Abstract Introduction Past studies on plant metabolomes have highlighted the influence of growing environments and varietal differences in variation of levels of metabolites yet there remains continued interest in evaluating the effect of genetic modification (GM). Objectives Here we test the hypothesis that metabolomics differences in grain from maize hybrids derived from a series of GM (NK603, herbicide tolerance) inbreds and corresponding negative segregants can arise from residual genetic variation associated with backcrossing and that the effect of insertion of the GM trait is negligible. Methods Four NK603-positive and negative segregant inbred males were crossed with two different females (testers). The resultant hybrids, as well as conventional comparator hybrids, were then grown at three replicated field sites in Illinois, Minnesota, and Nebraska during the 2013 season. Metabolomics data acquisition using gas chromatography–time of flight-mass spectrometry (GC–TOF-MS) allowed the measurement of 367 unique metabolite features in harvested grain, of which 153 were identified with small molecule standards. Multivariate analyses of these data included multi-block principal component analysis and ANOVA-simultaneous component analysis. Univariate analyses of all 153 identified metabolites was conducted based on significance testing (α = 0.05), effect size evaluation (assessing magnitudes of differences), and variance component analysis. Results Results demonstrated that the largest effects on metabolomic variation were associated with different growing locations and the female tester. They further demonstrated that differences observed between GM and non-GM comparators, even in stringent tests utilizing near-isogenic positive and negative segregants, can simply reflect minor genomic differences associated with conventional back-crossing practices. Conclusion The effect of GM on metabolomics variation was determined to be negligible and supports that there is no scientific rationale for prioritizing GM as a source of variation.

Abstract Six new psychoactive substances were identified together with two other substances (compounds 1–8) in illegal products by our ongoing survey in Japan between January and July 2014. A new synthetic cannabinoid, FDU-NNEI [1-(4-fluorobenzyl)-N-(naphthalen-1-yl)-1H-indole-3-carboxamide, 2], was detected with the newly distributed synthetic cannabinoid FDU-PB-22 (1). Two 2H-indazole isomers of synthetic cannabinoids, AB-CHMINACA 2H-indazole analog (3) and NNEI 2H-indazole analog (4), were newly identified with 1H-indazoles [AB-CHMINACA and NNEI indazole analog (MN-18)]. In addition, 2-methylpropyl N-(naphthalen-1-yl) carbamate (5) and isobutyl 1-pentyl-1H-indazole-3-carboxylate (6) were detected in illegal products. Compound 6 is considered to be a by-product of the preparation of NNEI indazole analog from compound 5 and 1-pentyl-1H-indazole. A phenethylamine derivative, N–OH-EDMA [N-hydroxy-3,4-ethylenedioxy-N-methylamphetamine, 7], and a cathinone derivative, dimethoxy-α-PHP (dimethoxy-α-pyrrolidinohexanophenone, 8), were newly identified in illegal products. Among them, compounds 1 and 8 have been controlled as designated substances (Shitei-Yakubutsu) under the Pharmaceutical Affairs Law in Japan since August and November 2014, respectively.

The aim of this study is to compare two sheep farming practices (lamb farming with or without finishing period on concentrate), on carcass characteristics, fatty acid profile and organoleptic quality of the meat. The study was performed on 24 Beni-Guil lambs. Finished animals (F) had access to creep feeding until weaning and were fed a mixed ration of barley and alfalfa hay during a finishing period of 45 days; however, nonfinished animals (NF) remained on the pasture. This comparison was carried out by analyzing the carcass characteristics, proximate composition and fatty acids profile of the meat. Ultimate pH and meat lightness were lower for F-lambs than NF-lambs who didn’t go through the finishing period. The concentrate-based finished practice produced carcasses with better fatness state and conformation. The intramuscular fat content is more important in finished-lambs’ meats (3.81 vs 1.82) which show more juiciness and a high meat’s color lightness. However, meats of NF-lambs presented a high proportion of polyunsaturated fatty acids (PUFA: 20.70 vs 16.82), particularly the PUFA n-3 (3.97 vs 1.17) and consequently a low n-6/n-3 ratio (3.92 vs 12.72). Finally, we recommend, finished lamb meats for the large-scale market and meat of pasture-raised lamb for the niche market.

AbstractThe development of highly efficient C–C bond formation methods for the synthesis of ethyl 2-(2,4-dichloro-5-fluorobenzoyl)-3-(dimethylamino)acrylate 1 in continuous flow processes has been described, which is based on the concept of rapid and efficient activation of carboxylic acid. 2,4-Dichloro-5-fluorobenzoic acid is rapidly converted into highly reactive 2,4-dichloro-5-fluorobenzoyl chloride by treating with inexpensive and less-toxic solid bis(trichloromethyl)carbonate. And then it rapidly reacts with ethyl 3-(dimethylamino)acrylate to afford the desired 1. This process can be performed under mild conditions. Compared with the traditional tank reactor process, less raw material consumption, higher product yield, less reaction time, higher operation safety ensured by more the environmentally friendly procedure, and process continuity are achieved in the continuous-flow system.

AbstractThe biosynthesis of the tetracyclic diterpene ent-kaurene is a critical step in the general (primary) metabolism of gibberellin hormones. ent-Kaurene is formed by a two-step cyclization of geranylgeranyl diphosphate via the intermediate ent-copalyl diphosphate. In a lower land plant, the moss Physcomitrella patens, a single bifunctional diterpene synthase (diTPS) catalyzes both steps. In contrast, in angiosperms, the two consecutive cyclizations are catalyzed by two distinct monofunctional enzymes, ent-copalyl diphosphate synthase (CPS) and ent-kaurene synthase (KS). The enzyme, or enzymes, responsible for ent-kaurene biosynthesis in gymnosperms has been elusive. However, several bifunctional diTPS of specialized (secondary) metabolism have previously been characterized in gymnosperms, and all known diTPSs for resin acid biosynthesis in conifers are bifunctional. To further understand the evolution of ent-kaurene biosynthesis as well as the evolution of general and specialized diterpenoid metabolisms in gymnosperms, we set out to determine whether conifers use a single bifunctional diTPS or two monofunctional diTPSs in the ent-kaurene pathway. Using a combination of expressed sequence tag, full-length cDNA, genomic DNA, and targeted bacterial artificial chromosome sequencing, we identified two candidate CPS and KS genes from white spruce (Picea glauca) and their orthologs in Sitka spruce (Picea sitchensis). Functional characterization of the recombinant enzymes established that ent-kaurene biosynthesis in white spruce is catalyzed by two monofunctional diTPSs, PgCPS and PgKS. Comparative analysis of gene structures and enzyme functions highlights the molecular evolution of these diTPSs as conserved between gymnosperms and angiosperms. In contrast, diTPSs for specialized metabolism have evolved differently in angiosperms and gymnosperms.

Abstract Compounds of the terpenoid class play numerous roles in the interactions of plants with their environment, such as attracting pollinators and defending the plant against pests. We show here that the genome of cultivated tomato (Solanum lycopersicum) contains 44 terpene synthase (TPS) genes, including 29 that are functional or potentially functional. Of these 29 TPS genes, 26 were expressed in at least some organs or tissues of the plant. The enzymatic functions of eight of the TPS proteins were previously reported, and here we report the specific in vitro catalytic activity of 10 additional tomato terpene synthases. Many of the tomato TPS genes are found in clusters, notably on chromosomes 1, 2, 6, 8, and 10. All TPS family clades previously identified in angiosperms are also present in tomato. The largest clade of functional TPS genes found in tomato, with 12 members, is the TPS-a clade, and it appears to encode only sesquiterpene synthases, one of which is localized to the mitochondria, while the rest are likely cytosolic. A few additional sesquiterpene synthases are encoded by TPS-b clade genes. Some of the tomato sesquiterpene synthases use z,z-farnesyl diphosphate in vitro as well, or more efficiently than, the e,e-farnesyl diphosphate substrate. Genes encoding monoterpene synthases are also prevalent, and they fall into three clades: TPS-b, TPS-g, and TPS-e/f. With the exception of two enzymes involved in the synthesis of ent-kaurene, the precursor of gibberellins, no other tomato TPS genes could be demonstrated to encode diterpene synthases so far.

Abstract Diterpene resin acids (DRAs) are specialized (secondary) metabolites of the oleoresin defense of conifers produced by diterpene synthases and cytochrome P450s of the CYP720B family. The evolution of DRA metabolism shares common origins with the biosynthesis of ent-kaurenoic acid, which is highly conserved in general (primary) metabolism of gibberellin biosynthesis. Transcriptome mining in species of spruce (Picea) and pine (Pinus) revealed CYP720Bs of four distinct clades. We cloned a comprehensive set of 12 different Sitka spruce (Picea sitchensis) CYP720Bs as full-length cDNAs. Spatial expression profiles, methyl jasmonate induction, and transcript enrichment in terpenoid-producing resin ducts suggested a role of CYP720B4 in DRA biosynthesis. CYP720B4 was characterized as a multisubstrate, multifunctional enzyme by the formation of oxygenated diterpenoids in metabolically engineered yeast, yeast in vivo transformation of diterpene substrates, in vitro assays with CYP720B4 protein produced in Escherichia coli, and alteration of DRA profiles in RNA interference-suppressed spruce seedlings. CYP720B4 was active with 24 different diterpenoid substrates, catalyzing consecutive C-18 oxidations in the biosynthesis of an array of diterpene alcohols, aldehydes, and acids. CYP720B4 was most active in the formation of dehydroabietic acid, a compound associated with insect resistance of Sitka spruce. We identified patterns of convergent evolution of CYP720B4 in DRA metabolism and ent-kaurene oxidase CYP701 in gibberellin metabolism and revealed differences in the evolution of specialized and general diterpene metabolism in a gymnosperm. The genomic and functional characterization of the gymnosperm CYP720B family highlights that the evolution of specialized metabolism involves substantial diversification relative to conserved, general metabolism.

Abstract Cannabis (Cannabis sativa) resin is the foundation of a multibillion dollar medicinal and recreational plant bioproducts industry. Major components of the cannabis resin are the cannabinoids and terpenes. Variations of cannabis terpene profiles contribute much to the different flavor and fragrance phenotypes that affect consumer preferences. A major problem in the cannabis industry is the lack of proper metabolic characterization of many of the existing cultivars, combined with sometimes incorrect cultivar labeling. We characterized foliar terpene profiles of plants grown from 32 seed sources and found large variation both within and between sets of plants labeled as the same cultivar. We selected five plants representing different cultivars with contrasting terpene profiles for clonal propagation, floral metabolite profiling, and trichome-specific transcriptome sequencing. Sequence analysis of these five cultivars and the reference genome of cv Purple Kush revealed a total of 33 different cannabis terpene synthase (CsTPS) genes, as well as variations of the CsTPS gene family and differential expression of terpenoid and cannabinoid pathway genes between cultivars. Our annotation of the cv Purple Kush reference genome identified 19 complete CsTPS gene models, and tandem arrays of isoprenoid and cannabinoid biosynthetic genes. An updated phylogeny of the CsTPS gene family showed three cannabis-specific clades, including a clade of sesquiterpene synthases within the TPS-b subfamily that typically contains mostly monoterpene synthases. The CsTPSs described and functionally characterized here include 13 that had not been previously characterized and that collectively explain a diverse range of cannabis terpenes.

Introduction. Propolis has been used traditionally in several countries for treating various diseases as it possessed healing properties including antioxidant and anticancer qualities. In Peninsular Malaysia, Tetrigona apicalis is one of the species of stingless bees mainly found in virgin jungle reserves which largely contribute to propolis production. Therefore, this study is designed to evaluate the phytochemical contents, antioxidant properties, and the cytotoxic effect of ethanolic crude of propolis extract against MCF7 and MCF 10A cell lines. Method. The ethanolic extract of propolis (EEP) was extracted using 80% ethanol. Identification of phytochemical contents and antioxidant properties of EEP was analysed by gas chromatography-mass spectrometry (GC-MS) and using 2, 2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) method, respectively. The EEP cytotoxic activity was evaluated on MCF7 and MCF 10A using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Results. Phytochemical contents of EEP demonstrated 28 compounds in which caryophyllene (99%), β-amyrin (96%), α-amyrin (93%), and caryophyllene oxide (93%) were the main compounds. The percentage of ABTS+ scavenging activity of EEP showed an inhibition of 9.5% with half-inhibitory concentration (IC50) value of 1.68 mg/mL. The EEP reduced MCF7 cells viability at IC50 value of 62.24 μg/mL, 44.15 μg/mL, and 32.70 μg/mL at 24, 48, and 72 hours, respectively. The IC50 value of MCF 10A was 49.55 μg/mL, 56.05 μg/mL, and 72.10 μg/mL at 24, 48, and 72 hours, respectively. The EEP cytotoxic effect of T. apicalis was more selective towards MCF7 at 72-hour incubation with a selectivity index (SI) of 2.20. Conclusion. The EEP has been shown to have antioxidants and potential bioactive compounds and inhibited proliferation of the MCF7 cells. Further studies on the EEP role in the apoptosis pathway and its screening towards other cell lines will be evaluated.

Rhynchanthus beesianus is a medicinal, ornamental, and edible plant, and its essential oil has been used as an aromatic stomachic in China. In this study, the chemical constituents, antibacterial, and anti-inflammatory properties of flower essential oil (F-EO), leaf essential oil (L-EO), and stem essential oil (S-EO) of R. beesianus were investigated for the first time. According to the GC-FID/MS assay, the F-EO was mainly composed of bornyl formate (21.7%), 1,8-cineole (21.6%), borneol (9.7%), methyleugenol (7.7%), β-myrcene (5.4%), limonene (4.7%), camphene (4.5%), linalool (3.4%), and α-pinene (3.1%). The predominant components of L-EO were bornyl formate (33.9%), borneol (13.2%), 1,8-cineole (12.1%), methyleugenol (8.0%), camphene (7.8%), bornyl acetate (6.2%), and α-pinene (4.3%). The main components of S-EO were borneol (22.5%), 1,8-cineole (21.3%), methyleugenol (14.6%), bornyl formate (11.6%), and bornyl acetate (3.9%). For the bioactivities, the F-EO, L-EO, and S-EO exhibited significant antibacterial property against Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, Proteus vulgaris, Pseudomonas aeruginosa, and Escherichia coli with the inhibition zones (7.28–9.69 mm), MIC (3.13–12.50 mg/mL), and MBC (6.25–12.50 mg/mL). Besides, the F-EO, L-EO, and S-EO significantly inhibited the production of proinflammatory mediator nitric oxide (NO) (93.15–94.72%) and cytokines interleukin-6 (IL-6) (23.99–77.81%) and tumor necrosis factor-α (TNF-α) (17.69–24.93%) in LPS-stimulated RAW264.7 cells at the dose of 128 μg/mL in the absence of cytotoxicity. Hence, the essential oils of R. beesianus flower, leaf, and stem could be used as natural antibacterial and anti-inflammatory agents with a high application potential in the pharmaceutical and cosmetic fields.

Twenty Beni-Guil-PGI female lambs were used to study the effects of rearing season on meat quality characteristics, fatty acids profile, and lipid content. The animals were reared according to the pastoral-transhumant system in the eastern region of Morocco. The treatments consisted of 10 female lambs reared in summer-autumn (SA) and slaughtered at winter season and 10 female lambs reared in winter-spring (WS) and slaughtered at spring season. After the slaughter, the longissimus lumborum was collected for each animal for meat quality analysis. Compared to lambs reared in SA, the meat from the WS group showed higher ( $p<0.01$ ) pH, chroma, and lightness values (5.79 vs. 5.72, 23.97 vs. 18.46, and 47.03 vs. 41.04, respectively). On the other hand, the meat from WS presented higher ( $p<0.05$ ) intramuscular fat content (5.14 % vs. 3.82%, respectively). However, the intramuscular fat of the lambs reared in SA was characterized by greater ( $p<0.01$ ) PUFA percentage (16.82% vs. 12.40%, respectively), thrombogenic ( $p<0.001$ ) and atherogenic index ( $p<0.001$ ), and PUFA/SFA ratio ( $p<0.01$ ; 0.42 vs. 0.25, respectively). Nevertheless, those reared in WS season have a higher ( $p<0.001$ ) PUFA n − 3 (2.58% vs. 1.14%, respectively) content, and therefore favorable ( $p<0.001$ ) n − 6/n − 3 ratio (3.78 vs. 12.98, respectively).

As a popular fermented condiment in oriental countries, soy sauce plays a more and more important role in modern food culture due to its unique smell and delicious taste. With the help of microwave extraction and gas chromatography-tandem mass spectrometry, the sample preparation method is aimed to determine the content of cyclohexane, benzene, toluene, chlorobenzene, and styrene in soy sauce. The method was validated by examining the linearity, accuracy, specificity, precision, the limit of detection, and quantitation. Meanwhile, three key factors have an impact on the efficiency and accuracy of the method including extracting solvent, temperature, and time which were optimized. The result shows that the recoveries of spiked analytes ranged from 80.86% to 105.71%, the relative standard deviation of intraday and interday precision was no more than 12.1% and 12.5%, and the limit of detection and quantitation were 0.25–1.00 ng/mL and 0.50–2.00 ng/mL, respectively. The results also indicated that the proposed method was a simple, reliable, and sensitive approach for the determination trace amount of five harmful volatile organic compounds from soy sauce.

Visual detection of meat spoilage was performed based on hydrolysis-induced silver metallization on gold nanoparticles (Au NPs). The hydrolysis of 4-I-benzene-bounded Wang resin was induced by the release of a biogenic amine followed by Au-catalyzed Heck cross-coupling reaction that made silver-coated gold core-shell NPs (Au@Ag) in the presence of Ag ions (Ag metallization). A portable sensory cap was designed by this hypothesis and the successful results were obtained for histamine, trimethylamine, and a spoilage sheep meat. With this protocol, the localized surface plasmon resonance (LSPR) is tuned for absorption of Au NPs and leads to LSPR peak blue shift of gold nanoparticles due to the Ag metallization and the preparation of Au@Ag core-shell NPs. Au NPs and the resulting Au@Ag NPs were characterized by transmission electron microscopy (TEM), BET, ultraviolet-visible (UV-Vis), X-ray diffraction (XRD), energy dispersive X-ray (EDX), and dynamic light scattering (DLS) analyses. Also, various control experiments were set up to credit the portable sensory tube.

Acinetobacter baumannii (A. baumannii) is a dangerous nosocomial pathogen in intensive care units, causing fatal clinical challenges and mortality. In this study, the green synthesis of silver nanoparticles (AgNPs) using the extract of Ferula asafetida and the chemical synthesis of AgNPs were carried out to evaluate their effects on A. baumannii bacterial strain and a human adenocarcinoma cell line. The NPs were characterized using several techniques, including field emission-scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray spectrometry, UV-visible spectroscopy, and Fourier-transform infrared spectroscopy. After synthesis, the arrangement of AgNPs was confirmed based on the maximum absorption peak at 450 nm. The results showed that the AgNPs had a hexagonal structure. The antimicrobial activity of biogenic NPs significantly increased and reached a minimum inhibitory concentration of 2 μg/mL. The nanomaterials did not exhibit any toxic effects on the human cell line at certain concentrations and showed improvements compared to chemically synthesized AgNPs. However, at higher concentrations (100 μg/mL), the cytotoxicity increased. Finally, it was concluded that biosynthesized AgNPs had significant antimicrobial effects on A. baumannii isolated from intensive care units.

This study aimed to synthesize alumina from an inorganic aluminum nitrate precursor in various binary solvent systems of ethanol and water using the sol-gel self-assembly (SSA) method, employing a triblock copolymer, pluronic P123, as the pore-directing agent. The resulting materials were implemented as a support for the cobalt (Co) catalyst in a methane dry reforming (MDR) reaction at 1073 K under 1 atm. Regardless of the water percentage used in the support synthesis, the methane dry reforming reaction over Co catalysts on alumina supports showed the negligible change in conversion during the 12 h reaction. Moreover, there was evidence of large quantities of amorphous carbon and graphitic carbon on the spent catalyst surface. However, the low oxidation temperature of these deposited carbons could help maintain the balance between the carbon formation and the carbon elimination processes on the catalyst surface during the reforming reaction, hence prolonging the lifetime of the catalyst. The high conversion of methane (CH4) from 64.6% to 82.8% and carbon dioxide (CO2) from 70.7% to 86.6% for the MDR reaction over the as-prepared alumina-supported Co catalyst demonstrated a significant improvement in catalyst production for the MDR reaction from the viewpoint of large-scale applications.

This article reports on the preparation of iron nanoparticles (FeNPs) supported in chitosan beads (Chi-EDGE-Fe) for removing aldrin from aqueous solutions. The FeNPs and Chi-EDGE-Fe beads were characterized by means of scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), and the Mössbauer spectroscopy (MS) techniques. TEM, XRD, and MS showed that the FeNPs had core-shell structures consisting of a core of either Fe0 or Fe2B and a shell of magnetite. Furthermore, SEM images showed that Chi-EDGE-Fe beads were spherical with irregular surfaces and certain degrees of roughness and porosity, whilst the sorbent mean pore size was 204 nm, and the occluded iron nanoparticles in the chitosan material had diameters of 70 nm and formed agglomerates. The sorbent beads consisted of carbon, oxygen, chlorine, aluminum, silicon, and iron according to the SEM-EDS analysis. Functional groups such as O-H, C-H, -CH2, N-H, C-O, C-OH, and Fe-OH were detected in the FTIR spectra. In addition, a characteristic band appeared at about 1700 cm−1 after the sorption process involving aldrin. MS also showed that the iron nanoparticles in the beads probably oxidized into NPs of α-Fe2O3 as a result of the supporting process. The isotherm of the aldrin removal followed the Langmuir–Freundlich model and presented a maximum adsorption capacity of 74.84 mg/g, demonstrating that chitosan-Fe beads are promising sorbents for the removal of toxic pollutants in aqueous solutions.

Objective. The aim of the study was to investigate the potential effects of waste anesthetic gas (WAG) on oxidative stress, DNA damage, and vital organs. Methods. A total of 150 members of the staff at a hospital were assigned to an exposure group or control group. The 68 operating room (OR) staff in the exposure group were exposed to WAG, and the 82 non-OR staff in the control group were not exposed to WAG. Air samples were collected in the OR, and the sevoflurane concentrations in the samples were determined. Superoxide dismutase (SOD), glutathione peroxidase (GSH-px), and malondialdehyde (MDA) in plasma from the participants were determined to assess oxidative stress. Western blot analysis was used to detect γH2AX in peripheral blood to assess DNA damage. Hematopoietic parameters, liver function, kidney function, and changes in electrophysiology were assessed to identify the effects on the vital organs. Results. The mean (±standard deviation) sevoflurane concentration in 172 air samples from 22 operating rooms was $1.11\pm 0.65\text{ppm}$ . The superoxide dismutase activity and vital organ parameters (lymphocyte, hemoglobin, and total protein concentrations and heart rate) were significantly lower ( $P<0.05$ ) in the exposed group than the control group. The malondialdehyde, total bilirubin, and creatinine concentrations and QT and QTc intervals were significantly higher ( $P<0.05$ ) in the exposed group than the control group. There were no significant differences between the glutathione peroxidase activities and γH2AX concentrations for the exposed and control groups. Conclusions. Long-term occupational exposure to waste anesthetic gas may affect the antioxidant defense system and probably affects vital organ functions to some extent. No correlation between DNA damage and chronic exposure to WAG was observed.

Fruit of salak (Salaaca zalacca) is traditionally used and commercialized as an antidiabetic agent. However, scientific evidence to prove this folk claim is quite lacking. Therefore, this research was aimed to evaluate the α-glucosidase inhibition activity of S. zalacca fruit and identify the bioactive compounds. The fruits were extracted by different ratios of ethanol and water (0, 20, 40, 60, 80, 100%, v/v) to get E0 (100% water), E20 (20% ethanol), E40 (40% ethanol), E60 (60% ethanol), E80 (80% ethanol), and E100 (100% ethanol) extracts. The extracts obtained were subjected to the α-glucosidase inhibitory assay. Gas chromatography-mass spectrometry- (GC-MS-) based metabolomics approach was used in profiling the bioactive metabolites present in the extracts. Orthogonal partial least square (OPLS) was used to correlate GC-MS data and α-glucosidase assay results to identify the possible chemical markers. All active compounds identified were subjected to molecular docking. The extracts from the S. zalacca fruit showed potent inhibition activity against α-glucosidase. The IC50 values from the α-glucosidase inhibitory assay ranged between 16 and 275 µg/ml. Overall, E60 displayed significantly higher α-glucosidase inhibition activity, while E0 showed the lowest α-glucosidase inhibition activity. Major compounds detected in S. zalacca fruits were sugars, fatty acids, and sterols, including myo-inositol, palmitic acid, stearic acid, and β-sitosterol. Moreover, the results obtained from molecular docking indicated that palmitic acid and β-sitosterol were close to the active side of the enzyme. Some of the residues that interacted include HID295, ASN259, LEU313, LYS125, PHE159, VAL216, PHE178, TYR72, TYR158, HIE315, ARG315, and PHE303. The bioassay result strongly suggests that E60 extract from S. zalacca fruits has potential α-glucosidase inhibitory activity. The hydrophobic compounds, including palmitic acid and β-sitosterol, were found to induce the α-glucosidase inhibition activity.

Chemical contamination and safe work practices of workers in automotive refinishing shops have been extensively studied in industrialized countries, but the evidence in developing countries is limited. This study aimed to evaluate chemical contamination and the use of personal protective equipment (PPE) of workers in local small-scale automotive refinishing shops in Nakhon Si Thammarat, Thailand. Airborne toluene and heavy metals, i.e., lead, chromium, and cadmium, were measured in 3 automotive refinishing shops. Toluene exposure assessed by urinary hippuric acid (n = 27) and metal contamination on workers’ hands (n = 24) were also determined. Information on the use of PPE and personal hygiene practices of the workers was collected by questionnaires. Average ambient levels of toluene (0.04–18.26 ppm) and the metals (Pb: ND-26.34, Cr: 0.02–4.46, and Cd: ND-1.44 µg/m3) in all sites did not exceed the national standard levels of 200 ppm for toluene (1998) and 50, 12, and 5 µg/m3 for Pb, Cr, and Cd, respectively (2017). The mean ambient levels of these chemicals were highest in paint spray booths followed by nonpainting areas and office rooms, respectively. The highest level of urinary hippuric acid (1.13 g/g creatinine) was found in a painter but did not exceed the recommended biological exposure index of 1.6 g/g creatinine (2014). In contrast, the highest levels of lead and chromium detected on the workers’ hands were found in body repair technicians. Direct hand contact without using gloves was suggested as a primary cause of metal contamination.

Ochratoxin A (OTA) is a common environmental pollutant found in a variety of foods and grains, and excessive OTA consumption causes serious global health effects on animals and humans. Astaxanthin (AST) is a natural carotenoid that has anti-inflammatory, antiapoptotic, immunomodulatory, antitumor, antidiabetes, and other biological activities. The present study is aimed at investigating the effects of AST on OTA-induced cecum injury and its mechanism of action. Eighty C57 mice were randomly divided into four groups, including the control group, OTA group (5 mg/kg body weight), AST group (100 mg/kg body weight), and AST intervention group (100 mg/kg body weight AST+5 mg/kg body weight OTA). It was found that AST decreased the endotoxin content, effectively prevented the shortening of mouse cecum villi, and increased the expression levels of tight junction (TJ) proteins, consisting of occludin, claudin-1, and zonula occludens-1 (ZO-1). AST increased the number of goblet cells, the contents of mucin-2 (MUC2), and defensins (Defa5 and β-pD2) significantly, while the expression of mucin-1 (MUC1) decreased significantly. The 16S rRNA sequencing showed that AST affected the richness and diversity of cecum flora, decreased the proportion of lactobacillus, and also decreased the contents of short-chain fatty acids (SCFAs) (acetate and butyrate). In addition, AST significantly decreased the expression of TLR4, MyD88, and p-p65, while increasing the expression of p65. Meanwhile, the expression of inflammatory factors including TNF-α and INF-γ decreased, while the expression of IL-10 increased. In conclusion, AST reduced OTA-induced cecum injury by regulating the cecum barrier function and TLR4/MyD88/NF-κB signaling pathway.