Innovative utilization of herbal residues: Exploring the diversity of mechanisms beneficial to regulate anaerobic fermentation of alfalfa.

In order to increase the utilization of herbal residues, realize efficient utilization of resources, the bacterial community and anaerobic fermentation characteristics of alfalfa ensiling treated with 36 kinds of herbal residues were studied. All the herbal residues improved the anaerobic fermentation quality in different degrees, indicated by lower pH, NH3-N and butyric acid concentrations. However, the contents of lactic and acetic acids varied widely in silage with different herbal residues. Pearson's correlation analysis showed that the improved fermentation quality was closely associated with the variation of lactic acid bacteria community. Consequently, the herbal residues could improve anaerobic fermentation quality by stimulating desirable Lactobacillus species and inhibiting undesirable microbes. This study provides new insights for efficient utilization of herbal residues.

The Preventive Value of Acupoint Sensitization for Patients with Stable Angina Pectoris: A Randomized, Double-Blind, Positive-Controlled, Multicentre Trial.

BACKGROUND: Acupoint sensitization is considered an important factor in the efficacy of acupoint therapy. This study aimed to evaluate the efficacy of acupressure in the prevention of stable angina pectoris using acupoints with different pressure-pain sensitivities. METHODS: A total of 202 patients were enrolled and randomly assigned to a high-sensitivity group (HSG) (n = 109) in which patients received acupressure at the five acupoints with the highest sensitivity to pain and a low-sensitivity group (LSG) (n = 93) in which patients received acupressure at the five acupoints with the lowest sensitivity to pain. The duration of acupressure treatment was 4 weeks, and the patients were evaluated at baseline, week 4, and week 8. The primary outcome was a change in the frequency of angina attacks from baseline. The secondary outcomes included nitroglycerin consumption, the Canadian Cardiovascular Society classification, and the Seattle Angina Questionnaire score. Adverse events such as bleeding and subcutaneous haemorrhage were recorded in both groups. RESULTS: The effect of acupressure compared with baseline on the prevention of angina pectoris in HSG was better than that in LSG at week 4 (incidence rate ratio (IRR): 0.691 and 95% confidence interval (CI): [0.569, 0.839]) and week 8 (IRR: 0.692 and 95% CI: [0.569, 0.839]). No significant difference between groups was found in the frequency of nitroglycerin consumption at week 4 (odds ratio (OR) = 0.863 and 95% CI: [0.147, 5.077]) or week 8 (OR = 1.426 and 95% CI: [0.211, 9.661]). Two themes in the questionnaire showed significantly different changes from baseline between the two groups. Scores on the angina frequency (AF) subscale had changed more from the baseline in the HSG at week 8 than in the LSG (mean difference (MD) = 3.807 and 95% CI: [0.673, 6.942]). Scores on the treatment satisfaction (TS) subscale had also changed more in the HSG than in the LSG at week 4 (MD = 3.651 and 95% CI: [0.327, 7.327]) and week 8 (MD = 4.220 and 95% CI: [0.347, 7.346]). One patient in the LSG reported bruising at the acupoint. No unexpected safety problems arose. CONCLUSIONS: This study showed that acupressure at acupoints with high sensitivity to pain may effectively reduce the frequency of stable angina pectoris episodes. This trial is registered with NCT03975140.

Exploring the Addition of Herbal Residues on Fermentation Quality, Bacterial Communities, and Ruminal Greenhouse Gas Emissions of Paper Mulberry Silage.

This study aimed to investigate the influence of herbal residues on the fermentation quality and ruminal fermentation of paper mulberry silage. Clove, mint, and purple perilla residues were used as additives. Silage treatments were designed as control (no additives), 5% of clove, 5% of mint, and 5% of purple perilla. After 21 and 75 days of fermentation, the fermentation characteristics, bacterial communities, and ruminal greenhouse gas emissions in vitro incubation of paper mulberry were analyzed. The results showed that the used herbal residues could reduce the protein losses in paper mulberry silage based on the lower contents of ammoniacal nitrogen and nonprotein nitrogen. Compared with control, higher lactic acid and propionic acid contents were observed in the silages treated with mint and purple perilla but with a higher acetic acid content in clove treatment. Real-time sequencing technology (single-molecule real-time) revealed that Lactobacillus was the dominant bacteria in all silages at the genus level, whereas the bacterial abundance in the treated silages differed greatly from control at the species level. Lactobacillus hammesii abundance was the highest in control, whereas Lactobacillus acetotolerans was the first predominant in the treated silages. All the additives enhanced the digestibility of in vitro dry matter significantly. However, purple perilla decreased the production of total gas, methane, and carbon dioxide. The findings discussed earlier suggested that herbal residues have potential effects in improving fermentation quality, reducing protein loss, and modulating greenhouse gas emissions in the rumen of paper mulberry silage by shifting bacterial community composition.

Antioxidant evaluation-guided chemical profiling and structure-activity analysis of leaf extracts from five trees in Broussonetia and Morus (Moraceae).

Morus and Broussonetia trees are widely used as food and/or feed. Among 23 phenolics identified from leaves of five Moraceae species using UPLC-QTOF-MS/MS, 15 were screened using DPPH/ABTS-guided HPLCs, including seven weak (flavonoids with one hydroxyl on B-ring) and eight strong (four caffeoylquinic acids and four flavonoids, each with a double hydroxyl on B-ring) antioxidants. We then determined the activity and synergistic effects of individual antioxidants and a mixture of the eight strongest antioxidants using DPPH-guided HPLC. Our findings revealed that (1) flavonoid glucuronide may have a more negative effect on antioxidant activity than glucoside, and (2) other compounds in the mixture may exert a negative synergistic effect on antioxidant activity of the four flavonoids with B-ring double hydroxyls but not the four caffeoylquinic acids. In conclusion, the eight phenolics with the strongest antioxidant ability reliably represented the bioactivity of the five extracts examined in this study. Moreover, the Morus alba hybrid had more phenolic biosynthesis machinery than its cross-parent M. alba, whereas the Broussonetia papyrifera hybrid had significantly less phenolic machinery than B. papyrifera. This difference is probably the main reason for livestock preference for the hybrid of B. papyrifera over B. papyrifera in feed.

Water-induced self-assembly of mixed gelator system (ceramide and lecithin) for edible oil structuring.

Ceramide (CER) is a highly effective organogelator with numerous health implications. Lecithin (LEC) is often used as a crystal modifier to improve gel properties. It was found in this study that in the presence of a small amount of water, CER and LEC self-assembled at specific ratios and form oleogels in sunflower oil (SFO), and the structure of which differed from any of the mono-component gel structures. To the best of our knowledge, this is the first water-induced multi-component oleogelation reported to date. To clarify when gelation occurs, a phase diagram of the four-component system SFO/CER/LEC/water was established and characterized. In the gel region, confocal laser scanning microscopy revealed that water was evenly distributed in the self-assembled microstructure. To clarify the vital role of water in formation of stable gel, a series of samples with varying ratios in CER and LEC with and without water were explored. The thermal properties, viscoelastic properties, microstructures, as well as crystal packing were investigated. With water added, the microscopic structures of CER/LEC gels changed from three-dimensional dendritic crystals to two-dimensional rod-like or platelet-like crystals, which are more effective at structuring liquid oil. A different reflection was observed (d-spacing of 14.9 Å), suggesting the rearrangement in crystal packing of ceramide and lecithin molecules, compared to the d-spacing in oleogels with single gelator (13.1 Å and 15.3 Å, respectively for ceramide and lecithin). Fourier transform infrared spectroscopy results indicated the presence of interaction between water and gelators, which confirmed the importance of water in self-assembly of this mixed-gelator system. Water-induced conversion of oleogelation provides alternatives in producing hard-stock fat replacers as well as possibilities to incorporate water-soluble nutrients into oil-based products.

Jasmonate-responsive MYB factors spatially repress rutin biosynthesis in Fagopyrum tataricum.

Jasmonates are plant hormones that induce the accumulation of many secondary metabolites, such as rutin in buckwheat, via regulation of jasmonate-responsive transcription factors. Here, we report on the identification of a clade of jasmonate-responsive subgroup 4 MYB transcription factors, FtMYB13, FtMYB14, FtMYB15, and FtMYB16, which directly repress rutin biosynthesis in Fagopyrum tataricum. Immunoblot analysis showed that FtMYB13, FtMYB14, and FtMYB15 could be degraded via the 26S proteasome in the COI1-dependent jasmonate signaling pathway, and that this degradation is due to the SID motif in their C-terminus. Yeast two-hybrid and bimolecular fluorescence complementation assays revealed that FtMYB13, FtMYB14, and FtMYB15 interact with the importin protein Sensitive to ABA and Drought 2 (FtSAD2) in stem and inflorescence. Furthermore, the key repressor of jasmonate signaling FtJAZ1 specifically interacts with FtMYB13. Point mutation analysis showed that the conserved Asp residue of the SID domain contributes to mediating protein-protein interaction. Protoplast transient activation assays demonstrated that FtMYB13, FtMYB14, and FtMYB15 directly repress phenylalanine ammonia lyase (FtPAL) gene expression, and FtSAD2 and FtJAZ1 significantly promote the repressing activity of FtMYBs. These findings may ultimately be promising for further engineering of plant secondary metabolism.

Antihyperglycemic drugs use and new-onset atrial fibrillation in elderly patients.

BACKGROUND: Antihyperglycemic drugs have been linked to new-onset atrial fibrillation (NAF). However, the effect of the different classes of antihyperglycemic drugs on the development of NAF in elderly patients has not been well studied. In this study, we investigated the association between different classes of antihyperglycemic drugs and NAF in elderly patients. MATERIALS AND METHODS: This was a nested case-control study performed using the database of National Health Insurance programme in Taiwan. Each participant aged 65 years and older who were NAF from 2005 to 2012 were assigned to the NAF group, whereas case was sex-, age-, diabetes duration-, index date-matched, and Charlson Comorbidity Index score-matched randomly selected participant without NAF were assigned to the non-NAF group. Multivariable logistic regression model was used for the estimation of odds ratios (ORs) and 95% confidence intervals (CIs) of NAF associated with use of different classes of antihyperglycemic agents. Nonusers served as the reference group. RESULTS: We identified 1958 cases and 7832 controls. The risk of NAF after adjusting for sex, age, comorbidities and concurrent medication was higher among the users of insulin than among the nonusers (OR, 1·58; 95% CI, 1·37-1·82). Patients who took dipeptidyl peptidase 4 inhibitors were at lower risk of developing NAF than the nonusers (OR, 0·65; 95% CI, 0·45-0·93). CONCLUSIONS: In this population, use of dipeptidyl peptidase 4 inhibitor was associated with a low risk of NAF. Insulin use was associated with a significant increase in the risk of NAF during the long-term follow-up.

Antimicrobial and antioxidant activities of the flower essential oil of Halimodendron halodendron.

The essential oil obtained by hydro-distillation from the flowers of Halimondendron halodendron (Leguminosae) was analyzed for its chemical composition by gas chromatography-mass spectrometry (GC-MS). Undecane (16.4%), dodecane (15.3%), tridecane (12.5%), decane (8.2%), 6,10,14-trimethyl-pentadecan-2-one (6.3%), methyl palmitate (6.0%), methyl linolenate (4.1%) and ethylcyclohexane (4.1%) were the major compounds of the thirty-five identified components of the oil. The essential oil was shown to have a broad spectrum of antimicrobial activity with MIC values ranging from 100 to 250 microg/mL, and IC50 values from 40.4 to 193.8 microg/mL. The oil also showed strong antioxidant activity, with an especially high metal chelating capacity of ferrous ions with an IC50 value of 7.4 microg/mL on ferrozine-Fe2+ complex formation.

Antimicrobial activity of sphingolipids isolated from the stems of cucumber (Cucumis sativus L.).

Three antimicrobial sphingolipids were separated by bioassay-guided isolation from the chloroform fraction of the crude methanol extract of cucumber (Cucumis sativus L.) stems and identified as (2S,3S,4R,10E)-2-[(2'R)-2-hydroxytetra-cosanoylamino]-1,3,4-octadecanetriol-10-ene (1), 1-O-ß-D-glucopyranosyl(2S,3S,4R,10E)-2-[(2'R)-2-hydroxy-tetracosanoylamino]-1,3,4-octadecanetriol-10-ene (2) and soya-cerebroside I (3) by their physicochemical properties and spectroscopic analysis. They were evaluated to show antifungal and antibacterial activity on test microorganisms including four fungal and three bacterial species. Among them, compound 1, a relatively low polarity aglycone,  exhibited stronger antimicrobial activity than its corresponding glycoside 2. The results indicated that sphingolipids could be the main antimicrobial compounds in the crude methanol extract of cucumber stems.

Preparative separation of phenolic compounds from Halimodendron halodendron by high-speed counter-current chromatography.

Three phenolic compounds, p-hydroxybenzoic acid (1), isorhamnetin-3-O-ß-D-rutinoside (2), and 3,3'-di-O-methylquercetin (5), along with a phenolic mixture were successfully separated from the ethyl acetate crude extract of Halimodendron halodendron by high-speed counter-current chromatography (HSCCC) with chloroform-methanol-water-acetic acid (4:3:2:0.05, v/v) as the two-phase solvent system. The phenolic mixture from HSCCC was further separated by preparative HPLC and purified by Sephadex LH-20 to afford quercetin (3) and 3-O-methylquercetin (4). Seven hundred mg of ethyl acetate crude extract was separated by HSCCC to obtain six fractions which were then analyzed by high performance liquid chromatography (HPLC). The HSCCC separation obtained total of 80 mg of the mixture of quercetin (3) and 3-O-methylquercetin (4) (26.43% and 71.89%, respectively) in fraction 2, 14 mg of 3,3'-di-O-methylquercetin (5) at 95.14% of purity in fraction 3, 15 mg of p-hydroxybenzoic acid (1) at 92.83% of purity in fraction 5, 12 mg of isorhamnetin-3-O-ß-D-rutinoside (2) at 97.99% of purity in fraction 6. This is the first time these phenolic compounds have been obtained from H. halodendron, and their chemical structures identified by means of physicochemical and spectrometric analysis.

Chemical analysis and biological activity of the essential oils of two valerianaceous species from China: Nardostachys chinensis and Valeriana officinalis.

In order to investigate essential oils with biological activity from local wild plants, two valerianaceous species, Nardostachys chinensis and Valeriana officinalis, were screened for their antimicrobial and antioxidant activity. The essential oils were obtained from the roots and rhizomes of the two plants by hydro-distillation, and were analyzed for their chemical composition by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Calarene (25.31%), aristolone (13.35%), α-selinene (7.32%) and ß-maaliene (6.70%) were the major compounds of the 23 identified components which accounted for 92.76% of the total oil of N. chinensis. Patchoulol (16.75%), α-pinene (14.81%), and ß-humulene (8.19%) were the major compounds among the 20 identified components, which accounted for 88.11% of the total oil of V. officinalis. Both oils were rich in sesquiterpene hydrocarbons as well as their oxygenated derivatives. Essential oils were shown to have broad spectrum antibacterial activity with MIC values that ranged from 62.5 µg/mL to 400 µg/mL, and IC(50) values from 36.93 µg/mL to 374.72 µg/mL. The oils were also shown to have moderate antifungal activity to Candida albicans growth as well as inhibition of spore germination of Magnaporthe oryzae. Two essential oils were assessed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, ß-carotene bleaching and ferrozine-ferrous ions assays, respectively, to show moderate antioxidant activity. Results suggest that the isolated essential oils could be used for future development of antimicrobial and antioxidant agents.

Inhibition of mitochondrial bioenergetics: the effects on structure of mitochondria in the cell and on apoptosis.

The effects of specific inhibitors of respiratory chain, F(o)F(1)ATP synthase and uncouplers of oxidative phosphorylation on survival of carcinoma HeLa cells and on the structure of mitochondria in the cells were studied. The inhibitors of respiration (piericidin, antimycin, myxothiazol), the F(1)-component of ATP synthase (aurovertin) and uncouplers (DNP, FCCP) did not affect viability of HeLa cells, apoptosis induced by TNF or staurosporin and the anti-apoptotic action of Bcl-2. Apoptosis was induced by combined action of respiratory inhibitors and uncouplers indicating possible pro-apoptotic action of reactive oxygen species (ROS) generated by mitochondria. Short-term incubation of HeLa cells with the mitochondrial inhibitors and 2-deoxyglucose followed by 24-48 h recovery resulted in massive apoptosis. Apoptosis correlated to transient (3-4 h) and limited (60-70%) depletion of ATP. More prolonged or more complete transient ATP depletion induced pronounced necrosis. The inhibitors of respiration and uncouplers caused fragmentation of tubular mitochondria and formation of small round bodies followed by swelling. These transitions were not accompanied with release of cytochrome c into the cytosol and were fully reversible. The combined effect of respiratory inhibitors and uncouplers developed more rapidly indicating possible involvement of ROS generated by mitochondria. More prolonged (48-72 h) incubation with this combination of inhibitors caused clustering and degradation of mitochondria.