Chromatography-mass Spectrometry Research Articles

Integrated metabolomic and transcriptomic analyses of the synergistic effect of polymyxin–rifampicin combination against Pseudomonas aeruginosa

BackgroundUnderstanding the mechanism of antimicrobial action is critical for improving antibiotic therapy. For the first time, we integrated correlative metabolomics and transcriptomics of Pseudomonas aeruginosa to elucidate the mechanism of synergistic killing of polymyxin–rifampicin combination.MethodsLiquid chromatography-mass spectrometry and RNA-seq analyses were conducted to identify the significant changes in the metabolome and transcriptome of P. aeruginosa PAO1 after exposure to polymyxin B (1 mg/L) and rifampicin (2 mg/L) alone, or in combination over 24 h. A genome-scale metabolic network was employed for integrative analysis.ResultsIn the first 4-h treatment, polymyxin B monotherapy induced significant lipid perturbations, predominantly to fatty acids and glycerophospholipids, indicating a substantial disorganization of the bacterial outer membrane. Expression of ParRS, a two-component regulatory system involved in polymyxin resistance, was increased by polymyxin B alone. Rifampicin alone caused marginal metabolic perturbations but significantly affected gene expression at 24 h. The combination decreased the gene expression of quorum sensing regulated virulence factors at 1 h (e.g. key genes involved in phenazine biosynthesis, secretion system and biofilm formation); and increased the expression of peptidoglycan biosynthesis genes at 4 h. Notably, the combination caused substantial accumulation of nucleotides and amino acids that last at least 4 h, indicating that bacterial cells were in a state of metabolic arrest.ConclusionThis study underscores the substantial potential of integrative systems pharmacology to determine mechanisms of synergistic bacterial killing by antibiotic combinations, which will help optimize their use in patients.Graphical

ABOUT OIL AND GAS POTENTIAL OF THE ARYSKUM DEPRESSION OF THE SOUTH TORGAI SEDIMENTARY BASIN

The article discusses laboratory studies carried out to compare the composition of oils from Mesozoic and pre-Mesozoic formations: a comprehensive analysis of the molecular composition of oils by chromatography-mass spectrometry; separation and identification of the composition of alkanes by gas-liquid chromatography; NMR-spectroscopic determination of the fragment composition of oils; mass spectrometry was used to determine the isotopic composition of carbon, which identifies the genesis of hydrocarbons in relation to the zones of oil and gas formation. As a result of the study, the laboratory data were obtained, which made it possible to reveal the similarity between oils from deposits of different ages within the graben-synclines, namely, close values of geochemical parameters and the relative content of individual classes of biomarkers made it possible to assume the genetic unity of oils from the Lower Cretaceous and Proterozoic of the Akshabulak graben-syncline and genetic unity of oils from the Lower Cretaceous and Paleozoic of the Aksai horst anticline. The value of the pristane/ phytane ratios indicates a common environment for the formation of the initial organic matter within the structures - suboxidizing conditions in the Akshabulak graben syncline and oxidizing conditions in the Aksai horst anticline, and the existence of a higher redox potential in the Bosingen graben-syncline. Gas- liquid chromatography data showed a slight difference in the thermal transformation of the studied oils, which were classified as "mature". The results of NMR spectrometry in combination with other methods indicate the genetic unity of oils from the Lower Cretaceous and Proterozoic of the Akshabulak graben- syncline and the genetic unity of oils from the Lower Cretaceous and Paleozoic of the Aksai horst anticline. A comparative analysis of the carbon isotopic composition of oils from the Jurassic and Cretaceous deposits of the Aryskum depression and oils from various basins of the world showed a similar range of their variations and a possible genetic relationship of these oils with organic matter of the sapropel type.

Analysis of volatile compounds contributing to distinctive odour of silver carp (Hypophthalmichthys molitrix) surimi

SummaryAlthough the characterisation of volatile compounds in silver carp (Hypophthalmichthys molitrix) have been studied, their respective contribution to specific odour feeling was unclear. Therefore, an investigation was made in this study by big data mining, solid phase microextraction‐gas chromatography–mass spectrometry (SPME‐GC–MS), odour activity values (OAVs), odour recombination in surimi models and omission experiment. The results demonstrated that 91 volatile compounds were obtained by data mining and 62 volatile compounds were identified by instrumental analysis. Among them, twenty‐one volatile compounds were initially selected for the construction of the surimi model. The odour recombination and omission experiment confirmed that characteristic odour of silver carp surimi were hexanal (fishy, grassy), octanal (grassy, fatty), (E)‐2‐octenal (fishy), nonanal (grassy, fatty), (E)‐2‐nonanal (fatty, grass), (E,E)‐2,4‐heptadienal (fishy), 1‐hexanol (grassy) and 1‐octen‐3‐ol (mushroom). This study specified distinct silver carp surimi odour with definite compounds.

Occurrence of 3-monochloropropane-1,2-Diol (3-MCPD) in canned vegetables: A probabilistic assessment of health risk for Iranian consumers

The present study assessed the potential risk to Iranian consumers posed by oral exposure to 3-MCPD in canned vegetables. Ten brands of canned vegetables (i.e. cucumber, pea, sweet corn, pinto bean, lentil, and mixed vegetables) were collected from retail markets in Iran. Overall, 3-MCPD in 180 canned vegetable samples was quantified by chromatography-mass spectrometry (GC-MS). Our findings showed statistically significant associations between 3-MCPD level and the type of canned vegetables as well as significant variations in 3-MCPD mean level among the brands. The highest level of 3-MCPD was measured in pea samples, while pinto beans showed the lowest content. Hazard Index (HI) values calculated based on 3-MCPD levels measured in the collected samples indicated no major risk in both deterministic and probabilistic scenarios (HI values <1.0) and no health risk based on the calculated margin of exposure (MOE) for 3-MCPD to Iranian consumers. Based on the Monte Carlo Simulation (MCS) sensitivity analysis, the concentration of 3-MCPD was the most influential variable. Nonetheless, industrial policies should maintain reduced 3-MCPD levels in canned foods.

Yin Huo Tang, a traditional Chinese herbal formula, relives ovariectomy and empty bottle stimulation-induced menopause-like symptoms in mice.

Yin Huo Tang (YHT), a traditional Chinese herbal formula, is effectively used for the clinical treatment of menopause-like symptoms in China. This study aimed to investigate its efficacy on menopause-like symptoms in mice using behavioral tests and histopathological assessment, and to determine its possible mechanism of action based on network pharmacology. Liquid chromatography-mass spectrometry (LC-MS) technology was used to identify the potential active ingredients of YHT. In mice, menopause-like symptoms were induced by combination of bilateral ovariectomy and empty bottle stimulation. The mice were then treated with the YHT aqueous extract for three weeks. Behavior, sleep state, body weight, organ index, and histomorphology were analyzed separately. Additionally, network pharmacology and molecular docking were used to predict the mechanisms underlying the action of YHT. Finally, serum estradiol was quantified to preliminarily verify the results of network pharmacology. YHT not only improved the behavior of mice (attack and explore behavior reduced; modify behavior increased) but also ameliorated the sleep state (sleep time increased and incubation time reduced). YHT reduced body weight, increased uterine weight, and improved the histomorphology of some organs. Network pharmacology and molecular docking analyses revealed that the estrogen signaling pathway might play a key role in attenuating menopause-like symptoms. Furthermore, YHT treatment reversed the reduction in serum estradiol levels. YHT alleviates menopause-like symptoms in a mouse model, providing a rationale for using it as a potential therapeutic strategy.

Identification of off-flavor compounds and deodorizing of cattle by-products.

An unnatural flavor in a food or drink product caused by the presence of undesirable compounds due to contamination or deterioration is called off-flavor. This study determined the characteristics of cattle by-products off-flavor (heart, liver, lung, rumen, and intestine). We identified 25, 34, 26, 22, and 26 volatile compounds from the heart, liver, lung, rumen, and intestine, respectively, in the bovine via headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). Based on the relative odor activity value (ROAV ≥ 1), 16 volatile compounds were labeled as characteristic off-flavor by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). The compounds involved in the characteristic off-flavor in bovine heart were E,E-2,4-nonadienal, E,E-2,4-decadien-1-al, hexanal, (E)-2-octenal, and decyl aldehyde. In the bovine liver, the off-flavor compounds were 1-nonanol, ethyl hexanoate, 2-octanone, and dodecyl aldehyde and in bovine lung 3-heptylacrolein was the off-flavor compound. In bovine rumen, heptaldehyde, octanal, p-cresol, and 1-nonanal were off-flavor compounds, and lastly, 1-octen-3-ol and E-2-nonenal were off-flavor compounds with bovine intestine. The cattle by-products were deodorized by shallot-ginger extract masking, baker's yeast fermentation, active dry yeast + β-cyclodextrin (β-CD) composite, and ultrasound + chitosan composite. The above 16 labeled characteristic compounds decreased in concentration. The ultrasound + chitosan composite method showed a significantly better effect than the other methods (p< .05). The aim of this study was to determine the characteristic flavor information of cattle by-products and provide idea on how to improve the flavor by various deodorization methods. PRACTICAL APPLICATIONS: This study investigated the volatile flavor compounds of cattle by-products from five organs (heart, liver, lung, rumen, and intestine) by headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). The 16 volatile compounds were labeled as the major characteristic off-flavor compounds by relative odor activity values and principal component analysis. Four different deodorization methods were adopted, and among them, ultrasound + chitosan composite method showed best results. This study has provided useful information about the characteristic off-flavor compounds and suggests how to improve the flavor of cattle by-products through various deodorization methods.

Antifungal Activity of Citronella Oil Against Clinical Isolate Candida albicans

Citronella (Cymbopogon nardus (L.) Rendle) is commonly claimed as a multifunction plant. Citronella produces an essential oil whose quality standard was set in SNI 06-3953-1995. Citronella oil is used for traditional medication, such as an antimicrobial, antipyretic, analgesic, and many other functions. This study evaluated the in-vitro antifungal activity of citronella oil obtained from the farm in Menderek, Pintu Rime Gayo, Bener Meriah, Aceh, against clinical isolates of Candida albicans. The citronella oil analyzed using chromatography-mass spectrometry (GC-MS) showed chemical components of citronellal (26.06%), β-citronellol (26.314%), and geraniol (17.90%). Antifungal activity test of citronella oil against Candida albicans by well diffusion method showed the highest activity at 40% v/v concentration with the diameter of inhibition zone of 23.31±0.04 mm. The result of this study indicated that the citronella oil has a good potential to inhibit clinical isolate Candida albicans growth.

Monitoring of pesticide and some metal residues in Mazafati date fruit cultivar and risk assessment to the health

Metals and pesticides pose risk to human health if accumulated in date fruits eaten for their distinctive sweetness and beneficial minerals. The present study evaluated the concentrations of metals and pesticides in Mazafati date fruit, soil, and irrigation water from three locations of Kerman province in Iran to show potential pollution hazards to consumers of date fruit in Iran. Levels of pesticides and metals were measured using chromatography-mass spectrometry (GC-MS) and atomic absorption spectrophotometry, respectively. The significant difference between the levels of Cu, Fe, chlorpyrifos, and malathion from three various locations was identified. The majority of metals except for Pb and Cd did not meet the safety limits concerning maximum allowable levels (MAL) in the Mazafati date cultivar. Estimated daily intake (EDI) of metals through date consumption (g/person/day) for all metals was above the tolerable daily intake in all studied locations. Furthermore, the concentrations of identified residues of chlorpyrifos, HCH total, and malathion are considerably upper than the maximum residue limits (MRL), showing a risky trend in the cultivation of date palm. As a result, the public consciousness of Iranian farmers should be raised to use pesticides more prudently. Also, consuming date fruits cultivated close to the industrial zones is hazardous and must be prevented.

Selection of Non-Saccharomyces Wine Yeasts for the Production of Leavened Doughs

Background: Non-conventional yeasts (NCY) (i.e., non-Saccharomyces) may be used as alternative starters to promote biodiversity and quality of fermented foods and beverages (e.g., wine, beer, bakery products). Methods: A total of 32 wine-associated yeasts (Campania region, Italy) were genetically identified and screened for decarboxylase activity and leavening ability. The best selected strains were used to study the leavening kinetics in model doughs (MDs). A commercial strain of Saccharomyces cerevisiae was used as the control. The volatile organic profiles of the inoculated MDs were analyzed by solid phase microextraction/gas chromatography-mass spectrometry (SPME/GC-MS). Results: Most of strains belonged to the NCY species Hanseniaspora uvarum, Metschnikowia pulcherrima, Pichia kudriavzevii, Torulaspora delbruekii, and Zygotorulaspora florentina, while a few strains were S. cerevisiae. Most strains of H. uvarum lacked decarboxylase activity and showed a high leaving activity after 24 h of incubation that was comparable to the S. cerevisiae strains. The selected H. uvarum strains generated a different flavor profile of the doughs compared to the S. cerevisiae strains. In particular, NCY reduced the fraction of aldehydes that were potentially involved in oxidative phenomena. Conclusions: The use of NCY could be advantageous in the bakery industry, as they can provide greater diversity than S. cerevisiae-based products, and may be useful in reducing and avoiding yeast intolerance.

The Human Skin Volatolome: A Systematic Review of Untargeted Mass Spectrometry Analysis.

The analysis of volatile organic compounds (VOCs) can provide important clinical information (entirely non-invasively); however, the exact extent to which VOCs from human skin can be signatures of health and disease is unknown. This systematic review summarises the published literature concerning the methodology, application, and volatile profiles of skin VOC studies. An online literature search was conducted in accordance with the preferred reporting items for systematic reviews and meta-analysis, to identify human skin VOC studies using untargeted mass spectrometry (MS) methods. The principal outcome was chemically verified VOCs detected from the skin. Each VOC was cross-referenced using the CAS number against the Human Metabolome and KEGG databases to evaluate biological origins. A total of 29 studies identified 822 skin VOCs from 935 participants. Skin VOCs were commonly sampled from the hand (n = 9) or forearm (n = 7) using an absorbent patch (n = 15) with analysis by gas chromatography MS (n = 23). Twenty-two studies profiled the skin VOCs of healthy subjects, demonstrating a volatolome consisting of aldehydes (18%), carboxylic acids (12%), alkanes (12%), fatty alcohols (9%), ketones (7%), benzenes and derivatives (6%), alkenes (2%), and menthane monoterpenoids (2%). Of the VOCs identified, 13% had putative endogenous origins, 46% had tentative exogenous origins, and 40% were metabolites from mixed metabolic pathways. This review has comprehensively profiled the human skin volatolome, demonstrating the presence of a distinct VOC signature of healthy skin, which can be used as a reference for future researchers seeking to unlock the clinical potential of skin volatolomics. As significant proportions of identified VOCs have putative exogenous origins, strategies to minimise their presence through methodological refinements and identifying confounding compounds are discussed.

Amphetamine, methamphetamine, and MDMA in hair samples from a rehabilitation facility: Validation and applicability of HF-LPME-GC-MS

IntroductionIt is known that drug abuse jeopardizes economic and social development. Toxicological analyses can guide prevention and treatment strategies in rehabilitation facilities. The current greatest challenge is finding easily adaptable and less costly sensitive methods that meet the principles of green chemistry. Hair, as a biological matrix, has several advantages, and its ability to detect consumption for longer periods keeping the matrix stable and unaltered stands out. This manuscript addresses the use of a miniaturized technique in an alternative matrix, by making use of a reduced amount of solvents to quantify amphetamines, aiming to guide prevention and treatment strategies in rehabilitation facilities. MethodsA Hollow Fiber Liquid-phase Microextraction (HF-LPME) technique for extracting amphetamines from hair samples with Gas Chromatography-Mass Spectrometry (CG-MS) was validated, adapted, and applied to ten samples from patients of a rehabilitation facility. ResultsThe technique proved to be sensitive, accurate, precise, and not affected by interference from the biological matrix and the linear range for the analytes was 0.2 to 20 ng mg −1. The three analytes were quantified in the samples analyzed. It is worth stressing that the patients were young. ConclusionThe HF-LPME-GC-MS technique complied with the principles of green chemistry, and proved to be a sensitive technique, adaptable to the routine of common laboratories. Validation in the analysis phase with authentic samples, thus, showed that it can be an important tool for preventing and controlling drug addiction.

Application of Mass Spectrometry for Determining the Geographic Production Area of Wagyu Beef.

Japanese Black cattle (Japanese Wagyu) beef is attracting attention for its aroma and marbling, and its handling is increasing worldwide. Here, we focused on the origin discrimination of Wagyu beef and analyzed the nutritional components of Japanese Wagyu (produced in multiple prefectures of Japan), Hybrid Wagyu (a cross between Angus and Wagyu cattle born in Australia and transported to Japan), and Australian Wagyu beef using mass spectrometry (MS). Triple-quadrupole liquid chromatography–MS was used to clarify the molecular species of lipids in Wagyu beef. Fourteen classes of lipids were separated, and 128 different triacylglycerides (TGs) were detected. A simple comparative analysis of these TGs using high-performance liquid chromatography revealed significantly higher levels of triolein (C18:1/C18:1/C18:1; abbreviated OOO) and C18:1/C18:1/C16:1 (OOPo) in Japanese Wagyu. Wagyu elements beef were comprehensively analyzed using inductively coupled plasma (ICP)–MS and ICP–optical emission spectrometry. We found significant differences in the rubidium, cesium, and lithium levels of Japanese and Australian Wagyu beef. On comparing metabolites using gas chromatography–MS, we identified significant differences in the levels of amino acids and other components of the Japanese and Australian Wagyu beef. These results suggest the possibility of determining the origin of Wagyu cattle breeds using MS and genetic discrimination.

A phosphaphenanthrene-based derivative as multifunctional flame retardant for epoxy resins

In this work, the transparent, mechanically strengthened, flame retardant and smoke suppression epoxy resin (EP) composites was prepared by the addition of an indole/phosphaphenanthrene-based compound (IDT). The results presented that limiting oxygen index (LOI) of EP composite containing 5 wt% IDT raised to 32% and it passed a UL-94 V-0 rating. In the meantime, compared with EP, total smoke production (TSP) and the peak of heat release (PHRR) of EP/IDT5 were decreased by 51.9% and 32.1%, respectively. The flame-retardant mechanism of EP/IDT was also explored by pyrolysis/gas chromatography–mass spectrometry (Py-GC/MS), scanning electron microscope (SEM) and X-ray photoelectron spectroscopy (XPS). Besides, the bending and tensile strengths were improved thanks to the presence of π-π conjugation and hydrogen bonds between IDT and EP. This work provided a promising way to meet the increasing multifunctional requirements of EP composites.

Combined high-pressure experimental and kinetic modeling study of cyclopentene pyrolysis and its reactions with acetylene

A combined experimental and kinetic modeling study is presented to improve the understanding of the formation of polycyclic aromatic hydrocarbons (PAHs) from neat cyclopentene and cyclopentene/acetylene mixtures. High-pressure experiments are conducted for the first time over a temperature range covering 930–1650 K using a single-pulse shock tube coupled to gas chromatography/gas chromatography-mass spectrometry (GC/GC–MS) techniques. Several updates and inclusions, mainly regarding the reactions involving C5 molecules and radicals, are made in our on-going PAH kinetic model, which shows satisfactory predictive performances for the speciation measurements obtained in the current work and in the literature. On the basis of the experimental observations and modeling analyses, the reaction pathways active during the pyrolysis of cyclopentene are illustrated and the effects of acetylene addition as co-reactant on the PAH chemistry are assessed. In all of the cases investigated, it is noted that the cyclopentadienyl radical largely participate in the formation of mono-aromatic hydrocarbons (benzene and styrene) and PAHs (indene, naphthalene and phenanthrene).

Binding of dual‐flavour compounds by soy protein isolate in aqueous model systems

SummaryBy the technique of headspace solid‐phase microextraction/gas chromatography–mass spectrometry (SPME‐GC/MS) and static headspace/gas chromatography (HS/GC), the interaction of dual‐flavours and soy protein isolates (SPI) was determinated at low flavour concentration (0.04–0.16 mm) and high flavour concentration (0.4–1.6 mm). The results show that the mixture of ethyl caprylate and undecanal that has elongated structure possesses the same binding sites on SPI and shows competitive binding; the mixture of carvone that has compact structure, at low flavour concentration, possesses different and independent binding sites on SPI, and there maybe existed noncompetitive binding. If high concentration multi‐flavour binding with SPI induces the conformational change and increased more binding sites for another single flavour, there may be existed anticompetitive binding. ‘Salt out’ effects of interaction model were observed for terpene flavours.

A Metabolomic Approach and Traditional Physical Assessments to Compare U22 Soccer Players According to Their Competitive Level.

Simple SummaryDifferent physical tests are applied in the sports context for athlete selection and training prescriptions. Recently, the metabolomics approach has been used to understand biological systems; it has shown high potential to answer integrated questions, including with regard to soccer. Our study is the first to combine traditional physical assessment protocols with metabolomics to detail and compare the physiological profiles of two soccer teams (under-22 soccer players) at different competitive levels. Using anthropometric measurements and the critical velocity model, no differences were observed between elite and non-elite soccer players. However, the metabolomic analysis was able to differentiate among the fasting serum metabolic profiles of athletes of different competitive levels. Our results suggest that the application of metabolomics combined with traditional physical assessment protocols can improve characterizations of athletes, strengthening the differentiation of the physiological profiles of soccer teams. Thus, the metabolomic approach can contribute by offering additional data in individual characterizations of athletes, improving training programs, and consequently, increasing the performance of players during competitions.The purpose of this study was to use traditional physical assessments combined with a metabolomic approach to compare the anthropometric, physical fitness level, and serum fasting metabolic profile among U22 soccer players at different competitive levels. In the experimental design, two teams of male U22 soccer were evaluated (non-elite = 20 athletes, competing in a regional division; elite = 16 athletes, competing in the first division of the national U22 youth league). Earlobe blood samples were collected, and metabolites were extracted after overnight fasting (12 h). Untargeted metabolomics through Liquid Chromatograph Mass Spectrometry (LC-MS) analysis and anthropometric evaluation were performed. Critical velocity was applied to determine aerobic (CV) and anaerobic (ARC) capacity. Height (non-elite = 174.4 ± 7.0 cm; elite = 176.5 ± 7.0 cm), body mass index (non-elite = 22.1 ± 2.4 kg/m2; elite = 21.9 ± 2.3 kg/m2), body mass (non-elite = 67.1 ± 8.8 kg; elite = 68.5 ± 10.1 kg), lean body mass (non-elite = 59.3 ± 7.1 kg; elite = 61.1 ± 7.9 kg), body fat (non-elite = 7.8 ± 2.4 kg; elite = 7.3 ± 2.4 kg), body fat percentage (non-elite = 11.4 ± 2.4%; elite = 10.5 ± 1.7%), hematocrit (non-elite = 50.2 ± 4.0%; elite = 51.0 ± 4.0%), CV (non-elite = 3.1 ± 0.4 m/s; elite = 3.0 ± 0.2 m/s), and ARC (non-elite = 129.6 ± 55.7 m; elite = 161.5 ± 61.0 m) showed no significant differences between the elite and non-elite teams, while the multivariate Partial Least Squares Discriminant Analysis (PLS-DA) model revealed a separation between the elite and non-elite athletes. Nineteen metabolites with importance for projection (VIP) >1.0 were annotated as belonging to the glycerolipid, sterol lipid, fatty acyl, flavonoid, and glycerophospholipid classes. Metabolites with a high relative abundance in the elite group were related in the literature to a better level of aerobic power, greater efficiency in the recovery process, and improvement of mood, immunity, decision making, and accuracy, in addition to acting in mitochondrial preservation and electron transport chain maintenance. In conclusion, although classical physical assessments were not able to distinguish the teams at different competitive levels, the metabolomics approach successfully indicated differences between the fasting metabolic profiles of elite and non-elite teams.

Study on the Interaction Mechanism Between Soybean Protein Isolates and Lemon Flavor: Isomerization and Degradation of Citral.

By headspace solid-phase microextraction/gas chromatography–mass spectrometry, the effects of 1% (w/v) alcohol denatured soybean protein isolates (L-SPI), native soybean protein isolates (N-SPI), as well as the thermal denaturation of soybean protein isolates (H-SPI) on low concentration (24 μmol/L) of citral was studied in aqueous. The results shows that the SPI could catalyze citral isomerization and yield methyl heptenone and acetaldehyde by inverse aldol condensation degradation. 3-Hydroxycitronelloal was formed as an intermediate in this reaction. The catalytic efficiency of the L-SPI was higher than that of N-SPI, whereas the catalytic efficiency of H-SPI was the lowest. Additionally, it shows that the catalytic efficiency increased as the pH increased. The catalytic efficiency of 7S (Soybean β-Conglycinin) was greater than that of 11S (Soy bean Proglycinin).

A novel lncRNA RP11-386G11.10 reprograms lipid metabolism to promote hepatocellular carcinoma progression

ObjectiveEmerging studies suggest that long non-coding RNAs (lncRNAs) play crucial roles in hepatocellular carcinoma (HCC). A rapidly increasing number of studies have shown that metabolic changes including lipid metabolic reprogramming play a significant role in the progression of HCC. But it remains to be elucidated how lncRNAs affect tumor cell metabolism. MethodsThrough analysis and screening of The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) dataset, we found a novel lncRNA RP11-386G11.10 was overexpressed, related to prognosis, conserved and non-protein-coding in HCC and related to poor prognosis. Then, CCK-8, colony formation, Transwell invasion, wound healing assays were performed and nude mouse subcutaneous tumour formation and lung metastasis models were established to explore the effect of RP11-386G11.10 on HCC tumour growth and metastasis. Chromatography-mass spectrometry (GC-MS) and Nile red staining detected the effect of RP11-386G11.10 on lipid metabolism in HCC. Mechanistically, we clarified the RP11-386G11.10/miR-345-3p/HNRNPU signalling pathway through dual luciferase reporter, RNA immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP) assays and identified ZBTB7A as a transcription factor of RP11-386G11.10. ResultsRP11-386G11.10 was overexpressed in HCC and positively correlated with tumour size, TNM stage, and poor prognosis in HCC patients. RP11-386G11.10 promoted the proliferation and metastasis of HCC cells in vitro and in vivo. Mechanistically, RP11-386G11.10 acted as a competing endogenous RNA (ceRNA) for miR-345-3p to regulate the expression of HNRNPU and its downstream lipogenic enzymes, leading to lipid accumulation in HCC cells and promoting their growth and metastasis. In addition, we identified ZBTB7A as a transcription factor of RP11-386G11.10. Moreover, HNRNPU promoted the expression of ZBTB7A in HCC cells, thereby increasing the transcriptional activity of RP11-386G11.10, and forming a positive feedback loop, ultimately leading continuous lipid accumulation, growth and metastasis in HCC cells. ConclusionsOur results indicated that the lncRNA RP11-386G11.10 was a novel oncogenic lncRNA that was strongly correlated with the poor prognosis of HCC. The ZBTB7A-RP11-386G11.10-HNRNPU positive feedback loop promoted the progression of HCC by regulating lipid anabolism. RP11-386G11.10 may become a new diagnostic and prognostic biomarker and therapy target for HCC.

Chocolates with Brazilian cocoa: Tracking volatile compounds according to consumers’ preference

This study aimed to evaluate the volatile compounds of chocolates made of Brazilian cocoas and statistically track them according to the products’ sensorial profile in order to relate them to consumers’ acceptance by preference map methodology. The intensity of the chocolate, acidity, woody, smoked, green, floral, burned, musty, and cocoa notes from chocolates produced with cocoa from different Brazilian states were analyzed by a trained panel and by 128 consumers. Samples from Côte d’Ivoire, which is known for its high-quality chocolate, were evaluated for comparison. Solid-phase microextraction headspace sampling/gas chromatography-mass spectrometry was employed to evaluate the samples’ volatile compounds. One hundred volatile compounds were identified within the samples. The results from the preference maps showed that the maximum preference was found for chocolate made of cocoa from Rondônia, Bahia, and Espírito Santo and Côte d’Ivoire and organic samples from Pará. The ideal sample point was characterized by intense chocolate, floral, and woody notes and mild green and burned notes. The presence of furfural, 3-methyl butanal, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, methyl pyrazine, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, and tetramethyl pyrazine were shown to be important for consumer acceptance in the ideal product, whereas the presence of (Z)-2-heptenal and 2-pentyl furan may increase consumer rejection. 2,3-Methyl pyrazine, methyl pyrazine, and 2,3-butanediol, which are important volatile compounds previously reported in the literature, were statistically tracked to both positive and negative sample attributes and must be better explored concerning consumers’ acceptance of chocolates.

Association between lipid profiles and viral respiratory infections in human sputum samples

BackgroundRespiratory infections such as influenza account for significant global mortality each year. Generating lipid profiles is a novel and emerging research approach that may provide new insights regarding the development and progression of priority respiratory infections. We hypothesized that select clusters of lipids in human sputum would be associated with specific viral infections (Influenza (H1N1, H3N2) or Rhinovirus).MethodsLipid identification and semi-quantitation was determined with liquid chromatography and high-resolution mass spectrometry in induced sputum from individuals with confirmed respiratory infections (influenza (H1N1, H3N2) or rhinovirus). Clusters of lipid species and associations between lipid profiles and the type of respiratory viral agent was determined using Bayesian profile regression and multinomial logistic regression.ResultsMore than 600 lipid compounds were identified across the sputum samples with the most abundant lipid classes identified as triglycerides (TG), phosphatidylethanolamines (PE), phosphatidylcholines (PC), Sphingomyelins (SM), ether-PC, and ether-PE. A total of 12 lipid species were significantly different when stratified by infection type and included acylcarnitine (AcCar) (10:1, 16:1, 18:2), diacylglycerols (DG) (16:0_18:0, 18:0_18:0), Lysophosphatidylcholine (LPC) (12:0, 20:5), PE (18:0_18:0), and TG (14:1_16:0_18:2, 15:0_17:0_19:0, 16:0_17:0_18:0, 19:0_19:0_19:0). Cluster analysis yielded three clusters of lipid profiles that were driven by just 10 lipid species (TGs and DGs). Cluster 1 had the highest levels of each lipid species and the highest prevalence of influenza A H3 infection (56%, n = 5) whereas cluster 3 had lower levels of each lipid species and the highest prevalence of rhinovirus (60%; n = 6). Using cluster 3 as the reference group, the crude odds of influenza A H3 infection compared to rhinovirus in cluster 1 was significantly (p = 0.047) higher (OR = 15.00 [95% CI: 1.03, 218.29]). After adjustment for confounders (smoking status and pulmonary comorbidities), the odds ratio (OR) became only marginally significant (p = 0.099), but the magnitude of the effect estimate was similar (OR = 16.00 [0.59, 433.03]).ConclusionsIn this study, human sputum lipid profiles were shown to be associated with distinct types of viral infection. Better understanding the relationship between respiratory infections of global importance and lipids contributes to advancing knowledge of pathogenesis of infections including identifying populations with increased susceptibility and developing effective therapeutics and biomarkers of health status.