Impacts of Ultrasonic Treatment for Black Soybean Okara Culture Medium Containing Choline Chloride on the ß-Glucosidase Activity of Lactiplantibacillus plantarum BCRC 10357.

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.

Ultrasound-Assisted Aqueous Extraction of Chlorogenic Acid and Cynarin with the Impact of Inulin from Burdock (Arctium lappa L.) Roots.

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.

Quantitative Proteomics Reveals That ADAM15 Can Have Proteolytic-Independent Functions in the Steady State.

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.

Assessment of Ultrasonic Stress on Survival and ß-Glucosidase Activity of Encapsulated Lactiplantibacillus plantarum BCRC 10357 in Fermentation of Black Soymilk.

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.

Evaluation of Semi-Solid-State Fermentation of Elaeocarpus serratus L. Leaves and Black Soymilk by Lactobacillus plantarum on Bioactive Compounds and Antioxidant Capacity.

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.

Quantitative Mass Spectrometry-Based Secretome Analysis as a Tool to Investigate Metalloprotease and TIMP Activity.

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.

Role of estradiol in mediation of etomidate-caused seizure-like activity in neonatal rats.

BACKGROUND: The goal of this study was to investigate the effect of estradiol in mediation of electroencephalogram (EEG) abnormality induced by etomidate in neonatal rats. METHODS: Sprague-Dawley rats were anesthetized using intraperitoneal etomidate for 2 h on postnatal days (P) 4, 5, or 6 and recorded electroencephalogram in two ways. First, pups were recorded EEG two and a half hours under etomidate anesthesia, in subgroups, estradiol receptor antagonist ICI182780 and estradiol synthase inhibitor formestane were given subcutaneously in male rats 15 min prior to etomidate. Second, pups were anesthetized with etomidate for 2 h on P4,5 or 6 and then recovered from anesthesia, EEG were recorded for one hour in two postnatal periods of P9-P11 and P14-P16. Subgroup rats that received bumetanide, NKCC1 inhibitor, to test the NKCC1-GABAAR signaling effect on neonatal brain development, negative control groups and maternally separated for 2 h on P4, 5, or 6 were studied in 16 groups. Each group's n was = 8. RESULTS: Male pups showed more severe seizure-like activities than female pups in P4-P6 under etomidate anesthesia. Pups pretreated with ICI182780 and formestane showed a less abnormalities of EEG in male rats. Etomidate caused seizure-like activity in P4-P6 could extend to P9-P11, but not seen in P14-P16, Pretreated with bumetanide only alleviated abnormalities in male pups other than female in P9-P11. CONCLUSIONS: Estradiol involves in the NKCC1-GABAAR mediated seizure-like activity caused by etomidte in neonatal rats and these the abnormality lasts near two weeks.

Bio-ionic liquid pretreatment and ultrasound-promoted enzymatic hydrolysis of black soybean okara.

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.

Effects of combined brief etomidate anesthesia and postnatal stress on amygdala expression of Cl- cotransporters and corticotropin-releasing hormone and alcohol intake in adult rats.

Early life stressors, including general anesthesia, can have adverse effects on adult neural and behavioral outcomes, such as disruptions in inhibitory signaling, stress responsivity and increased risk of psychiatric disorders. Here we used a rat model to determine the effects of combined exposure to etomidate (ET) neonatal anesthesia and maternal separation on adult amygdala expression of genes for corticotropin-releasing hormone (Crh) and the chloride co-transporters Nkcc1 and Kcc2, as well as ethanol intake. Male and female Sprague-Dawley rats were subjected to 2 h of ET anesthesia on postnatal days (P) 4, 5, or 6 followed by maternal separation for 3 h on P10 (ET + SEP). During the P91-P120 period rats had daily 2 h access to three 0.05% saccharin solutions containing 0%, 5%, or 10% ethanol, followed by gene expression analyses. The ET + SEP group had increased Crh mRNA levels and Nkcc1/Kcc2 mRNA ratios in the amygdala, with greater increases in Nkcc1/Kcc2 mRNA ratios in males. A moderate increase in 5% ethanol intake was evident in the ET + SEP males, but not females, after calculation of the ratio of alcohol intake between the last week and first week of exposure. In contrast, control males tended to decrease alcohol consumption during the same period. A brief exposure to ET combined with a subsequent episode of stress early in life induced significant alterations in expression of amygdala Crh, Nkcc1 and Kcc2 with greater changes in the Cl- transporter expression in males. The possibility of increased alcohol intake in the exposed males requires further confirmation using different alcohol intake paradigms.

Strategic ultrasound-induced stress response of lactic acid bacteria on enhancement of ß-glucosidase activity for bioconversion of isoflavones in soymilk.

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 (20 kHz, amplitude at 20%) to irradiate on L. acidophilus BCRC 10695 at stationary phase of growth for 2 min and 24 h of re-incubation, the ß-glucosidase activity was enhanced to 3.91 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 24 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.

Parametric down-conversion with nonideal and random quasi-phase-matching.

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.

Effects of mechanical ventilation with different tidal volume on oxidative stress and antioxidant in lung.

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.

Combination of ultrasonic irradiation with ionic liquid pretreatment for enzymatic hydrolysis of rice straw.

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.

Fermentation of rice hull by Aspergillus japonicus under ultrasonic pretreatment.

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.

Effect of intravenous dezocine on fentanyl-induced cough during general anesthesia induction: a double-blinded, prospective, randomized, controlled trial.

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.

Gold-catalyzed hydrative carbocyclization of 1,5- and 1,7-allenynes mediated by pi-allene complex: mechanistic evidence supported by the chirality transfer of allenyne substrates.

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.

Gold-catalyzed synthesis of bicyclo[4.3.0]nonadiene derivatives via tandem 6-endo-dig/Nazarov cyclization of 1,6-allenynes.

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.