative correlated with high-density lipoprotein (r=-0,378 and r=-0,543, respectively, p&lt;0,05), and the level of Tyr with LDL (r=0,349, p&lt;0,05). When isolating the group with proven atherosclerosis of peripheral and/or coronary arteries, a statistically significant difference was revealed between the group of patients with CVD and clinical and instrumental signs of atherosclerosis and the group of patients with CVD without proven atherosclerosis in Phe level (p=0,019).Conclusion. Concentrations of AAA were higher in patients with CVD, comparing with the control group. At the same time, an increase of the Phe level was associated with the presence of peripheral or coronary atherosclerosis. The revealed correlations of AAA with EchoCG parameters and lipid spectrum parameters require further study to understand the involvement of AAA in pathogenesis of CVD and its potential role as treatment target.

Blood and faecal biomarkers to assess dietary energy, protein and amino acid efficiency of utilization by growing and finishing pigs

BackgroundDiet evaluation and optimization is a slow and expensive process and it is not possible to do it at a farm level. This study aimed to use the blood serum metabolite (BSM) and faecal volatile fatty acid (VFA) profiles as potential biomarkers to identify changes in protein, amino acid and energy dietary content in growing and finishing pig diets at farm level.ResultsTwo studies were conducted. The first study (S1) included 20 pens of 11 pigs (87.0 ± 4.10 kg; 18 weeks old) assigned to 5 diets: control (C1), high or low crude protein (HP1 and LP1, respectively), and high or low net energy (HE1 and LE1, respectively). The second study (S2) included 28 pens of 11 pigs (41.3 ± 2.60 kg; 12 weeks old) assigned to 7 diets: control (C2), high or low crude protein (HP2 and LP2, respectively), high or low amino acid (HA2 and LA2, respectively), and high or low net energy (HE2 and LE2, respectively). Pigs were followed for 10 (S1) and 20 (S2) days, and blood and faecal samples were collected at 20 (S1) and 14 (S2) weeks of age. Data were analysed using general linear models and receiver operating characteristic curve analysis. Urea nitrogen showed the best results as a biomarker. Urea nitrogen was higher in pigs fed high protein diets, HP1 (13.6 ± 0.95 mg/dL) and HP2 (11.6 ± 0.61), compared to those fed low protein diets, LP1 (6.0 ± 0.95) and LP2 (5.2 ± 0.61; P < 0.001), showing good discrimination ability (Area under the curve (AUC) = 98.4 and 100%, respectively). These differences were not observed between diets LA2 (6.5 ± 0.61) and HA2 (8.7 ± 0.61; P > 0.05; AUC = 71.9%), which were formulated based on the ideal protein profile but with no excess of protein. Creatinine, triglycerides, branched-chain fatty acids, albumin, propionic acid, and cholesterol showed differences between at least 2 diets but only in one of the studies.ConclusionsUrea nitrogen showed high accuracy to detect excess of crude protein in growing and finishing pig diets. Other biomarkers like BCFA showed promising results and need to be further studied.

Chemical Composition of Cactus Pear Seed Oil: phenolics identification and antioxidant activity.

ObjectivesThe chemical composition of cactus pear seed oil (Opuntia ficus-indica [L.] Mill.) was analyzed in terms of its fatty acid composition, tocopherol content, phenolic identification, and the oil’s phenolic-rich fraction antioxidant power was determined.MethodsFatty acid profiling was performed by gas chromatography coupled to an FI detector. Tocopherols and phenolic compounds were analyzed by LC-FLD/UV, and the oil’s phenolic-rich fraction antioxidant power was determined by phosphomolybdenum, DPPH assay and β-carotene bleaching test.ResultsFatty acid composition was marked by a high unsaturation level (83.22 ± 0.34%). The predominant fatty acid was linoleic acid (66.79 ± 0.78%), followed by oleic acid (15.16 ± 0.42%) and palmitic acid (12.70 ± 0.03%). The main tocopherol was γ-tocopherol (172.59 ± 7.59 mg/kg. In addition, Tyrosol, vanillic acid, vanillin, ferulic acid, pinoresinol, and cinnamic acid were identified as phenolic compounds in the analyzed seed oil. Moreover, the oil’s phenolics-rich fraction showed a significant total antioxidant activity, scavenged DPPH up to 97.85%, and effectively protected β-carotene against bleaching (97.56%).ConclusionThe results support the potential use of cactus pear seed oil as a functional food.

Influence of In Ovo Leptin Injection into Yolk Sac on Embryo Development, Blood Biochemical Parameters and Lipid Metabolism of Broiler Chicks during Early Post-Hatching Period

The study aimed to investigate the effect of in ovo (IO) leptin injection on yolk sac leptin concentration, embryo development, blood lipid metabolism, and biochemical parameters of broiler chicks during the early post-hatching period. Hatching eggs were weighed and placed in an incubator with 100 eggs/4 replications/group. The groups were: non-injected group (C); and 3 injected groups with 100 μl of phosphate buffer solution (PBS); 0.5 μl leptin (L0.5), and 1 μl leptin (L1) in 100 μl of PBS. Pure leptin hormone dissolved in PBS was injected into the yolk sac on day 7 of incubation. Yolk/yolk sac leptin concentrations at the onset of incubation, on days 7 and 15 of incubation, and at hatching were determined. Embryo/chick development was determined at all ages. Serum leptin, tri-iodothyronine (T3), thyroxine (T4), creatine kinase, total protein, uric acid, and lipid profiles were measured at hatching and day 7 of chick age. The leptin level of the yolk was similar to the leptin level of the yolk sac at hatching. Leptin level of a yolk sac was higher on 7th day than at other ages. IO injection* age was significant for serum leptin level, which was similar to one-day chicks in the L0.5 at day 7 of chick age. T3, T4, creatine kinase, uric acid, lipid profiles, and embryo/chick development did not change with IO leptin. Serum HDL level was higher in the leptin groups than C and PBS groups at hatching. IO leptin application affects blood lipid metabolism depending on dose level for male broiler chicks at the early post-hatch period, without affecting embryo/chick development.Leptin level of yolk was similar to leptin level of yolk sac at hatching. Leptin level of yolk sac was higher on 7th day than other ages. IO injection* age was significant for serum leptin level, which was similar to one-day chicks in the L0.5 at day 7 of chick age. T3, T4, creatine kinase, uric acid, lipid profiles, and embryo/chick development did not change with IO leptin. Serum HDL level was higher in the leptin groups than C and PBS groups at hatching. IO leptin application affects blood lipid metabolism depending on dose level for male broiler chicks at early post-hatch period, without affecting embryo/chick development.

P-052 Targeted metabolomics (amino acids) profiling of seminal plasma distinguished teratozoospermia from normozoospermic samples

Abstract Study question Is there a difference in metabolic composition between teratozoospermia and normozoospermic seminal samples? Summary answer Teratozoospermia and normozoospermia in humans are distinguished by seminal fluid metabolome markers especially amino acid content. What is known already Metabolomic profiling is the study of metabolites as downstream gene expression products in physiological and pathological circumstances. Identifying the metabolome of human seminal plasma (HSP) is a new research area to screen putative (in)fertility biomarkers. Knowing the semen metabolic signature and whether there are differences in metabolic composition between the teratozoospermia sample and semen with normal morphology could enhance our understanding of aspects associated with poor sperm quality and consequent male infertility. Study design, size, duration The Ethics Committee of Islamic Azad Tehran Medical Sciences University-Pharmacy and Pharmaceutical Branches approved the case-control study. Teratozoospermia was defined based on 5th WHO (2010) as normal morphology&amp;lt;4%, count&amp;gt;15*106/ml, progressive and non-progressive motility≥40%. In contrast, in normozoospermic samples from fertile men (who had at least one child), normal morphology was greater than 4%. HSP specimens were obtained from 15 infertile patients with teratozoospermia, and 12 confirmed fertile men as the control group. Participants/materials, setting, methods Fresh semen samples were obtained through masturbation following 3–4 days of abstention. A portion of each sample was used for routine semen analysis. Remained seminal plasma was isolated by centrifugation and stored; following post-thaw, HSPs were subjected to LC-MS-MSanalysis. MetaboAnalyst 5.0 software(www.metaboanalyst.ca) was utilized to identify fundamental metabolism alterations related to the morphology status of spermatozoa. In addition, different uni/multivariate analysis was performed to ascertain the group of metabolites responsible for the overall discrimination ability. Main results and the role of chance Seventeen significantly differentially metabolites (14 decreased and three increased) were observed in teratozoospermia samples vs. fertile control. Down-regulated metabolites include Cysteine, Gamma-aminobutyric acid, Glycine, Histidine, Hydroxylysine, Hydroxyproline, Methionine, Ornithine, Proline, Tryptophan, Aspartic acid, Argininosuccinic acid, Alpha-aminoadipic acid, and Beta-aminoisobutyric acid; whereas up. In contrast, up-regulatedlutamine, Asparagine, and Glycylproline. The PLS-DA score plot demonstrated the clear separation obtained according to the sperm morphological status. The two distinct groups were well-clustered, with distinct metabolic profiles for each group. The random forest (RF) algorithm was utilized to select relevant variables for teratozoospermia status classification by estimating the importance of each metabolite to teratozoospermia. The RF analysis defined a set of 15 metabolites that constitute the best predictors of teratozoospermia. In particular, increased Glutamine, Asparagine, and decreased Cysteine, Tryptophan, Glycine, and Valine were strong predictors of teratozoospermia. Interestingly, pathway enrichment analysis revealed that the most affected metabolic pathways in teratozoospermia men were the Aminoacyle-tRANA, arginine, valine-leucine, and isoleucine biosynthesis. Among 34 metabolites, Asparagine, Glutamine, and Glycylproline negatively correlated with sperm morphology status; whereas 21 metabolites were positively correlated with morphology, including Alanine, Argininesuccinic, Aspartic, Cystine, Gamma-aminobutyric acid, Glutamic, Glycine, Histidine, Homocitrulline, Hydroxylysine, Hydroxyproline, Isoleucine, Leucine, Methionine, Ornithine, Phenylalanine, Proline, Threonine, Tryptophan, Tyrosine, and Valine. Limitations, reasons for caution Since the strict criteria were considered for teratozoospermia patients, these results are preliminary and need to be confirmed with more sample size. Wider implications of the findings The possible cause of metabolite difference in seminal plasma of teratozoospermia patients comes from the disability of spermatozoa for selectively absorption or secretion of specific amino acids into/out of the cell, which is critical for sperm function and maturation and reflects the potential of fertility. Trial registration number Not Applicable

Abstract 2522: Pathway and immune profile analysis of cyst-derived versus PanIN-derived pancreatic ductal adenocarcinomas

Abstract Introduction: Pancreatic cystic lesions (PCL) are common and a subset of mucinous cysts will transform into pancreatic ductal adenocarcinoma (PDAC). However, predicting which of these mucinous PCL may progress to PDAC and warrant surgery remains a clinical challenge. Moreover, identifying those clinically emergent mucinous PCL for which a surveillance approach is best is a dire clinical need. Therefore, we aimed to identify molecular signatures that distinguished between PDAC with and without clinical evidence of a PCL to identify novel biomarkers. Methods: We leveraged data from the Oncology Research Information Exchange Network (ORIEN) multi-institute sequencing project and analyzed 66 PDAC cases recruited to ORIEN from The Ohio State University Wexner Medical Center and Moffitt Cancer Center for which tumor whole transcriptome sequencing datasets were generated. We separated the cases based on whether a tumor had originated from a cystic lesion (n=16) or presumably through the pancreatic intraepithelial neoplasia (PanIN) pathway (n=50). We then performed differential expression and pathway analysis using both Gene-Set Enrichment Analysis (GSEA) and Pathway Analysis with Down-weighted Genes (PADOG) algorithms. Based on the emerging importance of the immune landscape in PDAC development, we also analyzed immune profiles using a novel tool, Tumor-immune Microenvironment Deconvolution Web-portal for Bulk Transcriptomics (TIMEx). Results: When grouped by tumor origin, cyst-derived PDAC gene expression sets are enriched in immune signaling pathways, specifically NOTCH signaling (p=0.04), and demonstrate significant downregulation in amino acid metabolism, mitochondrial import and Gsα signaling pathways. Furthermore, GSEA based on TIMEx signatures indicated that multiple immune cell-specific profiles had significant enrichment scores in either the cyst-derived (for example, plasma cell: normalized enrichment score=-1.53; p=0.007) or non-cyst-derived (for example, neutrophil: normalized enrichment score=2.24; p=0.0001) PDAC cohorts. Conclusions: Our data suggest that cyst-derived and non-cyst-derived PDACs differ by immune profile, enhanced NOTCH pathway usage and in the metabolic processing of multiple amino acids. These initial findings support future studies to assess the accuracy of risk stratifying PCLs based on their amino acid, metabolic, or immune profiles, and exploration into mechanisms to explain these findings. Citation Format: Margaret A. Park, Somashekar G. Krishna, Maria C. Genilo-Delgado, Kristyn Gumpper-Fedus, Darwin L. Conwell, Phil A. Hart, Mary E. Dillhoff, Maria F. Gomez, Toni L. Basinski, Aamir N. Dam, Jason B. Klapman, Jason B. Fleming, Mokenge Malafa, Amir Mohammadi, Barbara A. Centeno, Kun Jiang, Daniel Jeong, Dung-Tsa Chen, Mengyu Xie, Aik Choon Tan, Brooke L. Fridley, Jamie K. Teer, Zobeida Cruz-Monserrate, Jennifer B. Permuth. Pathway and immune profile analysis of cyst-derived versus PanIN-derived pancreatic ductal adenocarcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2522.

Modelling Nannochloropsis gaditana Growth in Reactors with Different Geometries, Determination of Kinetic Parameters and Biochemical Analysis in Response to Light Intensity

Microalgae are unicellular and photosynthetic microorganisms which grow thanks to inorganic salts, CO2 and light, and find applications in several fields thanks to their variety. The industrial application of microalgae has not often been fully exploited because of a lack of information about how microalgae respond to inputs and to different growth environments. In the present work a model able to predict the microalgae growth in reactors with different geometries was developed. We combined a Monod-like model for the specific growth rate with the Lambert-Beer law of homogeneous light distribution in thick photobioreactors. Kinetic parameters related to the cultivation of the microalga Nannochloropsis gaditana were obtained, for the first time through batch cultivation under different photon flux densities inside a quasi-isoactinic photobioreactor, in order to obtain a practically homogeneous light distribution. The maximum specific growth rate and saturation constant resulted, respectively as µmax = 0.0256 h−1 and Ik = 15.28 µE s−1m−2. These parameters were applied to the model to obtain data on microalgae growth in different geometries. Model simulation results are presented and discussed. Furthermore, biochemical analysis was performed on the biomass obtained at the end of each batch cultivation, grown both under different light intensities and in reactors with different configurations. Results indicated that lipid content increases with increasing average photon flux density. The fatty acid and carotenoids profiles markedly shift when the average light intensity varies: the PUFA content decreases and the SFA content increases when the average light intensity rises, and an accumulation of carotenoids at lower photon flux densities is observed. In conclusion, the model resulted in a useful tool, able to predict the growth of the microalga Nannochloropsis gaditana in reactors with different configurations.

Silicone hydrogel contact lenses retain and document ocular surface lipid mediator profiles

ABSTRACT Clinical relevance A leading reason for patients to abandon their contact lenses is discomfort. Mechanisms and biomarkers for lens discomfort remain to be elucidated. Background Physical stress and tear film interaction are likely factors for lens discomfort. Lipid mediators are generated from polyunsaturated fatty acids. They regulate ocular surface physiology and pathophysiology, are constituents of human tears and may interact with contact lenses. This study set out to determine if hydrogel lenses and silicone hydrogel lenses interact with tear film polyunsaturated fatty acids and polyunsaturated fatty acids-derived mediators. Methods In vitro incubations, rat experiments and analysis of worn human lenses assessed polyunsaturated fatty acids and lipid mediator interactions with lenses. Silicone hydrogel and hydrogel lenses were incubated with lipid mediators and polyunsaturated fatty acids up to 24 hours. Rats were fitted with custom silicone hydrogel lenses and basal tears collected. Silicone hydrogel lenses worn for 2 weeks were obtained from 57 human subjects. Tear and lens lipidomes were quantified by mass spectrometry. Results Silicone hydrogel lenses retained polyunsaturated fatty acids and lipid mediators within 15 minutes in vitro. Lenses contained 90% of total polyunsaturated fatty acids and 83–89% of total monohydroxy fatty acids by 12 hours. Retention correlated with polarity of lipid mediators and lipophilic properties of silicone hydrogel lenses. Polyunsaturated fatty acids and lipid mediators such as lipoxygenase- and cyclooxygenase-derived eicosanoids were present in tears and worn lenses from rats. Worn silicone hydrogel lenses from human subjects established robust and lens-type specific lipidomes with high levels of polyunsaturated fatty acids, lipoxygenase-pathway markers and subject-specific differences in lipoxin A4 and leukotriene B4. Conclusion Worn silicone hydrogel lenses rapidly retain and accumulate tear polyunsaturated fatty acids and lipid mediators. Marked subject and lens type differences in the lipidome may document changes in ocular surface physiology, cell activation or infection that are associated with lens wear. If contact lens discomfort and adverse events induce specific tear and lens fatty acid and lipid mediator profiles warrants further studies.

A Detailed Comparative Study on Some Physicochemical Properties, Volatile Composition, Fatty Acid, and Mineral Profile of Different Almond (Prunus dulcis L.) Varieties

In the present investigation, the main purpose of the research was to reveal the differences among the almond genotypes in terms of their physicochemical properties, volatile composition, fatty acid, and mineral profile. For that reason, ten different almond genotypes originated from different countries were subjected to relevant analysis. The results showed that the total oil, protein, and ash levels of the almond samples ranged between 30.84–41.43%, 17.43–22.72%, and 2.90–3.40%, respectively. Additionally, total phenolic content of the samples was in the range of 38.7–101.03 mg GAE/100 g sample. It was revealed that the almond samples were rich in monounsaturated fatty acids, and oleic acid was the major one with levels of 61.22–77.63%. For all samples, potassium, magnesium, and phosphorus were the major minerals, and the highest concentration was for potassium with levels of 6192.08–11,046.05 mg/kg. Volatile profile analysis showed that the toluene, 4-octanone, pinacol, and 2-methylpentanal were the dominant volatile compounds for all almond genotypes. The results revealed that the different almond varieties showed significant differences depending on the origin.

Treatment With LAU-7b Complements CFTR Modulator Therapy by Improving Lung Physiology and Normalizing Lipid Imbalance Associated With CF Lung Disease.

Cystic fibrosis (CF) is the most common autosomal recessive genetic disease in Caucasians, affecting more than 100,000 individuals worldwide. It is caused by pathogenic variants in the gene encoding CFTR, an anion channel at the plasma membrane of epithelial and other cells. Many CF pathogenic variants disrupt the biosynthesis and trafficking of CFTR or reduce its ion channel function. The most frequent mutation, loss of a phenylalanine at position 508 (F508del), leads to misfolding, retention in the endoplasmic reticulum, and premature degradation of the protein. The therapeutics available for treating CF lung disease include antibiotics, mucolytics, bronchodilators, physiotherapy, and most recently CFTR modulators. To date, no cure for this life shortening disease has been found. Treatment with the Triple combination drug therapy, TRIKAFTA®, is composed of three drugs: Elexacaftor (VX-445), Tezacaftor (VX-661) and Ivacaftor (VX-770). This therapy, benefits persons with CF, improving their weight, lung function, energy levels (as defined by reduced fatigue), and overall quality of life. We examined the effect of combining LAU-7b oral treatment and Triple therapy combination on lung function in a F508deltm1EUR mouse model that displays lung abnormalities relevant to human CF. We assessed lung function, lung histopathology, protein oxidation, lipid oxidation, and fatty acid and lipid profiles in F508deltm1EUR mice.