Qualitative and quantitative mass spectrometry comparison of characteristic galactosyl lactose isomers from goat milk at different lactation stages.

Galactooligosaccharides are composed mainly of galactosyl lactose, which is important for infant growth and as a functional food additive. Although galactosyl lactose is abundant in goat milk, its complex structure has hindered the separation and analysis of its isomers. In this study, 5 isomers of goat milk galactosyl lactose were separated by HPLC: ß6'-galactosyl lactose (ß6'-GL), α6'-galactosyl lactose (α6'-GL), ß4'-galactosyl lactose (ß4'-GL), α3'-galactosyl lactose (α3'-GL), and ß3'-galactosyl lactose (ß3'-GL). This composition differs from that of commercial galactooligosaccharide products, which comprise mainly ß-configuration oligosaccharides. The isomers were then qualitatively and quantitatively compared at different lactation stages using online HPLC-mass spectrometry. Relative quantitative analysis showed that the total content of the 5 galactosyl lactose isomers was highest in transitional goat milk. Specifically, ß3'-GL was the main isomer in colostrum and α3'-GL was the main isomer in transitional and mature milk. ß6'-Galactosyl lactose and ß4'-GL tended to increase and then decrease during lactation. Moreover, α3'-GL content was 2 times higher than in colostrum and 10 times higher in transitional milk than in mature milk; in contrast, for ß3'-GL, the values were 5 and 2 times higher, respectively. Absolute quantitative analysis revealed that ß3'-GL was the most abundant isomers in colostrum (32.3 mg/L), and α3'-GL was the most abundant in transitional milk (88.1 mg/L) and mature milk (36.3 mg/L). These findings provide an important quantitative basis for understanding the relationship between structure and function of galactosyl lactose in goat milk, as well as its exploitation as a functional food.

Assessing the Role of Nrf2/GPX4-Mediated Oxidative Stress in Arsenic-Induced Liver Damage and the Potential Application Value of Rosa roxburghii Tratt [Rosaceae].

Arsenic poisoning is a geochemical disease that seriously endangers human health. The liver is one of the important target organs for arsenic poisoning, several studies have shown that oxidative stress plays an important role in arsenic-induced liver damage. However, the specific mechanism of arsenic-induced oxidative stress has not yet been fully elucidated, and currently, there are no effective intervention measures for the prevention and treatment of arsenic-induced liver damage. In this study, the effect of the Nrf2/GPX4 signaling pathway and oxidative stress in the arsenic-induced liver damage was first evaluated. The results show that arsenic can activate the Nrf2/GPX4 signaling pathway and increase the oxidative stress, which in turn promotes arsenic-induced liver damage in MIHA cells. Moreover, when we applied the Nrf2 inhibitor, the promoting effect of arsenic on liver damage was alleviated by inhibiting the activation of the Nrf2/GPX4 signaling pathway. Subsequently, the Rosa roxburghii Tratt [Rosaceae] (RRT) intervention experiments in cells and arsenic poisoning population were designed. The results revealed that RRT can inhibit Nrf2/GPX4 signaling pathway to reduce oxidative stress, thereby alleviates arsenic-induced liver damage. This study provides some limited evidence that arsenite can activate Nrf2/GPX4 signaling pathway to induce oxidative stress, which in turn promotes arsenic-induced liver damage in MIHA cells. The second major finding was that Kaji-ichigoside F1 may be a potential bioactive compound of RRT, which can inhibit Nrf2/GPX4 signaling pathway to reduce oxidative stress, thereby alleviates arsenic-induced liver damage. Our study will contribute to a deeper understanding of the mechanisms in arsenic-induced liver damage, these findings will identify a possible natural medicinal food dual-purpose fruit, RRT, as a more effective prevention and control strategies for arsenic poisoning.

Guishaozichuan granules can attenuate asthma in rats via the MUC5AC/EGFR signaling pathway.

Background: Guishaozichuan (GSZC) granules are a traditional Chinese medicine formulation created by Professor Li (Chinese-Japanese Friendship Hospital, Beijing, China) we studied the effect of GSZC granules in rats suffering from asthma. Methods: Specific pathogen-free Sprague-Dawley rats were divided randomly into seven groups. Ovalbumin (OVA) and Al (OH)3 gel were used to create an asthma model. On day 1, rats were injected with OVA (10 mg) and an Al(OH)3 gel suspension (100 mg). One week later, rats were sensitized again. On day 15, rats were given aerosolized OVA (1%) for 30 min/day for 10 days. Gastric administration of OVA was 1 h before nebulization. At 24 h after the last stimulation, changes in airway resistance (RI) and dynamic compliance (Cdyn) in rat lungs were measured after challenge with methacholine at increasing concentrations. The contents of immunoglobulin (Ig)E, interleukin (IL)-4, IL-5, IL-13, and IL-17 in serum were measured by enzyme-linked immunosorbent assays. The percentage of eosinophils (EOS) and the white blood cell (WBC) count in bronchoalveolar lavage fluid (BALF) were counted under an optical microscope. Pathologic alterations in lung tissue were evaluated by optical microscopy, and lung injury score calculated. Expression of mucin 5AC, oligomeric mucus/gel-forming (MUC5AC) and epidermal growth factor receptor (EGFR) in lung tissue was measured by immunohistochemistry. mRNA expression of MUC5AC and EGFR in lung tissue was measured by real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Results: GSZC granules reduced RI markedly and improved Cdyn, decreased serum levels of IgE, IL-4, IL-5, IL-13, IL-17, %EOS and the WBC count in BALF. GSZC granules alleviated lung-tissue damage, diminished the Inflammation Score, and reduced mRNA and protein expression of MUC5AC and EGFR in lung tissue. Conclusion: GSZC granules could improve bronchial hyperresponsiveness, bronchial inflammation, and histopathologic damage in the lungs of rats suffering from asthma. This phenomenon may be related to its regulation of cytokine levels and the MUC5AC/EGFR signaling pathway.

Hydrogen peroxide combined with surfactant leaching and microbial community recovery from oil sludge.

The remediation effects of hydrogen peroxide (H2O2) oxidation and surfactant-leaching alone or in combination on three typical oilfield sludges were studied. The removal efficiency of total petroleum hydrocarbons (TPHs) of Jidong, Liaohe and Jiangsu oil sludges by hydrogen peroxide oxidation alone was very poor (6.5, 6.8, and 3.4 %, respectively) but increased significantly (p < 0.05), especially of long-chain hydrocarbons, by combining the use of H2O2 with surfactants (80.0, 79.8 and 82.2 %, respectively). Oxidation combined with leaching may impair microbial activity and organic manure was therefore added to the treated sludges for biostimulation and the composition and function of the microbial community were studied. The addition of manure rapidly restored sludge microbial activity and significantly increased the relative abundance of some salt-tolerant and alkali-tolerant petroleum-degrading bacteria such as Corynebacterium, Pseudomonas, Dietzia and Jeotgalicoccus. Moreover, the relative abundance of two classic petroleum-degrading enzyme genes, alkane 1-monooxygenase and catechol 1, 2-dioxygenase, increased significantly.

Myofibrillar protein composite gels: effect of esterified potato starch, lard and peanut oil on the gel properties.

BACKGROUND: Heat-induced composite gels were prepared with 20 g kg-1 myofibrillar protein (MP) sol, 20 g kg-1 modified starch and 100 g kg-1 lipid pre-emulsified by MP in 0.6 mol L-1 NaCl, at pH 6.2. The effects of esterified potato starch (EPS) and emulsified lipid (lard or peanut oil) on the rheology, texture properties and nuclear magnetic resonance characterization of MP gel were evaluated. RESULTS: The addition of starch and lipid significantly improved the gel strength and water holding capacity (WHC) of the MP gel. Analysis of the relaxation time compared with the WHC tests showed that the variation range of the transverse T22 relaxation time of a gel was positively proportional to changes in WHC of the composite gel, and the lower the T22 relaxation time, the better the WHC of composite gel. Moreover, MP gel with starch and emulsified lard added at the same time has the lowest T2 relaxation time, and also the best WHC of the gel. Environmental scanning electron microscopy showed that emulsified oil droplets embedded the gaps in the protein network, and the gelatinized starch contributed to restrict the oil droplet size, resulting in thicker MP gel. CONCLUSION: Emulsified lipid and modified starch have an important influence on the rheology and microstructure of MP gels, indicating the subtle interaction between starch, lipid and protein. The results suggest the potential feasibility of modified starch and vegetable oil to improve the textural properties in comminuted meat products. © 2021 Society of Chemical Industry.

Mechanism of Microbial Metabolite Leupeptin in the Treatment of COVID-19 by Traditional Chinese Medicine Herbs.

Coronavirus disease 2019 (COVID-19) has caused huge deaths and economic losses worldwide in the current pandemic. The main protease (Mpro) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is thought to be an ideal drug target for treating COVID-19. Leupeptin, a broad-spectrum covalent inhibitor of serine, cysteine, and threonine proteases, showed inhibitory activity against Mpro, with a 50% inhibitory concentration (IC50) value of 127.2 µM in vitro in our study here. In addition, leupeptin can also inhibit SARS-CoV-2 in Vero cells, with 50% effective concentration (EC50) values of 42.34 µM. More importantly, various strains of streptomyces that have a broad symbiotic relationship with medicinal plants can produce leupeptin and leupeptin analogs to regulate autogenous proteases. Fingerprinting and structure elucidation using high-performance liquid chromatography (HPLC) and high-resolution mass spectrometry (HRMS), respectively, further proved that the Qing-Fei-Pai-Du (QFPD) decoction, a traditional Chinese medicine (TCM) formula for the effective treatment of COVID-19 during the period of the Wuhan outbreak, contains leupeptin. All these results indicate that leupeptin at least contributes to the antiviral activity of the QFPD decoction against SARS-CoV-2. This also reminds us to pay attention to the microbiomes in TCM herbs as streptomyces in the soil might produce leupeptin that will later infiltrate the medicinal plant. We propose that plants, microbiome, and microbial metabolites form an ecosystem for the effective components of TCM herbs. IMPORTANCE A TCM formula has played an important role in the treatment of COVID-19 in China. However, the mechanism of TCM action is still unclear. In this study, we identified leupeptin, a metabolite produced by plant-symbiotic actinomyces (PSA), which showed antiviral activity in both cell culture and enzyme assays. Moreover, leupeptin found in the QFPD decoction was confirmed by both HPLC fingerprinting and HRMS. These results suggest that leupeptin likely contributes to the antiviral activity of the QFPD decoction against SARS-CoV-2. This result gives us important insight into further studies of the PSA metabolite and medicinal plant ecosystem for future TCM modernization research.

Soil microbial community and association network shift induced by several tall fescue cultivars during the phytoremediation of a petroleum hydrocarbon-contaminated soil.

Biodegradation of soil contaminants may be promoted near plant roots due to the "rhizosphere effect" which may enhance microbial growth and activity. However, the effects of different plant cultivars within a single species on degradation remains unclear. Here, we evaluated the removal of soil total petroleum hydrocarbons (TPHs) by ten different cultivars of tall fescue grass (Festuca arundinacea L.) and their associated rhizosphere microbiomes. TPH removal efficiency across the ten different cultivars was not significantly correlated with plant biomass. Rhizing Star and Greenbrooks cultivars showed the maximum (76.6%) and minimum (62.2%) TPH removal efficiencies, respectively, after 120 days. Significant differences were observed between these two cultivars in the composition of rhizosphere bacterial and fungal communities, especially during the early stages (day 30) of remediation but the differences decreased later (day 90). Putative petroleum-degrading bacterial and fungal guilds were enriched in the presence of tall fescue. Moreover, the complexity of microbial networks declined in treatments with higher TPH removal efficiency. The relative abundances of saprotrophic fungi and putative genes alkB and C12O in bacetria involved in petroleum degradation increased, especially in the presence of Rhizing Star cultivar, and this was consistent with the TPH removal efficiency results. These results indicate the potential of tall fescue grass cultivars and their associated rhizosphere microbiomes to phytoremediate petroleum hydrocarbon-contaminated soils.

Inclusion complexes of tea polyphenols with HP-ß-cyclodextrin:Preparation, characterization, molecular docking, and antioxidant activity.

The purpose of this study was to prepare and characterize inclusion complexes between tea polyphenol (TP) and hydroxypropyl-ß-cyclodextrin (HP-ß-CD), and to evaluate their antioxidant properties. Freeze-drying was used to prepare the inclusion complex of TP/HP-ß-CD at different component ratios (1:0.5, 1:1, and 1:2). The supermolecular structure of the TP/HP-ß-CD complex was characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). Molecular docking was used to simulate the positions and interactions of the binding sites of TP/HP-ß-CD inclusion complexes and target protein receptors. In addition, the effects of TP/HP-ß-CD inclusion complexes on myofibrillar protein (MP) from lamb tripe were observed under oxidative conditions. Results showed that TP was encapsulated in the cavity of HP-ß-CD to form an optimal complex with 1:2 molar ratio of stoichiometry, while the FTIR, TGA, and SEM studies also support the inclusion process. Molecular modeling results were systematically analyzed to determine the stability of inclusion complexes and protein. Furthermore, the addition of an appropriate concentration (5 to 105 µmol/g) of TP/HP-ß-CD inclusion complex decreased the carbonyl content, hydrophobicity, and protein aggregation of MP from lamb tripe, whereas it increased the sulfhydryl content. This improved antioxidant activity and bioavailability of the inclusion complexes will be beneficial for its potential applications in food. PRACTICAL APPLICATION: Tea polyphenol was an antioxidant with potential for the field of food. In this study, the unstable properties of tea polyphenols were evaluated and were improved by inclusion of HP-ß-cyclodextrin. The binding mode of the inclusion complex with protein was revealed via the molecular docking method, and the application of inclusion complex to control protein oxidation was studied. Results showed that the inclusion complex could effectively inhibit protein oxidation, which can provide a reference for the application of polyphenols in meat products and the improvement of protein properties.

Enrichment of the soil microbial community in the bioremediation of a petroleum-contaminated soil amended with rice straw or sawdust.

Two common organic wastes from agriculture (rice straw) and forestry (sawdust) were applied to a petroleum-contaminated soil to estimate their effectiveness in the removal of total petroleum hydrocarbons (TPHs) and polycyclic aromatic hydrocarbons (PAHs). Rice straw was the more effective amendment than the other treatments in reducing TPH contents and addition of sawdust resulted in a significant decrease in PAH removal, particularly high-molecular-weight (5-6 ring) PAHs. Principal coordinates analysis (PCoA) indicates that rice straw treatment separated only the bacterial community but sawdust greatly affected both the soil bacterial and fungal communities. Moreover, the abundance of some petroleum degraders such as the bacteria Sphingomonas, Idiomarina and Phenylobacterium and the fungi Humicola, Wallemia and Graphium was promoted by inputs of the two agricultural and forestry wastes. These results highlight the potential of waste applications in accelerating hydrocarbon biodegradation which may be attributed to the enrichment of keystone taxa that show strong positive associations with hydrocarbon degradation.

High pressure homogenization combined with pH shift treatment: A process to produce physically and oxidatively stable hemp milk.

Hemp milk, an emerging beverage with high nutritional value and low allergenicity, is an attractive alternative to dairy, soy, and nut milks. To obtain a non-thermally processed, physically and oxidatively stable hemp milk, high pressure homogenization (HPH) combined with pH shift treatment was investigated. For hemp milk (4% protein, 5% fat) without pH shift, increasing the homogenization pressure (up to 60 MPa) resulted in a more uniform distribution of emulsion droplets (2.2-2.7 µm). When pH shift was applied prior to HPH, large clusters and aggregates of oil droplets (3.5-8.2 µm) were formed. Interestingly, hemp milk with such interactive structures was remarkably stable, showing negligible phase separation within 3-day storage at 4 °C. Moreover, hemp milk made by combined pH shift and HPH exhibited delayed hydroperoxides (expressed as peroxide value, PV) and malondialdehyde (expressed as thiobarbituric acid-reactive substances, TBARS) production, suggesting the resistance of such emulsion cluster structures to radicals. On the other hand, a significant reduction of microbial population was observed in hemp milk prepared by pH shift combined with HPH. The results indicate that the pH shift + HPH combination treatment may potentially be employed for the production of non-thermally processed hemp milk.

Cassiae semen: A review of its phytochemistry and pharmacology (Review).

Cassiae semen (Leguminosae), a well­known traditional Chinese medicine, has been used for a number of centuries in areas of Southeast Asia, including Korea, Japan and China. The present review aims to provide updated and comprehensive information, on the botany, phytochemistry and pharmacology of Cassiae semen. The available information on Cassiae semen was collected using several different resources, including classic books on Chinese herbal medicine and a number of scientific databases, including the China Academic Journals full­text database, PubMed, SciFinder, the Web of Science and Science Direct. To date >70 chemical compounds have been isolated from Cassiae semen, and the major components have been determined to be anthraquinones, naphthopyrones and volatile oil. The crude extracts and pure compounds of Cassiae semen have been used as effective agents in preclinical and clinical practice due to their beneficial activities, including antihyperlipidemic, antidiabetic, neuroprotective, hepatoprotective, antibacterial, antioxidant and hypotensive activities. With the body of reported data, it has been suggested that Cassiae semen has convincing medicinal potential. However, the pharmacological mechanisms of the main bioactive compounds and the association between structure and activity require further investigation.

Characteristics of metal-tolerant plant growth-promoting yeast (Cryptococcus sp. NSE1) and its influence on Cd hyperaccumulator Sedum plumbizincicola.

Plant growth-promoting yeasts are often over looked as a mechanism to improve phytoremediation of heavy metals. In this study, Cryptococcus sp. NSE1, a Cd-tolerant yeast with plant growth capabilities, was isolated from the rhizosphere of the heavy metal hyperaccumulator Sedum plumbizincicola. The yeast exhibited strong tolerance to a range of heavy metals including Cd, Cu, and Zn on plate assays. The adsorption rate Cd, Cu, Zn by NSE1 was 26.1, 13.2, and 25.2 %, respectively. Irregular spines were formed on the surface of NSE1 when grown in MSM medium supplemented with 200 mg L(-1) Cd. NSE1 was capable of utilizing 1-aminocyclopropane-1-carboxylate (ACC) as a sole nitrogen source and was capable of solubilization of inorganic phosphate at rates of 195.2 mg L(-1). Field experiments demonstrated that NSE1 increased phytoremediation by increasing the biomass of Cd hyperaccumulator S. plumbizincicola (46 %, p < 0.05) during phytoremediation. Overall, Cd accumulation by S. plumbizincicola was increased from 19.6 to 31.1 mg m(-2) though no difference in the concentration of Cd in the shoot biomass was observed between NSE1 and control. A Cd accumulation ratio of 38.0 % for NSE1 and 17.2 % for control was observed. The HCl-extractable Cd and CaCl2-extractable Cd concentration in the soil of the NSE1 treatment were reduced by 39.2 and 29.5 %, respectively. Community-level physiology profiling, assessed using Biolog Eco plates, indicated functional changes to the rhizosphere community inoculated with NSE1 by average well color development (AWCD) and measurement of richness (diversity). Values of Shannon-Weiner index, Simpson index, and McIntosh index showed a slight but no significant increases. These results indicate that inoculation of NSE1 could increase the shoot biomass of S. plumbizincicola, enhance the Cd accumulation in S. plumbizincicola, and decrease the available heavy metal content in soils significantly without overall significant changes to the microbial community.