The effects of ultrasonic treatment for the culture medium of solid black soybean okara with choline chloride (ChCl) on the survival and ß-glucosidase activity of Lactiplantibacillus plantarum BCRC 10357 (Lp-BCRC10357) were investigated. A mixture of 3% dried black soybean okara in de Man-Rogosa-Sharpe (w/v) was used as the Oka medium. With ultrasonic treatment (40 kHz/300 W) of the Oka medium at 60 °C for 3 h before inoculation, the ß-glucosidase activity of Lp-BCRC10357 at 12 h and 24 h of incubation amounted to 13.35 and 15.50 U/mL, respectively, which was significantly larger than that (12.58 U/mL at 12 h and 2.86 U/mL at 24 h) without ultrasonic treatment of the medium. This indicated that ultrasonic treatment could cause the microstructure of the solid black soybean okara to be broken, facilitating the transport of ingredients and Lp-BCRC10357 into the internal structure of the okara for utilization. For the effect of ChCl (1, 3, or 5%) added to the Oka medium (w/v) with ultrasonic treatment before inoculation, using 1% ChCl in the Oka medium could stimulate the best response of Lp-BCRC10357 with the highest ß-glucosidase activity of 19.47 U/mL in 12 h of incubation, showing that Lp-BCRC10357 had a positive response when confronting the extra ChCl that acted as an osmoprotectant and nano-crowder in the extracellular environment. Furthermore, the Oka medium containing 1% ChCl with ultrasonic treatment led to higher ß-glucosidase activity of Lp-BCRC10357 than that without ultrasonic treatment, demonstrating that the ultrasonic treatment could enhance the contact of ChCl and Lp-BCRC10357 to regulate the physiological behavior for the release of enzymes. In addition, the analysis of the isoflavone content and antioxidant activity of the fermented product revealed that the addition of 1% ChCl in the Oka medium with ultrasonic treatment before inoculation allowed a higher enhancement ratio for the biotransformation of isoflavone glycosides to their aglycones, with a slight enhancement in the antioxidant activity at 24 h of fermentation. This study developed a methodology by combining ultrasonic treatment with a limited amount of ChCl to allow the culture medium to acclimate Lp-BCRC10357 and release high levels of ß-glucosidase, and this approach has the potential to be used in the fermentation of okara-related products as nutritional supplements in foods.
The ultrasound-assisted aqueous extraction of chlorogenic acid (CGA) and cynarin with the impact of inulin from burdock (Arctium lappa L.) roots was investigated. Three extraction modes, ultrasound at 40 kHz/300 W (U-40), ultrasound at 120 kHz/300 W (U-120), and shaking at 120 rpm (S-120), were compared. The effects of process parameters on the extraction of polyphenols, CGA, cynarin, inulin, and antioxidant activity using U-40 were evaluated. In 10 min, 50 °C, and 1/30 (g/mL-water) of solid-to-liquid ratio, the order of CGA content in the dried burdock root powder (DBR) was U-40 (484.65 µg/g-DBR) > U-120 (369.93 µg/g-DBR) > S-120 (176.99 µg/g-DBR), while the order of cynarin content in DBR was U-120 (376.47 µg/g-DBR) > U-40 (341.54 µg/g-DBR) > S-120 (330.44 µg/g-DBR), showing the selective extraction of CGA and cynarin between using 40 and 120 kHz of ultrasound. The profiles of increase and then decrease in free CGA and cynarin concentrations against time revealed their degradation, including their interactions with the abundant inulin. The kinetic model, considering extraction followed by degradation, was proposed to describe the variations of free CGA and cynarin against time. This study provides an effective method using water to extract CGA, cynarin, and inulin from burdock roots.
A disintegrin and metalloproteinase 15 (ADAM15) is a member of the ADAM family of sheddases. Its genetic ablation in mice suggests that ADAM15 plays an important role in a wide variety of biological functions, including cartilage homeostasis. Nevertheless, while the substrate repertoire of other members of the ADAM family, including ADAM10 and ADAM17, is largely established, little is known about the substrates of ADAM15 and how it exerts its biological functions. Herein, we used unbiased proteomics to identify ADAM15 substrates and proteins regulated by the proteinase in chondrocyte-like HTB94 cells. ADAM15 silencing did not induce major changes in the secretome composition of HTB94 cells, as revealed by two different proteomic approaches. Conversely, overexpression of ADAM15 remodeled the secretome, with levels of several secreted proteins being altered compared to GFP-overexpressing controls. However, the analysis did not identify potential substrates of the sheddase, i.e., transmembrane proteins released by ADAM15 in the extracellular milieu. Intriguingly, secretome analysis and immunoblotting demonstrated that ADAM15 overexpression increased secreted levels of tissue inhibitor of metalloproteinases 3 (TIMP-3), a major regulator of extracellular matrix turnover. An inactive form of ADAM15 led to a similar increase in the inhibitor, indicating that ADAM15 regulates TIMP-3 secretion by an unknown mechanism independent of its catalytic activity. In conclusion, high-resolution quantitative proteomics of HTB94 cells manipulated to have increased or decreased ADAM15 expression did not identify canonical substrates of the proteinase in the steady state, but it revealed that ADAM15 can modulate the secretome in a catalytically-independent manner.
The enhanced ß-glucosidase activity of encapsulated Lactiplantibacillus plantarum BCRC 10357 within calcium alginate capsules was investigated by ultrasonic stimulation to induce the stress response of the bacteria for the biotransformation of isoflavones in black soymilk. The effects of various ultrasound durations, sodium alginate concentrations (% ALG), and cell suspensions on the ß-glucosidase activity of encapsulated bacteria were explored. The ß-glucosidase activity of encapsulated L. plantarum BCRC 10357 with ultrasonic stimulation (40 kHz/300 W) was greater than that without ultrasound. With 20 min of ultrasonic treatment, the ß-glucosidase activity of encapsulated L. plantarum BCRC 10357 from 2% ALG/0.85% NaCl cell suspension was 11.47 U/mL at 12 h, then increased to 27.43 U/mL at 36 h and to 26.25 U/mL at 48 h in black soymilk at 37 °C, showing the high adaptation of encapsulated L. plantarum BCRC 10357 encountering ultrasonic stress to release high ß-glucosidase until 48 h, at which point the ratio of isoflavone aglycones (daidzein and genistein) in total isoflavones (daidzin, genistin, daidzein, and genistein) was 98.65%, reflecting the effective biotransformation of isoflavone glycosides into aglycones by ß-glucosidase. In this study, the survivability and ß-glucosidase activity of encapsulated L. plantarum BCRC 10357 were enhanced under ultrasonic stimulation, and were favorably used in the fermentation of black soymilk.
Elaeocarpus serratus L. leaves (EL) containing phenolic compounds and flavonoids, including myricitrin with pharmacological properties, could be valorized as nutritional additive in foods. In this study, the semi-solid-state fermentation of EL and black soymilk (BS) by Lactobacillus plantarum BCRC 10357 was investigated. Without adding EL in MRS medium, the ß-glucosidase activity of L. plantarum quickly reduced to 2.33 ± 0.15 U/mL in 36 h of fermentation; by using 3% EL, the stability period of ß-glucosidase activity was prolonged as 12.94 ± 0.69 U/mL in 12 h to 13.71 ± 0.94 in 36 h, showing positive response of the bacteria encountering EL. Using L. plantarum to ferment BS with 3% EL, the ß-glucosidase activity increased to 23.78 ± 1.34 U/mL in 24 h, and in the fermented product extract (FPE), the content of myricitrin (2297.06 µg/g-FPE) and isoflavone aglycones (daidzein and genistein, 474.47 µg/g-FPE) at 48 h of fermentation were 1.61-fold and 1.95-fold of that before fermentation (at 0 h), respectively. Total flavonoid content, myricitrin, and ferric reducing antioxidant power in FPE using BS and EL were higher than that using EL alone. This study developed the potential fermented product of black soymilk using EL as a nutritional supplement with probiotics.
Cell surface proteolysis controls numerous biological processes including cell-cell attachment and the communication between cells. The membrane-tethered families of matrix metalloproteinases (MT-MMPs) and disintegrin metalloproteinases (ADAMs) are major enzymes involved in the cleavage of molecules at the cell surface, and their activity is finely regulated by their endogenous inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). The biological function of a metalloproteinase closely depends on the subset of substrates that it cleaves. Similarly, molecular processes that are regulated by a specific TIMP strictly depend on its unique inhibitory profile.Herein, we describe a mass spectrometry-based method for the quantitative analysis of protein abundance in conditioned media of cultured cells that is particularly suited for substrate identification of membrane-tethered metalloproteinases and for the identification of membrane proteins whose cleavage is regulated by TIMPs. This unbiased proteomic method represents a valuable tool to investigate biological functions of metalloproteinases and TIMPs at the "omic" level.
Matrix Metalloproteinases/metabolism , Proteomics/methods , Tissue Inhibitor of Metalloproteinases/metabolism , Animals , Cell Culture Techniques , Cells, Cultured , Culture Media, Conditioned/chemistry , Humans , Mass Spectrometry
The effective processing method to produce fermentable sugars and modify the microstructure of black soybean okara using bio-ionic liquid (bio-IL) pretreatment and ultrasound-promoted enzymatic hydrolysis was investigated. The morphology and structural characteristics of okara before and after bio-IL pretreatment and enzymatic hydrolysis under different ultrasonic frequencies were analyzed by field emission scanning electron microscope (FE-SEM), X-ray energy dispersive spectrometer (EDS), and Fourier transform infrared spectroscopy (FTIR). Without pretreatment, the production of total reducing sugar (TRS) under ultrasound (40 kHz/300 W) was 3.4 times of that without ultrasound. Using the bio-IL choline acetate ([Ch][OAc]) in water for the pretreatment of black soybean okara, the TRS production of enzymatic hydrolysis was further increased to 5.2 times of that without ultrasound in 4 h of reaction. The analysis by FTIR and EDS showed that the highly structured matrix of okara was unfolded and broken by the action of combining ultrasound and choline acetate pretreatment, due to which the surface structures with large pores were presented to facilitate the reduction of unfavorable hindrance for enzymatic hydrolysis. The simplified kinetic model was proposed to describe the transport and reaction phenomena of enzymes in a solid-liquid system by using two kinetic parameters to show the impeded behavior of enzyme within the matrix of okara. The combination of bio-IL pretreatment and ultrasound-promoted enzymatic hydrolysis was able to make the efficient structural changes of black soybean okara to enhance the digestion of enzymes, and the okara could be valorized for use in foods.
Cellulase/metabolism , Glycine max/chemistry , Ionic Liquids/chemistry , Ultrasonic Waves , Waste Products , Biomass , Hydrolysis , Trichoderma/enzymology
The purpose of the study is to develop the new methodology of strategic ultrasound treatment on lactic acid bacteria (LAB) to induce stress response for the enhancement of ß-glucosidase activity that can be used for the biotransformation of glucosides into aglycones isoflavones in soymilk. Among the five LAB tested, Lactobacillus acidophilus BCRC 10695 showed the best ability to release ß-glucosidase for further ultrasonic stimulation to induce proper stress response. With ultrasound (20â¯kHz, amplitude at 20%) to irradiate on L. acidophilus BCRC 10695 at stationary phase of growth for 2â¯min and 24â¯h of re-incubation, the ß-glucosidase activity was enhanced to 3.91â¯U/ml, which was 1.82 times of that without ultrasound treatment. Using the ultrasound-treated L. acidophilus BCRC 10695 to ferment soymilk, the fraction of aglycones in total isoflavones in soymilk was effectively increased from 21.8% initially to 97.9% in 24â¯h. The strategic ultrasound treatment on L. acidophilus BCRC 10695 demonstrated promotion of ß-glucosidase activity, and this methodology had the potential to be applied in the production of functional soymilk by adding probiotics LAB to increase the bioactive isoflavones and nutritional values for human health.
Isoflavones/metabolism , Lactobacillus acidophilus/metabolism , Lactobacillus acidophilus/radiation effects , Sonication/methods , beta-Glucosidase/metabolism , Biotransformation , Fermentation , Lactobacillus acidophilus/enzymology , Lactobacillus acidophilus/growth & development , Soy Milk/metabolism
Quasi-phase-matching (QPM) has enriched the capacity of parametric down-conversion (PDC) in generating biphotons for many fundamental tests and advanced applications. However, it is not clear how the nonidealities and randomness in the QPM grating of a parametric down-converter may affect the quantum properties of the biphotons. This paper intends to provide insights into the interplay between PDC and nonideal or random QPM structures. Using a periodically poled nonlinear crystal with short periodicity, we conduct experimental and theoretical studies of PDC subject to nonideal duty cycle and random errors in domain lengths. We report the observation of biphotons emerging through noncritical birefringent-phasematching, which is impossible to occur in PDC with an ideal QPM grating, and a biphoton spectrum determined by the details of nonidealities and randomness. We also observed QPM biphotons with a diminished strength. These features are both confirmed by our theory. Our work provides new perspectives for biphoton engineering with QPM.
PURPOSE: The aim of this study was to investigate the changes in oxidative stress and antioxidants in lung tissue under different tidal volume ventilation conditions. METHODS: Forty-eight male Wistar rats were randomized into four groups, namely, group C, the control group, which was not ventilated, and groups C1, C2 and C3, the treatment groups, which were ventilated for 2 h with tidal volumes of 8, 30 and 42 ml/kg, respectively. The right middle lobe was assayed for malondialdehyde (MDA), the right posterior lobe was assayed using Western blotting for Nrf2, GCLm and SrX1 and the left lobe was assayed for Nrf2, GCLm and SrX1 mRNA. RESULTS: The MDA levels were increased in the three treatment groups, with MDA levels highest in group C3 and lowest in group C1 (C3 > C2 > C1) (all P < 0.05). The mRNA expression of Nrf2, GCLm and SrX1 was highest in group C3 and lowest in group 1 (C3 > C2 > C1) (all P < 0.05). No significant difference was observed between group C1 and group C (P > 0.05). A Western blot analysis showed that Nrf2, GCLm and SrX1 expression was highest in group C3 and lowest in group C1 (C3 > C2 > C1) (all P < 0.05). No significant difference was observed between group C1 and group C (P > 0.05). CONCLUSIONS: Oxidative stress and antioxidant enzyme levels in the lungs of rats were positively associated with the tidal volumes of mechanical ventilation, suggesting that higher tidal volumes cause more severe oxidative stress and increased antioxidant responses.
Antioxidants/metabolism , Lung/metabolism , Oxidative Stress/physiology , Respiration, Artificial/methods , Animals , Male , Malondialdehyde/metabolism , Rats , Rats, Wistar , Tidal Volume/physiology
The application of ultrasonic irradiation and ionic liquids (ILs) in the degradation of rice straw under different processes of pretreatment and enzymatic hydrolysis was investigated. Various substrates for enzymatic hydrolysis by cellulase with and without ultrasound were as follows: untreated rice-straw powder (RS); RS treated by ILs of 1-ethyl-3-methylimidazolium ethylsulfate and trihexyl (tetradecyl) phosphonium decanoate with ultrasound at 300 W/(40 kHz, 28 kHz); RS treated by IL of choline hydroxide ([Ch][OH]) with ultrasound at 300 W/40 kHz (CHRS). In ultrasound-mediated enzymatic hydrolysis, the yield of total reducing sugar (TRS) converted from CHRS was up to 96.22% at 240 min and was greater than that from the other substrates; the TRS yield for CHRS with ultrasound was 19.5% greater than that without irradiation. Lignocellulosic biomass pretreated with [Ch][OH] showed the highest efficiency among the tested ILs, and ultrasound can be applied effectively in rice-straw pretreatment and enzymatic hydrolysis.
Biotechnology/methods , Cellulase/metabolism , Ionic Liquids/pharmacology , Oryza/chemistry , Ultrasonics/methods , Waste Products , Biomass , Hydrolysis/drug effects , Substrate Specificity/drug effects , Time Factors
The application of ultrasound for treating rice hull used as the fermentation substrate for xylooligosaccharides production was investigated. Aspergillus japonicus CY6-1 was selected to produce cellulolytic and xylanolytic enzymes from untreated rice hull (RH) and rice hull treated with ultrasound (USRH-M). The hemicellulose yield was increased to 1.4-fold with ultrasound, and treatment time was greatly shortened from 24h to 1.5h at 80 °C and 300 W/28 kHz. The morphology of RH from various pretreatments was observed with field emission scanning electron microscope (FESEM), revealing the surface structure of USRH-M smoother than that of RH. USRH-M was much easier to be utilized by fungi, to extend the stability of enzyme activity and to increase activities of CMCase, ß-glucosidase, and xylanase compared with those of untreated RH. The final fermentative products were xylotetraose, xylohexaose, and higher molecular weight xylooligosaccharides, achieving xylohexaose yield for USRH-M 80% higher than that for RH group.
Aspergillus/metabolism , Aspergillus/radiation effects , Aspergillus/ultrastructure , Glucuronates/biosynthesis , Oligosaccharides/biosynthesis , Oryza/microbiology , Oryza/radiation effects , Sonication/methods , High-Energy Shock Waves , Plant Components, Aerial/microbiology , Plant Components, Aerial/radiation effects , Radiation Dosage
PURPOSE: To evaluate the suppressive effect of intravenous dezocine on fentanyl-induced cough during the induction of general anesthesia. METHODS: A total of 120 patients, American Society of Anesthesiologists (ASA) physical status I-II, were randomized into two equally sized groups (n = 60). These two groups were given either intravenous dezocine 0.1 mg/kg or a matching placebo (equal volume of 0.9% saline) 10 min before the induction of general anesthesia. Patients were induced with midazolam 0.1 mg/kg, fentanyl 5 µg/kg, propofol 1-1.5 mg/kg, and suxamethonium 1.5 mg/kg. The injection time of fentanyl was less than 2 s in all patients. The occurrence of cough was recorded 2 min after fentanyl bolus. RESULTS: No patient in the dezocine group had cough, and 42 patients in the control group had cough. This difference was statistically different between these two groups (P = 0.000). CONCLUSION: These results demonstrate that intravenous dezocine 0.1 mg/kg 10 min prior to induction was effective in suppressing fentanyl-induced cough in our patients.
Anesthetics, Intravenous/adverse effects , Bridged Bicyclo Compounds, Heterocyclic/therapeutic use , Cough/chemically induced , Cough/drug therapy , Fentanyl/adverse effects , Tetrahydronaphthalenes/therapeutic use , Anesthesia, General/methods , Anesthetics, Intravenous/administration & dosage , Double-Blind Method , Drug Interactions , Female , Fentanyl/administration & dosage , Humans , Male , Middle Aged
We report PPh3AuCl/AgOTf-catalyzed hydrative carbocyclization of 1,5- and 1,7-allenynes to give cyclized ketones chemoselectively. In this transformation, hydration occurrs regioselectively at the C[triple bond]CPh carbon, accompanied by addition of the C[triple bond]CPh carbon to the two terminal allenyl carbons. This method is effective for the construction of a quaternary carbon center. On the basis of the chirality transfer of allenyne substrates, control experiments, and theoretic calculations, we propose that this hydrative carbocyclization proceeds through an initial pi-allene complex with a small energy barrier.
Catalytic cyclization of 1,6-allenynes was achieved by AuPPh(3)SbF(6) (5 mol %) in cold CH(2)Cl(2) (0 degrees C, 0.5-4 h) to form bicyclo[4.3.0]nonadiene products; this cyclization proceeded more efficiently for a substrate bearing R = alkyl (yields >70%). We propose a reaction mechanism involving a 6-endo-dig cyclization of Au(I)-pi-alkyne, followed by Nazarov cyclization.
