Enhanced production of pinosylvin stilbene with aging of Pinus strobus callus and nematicidal activity of callus extracts against pinewood nematodes

Pinosylvin stilbenes are phenolic compounds mainly occurring in the Pinaceae family. We previously reported that the accumulation of two pinosylvin stilbene compounds, dihydropinosylvin methyl ether (DPME) and pinosylvin monomethyl ether (PME), in Pinus strobus trees was highly enhanced by infection with pine wood nematodes (PWNs: Bursaphelenchus xylophilus), and these two compounds showed strong nematicidal activity against PWNs. In this work, we established a system of pinosylvin stilbene (DPME and PME) production via the in vitro culture of P. strobus calli, and we examined the nematicidal activity of callus extracts. Calli were induced from the culture of mature zygotic embryos of P. strobus. Optimized growth of calli was obtained in 1/2 Litvay medium with 1.0 mg/L 2,4-D and 0.5 mg/L BA. DPME and PME accumulation did not occur in nonaged (one-month-old) calli but increased greatly with prolonged callus culture. The concentrations of DPME and PME in three-month-old dark-brown calli were 6.4 mg/g DW and 0.28 mg/g DW, respectively. The effect of methyl jasmonate treatment on the accumulation of DPME and PME was evaluated in cell suspension culture of P. strobus. However, the treatment appeared to show slight increase of DPME accumulation compared to callus browning. A test solution prepared from crude ethanol extracts from aged calli (three months old) containing 120 µg/ml DPME and 5.16 µg/ml PME treated with PWNs resulted in 100% immobilization of the adult PWNs and 66.7% immobilization of the juvenile PWNs within 24 h. However, nonaged callus extracts did not show any nematicidal activity against juvenile PWNs and showed less than 20% nematicidal activity against adult PWNs. These results indicate that pinosylvin stilbenes can be effectively produced by prolonged culture of P. strobus calli, can be isolated using simple ethanolic extraction, and are applicable as beneficial eco-friendly compounds with nematicidal activity against PWNs.

Extraction of Non-Structural Carbohydrates (Total Soluble Sugars + Starch) in Plant Tissues v1

Ethanolic extraction of total soluble sugars (liquid fraction) and starch (precipitated fraction) from leaf tissue.

In vitro pharmacological attributes and metabolite’s fingerprinting of Conocarpus lancifolius

Search for safe antioxidants and novel nutraceuticals urged to evaluate the antioxidant, anti-acetylcholine esterase and anti-lipoxygenase activity of various leaf extracts of Conocarpus lancifolius. Extraction was optimized from freeze dried plant extracts quenched with liquid nitrogen using water, ethanol, methanol, hexane, ethyl acetate and chloroform. Maximum extract yield, total phenolic contents and total flavonoid contents were obtained in case of ethanolic extraction. The highest 2,2-diphenyl-1-picrylhydrazylradical scavenging in terms of IC50 value of 55.26 µg/mL was observed for ethanolic leaf extract. The acetylcholine esterase and lipoxygenase inhibitory activities (IC50) were also observed for ethanolic extract. These findings for ethanolic extract were statistically significant when compared with other extracts (ρ<0.05). The haemolytic % values indicated that all extracts were associated with very low or negligible toxicity. The epicatechin, isorhamnetin, rutin, scopoleptin, skimmianine, quercetin-3-O-α-rhamnoside, quercetin-3-O-β-glucoside, cornoside, creatinine, choline, pyruvic acid, α-hydroxybutyric acid, phyllanthin and hypophyllanthin were identified as major functional metabolites in ethanolic leaf extract of C. lancifolius by 1H-NMR. The identified metabolites were probably responsible for the pharmacological properties of C. lancifolius. The findings may be utilized as pharmacological leads for drug development and food fortification.

Valorization of Arnica montana Wastes after Extraction of the Ethanol Tincture: Application in Polymer-Based Matrices

The waste solids remaining after the ethanolic extraction of arnica were proposed as raw material for the recovery of the remaining phenolic fraction. Greener processes based on intensification extraction, with microwave (MHG) and ultrasound (UAE) assistance and pressurized hot water extraction under subcritical conditions (AH), were studied. The entire process provided approximately 28% of phenolics for the sequence when MHG was used, 22% in the sequence where AH was employed, and the extracts showed up to 60% the ABTS radical scavenging capacity of Trolox. However, the cytotoxic effects on the cell growth of tumoral cells were only moderate. Therefore, considering a possible external topical application, extracts containing selected arnica extracts were further used to develop polymer-based gelled matrices formulated with either chitosan or polyvinyl alcohol. Rheological testing indicated that all proposed matrices exhibited relatively high mechanical features, even better than those determined with matrices prepared with their counterpart commercial arnica tinctures. Overall, the shear-thinning behavior of gelled matrices loaded with arnica extracts obtained by MHG or AH stages was enhanced compared to those containing ethanolic extracts, whereas the viscoelastic features became smoother for polymeric matrices incorporated with arnica extracts recovered at the highest MHG irradiation powers or at the highest set point temperatures of AH treatments.

Enhanced Production of Pinosylvin Stilbene with Aging of Pinus strobus Callus and Nematicidal Activity of Callus Extracts against Pinewood Nematodes

Pinosylvin stilbenes are phenolic compounds mainly occurring in the Pinaceae family. We previously reported that the accumulation of two pinosylvin stilbene compounds, dihydropinosylvin methyl ether (DPME) and pinosylvin monomethyl ether (PME), in Pinus strobus trees was highly enhanced by infection with pine wood nematodes (PWNs: Bursaphelenchus xylophilus), and these two compounds showed strong nematicidal activity against PWNs. In this work, we established a system of pinosylvin stilbene (DPME and PME) production via the in vitro culture of P. strobus calli, and we examined the nematicidal activity of callus extracts. Calli were induced from the culture of mature zygotic embryos of P. strobus. Optimized growth of calli was obtained in 1/2 Litvay medium with 1.0 mg/L 2,4-D and 0.5 mg/L BA. DPME and PME accumulation did not occur in nonaged (one-month-old) calli but increased greatly with prolonged callus culture. The concentrations of DPME and PME in three-month-old dark-brown calli were 6.4 mg/g DW and 0.28 mg/g DW, respectively. A test solution prepared from crude ethanol extracts from aged calli (three months old) containing 120 µg/ml DPME and 5.16 µg/ml PME treated with PWNs resulted in 100% immobilization of the adult PWNs and 66.7% immobilization of the juvenile PWNs within 24 hr. However, nonaged callus extracts did not show any nematicidal activity against juvenile PWNs and showed less than 20% nematicidal activity against adult PWNs. These results indicate that pinosylvin stilbenes can be effectively produced by prolonged culture of P. strobus calli, can be isolated using simple ethanolic extraction, and are applicable as beneficial eco-friendly compounds with nematicidal activity against PWNs.

Activity of Ethanolic and Supercritical Propolis Extracts in Corynebacterium pseudotuberculosis and Its Associated Biofilm

Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis in small ruminants, a chronic disease characterized by the development of granulomas in superficial and visceral lymph nodes as well as in several organs. An important characteristic of the infection with this bacterium is the formation of a biofilm and the absence of effective antibiotic therapy against the disease. From this scenario, the objective of this study was to evaluate the susceptibility of C. pseudotuberculosis to conventional antibiotics and to red, green, and brown propolis extracts obtained by the supercritical and ethanolic extraction methods as well as its activity in the bacterial biofilm. The results of the sensitivity test using antibiotics indicated a sensitivity of C. pseudotuberculosis strains to the antimicrobial agents. The ethanolic extract of green propolis and the supercritical red propolis extract showed the best antibacterial activities against planktonic C. pseudotuberculosis. A lower antimicrobial activity of the brown propolis extract was identified. Propolis extracts were effective in interfering with the formation of the C. pseudotuberculosis biofilm but had little activity on the consolidated biofilm. In conclusion, propolis extracts are more effective against C. pseudotuberculosis in the planktonic stage, being able to interfere with the formation of bacterial biofilm. However, the action of propolis extracts in a sessile and structured microbial biofilm is reduced.

Extraction and Microencapsulation of Bioactive Compounds from Muicle (Justicia spicigera) and Their Use in the Formulation of Functional Foods

Bioactive compounds (BC) present in muicle leaves were extracted using the best extraction conditions obtained with a Box–Behnken experimental design, extracting 95% of BC. Microencapsulation of muicle BC was carried out by spray drying using DE10 maltodextrin (MD) and soy protein isolate (SPI) as encapsulating agents. The best conditions for the ethanolic extraction of BC from muicle were 30 °C, 40% aqueous ethanol, and one extraction for 1 h. The best spray drying encapsulating conditions for BC and antioxidant capacity (AC) using MD as an encapsulating agent were: 160–80 °C and 10% MD in the feeding solution, and for SPI: 180–70 °C and 5% SPI in the feeding solution. Microcapsules were added to yogurt and a sensory evaluation and retention of BC during 15-day storage at 4 °C was performed. Sensory evaluation showed that yogurt with added MD microcapsules had better acceptance than that with SPI microcapsules. Based on this, a jelly with added muicle MD microcapsules was also prepared which obtained better acceptance by the judges. At the end of the storage period, yogurt with SPI microcapsules showed better retention of BC and AC than yogurts with MD microcapsules; however, products with MD microcapsules had better acceptance.

Combination of Phyllanthus amarus Schum. & Thonn. and Gymnema sylvestre R. Br. for treatment of diabetes and its long-term complications

Objectives. The amount of patients with diabetes is increasing, and such patients experience several long-term complications. Therefore, finding a method to treat the disease and its complications is an urgent issue worldwide. In Vietnam, Phyllanthus amarus Schum. & Thonn. (PA) and Gymnema sylvestre R. Br. (GS) are common herbs used in traditional therapy including diabetes treatment. This study aimed to combine PA and GS to extend their bioactivities in antidiabetes, antioxidant, and anti-inflammatory treatments.Methods. Here, PA and GS powders were mixed at different ratios for extraction. Ethanolic extract was used to detect bioactive compounds, bioactivities, and appropriate ratios of the mixtures.Results. The optimal ratio for the PA and GS combination was 2 : 1 (g/g). The ethanolic extraction of the 2 : 1 sample at 50°C over two hours with a solid/liquid ratio of 1 : 10 achieved a high yield of 14.37%. This sample exhibited good α-glucosidase inhibition activity with a half-maximal inhibitory concentration (IC50) of 9.74 µg/mL, antioxidant activity with an IC50 of 29.87 µg/mL, and anti-inflammatory activity with an IC15 of 400 µg/mL.Conclusions. The study confirmed that combining PA and GS can have high α-glucosidase inhibition as well as antioxidant and anti-inflammatory effects.