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When grapes are exposed to wildfire smoke, certain smoke-related volatile phenols (VPs) can be absorbed into the fruit, where they can be then converted into volatile-phenol (VP) glycosides through glycosylation. These volatile-phenol glycosides can be particularly problematic from a winemaking standpoint as they can be hydrolyzed, releasing volatile phenols, which can contribute to smoke-related off-flavors. Current methods for quantitating these volatile-phenol glycosides present several challenges, including the requirement of expensive capital equipment, limited accuracy due to the molecular complexity of the glycosides, and the utilization of harsh reagents. To address these challenges, we proposed an enzymatic hydrolysis method enabled by a tailored enzyme cocktail of novel glycosidases discovered through genome mining, and the generated VPs from VP glycosides can be quantitated by gas chromatography-mass spectrometry (GC-MS). The enzyme cocktails displayed high activities and a broad substrate scope when using commercially available VP glycosides as the substrates for testing. When evaluated in an industrially relevant matrix of Cabernet Sauvignon wine and grapes, this enzymatic cocktail consistently achieved a comparable efficacy of acid hydrolysis. The proposed method offers a simple, safe, and affordable option for smoke taint analysis.

Beer has over 600 flavor compounds and creates a positive tasting experience with acceptable sensory properties, which are essential for the best consumer experience. Spontaneous and mixed-culture fermentation beers, generally classified as sour beers, are gaining popularity compared to typical lager or ale styles, which have dominated in the USA for the last few decades. Unique and acceptable flavor compounds characterize sour beers, but some unfavorable aspects appear in conjunction. One such unfavorable flavor is called "mousy". This description is usually labeled as an unpleasant odor, identifying spoilage of fermented food and beverages. It is related as having the odor of mouse urine, cereal, corn tortilla chips, or freshly baked sour bread. The main compounds responsible for it are N-heterocyclic compounds: 2-acetyltetrahydropyridine, 2-acetyl-1-pyrroline, and 2-ethyltetrahydropyridine. The most common beverages associated with mousy off-flavor are identified in wines, sour beers, other grain-based beverages, and kombucha, which may contain heterofermentative lactic acid bacteria, acetic acid bacteria, and/or yeast/fungus cultures. In particular, the fungal species Brettanomyces bruxellensis are associated with mousy-off flavor occurrence in fermented beverages matrices. However, many factors for N-heterocycle formation are not well-understood. Currently, the research and development of mixed-cultured beer and non/low alcohol beverages (NABLAB) has increased to obtain the highest quality, sensory, functionality, and most notably safety standards, and also to meet consumers' demand for a balanced sourness in these beverages. This paper introduces mousy off-flavor expression in beers and beverages, which occurs in spontaneous or mixed-culture fermentations, with a focus on sour beers due to common inconsistency aspects in fermentation. We discuss and suggest possible pathways of mousy off-flavor development in the beer matrix, which also apply to other fermented beverages, including non/low alcohol drinks, e.g., kombucha and low/nonalcohol beers. Some precautions and modifications may prevent the occurrence of these off-flavor compounds in the beverage matrix: improving raw material quality, adjusting brewing processes, and using specific strains of yeast and bacteria that are less likely to produce the off-flavor. Conceivably, it is clear that spontaneous and mixed culture fermentation is gaining popularity in industrial, craft, and home brewing. The review discusses important elements to identify and understand metabolic pathways, following the prevention of spoilage targeted to off-flavor compounds development in beers and NABLABs.

The synthesis of a hapten and antigen for the preparation of a monoclonal antibody (mAb) for buprofezin is described. The recognition mechanism of hapten and buprofezin by monoclonal antibodies (mAb-19F2) is described. The effectiveness of the mAb-19F2 immunoassay technique was assessed, and the effective detection of buprofezin in tea samples was achieved through the establishment of indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and colloidal gold immunochromatography assay (GICA). The mAb-19F2 subtype was IgG1, with an IC50 of 1.8 ng/mL and a linear range (IC20-IC80) of 0.6-5.4 μg/L, and had a cross-reaction rate of less than 0.18% with 29 other pesticides (neonicotinoids and insect growth regulators). The study identified π-π stacking interactions between hapten and TYR-61 at the mAb-19F2 site and alkyl/phosphate interactions with TRP-105 and ARG-103. The ic-ELISA had an IC50 of 12.9 ng/mL in green tea and 5.65 ng/mL in black tea, with a recovery rate of 92.4%-101.0% and RSD of 2.1%-4.8%. The GICA had a limit of detection (LOD) was 500 ng/mL, with the complete disappearance of the test lines visible to the naked eye. The limit of quantitation (LOQ, IC20) was determined to be 16.8 ng/mL. Additionally, the developed GICA showed no cross-reactivity with neonicotinoid pesticides. The recovery rate of tea spiked recovered samples was 83.6%-92.2%, with an RSD of 5.3%-12.6%, and the results were consistent with the LC/MS method. This study is important for the real-time detection of buprofezin residues to ensure food safety and human health.

A heavy-chain antibody (VHH) library against procymidone (PRM) was constructed via immunizing Bactrian camels. Through careful biopanning, seven nanobodies (Nbs) with different sequences were obtained. The variability in their performance was primarily attributed to the amino acid differences in complementarity-determining region 3 (CDR3), as analyzed by molecular docking. The Nb exhibiting the highest sensitivity, named NbFM5, was biotinylated and conjugated to streptavidin-labeled gold nanoparticles to preserve the epitope's activity and prevent a decrease in sensitivity due to traditional random electrostatic adsorption. Subsequently, a simple and sensitive immunochromatographic assay (ICA) was developed for rapid detection of PRM based on biotinylated Nb (btNb). The developed btNb-ICA showed a cut-off value of 200 ng/mL for visual judgment and a half-inhibitory concentration (IC50) of 6.04 ng/mL for quantitative detection. The limit of detection (LOD) was as low as 0.88 ng/mL. The recoveries in actual samples of crops ranged from 82.2 to 117.3%, aligning well with the results obtained from GC-MS/MS (R2 = 0.995). In summary, the developed btNb-ICA demonstrated high specificity and good accuracy for the rapid detection of PRM residues in vegetables. The total analysis time from preparing the sample to obtaining the result was less than 25 min.

Natural products biosynthesized from biocontrol fungi in the rhizosphere can have both beneficial and deleterious effects on plants. Herein, we performed a comprehensive analysis of natural product biosynthetic gene clusters (BGCs) from the widely used biocontrol fungus Trichoderma afroharzianum T22 (ThT22). This fungus encodes at least 64 BGCs, yet only seven compounds and four BGCs were previously characterized or mined. We correlated 21 BGCs of ThT22 with known primary and secondary metabolites through homologous BGC comparison and characterized one unknown BGC involved in the biosynthesis of eujavanicol A using heterologous expression. In addition, we performed untargeted transcriptomics and metabolic analysis to demonstrate the activation of silent ThT22 BGCs via the "one strain many compound" (OSMAC) approach. Collectively, our analysis showcases the biosynthetic capacity of ThT22 and paves the way for fully exploring the roles of natural products of ThT22.

Research continues to provide compelling insights into potential health benefits associated with diets rich in plant-based natural products (PBNPs). Coupled with evidence from dietary intervention trials, dietary recommendations increasingly include higher intakes of PBNPs. In addition to health benefits, PBNPs can drive flavor and sensory perceptions in foods and beverages. Chardonnay marc (pomace) is a byproduct of winemaking obtained after fruit pressing that has not undergone fermentation. Recent research has revealed that PBNP diversity within Chardonnay marc has potential relevance to human health and desirable sensory attributes in food and beverage products. This review explores the potential of Chardonnay marc as a valuable new PBNP ingredient in the food system by combining health, sensory, and environmental sustainability benefits that serves as a model for development of future ingredients within a sustainable circular bioeconomy. This includes a discussion on the potential role of computational methods, including artificial intelligence (AI), in accelerating research and development required to discover and commercialize this new source of PBNPs.

As an important chemical pollutant affecting the safety of agricultural products, the on-site and efficient detection of pesticide residues has become a global trend and hotspot in research. These methodologies were developed for simplicity, high sensitivity, and multiresidue detection. This review introduces the currently available technologies based on electrochemistry, optical analysis, biotechnology, and some innovative and novel technologies for the rapid detection of pesticide residues, focusing on the characteristics, research status, and application of the most innovative and novel technologies in the past 10 years, and analyzes challenges and future development prospects. The current review could be a good reference for researchers to choose the appropriate research direction in pesticide residue detection.

Acrylamide is toxic aliphatic amide formed via the Maillard reaction between asparagine and reducing sugars during thermal processing of food. Herein, a specific nanobody termed Nb-7E against the acrylamide derivative xanthyl acrylamide (XAA) was isolated from an immunized phage display library and confirmed to be able to detect acrylamide. First, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established for acrylamide with a limit of detection (LOD) of 0.089 μg/mL and working range from 0.23 to 5.6 μg/mL. Furthermore, an enhanced electrochemical immunoassay (ECIA) was developed based on the optimized reaction conditions. The LOD was as low as 0.033 μg/mL, threefold improved compared to that of ic-ELISA, and a wider linear detection range from 0.39 to 50.0 μg/mL was achieved. The average recoveries ranged from 88.29 to 111.76% in spiked baked biscuits and potato crisps. Finally, the analytical performance of the ECIA was validated by standard ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS).

A simple and sensitive fluoroimmunoassay (FIA) based on a heavy-chain antibody (VHH) for rapid detection of fenitrothion was developed. A VHH library was constructed from an immunized alpaca, and one clone recognizing fenitrothion (namely, VHHjd8) was achieved after careful biopanning. It was biotinylated by fusing with the Avi tag and biotin ligase to obtain a fusion protein (VHHjd8-BT), showing both binding capacity to fenitrothion and the streptavidin poly-horseradish peroxidase conjugate (SA-polyHRP). Based on a competitive assay format, the absorbance spectrum of oxidized 3,3',5,5'-tetramethylbenzidine generated by SA-polyHRP overlapped the emission spectrum of carbon dots, which resulted in quenching of signals due to the inner-filter effect. The developed FIA showed an IC50 value of 1.4 ng/mL and a limit of detection of 0.03 ng/mL, which exhibited 15-fold improvement compared with conventional enzyme-linked immunosorbent assay. The recovery test of FIA was validated by standard GC-MS/MS, and the results showed good consistency, indicating that the assay is an ideal tool for rapid screening of fenitrothion in bulk food samples.

The stable nitrogen isotope ratio δ15N is used as a marker of dietary protein sources in blood. Crop fertilization strategies affect δ15N in plant foods. In a double-blinded randomized cross-over dietary intervention trial with 33 participants, we quantified the effect of fertilizer type (conventional: synthetic fertilizer and organic: animal or green manure) on δ15N in blood plasma. At study baseline, plasma δ15N was +9.34 ± 0.29‰ (mean ± standard deviation). After 12 days intervention with a diet based on crops fertilized with animal manure, plasma δ15N was shifted by +0.27 ± 0.04‰ (mean ± standard error) compared to synthetic fertilization and by +0.22 ± 0.04‰ compared to fertilization with green manure (both p < 0.0001). Accordingly, differences in the δ15N values between fertilizers are propagated to the blood plasma of human consumers. The results indicate a need to consider agricultural practices when using δ15N as a dietary biomarker.

Existing methods for the analysis of pesticides in human breast milk involve multiple extraction steps requiring large sample and solvent volumes, which can be a major obstacle in large epidemiologic studies. Here, we developed a simple, low-volume method for extracting organophosphates, pyrethroids, carbamates, atrazine, and imidacloprid from 100 to 200 μL of human breast milk. Multiple extraction protocols were tested including microwave-assisted acid/base digestion and double-solvent extraction with 2 or 20 mL of 2:1 (v/v) dichloromethane/hexane, with or without subsequent solid-phase extraction (SPE) cleanup. Samples were analyzed by liquid chromatography tandem mass spectrometry. Analyte recoveries and reproducibility were highest when 100-200 μL of milk were extracted with 2 mL of dichloromethane/hexane without subsequent SPE steps. Analysis of 79 breast milk samples using this method revealed the presence of carbamates, organophosphates, pyrethroids, and imidacloprid at detection frequencies of 79-96, 53-90, 1-7, and 61%, respectively. This study demonstrates the feasibility of a simple low-volume extraction method for measuring pesticides in human breast milk.

Saccharin and trans-resveratrol were incorporated into small quantity lipid-based nutritional supplements (SQ-LNS) to be evaluated as the markers of consumption for nutritional intervention studies. Forty-seven healthy women consumed a single supplement with either 8.6 mg of saccharin or 5 mg of trans-resveratrol, and urine was collected for 4 h. A rapid 11 min method employing multiple reaction monitoring and ultrahigh performance liquid chromatography coupled to a triple quadrupole mass spectrometer was developed to measure saccharin and resveratrol metabolites in urine simultaneously. The linear dynamic range of the method was from 3 to 1000 ng mL-1, with the correlation coefficient of 0.999 and limits of quantification from 15.28 to 53.03 ng mL-1. Sample preparation was simple dilution with an average recovery of 97.8%. Ion suppression was observed with urine concentrations >10%. Mean levels of saccharin and resveratrol-3-O-sulfate in urine were 5.481 ± 4.359 and 3.440 ± 4.160 nmol L-1, respectively. We developed and validated a method to measure saccharin and trans-resveratrol metabolites in urine to objectively corroborate the consumption of SQ-LNS for the first time in nutrition intervention studies.

Fish and algae oil supplements are enriched with eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are precursors to oxidized fatty acids, known as oxylipins. Here, we optimized a base hydrolysis method for measuring oxylipins in oil with ultrahigh-performance liquid chromatography coupled to tandem mass-spectrometry (UPLC-MS/MS) and quantified them in fish and algae oil supplements. Hydrolysis of 2 μL of oil with sodium carbonate resulted in greater oxylipin concentrations and minimal matrix effects, compared to higher oil volumes (10, 20, and 30 μL). Oxylipin yield was higher when oil was hydrolyzed in methanol containing 0.1% acetic acid and 0.1% butylated hydroxytoluene, compared to no methanol, and using sodium hydroxide versus sodium carbonate. Oxylipins extracted from 2 μL of oil using sodium hydroxide in solvent showed that EPA-derived oxylipins were most abundant in fish oil (84-87%), whereas DHA-oxylipins were abundant in algae oil (83%). This study shows that fish and algae oils are direct sources of EPA- and DHA-derived oxylipins.

Dietary intake of linoleic acid (LA, 18:2ω-6) has risen dramatically in recent decades. Previous studies have suggested a high intake of LA could increase tissue concentrations of proinflammatory and protumorigenic ω-6-series eicosanoid metabolites, increasing risks of inflammation and associated diseases. However, the effects of a LA-rich diet on in vivo profiles of eicosanoids and development of inflammatory diseases are understudied. Here, we treated spontaneous colitis-prone (Il-10-/-) mice with a control diet (∼3 Cal% LA) or a LA-rich diet (∼9 Cal% LA) for 18 weeks and analyzed the effects of the LA-rich diet on profiles of eicosanoids and development of colitis. We found that treatment with the LA-rich diet increased the tissue level of LA: the liver levels of LA were 5.8 ± 0.6% in the control diet-treated mice versus 11.7 ± 0.7% in the LA-rich diet-treated mice (P < 0.01). The plasma concentrations of a series of LA-derived metabolites, including 9-hydroxyoctadecadienoic acid (HODE), 9,10-dihydroxyoctadecenoic acid (DiHOME), 12,13-DiHOME, and 13-HODE were significantly increased by treatment with the LA-rich diet (P < 0.05). However, the LA-rich diet had little effect on the severity of colitis in the treated Il-10-/- mice. These results suggest a limited role of increased consumption of dietary LA on promoting colitis in the Il-10-/- model.

Limited reports on the use of nanobodies (Nbs) in fluorescence polarization immunoassay (FPIA) aroused us to explore if the small size of Nbs is a drawback for the development of sensitive FPIA to small molecular compounds, particularly since FPIA is a technology strongly dependent on molecular weight. In the present work, three different molecular weight Nbs against 3-phenoxybenzoic acid (3-PBA), an exposure biomarker of pyrethroid insecticides, including bare Nbs (15 kDa), Nbs-Avidin (Nbs-AV, 60 kDa), and Nbs-Alkaline phosphatase (Nbs-AP, 130 kDa) were specifically generated to cover distinct regions on the polarization and molecular weight relationship curve for a fluorescein tracer. In competitive FPIA, similar half-maximal inhibitory concentrations (IC50) of 3-PBA of 16.4, 12.2, and 14.8 ng mL-1 were obtained for Nbs, Nbs-AV, and Nbs-AP, respectively, indicating that the size of Nbs in the range tested had no significant effect on the sensitivity of the resulting competitive FPIA. An IC50 of 20.2 ng mL-1 for an anti-3-PBA polyconal antibody based FPIA further demonstrated the performance of Nbs, which was comparable to that of traditional antibodies in FPIA. Spike-recovery studies showed good and reproducible recovery of 3-PBA in urine samples, demonstrating the applicability of Nb-based FPIA. Overall, our results show that Nb-based FPIA achieves sensitivity levels of FPIA based on conventional antibodies and further indicate that Nb absolutely meets the sensitivity requirement of FPIA.

A monoclonal antibody (mAb) was raised against tebuconazole (TEB) using a hapten where the p-chloro substituent of the TEB molecule was replaced with a long-chain carboxylic acid. The resulting mAb showed high sensitivity and specificity against TEB characterized by ELISA with a half-maximal inhibitory concentration (IC50) of 0.19 ng mL-1 and with cross-reactivity (CR) values below 0.01% to several analogues of triazole fungicides. On the basis of the mAb produced, a quantum dot beads-based fluorescence immunochromatographic test strip assay (QBs-FITSA) was developed for rapid and sensitive detection of TEB in agricultural product samples. The QBs-FITSA exhibited a linear detection range from 0.02 to 1.25 ng mL-1 with a limit of detection (LOD) of 0.02 ng mL-1. Furthermore, using produced mAb, multiple high-throughput rapid immunoassay formats could be achieved as a convenient monitoring tool for evaluation of human and environmental exposure to TEB.

Although dicamba has long been one of the most widely used selective herbicides, some U.S. states have banned the sale and use of dicamba because of farmers complaints of drift and damage to nonresistant crops. To prevent illegal use of dicamba and allow monitoring of nonresistant crops, a rapid and sensitive method for detection of dicamba is critical. In this paper, three novel dicamba haptens with an aldehyde group were synthesized, conjugated to the carrier protein via a reductive-amination procedure and an indirect competitive chemiluminescent enzyme immunoassay (CLEIA) for dicamba was developed. The assay showed an IC50 of 0.874 ng/mL which was over 15 times lower than that of the conventional enzyme immunoassay. The immunoassay was used to quantify dicamba concentrations in field samples of soil and soybean obtained from fields sprayed with dicamba. The developed CLEIA showed an excellent correlation with LC-MS analysis in spike-and-recovery studies, as well as in real samples. The recovery of dicamba ranged from 86 to 108% in plant samples and from 105 to 107% in soil samples. Thus, this assay is a rapid and simple analytical tool for detecting and quantifying dicamba levels in environmental samples and potentially a great tool for on-site crop and field monitoring.

Nanoluciferase (Nluc), the smallest luciferase known, was used as the fusion partner with a nanobody against aflatoxin B1 to develop a bioluminescent enzyme immunoassay (BLEIA) for detection of the aflatoxin B1 in cereal. Nanobody (clone G8) against aflatoxin B1 was fused with nanoluciferase and cloned into a pET22b expression vector, and then transformed into Escherichia coli. The nanobody fusion gene contained a hexahistidine tag for purification by immobilized metal affinity chromatography, yielding a biologically active fusion protein. The fusion protein G8-Nluc retained binding properties of the original nanobody. Concentration of the coelenterazine substrate and buffer composition were also optimized to provide high intensity and long half-life of the luminescent signal. The G8-Nluc was used as a detection antibody to establish a competitive bioluminescent ELISA for the detection of aflatoxin B1 in cereals successfully. Compared to classical ELISA, this novel assay showed more than 20-fold improvement in detection sensitivity, with an IC50 value of 0.41 ng/mL and linear range from 0.10 to 1.64 ng/mL. In addition, the entire BLEIA detection procedure can be completed in one step within 2 h, from sample preparation to data analysis. These results suggest that nanobody fragments fused with nanoluciferase might serve as useful and highly sensitive dual functional reagents for the development of rapid and highly sensitive immunoanalytical methods.

Bovine milk oligosaccharides (BMOs) that resemble human milk oligosaccharides are found in whey permeate, indicating that dairy streams can be used as a potential source of bioactive oligosaccharides. Recovery of oligosaccharides from whey permeate is hindered by their low abundance and high concentration of lactose. In the present work, lactose in bovine colostrum whey permeate was hydrolyzed by Aspergillus oryzae β-galactosidase to facilitate subsequent monosaccharide removal by membrane separation. Chromatographic separation coupled with high-resolution mass spectrometry revealed β-galactosidase degradation of several β-linkage-containing BMOs and production of novel oligosaccharides that ranged in size from 5 to 11 monosaccharide units containing several galactose repeating units and N-acetylhexosamine at their reducing ends. Optimization of BMO hydrolysis and separation methodology could generate high amounts of hetero-oligosaccharides for improved recovery of potentially biotherapeutic oligosaccharides.

This study describes the sensory composition of commercial sweet almond varieties across two California growing seasons. It also discusses the relationship between sensory attributes and chemical and physical measures. Raw, whole almonds (43 samples each of 13 varieties in 2015 and 40 samples each of 10 varieties in 2016) were evaluated for their sensory profiles using descriptive sensory analysis. The 2016 samples were also analyzed for macro- and micronutrients, amygdalin, volatile composition (using gas chromatography-mass spectrometry), and physical properties, and the results were modeled with the sensory data. Independence, Sonora, and Wood Colony were harder, more fracturable, and crunchy, whereas Fritz and Monterey were more moist and chewy, reflecting their moisture contents. Aldrich and Fritz were higher in marzipan/benzaldehyde flavor, which is related to amygdalin, benzaldehyde, phenylethyl alcohol, and benzyl alcohol. New insights are provided into sweet-almond composition and the sensorial contribution of headspace volatiles. This assists almond growers and processors in describing and marketing almond varieties.

Understanding almond flavor, in terms of both sensory aspects and chemistry, is essential for processors to maintain almond quality and to correctly identify acceptable or unacceptable product. This overview of the sensory and chemical characteristics of almond flavor discusses raw and heat-processed almonds, the volatile compounds generated upon heating, the aroma qualities associated with various odorants, and the use of descriptive sensory analysis for sweet almonds. Flavor development and off-flavors in almonds due to rancidity is also explored. The review examines the existing methods used to assess common nonvolatile as well as volatile indicators of lipid oxidation in almonds and the correlation of these indicators with consumer acceptance. Recent research on the relationship among volatile profile, rancidity indicators, and consumer acceptance is presented.

Neonicotinoids (NN), first introduced in 1991, are found on environmental surfaces where they undergo photolytic degradation. Photolysis studies of thin films of NN were performed using two approaches: (1) transmission FTIR, in which solid films of NN and the gas-phase products were analyzed simultaneously, and (2) attenuated-total-reflectance FTIR combined with transmission FTIR, in which solid films of NN and the gas-phase products were probed in the same experiment but not at the same time. Photolysis quantum yields using broadband irradiation centered at 313 nm were (2.2 ± 0.9) × 10-3 for clothianidin (CLD), (3.9 ± 0.3) × 10-3 for thiamethoxam (TMX), and (3.3 ± 0.5) × 10-3 for dinotefuran (DNF), with all errors being ±1 s. At 254 nm, which was used to gain insight into the wavelength dependence, quantum yields were in the range of (0.8-20) × 10-3 for all NNs, including acetamiprid (ACM) and thiacloprid (TCD). Nitrous oxide (N2O), a potent greenhouse gas, was the only gas-phase product detected for the photolysis of nitroguanidines, with yields of ΔN2O/ΔNN > 0.5 in air at both 313 and 254 nm. The atmospheric lifetimes with respect to photolysis for CLD, TMX, and DNF, which absorb light in the actinic region, are estimated to be 15, 10, and 11 h, respectively, at a solar zenith angle of 35° and 12, 8, and 10 h at a solar zenith angle of 15°.

Olives are inedible because of high levels of bitter phenolics (e.g., oleuropein) which are removed during commercial olive processing. Current commercial processing methods are highly water-intensive, produce toxic wastewater, and are environmentally unsustainable. To address this, macroreticular polymeric resins were used to assist debittering and decrease water use. Amberlite resins XAD4, XAD16N, XAD7HP, and FPX66 were evaluated for the ability to adsorb bitter and/or high-value phenolic compounds (i.e., oleuropein, ligstroside, oleuropein aglycone, ligstroside aglycone, oleocanthal, oleacein, and hydroxytyrosol) from whole olives during typical brine storage. All resins effectively adsorbed oleuropein and ligstroside. FPX66 reduced oleuropein in whole olives suspended in a 1.0% acetic acid brine to 0.635 mg/kg wet weight in 2.5 months with no further processing. This concentration is below levels measured in commercial California-style black ripe olives (0.975 mg/kg wet weight). Resins in storage brines effectively decrease levels of bitter phenolic compounds without additional lye processing. Excellent recoveries of high-value phenolic compounds are obtained from resins (e.g., 80.2 ± 3.3% to 89.4 ± 8.9% hydroxytyrosol).

3-Phenoxybenzoic acid (3-PBA) is a human urinary metabolite of many pyrethroid insecticides and can be used as a biomarker to monitor human exposure to these pesticides. A rapid and sensitive direct competitive fluorescence enzyme immunoassay (dc-FEIA) for detecting 3-PBA on the basis of a nanobody (Nb)-alkaline phosphatase (AP) fusion protein was developed. The anti-3-PBA Nb-AP fusion protein was expressed and purified. The 50% inhibitory concentration (IC50) and linear range of dc-FEIA were 0.082 and 0.015-0.447 ng/mL, respectively, with a detection limit of 0.011 ng/mL. The IC50 of dc-FEIA was improved by nearly ten times compared with those of one-step and three-step direct competitive enzyme-linked immunosorbent assay (dc-ELISA). Spiked urine samples were detected by both dc-FEIA and liquid chromatography-mass spectrometry (LC-MS), and the results showed good consistency between the two analysis methods, indicating the reliability of dc-FEIA based on the Nb-AP fusion protein for detecting 3-PBA in urine.

Due to their unique properties, copper-based nanopesticides are emerging in the market. Thus, understanding their effect on crop plants is very important. Metabolomics can capture a snapshot of cellular metabolic responses to a stressor. We selected maize and cucumber as model plants for exposure to different doses of Cu(OH)2 nanopesticide. GC-TOF-MS-based metabolomics was employed to determine the metabolic responses of these two species. Results revealed significant differences in metabolite profile changes between maize and cucumber. Furthermore, the Cu(OH)2 nanopesticide induced metabolic reprogramming in both species, but in different manners. In maize, several intermediate metabolites of the glycolysis pathway and tricarboxylic acid cycle (TCA) were up-regulated, indicating the energy metabolism was activated. In addition, the levels of aromatic compounds (4-hydroxycinnamic acid and 1,2,4-benzenetriol) and their precursors (phenylalanine, tyrosine) were enhanced, indicating the activation of shikimate-phenylpropanoid biosynthesis in maize leaves, which is an antioxidant defense-related pathway. In cucumber, arginine and proline metabolic pathways were the most significantly altered pathway. Both species exhibited altered levels of fatty acids and polysaccharides, suggesting the cell membrane and cell wall composition may change in response to Cu(OH)2 nanopesticide. Thus, metabolomics helps to deeply understand the differential response of these plants to the same nanopesticide stressor.

Oleuropein, ligstroside, and related hydrolysis products are key contributors to olive bitterness, and several of these phenolics are implicated in the prevention of lifestyle age-related diseases. While table olive processing methods are designed to reduce oleuropein, the impact of processing on ligstroside and related hydrolysis products (e.g., oleacein, oleocanthal, hydroxytyrosol glucoside, ligstroside aglycone, and oleuropein aglycone) is relatively unknown. Herein, levels of these compounds were measured in Spanish-style green (SP), Californian-style black ripe (CA), and Greek-style natural fermentation (GK) olives using rapid ultrahigh-performance liquid chromatography (UHPLC) tandem mass spectrometry (MS/MS). GK olives had the highest concentration of all compounds measured, with the exception of oleocanthal, which was highest in SP olives (0.081 mg kg-1 wet weight (w.wt)). CA olives had the lowest levels of most compounds measured, including ligstroside (0.115 mg kg-1 w.wt) and oleuropein (0.974 mg kg-1 w.wt). Hydroxytyrosol was the predominate compound in all three styles of commercial olives, with similar concentrations observed for GK and SP olives (134.329 and 133.685 mg kg-1 w.wt, respectively) and significantly lower concentrations observed for CA olives (19.981 mg kg-1 w.wt).

Renal fibrosis is a common end point of the progression of chronic kidney disease (CKD). Suppressing the development and progression of renal fibrosis is essential in the treatment of kidney disease. Our previous study demonstrated that the ethyl acetate extract of the surface layer of Poria cocos exhibited beneficial antitubulointerstitial fibrosis. In this study, we isolated new diterpene (PZF) and triterpenes (PZG and PZH) and examined their antifibrotic effect. TGF-β1 upregulated the collagen I protein expression in HK-2 cells, and PZG and PZH treatment significantly inhibited the upregulated collagen I expression (TGF group 0.59 ± 0.08 vs TGF+PZG group 0.36 ± 0.08, P < 0.01; TGF+PZH group 0.39 ± 0.12, P < 0.01). Triterpenes, PZG and PZH, exhibited a stronger inhibitory effect on renal fibrosis and podocyte injury than PZF. PZG and PZH further showed a stronger inhibitory effect on the activation of the renin-angiotensin system (RAS) than PZF. Additionally, PZG and PZH markedly inhibited the activation of Wnt/β-catenin signaling, which played an important role in fibrogenesis. Interestingly, PZG and PZH suppressed the TGF-β/Smad pathway by selectively inhibiting the phosphorylation of Smad3 through blocking the interactions of SARA with TGFβI and Smad3. The analysis of the structure-activity relationship demonstrated that their antifibrotic effects were closely associated with the first six-membered ring structure and the number of carboxyl groups in this type of compounds. Additionally, fifteen known triterpenes were identified. These novel tetracyclic triterpenoid compounds provided the potential lead compounds for the research and development of antifibrosis drug, and they possessed the potential to be utilized as RAS inhibitors.

Monitoring oxidative flavor changes in almonds is possible only if the chemical and sensory profile during roasting and storage is first established. Herein, almonds roasted at two different temperatures (115 and 152 °C) were stored at 39 °C for 0 to 12 months and were analyzed by headspace solid-phase microextraction gas chromatography-mass spectrometry, descriptive analysis, and consumer hedonic analysis. Volatile profiles, descriptive sensory profiles, and consumer hedonic scores were analyzed for predictive relationships. Descriptive attributes involving Roasted and Nutty as well as consumer liking were highest in fresh almonds, while flavors typically associated with oxidative rancidity such as Cardboard, Painty/Solvent, Soapy, and Total Oxidized increased during storage. Compounds most important for predicting rancidity-related attributes were lipid oxidation products, including pentanal, hexanal, heptanal, and octanal. Consumer liking was best predicted by similar compounds to those predicting Clean Nutty flavor, including Maillard reaction products such as 2- and 3-methylbutanal, 2-methylpyrazine, and 2,5-dimethylpyrazine.

Cry toxins have been widely used in genetically modified organisms for pest control, raising public concern regarding their effects on the natural environment and food safety. In this work, a phage-mediated competitive chemiluminescent immunoassay (c-CLIA) was developed for determination of Cry1Ab toxin using anti-idiotypic camel nanobodies. By extracting RNA from camels' peripheral blood lymphocytes, a naive phage-displayed nanobody library was established. Using anti-Cry1Ab toxin monoclonal antibodies (mAbs) against the library for anti-idiotypic antibody screening, four anti-idiotypic nanobodies were selected and confirmed to be specific for anti-Cry1Ab mAb binding. Thereafter, a c-CLIA was developed for detection of Cry1Ab toxin based on anti-idiotypic camel nanobodies and employed for sample testing. The results revealed a half-inhibition concentration of developed assay to be 42.68 ± 2.54 ng/mL, in the linear range of 10.49-307.1 ng/mL. The established method is highly specific for Cry1Ab recognition, with negligible cross-reactivity for other Cry toxins. For spiked cereal samples, the recoveries of Cry1Ab toxin ranged from 77.4% to 127%, with coefficient of variation of less than 9%. This study demonstrated that the competitive format based on phage-displayed anti-idiotypic nanobodies can provide an alternative strategy for Cry toxin detection.

Concealed damage (CD), is a term used by the nut industry to describe a brown discoloration of kernel nutmeat that becomes visible after moderate heat treatments (e.g., roasting). CD can result in consumer rejection and product loss. Postharvest exposure of almonds to moisture (e.g., rain) is a key factor in the development of CD as it promotes hydrolysis of proteins, carbohydrates, and lipids. The effect of drying moisture-exposed almonds between 45 to 95 °C, prior to roasting was evaluated as a method for controlling CD in roasted almonds. Additionally, moisture-exposed almonds dried at 55 and 75 °C were stored under accelerated shelf life conditions (45 °C/80% RH) and evaluated for headspace volatiles. Results indicate that drying temperatures below 65 °C decreases brown discoloration of nutmeat up to 40% while drying temperatures above 75 °C produce significant increases in brown discoloration and volatiles related to lipid oxidation, and nonsignificant increases in Amadori compounds. Results also demonstrate that raw almonds exposed to moisture and dried at 55 °C prior to roasting, reduce the visual sign of CD and maintain headspace volatiles profiles similar to almonds without moisture damage during accelerated storage.

Changes of the abundance of caprine milk oligosaccharides (CMO) at different lactation stages have been evaluated by hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-Q MS) and nanoflow liquid chromatography-quadrupole-time-of-flight mass spectrometry (nano-LC-Chip-QTOF MS). Eight major oligosaccharides (OS) were quantified at different lactation stages by HILIC-Q MS, while the use of nano-LC-Chip-QToF MS allowed expanding the study to forty-nine different OS by monitoring neutral non- and fucosylated species, as well as acidic species containing not only N-acetyl-neuraminic acid or N-glycolyl-neuraminic acid residues but also the combination of both sialic acids. Overall, the most abundant OS decreased with lactation time, whereas different trends were observed for minor OS. 6'-Sialyl-lactose was the most abundant acidic OS while galactosyl-lactose isomers were identified as the most abundant neutral OS. This is the first time that a comprehensive study regarding the changes of the abundance of CMO, both neutral and acidic, at different lactation stages is carried out.

In almonds, there is no standard method for detecting oxidative changes and little data correlating consumer perception with chemical markers of rancidity. To address this, we measured peroxide values (PV), free fatty acid values (FFAs), conjugated dienes, tocopherols, headspace volatiles, and consumer hedonic response in light roasted (LR) and dark roasted (DR) almonds stored under conditions that promote rancidity development over 12 months. Results demonstrate that, although rancidity develops at different rates in LR and DR almonds, consumer liking was not significantly different between LR and DR almonds. Average hedonic ratings of almonds were found to fall below a designated acceptable score of 5 ("neither like nor dislike") by 6 months of storage. This did not correspond with recommended industry rejection standard of PV < 5 mequiv peroxide/kg oil and FFA < 1.5% oleic. FFAs remain well below

Linoleic acid (LA) and α-linolenic acid (ALA) in plant or algae oils are precursors to oxidized fatty acid metabolites known as oxylipins. Liquid chromatography tandem mass spectrometry was used to quantify oxylipins in soybean, corn, olive, canola, and four high-oleic acid algae oils at room temperature or after heating for 10 min at 100 °C. Flaxseed oil oxylipin concentrations were determined in a follow-up experiment that compared it to soybean, canola, corn, and olive oil. Published consumption data for soybean, canola, corn, and olive oil were used to estimate daily oxylipin intake. The LA and ALA fatty acid composition of the oils was generally related to their respective oxylipin metabolites, except for olive and flaxseed oil, which had higher LA derived monohydroxy and ketone oxylipins than other oils, despite their low LA content. Algae oils had the least amount of oxylipins. The change in oxylipin concentrations was not significantly different among the oils after short-term heating. The estimated oxylipin intake from nonheated soybean, canola, corn, and olive oil was 1.1 mg per person per day. These findings suggest that oils represent a dietary source of LA and ALA derived oxylipins and that the response of oils to short-term heating does not differ among the various oils.

Insect pests cause serious economic, yield, and food safety problems to managed crops worldwide. Compounding these problems, insect pests often vector pathogenic or toxigenic microbes to plants. Previous work has considered plant-insect and plant-microbe interactions separately. Although insects are well-understood to use plant volatiles to locate hosts, microorganisms can produce distinct and abundant volatile compounds that in some cases strongly attract insects. In this paper, we focus on the microbial contribution to plant volatile blends, highlighting the compounds emitted and the potential for variation in microbial emission. We suggest that these aspects of microbial volatile emission may make these compounds ideal for use in agricultural applications, as they may be more specific or enhance methods currently used in insect control or monitoring. Our survey of microbial volatiles in insect-plant interactions suggests that these emissions not only signal host suitability but may indicate a distinctive time frame for optimal conditions for both insect and microbe. Exploitation of these host-specific microbe semiochemicals may provide important microbe- and host-based attractants and a basis for future plant-insect-microbe chemical ecology investigations.

A bovine milk oligosaccharide (BMO) library, prepared from cow colostrum, with 34 structures was generated and used to rapidly screen oligosaccharides in domestic animal milks and a whey permeate powder. The novel library was entered into a custom Personal Compound Database and Library (PCDL) and included accurate mass, retention time, and tandem mass spectra. Oligosaccharides in minute-sized samples were separated using nanoliquid chromatography (nanoLC) coupled to a high resolution and sensitive quadrupole-Time of Flight (Q-ToF) MS system. Using the PCDL, 18 oligosaccharides were found in a BMO-enriched product obtained from whey permeate processing. The usefulness of the analytical system and BMO library was further validated using milks from domestic sheep and buffaloes. Through BMO PCDL searching, 15 and 13 oligosaccharides in the BMO library were assigned in sheep and buffalo milks, respectively, thus demonstrating significant overlap between oligosaccharides in bovine (cow and buffalo) and ovine (sheep) milks. This method was shown to be an efficient, reliable, and rapid tool to identify oligosaccharide structures using automated spectral matching.

Concealed damage (CD) is defined as a brown discoloration of the kernel interior (nutmeat) that appears only after moderate to high heat treatment (e.g., blanching, drying, roasting, etc.). Raw almonds with CD have no visible defects before heat treatment. Currently, there are no screening methods available for detecting CD in raw almonds. Herein, the feasibility of using near-infrared (NIR) spectroscopy between 1125 and 2153 nm for the detection of CD in almonds is demonstrated. Almond kernels with CD have less NIR absorbance in the region related with oil, protein, and carbohydrates. With the use of partial least squares discriminant analysis (PLS-DA) and selection of specific wavelengths, three classification models were developed. The calibration models have false-positive and false-negative error rates ranging between 12.4 and 16.1% and between 10.6 and 17.2%, respectively. The percent error rates ranged between 8.2 and 9.2%. Second-derivative preprocessing of the selected wavelength resulted in the most robust predictive model.

Hyperlipidemia is a major cause of atherosclerotic cardiovascular disease. Poria cocos (PC) is a medicinal product widely used in Asia. This study was undertaken to define the alterations of lipid metabolites in rats fed a high-fat diet to induce hyperlipidemia and to explore efficacy and mechanism of action of PC in the treatment of diet-induced hyperlipidemia. Plasma samples were then analyzed using UPLC-HDMS. The untreated rats fed a high-fat diet exhibited significant elevation of plasma triglyceride and total and low-density lipoprotein (LDL) cholesterol concentrations. This was associated with marked changes in plasma concentrations of seven fatty acids (palmitic acid, hexadecenoic acid, hexanoylcarnitine, tetracosahexaenoic acid, cervonoyl ethanolamide, 3-hydroxytetradecanoic acid, and 5,6-DHET) and five sterols [cholesterol ester (18:2), cholesterol, hydroxytestosterone, 19-hydroxydeoxycorticosterone, and cholic acid]. These changes represented disorders of biosynthesis and metabolism of the primary bile acids, steroids, and fatty acids and mitochondrial fatty acid elongation pathways in diet-induced hyperlipidemia. Treatment with PC resulted in significant improvements of hyperlipidemia and the associated abnormalities of the lipid metabolites.

The taste and quality of red wine are determined by its highly complex mixture of polyphenols and many other metabolites. No single method can fully cover the full metabolome, but even for polyphenols and related compounds, current methods proved inadequate. We optimized liquid chromatography resolution and sensitivity using 1 mm i.d. columns with microLC pumps and compared data-dependent to data-independent (SWATH) MS/MS acquisitions. A high-throughput microLC-MS method was developed with a 4 min gradient at 0.05 mL/min flow rate on a Kinetex C18 column and Sciex TripleTOF mass spectrometry. Using the novel software MS-DIAL, we structurally annotated 264 compounds including 165 polyphenols in six commercial red wines by accurate mass MS/MS matching. As proof of concept, multivariate statistics revealed the difference in the metabolite profiles of the six red wines, and regression analysis linked the polyphenol contents to the taste of the red wines.

Concealed damage (CD) is a brown discoloration of nutmeat that appears only after kernels are treated with moderate heat (e.g., roasting). Identifying factors that promote CD in almonds is of significant interest to the nut industry. Herein, the effect of temperature (35 and 45 °C) and moisture (

The chemical composition of finished table olive products is influenced by the olive variety and the processing method used to debitter or cure table olives. Herein, a rapid ultrahigh-pressure liquid chromatography triple-quadrupole tandem mass spectrometry method, using dynamic multiple reaction monitoring, was developed for the quantitation of 12 predominant phenolic and secoiridoid compounds in olive fruit, including hydroxytyrosol, oleuropein, hydroxytyrosol-4-O-glucoside, luteolin-7-O-glucoside, rutin, verbascoside, oleoside-11-methyl ester, 2,6-dimethoxy-p-benzoquinone, phenolic acids (chlorogenic and o-coumaric acids), oleuropein aglycone, and ligstroside aglycone. Levels of these compounds were measured in fresh and California-style black ripe processed Manzanilla olives and two dry salt-cured olive varieties (Mission from California and Throuba Thassos from Greece). Results indicate that the variety and debittering processing method have strong impact on the profile of phenolic and secoiridoid compounds in table olives. The dry salt-cured olives contained higher amounts of most compounds studied, especially oleuropein (1459.5 ± 100.1 μg/g), whereas California-style black ripe olives had a significant reduction or loss of these bioactive compounds (e.g., oleuropein level at 36.7 ± 3.1 μg/g).

The aim of this work was to examine agronomic, compositional, and functional changes in rice (Oryza sativa L. cv. Nipponbare) grains from plants grown under low-to-moderate salinity stress in the greenhouse. Plants were grown in sodium chloride-containing soil (2 or 4 dS/m(2) electrical conductivity), which was imposed 4-weeks after transplant (called Seedling EC2 and EC4) or after the appearance of the anthers (called Anthesis EC2 and EC4). The former simulates field conditions while the latter permits observation of the isolated effect of salt on grain filling processes. Key findings of this study are the following: (i) Plants showed adaptive responses to prolonged salt treatment with no negative effects on grain weight or fertility. Seedling EC2 plants had more panicles and enhanced caryopsis dimensions, while surprisingly, Seedling EC4 plants did not differ from the control group in the agronomic parameters measured. (ii) Grain starch increased in Seedling EC4 (32.6%) and Anthesis EC2 (39%), respectively, suggesting a stimulatory effect of salt on starch accumulation. (iii) The salinity treatment of 2 dS/m(2) was better tolerated at anthesis than the 4 dS/m(2) treatment as the latter led to reduced grain weight (28.8%) and seed fertility (19.4%) and compensatory increases in protein (20.1%) and nitrogen (19.8%) contents. (iv) Although some salinity treatments led to changes in starch content, these did not alter starch fine structure, morphology, or composition. We observed no differences in reducing sugar and amylose content or starch granule size distribution among any of the treatments. The only alterations in starch were limited to small changes in thermal properties and glucan chain distribution, which were only seen in the Anthesis EC4 treatment. This similarity of compositional and functional features was supported by multivariate analysis of all variables measured, which suggested that differences due to treatments were minimal. Overall, this study documents the specific response of rice under defined conditions, and illustrates that the plasticity of plant response to mild stress is complex and highly context-dependent, even under greenhouse conditions in which other potential environmental stress impacts are minimized.

Hexanal, peroxide value, and lipid hydroperoxides are common indicators of lipid oxidation in food products. However, these markers are not always reliable as levels are dynamic and often can be detected only after significant oxidation has occurred. Changes in the volatile composition of light- and dark-roast almonds were evaluated during storage over 24 weeks at 25 or 35 °C using headspace solid phase microextraction (HS-SPME) gas chromatography-mass spectrometry (GC-MS). Several volatile changes were identified in association with early oxidation events in roasted almonds. Hexenal decreased significantly during the first 6 weeks of storage and did not increase above initial levels until 20-24 weeks of storage depending upon the degree of roast. In contrast, levels of 1-heptanol and 1-octanol increased at 16-20 weeks, depending upon the degree of roast, and no initial losses were observed. Seventeen new compounds, absent in raw and freshly roasted almonds but detectable after 6 weeks of storage, were identified. Of these, 2-octanone, 2-nonanone, 3-octen-2-one, 2-decanone, (E)-2-decenal, 2,4-nonadienal, pentyl oxirane, and especially acetic acid increased significantly (that is, >10 ng/g). The degree of roasting did not correlate with the levels of these compounds. Significant decreases in roasting-related aroma volatiles such as 2-methylbutanal, 3-methylbutanal, furfural, 2-phenylacetaldehyde, 2,3-butanedione, 2-methylpyrazine, and 1-methylthio-2-propanol were observed by 4 weeks of storage independent of the degree of roast or storage conditions.

To develop a more sensitive immunoassay for malachite green (MG) and leucomalachite green (LMG), we identified the immunocomplex binding phage-borne peptides for use in the noncompetitive phage anti-immunocomplex assay (PHAIA). An anti-LMG monoclonal antibody (mAb) was used to select immunocomplex binding peptides from a circular random eight-amino-acid phage-displayed library. After three rounds of panning-elution, five peptides that bound the LMG-mAb immunocomplex were obtained. One of the phage-borne peptide clones that resulted in an assay with the highest sensitivity was chosen for further research. The concentration of LMG producing 50% of the saturated signal and the limit of detection of the assay were 7.02 and 0.55 ng/mL, respectively, with a linear range of 1.35 to 21.56 ng/mL. The PHAIA based on the same antibody was 16 times more sensitive compared to the competitive immunoassay. PHAIA was used to analyze LMG, MG, and two mixtures of spiked fish samples, with validation by high-performance liquid chromatography (HPLC) with fluorescence detector. Results showed a good correlation (R(2)LMG = 0.9841; R(2)MG = 0.993; R(2)Mixture = 0.9903) between the data of PHAIA and HPLC, thus the assay was an efficient method for monitoring food safety.

Human milk is known to contain several proteases, but little is known about whether these enzymes are active, which proteins they cleave, and their relative contribution to milk protein digestion in vivo. This study analyzed the mass spectrometry-identified protein fragments found in pooled human milk by comparing their cleavage sites with the enzyme specificity patterns of an array of enzymes. The results indicate that several enzymes are actively taking part in the digestion of human milk proteins within the mammary gland, including plasmin and/or trypsin, elastase, cathepsin D, pepsin, chymotrypsin, a glutamyl endopeptidase-like enzyme, and proline endopeptidase. Two proteins were most affected by enzyme hydrolysis: β-casein and polymeric immunoglobulin receptor. In contrast, other highly abundant milk proteins such as α-lactalbumin and lactoferrin appear to have undergone no proteolytic cleavage. A peptide sequence containing a known antimicrobial peptide is released in breast milk by elastase and cathepsin D.

Huanglongbing (HLB), also known as Citrus Greening Disease, is caused by the bacterium 'Candidatus Liberibacter asiaticus' (CLas) and is a serious threat to the citrus industry. To understand the effect of CLas infection on the citrus metabolome, juice from healthy (n = 18), HLB-asymptomatic (n = 18), and HLB-symptomatic Hamlin (n = 18), as well as from healthy (n = 18) and HLB-symptomatic (n = 18) Valencia sweet oranges (from southern and eastern Florida) were evaluated using (1)H NMR-based metabolomics. Differences in the concentration of several metabolites including phenylalanine, histidine, limonin, and synephrine between control or asymptomatic fruit and symptomatic fruit were observed regardless of the citrus variety or location. There were no clear differences between the metabolite profiles of Hamlin fruits classified by PCR as asymptomatic and control, suggesting that some of the control fruit may have been infected. Taken together, these data indicate that infection due to CLas presents a strong metabolic response that is observed across different cultivars and regions, suggesting the potential for generation of metabolite-based biomarkers of CLas infection.

In recent years attention has been focused on the utilization of microorganisms as alternatives for industrial and nutritional applications. Considerable research has been devoted to techniques for growth, extraction, and purification of high-value lipids for their use as biofuels and biosurfactants as well as high-value metabolites for nutrition and health. These successes argue that the elucidation of the mechanisms underlying the microbial biosynthesis of such molecules, which are far from being completely understood, now will yield spectacular opportunities for industrial scale biomolecular production. There are important additional questions to be solved to optimize the processing strategies to take advantage of the assets of microbial lipids. The present review describes the current state of knowledge regarding lipid biosynthesis, accumulation, and transport mechanisms present in single-cell organisms, specifically yeasts, microalgae, bacteria, and archaea. Similarities and differences in biochemical pathways and strategies of different microorganisms provide a diverse toolset to the expansion of biotechnologies for lipid production. This paper is intended to inspire a generation of lipid scientists to insights that will drive the biotechnologies of microbial production as uniquely enabling players of lipid biotherapeutics, biofuels, biomaterials, and other opportunity areas into the 21st century.

Phosphorus is an important natural nutrient, but high dietary phosphorus intake, including that sourced from added preservatives, is of great concern in renal patients. In this context a reliable analytical method able to quantify differential phosphorus in food could be a valuable tool for monitoring diet composition This paper presents a novel analytical procedure to quantify the following kinds of phosphorus in cooked ham: total (TP), inorganic (IP), from phospholipids (PL), and from phosphoproteins (PP). This technique is based on a suitable sample preparation followed by spectrophotometric analyses. Analytical performances of each method were evaluated, taking advantage also of food industry certified material and in-house reference samples. Limit of detection and limit of quantification values for TP, IP, PP-derived, and PL-derived phosphorus were 13 and 37, 11 and 33, 2 and 20, and 6 and 16 mg P/100 g fresh mass, respectively. Similar results were obtained when this procedure was used to quantify different types of phosphorus present in cooked ham samples. In conclusion, this procedure is effective for quantifying the content of different types of phosphorus present in cooked ham, which can be contributed by different phosphorus-containing ingredients and additives. To the best of the authors' knowledge, this is the first time that simultaneous determination of TP, IP, PL, and PP in cooked ham has been reported.

Bovine milk is known to contain naturally occurring peptides, but relatively few of their sequences have been determined. Human milk contains hundreds of endogenous peptides, and the ensemble has been documented for antimicrobial actions. Naturally occurring peptides from bovine milk were sequenced and compared with human milk peptides. Bovine milk samples from six cows in second-stage peak lactation at 78-121 days postpartum revealed 159 peptides. Most peptides (73%) were found in all six cows sampled, demonstrating the similarity of the intramammary peptide degradation across these cows. One peptide sequence, ALPIIQKLEPQIA from bovine perilipin 2, was identical to another found in human milk. Most peptides derived from β-casein, αs1-casein, and αs2-casein. No peptides derived from abundant bovine milk proteins such as lactoferrin, β-lactoglobulin, and secretory immunoglobulin A. The enzymatic cleavage analysis revealed that milk proteins were degraded by plasmin, cathepsins B and D, and elastase in all samples.

Prebiotics are nondigestible substrates that stimulate the growth of beneficial microbes in the human intestine. Galactooligosaccharides (GOS) are food ingredients that possess prebiotic properties, in particular, promoting the growth of bifidobacteria in situ. However, precise mechanistic details of GOS consumption by bifidobacteria remain poorly understood. Because GOS are mixtures of polymers of different lengths and linkages, there is interest in determining which specific structures provide prebiotic effects to potentially create better supplements. This paper presents a method comprising porous graphitic carbon separation, isotopic labeling, and mass spectrometry analysis for the structure-specific analysis of GOS isomers and their bacterial consumption rate. Using this strategy, the differential bacterial consumption of GOS by the bifidobacteria species Bifidobacterium longum subsp. infantis, Bifidobacterium animalis subsp. lactis, and Bifidobacterium adolescentis was determined, indicating that the use of specific GOS isomers in infant formula may provide enrichment of distinct species.

The human gastrointestinal tract contains a highly complex ecosystem that harbors various microorganisms, which together create a unique environment within each individual. There is growing awareness that dietary habits are one of the essential factors contributing to the microbial diversity and community configuration that ultimately affects human health. From an evolutionary perspective, human dietary history can be viewed as a central factor in the selection of the gut microbial community and stabilization of the mutualistic host-microbial interaction, that together drive host phenotype. Herein, current knowledge concerning the influence of major dietary macrostructure and individual food ingredients is presented. This knowledge will provide perspectives for personalized gut microbiota management and, ultimately, movement toward an era of personalized nutrition and medicine.