Neuronal splicing regulator RBFOX3 mediates seizures via regulating Vamp1 expression preferentially in NPY-expressing GABAergic neurons.

Epilepsy is a common neurological disorder, which has been linked to mutations or deletions of RNA binding protein, fox-1 homolog (Caenorhabditis elegans) 3 (RBFOX3)/NeuN, a neuronal splicing regulator. However, the mechanism of seizure mediation by RBFOX3 remains unknown. Here, we show that mice with deletion of Rbfox3 in gamma-aminobutyric acid (GABA) ergic neurons exhibit spontaneous seizures and high premature mortality due to increased presynaptic release, postsynaptic potential, neuronal excitability, and synaptic transmission in hippocampal dentate gyrus granule cells (DGGCs). Attenuating early excitatory gamma-aminobutyric acid (GABA) action by administering bumetanide, an inhibitor of early GABA depolarization, rescued premature mortality. Rbfox3 deletion reduced hippocampal expression of vesicle-associated membrane protein 1 (VAMP1), a GABAergic neuron-specific presynaptic protein. Postnatal restoration of VAMP1 rescued premature mortality and neuronal excitability in DGGCs. Furthermore, Rbfox3 deletion in GABAergic neurons showed fewer neuropeptide Y (NPY)-expressing GABAergic neurons. In addition, deletion of Rbfox3 in NPY-expressing GABAergic neurons lowered intrinsic excitability and increased seizure susceptibility. Our results establish RBFOX3 as a critical regulator and possible treatment path for epilepsy.

Subjective experience of parent-child relationship in adolescents with congenital heart disease: A qualitative study.

PURPOSE: To explore the parent-child relationship through the subjective experience of adolescents with congenital heart disease (CHD). DESIGN AND METHODS: A descriptive phenomenology approach was adopted. Twelve adolescents aged from 12 to 18 years with CHD were recruited from the pediatric cardiology clinics at two medical centers in Taiwan. Data were collected through in-depth interviews. Data were analyzed using Colaizzi's phenomenological analysis method, and results were reported in accordance with the Consolidated Criteria for Reporting Qualitative Research guidelines. RESULTS: The experiences of the adolescents with CHD revealed five themes: 1. the enhancement of self-worth through parents' love; 2. the importance of parental support in desperate situations; 3. the development of a sense of security through mutual understanding; 4. growth under parental expectations; and 5. parental overcontrol disguised as love. CONCLUSIONS: The parent-child relationship encompasses both positive and negative experiences. Adolescents prioritize their relationship with parents over that with peers. PRACTICE IMPLICATIONS: Nurses caring for adolescents with CHD can improve care by recognizing the influence of parental love, support in challenges, mutual understanding, parental expectations, and potential negative consequences of overcontrol. This insight guides effective guidance for adolescents, enhancing parent-child interactions and overall well-being.

Characterization and Structural Determination of CmnG-A, the Adenylation Domain That Activates the Nonproteinogenic Amino Acid Capreomycidine in Capreomycin Biosynthesis.

Capreomycidine (Cap) is a nonproteinogenic amino acid and building block of nonribosomal peptide (NRP) natural products. We report the formation and activation of Cap in capreomycin biosynthesis. CmnC and CmnD catalyzed hydroxylation and cyclization, respectively, of l-Arg to form l-Cap. l-Cap is then adenylated by CmnG-A before being incorporated into the nonribosomal peptide. The co-crystal structures of CmnG-A with l-Cap and adenosine nucleotides provide insights into the specificity and engineering opportunities of this unique adenylation domain.

Future outcomes and health needs of chronic hepatitis C patients after receiving antiviral therapy.

AIMS: To study the future outcomes and health needs of chronic hepatitis C (CHC) patients after receiving direct-acting antiviral (DAA) therapy based on the health promotion perspectives and cardiometabolic risks in a rural setting. DESIGN: This cross-sectional study was conducted from May to December 2019 in coastal western and southern Taiwan. METHODS: We included CHC patients who were diagnosed and transferred by the gastroenterologist and hepatologist in outpatient clinics and completed DAA treatments. Data on demographic characteristics, health-related behaviours and physiological biomarkers were collected through one-on-one interview using a questionnaire and from medical records obtained from a teaching hospital. RESULTS: In total, 124 participants were enrolled. Most participants (87.1%) had no side effects and 79.8% felt satisfied after treatment. However, 62.1% had metabolic syndrome, 48.4% had hypertension and 37.9% had diabetes. Furthermore, 71.8% patients were considered to have medium-to-high risk based on Framingham risk scores. In the multiple regression model, after adjusting for education level, other chronic diseases were negatively associated with health-promoting behaviours in participants. CONCLUSIONS: Although there were few side effects and most patients were satisfied after treatment, there was a high prevalence of cardiometabolic risk factors and cardiometabolic diseases and less adoption of healthy behaviours in CHC patients. Despite the small sample size, the study suggests that clinicians can reduce the burden of the aforementioned comorbidities by providing adequate treatment and individualized lifestyle modification. IMPACT: This study highlights that primary healthcare providers should consider the health needs of CHC patients after DAA treatment since many patients have high cardiometabolic risks, but only a few adopt a healthy lifestyle. Further studies are needed to initiate health-promoting programs for these patients to reduce further injury to vital organs.

Complex Coacervation of Soy Proteins, Isoflavones and Chitosan.

In this study, the chitosan-induced coacervation of soy protein-isoflavone complexes in soymilk was investigated. Most of the soymilk proteins, including ß-conglycinin (7S), glycinin (11S), and isoflavones, were found to coacervate into the soymilk pellet fraction (SPF) following the addition of 0.5% chitosan. The total protein in the soymilk supernatant fraction (SSF) decreased from 18.1 ± 0.3 mg/mL to 1.6 ± 0.1 mg/mL, and the pH values decreased slightly, from 6.6 ± 0.0 to 6.0 ± 0.0. The results of SDS-PAGE revealed that the 7S α', 7S α, 7S ß, 11S A3, and 11S acidic subunits, as well as the 11S basic proteins in the SSF, decreased to 0.7 ± 0.5%, 0.2 ± 0.1%, 0.1 ± 0.0%, 0.2 ± 0.2%, 0.2 ± 0.2% and 0.3 ± 0.2%, respectively. We also found that isoflavones in the SSF, including daidzein, glycitein, and genistein, decreased to 9.6 ± 2.3%, 5.7 ± 0.9% and 5.9 ± 1.5%, respectively. HPLC analysis indicated that isoflavones mixed with soy proteins formed soy protein-isoflavone complexes and were precipitated into the SPF by 0.5% chitosan.

The effect of three dimensional printing hinged ankle foot orthosis for equinovarus control in stroke patients.

BACKGROUND: Hemiplegic stroke patients often suffered from equinovarus foot which result in plantar pressure overload over lateral sole. 3D printing is a promising technique utilized in orthosis fabrication, and the effects on plantar pressure remains unclear. The aim in this study focus on the effect of 3D printed ankle foot orthosis, by measuring plantar parameters. METHODS: Ten patients with first-ever unilateral stroke were enrolled in this study. All patients performed 10-m walk test in 3 conditions, including ambulation with 3D printed hinged ankle foot orthosis, anterior ankle foot orthosis, and bare foot. The plantar contact area, maximum force, and peak pressure were collected and evaluate using Pedar X in-sole system. Plantar parameters symmetric analysis was conducted to assess the similarity between hemiplegic leg and unaffected leg. We used Quebec User Evaluation of Satisfaction with Assistive Technology questionnaire to survey patients' subjective satisfaction. FINDINGS: Walking with 3D printed ankle foot orthosis revealed significant increase in medial midfoot peak pressure compared to bare foot walking. Plantar parameters symmetric analysis illustrated more even medial midfoot contact area compared to bare foot walking. In satisfaction survey, walking with 3D printed ankle foot orthosis outweighs anterior ankle foot orthosis in fitting and durableness. INTERPRETATION: Dynamic plantar pressure measurement is useful for evaluation of equinovarus deformity in hemiplegic stroke patients. Wearing 3D printed ankle foot orthosis increase plantar pressure in medial midfoot area. And medial midfoot contact area is also more symmetry.

Crystal structure of the α-ketoglutarate-dependent non-heme iron oxygenase CmnC in capreomycin biosynthesis and its engineering to catalyze hydroxylation of the substrate enantiomer.

CmnC is an α-ketoglutarate (α-KG)-dependent non-heme iron oxygenase involved in the formation of the l-capreomycidine (l-Cap) moiety in capreomycin (CMN) biosynthesis. CmnC and its homologues, VioC in viomycin (VIO) biosynthesis and OrfP in streptothricin (STT) biosynthesis, catalyze hydroxylation of l-Arg to form ß-hydroxy l-Arg (CmnC and VioC) or ß,γ-dihydroxy l-Arg (OrfP). In this study, a combination of biochemical characterization and structural determination was performed to understand the substrate binding environment and substrate specificity of CmnC. Interestingly, despite having a high conservation of the substrate binding environment among CmnC, VioC, and OrfP, only OrfP can hydroxylate the substrate enantiomer d-Arg. Superposition of the structures of CmnC, VioC, and OrfP revealed a similar folds and overall structures. The active site residues of CmnC, VioC, and OrfP are almost conserved; however Leu136, Ser138, and Asp249 around the substrate binding pocket in CmnC are replaced by Gln, Gly, and Tyr in OrfP, respectively. These residues may play important roles for the substrate binding. The mutagenesis analysis revealed that the triple mutant CmnCL136Q,S138G,D249Y switches the substrate stereoselectivity from l-Arg to d-Arg with ∼6% relative activity. The crystal structure of CmnCL136Q,S138G,D249Y in complex with d-Arg revealed that the substrate loses partial interactions and adopts a different orientation in the binding site. This study provides insights into the enzyme engineering to α-KG non-heme iron oxygenases for adjustment to the substrate stereoselectivity and development of biocatalysts.

Assessment of Wheelchair Propulsion Performance in an Immersive Virtual Reality Simulator.

Maneuvering a wheelchair is an important necessity for the everyday life and social activities of people with a range of physical disabilities. However, in real life, wheelchair users face several common challenges: articulate steering, spatial relationships, and negotiating obstacles. Therefore, our research group has developed a head-mounted display (HMD)-based intuitive virtual reality (VR) stimulator for wheelchair propulsion. The aim of this study was to investigate the feasibility and efficacy of this VR stimulator for wheelchair propulsion performance. Twenty manual wheelchair users (16 men and 4 women) with spinal cord injuries ranging from T8 to L2 participated in this study. The differences in wheelchair propulsion kinematics between immersive and non-immersive VR environments were assessed using a 3D motion analysis system. Subjective data of the HMD-based intuitive VR stimulator were collected with a Presence Questionnaire and individual semi-structured interview at the end of the trial. Results indicated that propulsion performance was very similar in terms of start angle (p = 0.34), end angle (p = 0.46), stroke angle (p = 0.76), and shoulder movement (p = 0.66) between immersive and non-immersive VR environments. In the VR episode featuring an uphill journey, an increase in propulsion speed (p < 0.01) and cadence (p < 0.01) were found, as well as a greater trunk forward inclination (p = 0.01). Qualitative interviews showed that this VR simulator made an attractive, novel impression and therefore demonstrated the potential as a tool for stimulating training motivation. This HMD-based intuitive VR stimulator can be an effective resource to enhance wheelchair maneuverability experiences.

Molecular Mechanisms of the Anti-obesity Properties of Agardhiella subulata in Mice Fed a High-Fat Diet.

The overweight and obese population has skyrocketed, resulting in a high incidence of metabolic disorders. Agardhiella subulata (AS) contains a variety of beneficial components, such as sulfur-containing polysaccharides (dietary fiber) and astaxanthin, which is considered to have anti-obesity potential. In this study, we investigated the effects and possible mechanisms of dietary AS on high-fat diet (HFD)-induced obesity in mice. AS supplementation significantly reduced HFD-induced weight gain (19%) and the visceral adiposity index (4.1%). In addition, the level of total cholesterol, triglyceride, and low-density lipoprotein was significantly decreased; adiponectin was significantly increased in serum and fecal triglyceride excretion was significantly higher in mice fed AS compared with mice on an HFD. Preadipocyte factor 1 and Sry-box transcription factor 9 that were significantly higher than the levels found for the HFD group lead to reduced adipogenesis. Moreover, accompanying the lipolysis and fatty acid ß-oxidation that occur in the AS group, the concentration of non-esterified fatty acids was lowered to 0.4 ± 0.1 mEq/L. In addition, peroxisome proliferator-activated receptor γ and phosphorylation acetyl-CoA carboxylase increased 1.5- and 1-fold, thus increasing the expression of adiponectin and the activation of AMPK and ultimately resulting in lower blood glucose levels. The results of this study suggest that AS supplementation increases lipid excretion and improves energy metabolism to prevent obesity in mice fed a HFD.

Therapeutic Effects of BCG Vaccination on Type 1 Diabetes Mellitus: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: Type 1 diabetes mellitus (T1DM) causes the irreversible destruction of pancreatic beta cells. The Bacillus Calmette-Guerin (BCG) vaccine can modulate the immune response and decelerate disease progression. The aim of this study is to investigate the efficacy of the BCG vaccine for the treatment of T1DM. OBJECTIVE: Six databases were systematically searched from inception to the end of August 2019. The randomized controlled trials (RCTs) that evaluated glycemic control in response to the BCG vaccine for T1DM were enrolled. The primary outcome was glycated hemoglobin (HbA1c) level, and secondary outcomes included fasting and stimulated C-peptide level, daily insulin dosage, and clinical remission. The revised Cochrane risk of bias tool was used for quality assessment, and meta-analyses were conducted to evaluate the efficacy of the BCG vaccine. RESULTS: Four studies with a total of 198 subjects were included. The results of HbA1c and fasting C-peptide levels were extracted for further quantitative assessment. The pooled meta-analysis demonstrated no significant difference in HbA1c levels (mean difference [MD], -0.12; 95% confidence interval [CI], -0.53 to 0.30; I 2 = 56%) or fasting C-peptide levels (MD, -0.15; 95% CI, -0.35 to 0.06; I 2 = 0%) in the BCG intervention group as compared with that in the placebo group. CONCLUSIONS: There is no robust evidence to support the use of the BCG vaccine for the treatment of T1DM although the HbA1c levels tended to improve. Additional RCTs to assess the long-term effects of the BCG vaccine on glycemic control are warranted.

Pterostilbene Inhibits Adipocyte Conditioned-Medium-Induced Colorectal Cancer Cell Migration through Targeting FABP5-Related Signaling Pathway.

Pterostilbene (PTS) is a phenolic compound with diverse pharmacologic activities. However, its potential for inhibiting obesity-related colorectal cancer (CRC) remains unclear. Our study evaluated the mechanism of inhibitory effects of PTS on adipocyte conditioned-medium (aCM)-induced malignant transformation in HT-29 colorectal adenocarcinoma cells. The results demonstrated that PTS could downregulate the expression of aCM-induced fatty acid-binding protein 5 (FABP5) and prometastatic factors such as vascular endothelial growth factor, matrix metalloproteinase-2 (MMP2), MMP9, and extracellular tumor necrosis factor α via inhibiting aCM-induced nuclear factor-kappa B (NF-κB), ß-catenin, and peroxisome proliferator-activated receptor γ (PPAR-γ). Moreover, PTS can suppress aCM-stimulated phosphoinositide 3-kinase (PI3K), protein kinase B (Akt), p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinases 1/2 (JNK 1/2) signaling pathways activation that are upstream of NF-κB, ß-catenin, and PPAR-γ. Therefore, we suggest that PTS could alleviate adiposity-induced metastasis in CRC via inhibiting cell migration through downregulating FABP5 gene expression.

Music Interventions for Anxiety in Pregnant Women: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

Prenatal anxiety is extremely common and may result in adverse effects on both the mother and the baby. Music interventions have been used to reduce anxiety in various medical patients and in pregnant women during childbirth. This study aims to assess the clinical efficacy of music interventions in women during pregnancy rather than during labor. Seven databases were searched from inception to September 2019 without language restrictions. We included only randomized controlled trials that compared music intervention and control groups for anxiety reduction in pregnant women. We used the revised Cochrane risk-of-bias tool (RoB 2.0) for quality assessment. Finally, 11 studies with 1482 participants were included. The pooled meta-analysis results showed that music interventions significantly decreased anxiety levels (standardized mean difference (SMD), -0.42; 95% confidence interval (CI), -0.83 to -0.02; I2 = 91%). Moreover, subgroup analysis showed that listening to music at home had significant anxiolytic benefits (SMD, -0.28; 95% CI, -0.47 to -0.08; I2 = 0%). However, meta-regression revealed a nonsignificant trend for increase in the anxiety-reducing effects of music interventions with increasing maternal age. In conclusion, music interventions may be beneficial in reducing anxiety and may be applied in pregnant women.

Early Post-Operative Intervention of Whole-Body Vibration in Patients After Total Knee Arthroplasty: A Pilot Study.

(1) Background: Knee osteoarthritis causes pain, weakness, muscle atrophy, and disability. The application of whole-body vibration in patients with knee osteoarthritis can improve strength, balance, and functional activities. The purpose of the study is to evaluate the effects of early whole-body vibration intervention in patients after total knee arthroplasty. (2) Method: A single-blinded randomized control trial. Fifty-two patients with knee osteoarthritis post total knee replacement from a medical center in southern Taiwan were randomly assigned to either a whole-body vibration group or control group. Main outcome measures included pain severity, leg circumference, knee range of motion, knee extensor strength, a five-times sit to stand test, and a timed up and go test. (3) Results: Immediately post treatment, the patients in the vibration group showed a significant increase in knee extensor strength and improvement in calf swelling compared to the control group. A trend toward decrease in pain severity and improvement in functional performance were observed in both groups without a significant difference between the groups. There was no significant difference in knee range of motion (ROM) and functional performance between the groups. (4) Conclusions: The whole-body vibration intervention in patients early post total knee arthroplasty showed significant immediate effect in increasing knee extensor strength and decreasing calf swelling.

Site-Directed Alkylation Detected by In-Gel Fluorescence (SDAF) to Determine the Topology Map and Probe the Solvent Accessibility of Membrane Proteins.

The topology of helix-bundle membrane proteins provides low-resolution structural information with regard to the number and orientation of membrane-spanning helices, as well as the sidedness of intra/extra-cellular domains. In the past decades, several strategies have been developed to experimentally determine the topology of membrane proteins. However, generally, these methods are labour-intensive, time-consuming and difficult to implement for quantitative analysis. Here, we report a novel approach, site-directed alkylation detected by in-gel fluorescence (SDAF), which monitors the fluorescent band shift caused by alkylation of the EGFP-fused target membrane protein bearing one single introduced cysteine. In-gel fluorescence provides a unique readout of target membrane proteins with EGFP fusion from non-purified samples, revealing a distinct 5 kDa shift on SDS-PAGE gel due to conjugation with mPEG-MAL-5K. Using the structurally characterised bile acid transporter ASBTNM as an example, we demonstrate that SDAF generates a topology map consistent with the crystal structure. The efficiency of mPEG-MAL-5K modification at each introduced cysteine can easily be quantified and analysed, providing a useful tool for probing the solvent accessibility at a specific position of the target membrane protein.

Immediate responses of multi-focal low level laser therapy on quadriceps in knee osteoarthritis patients.

Low-level laser therapy (LLLT) applying on knee osteoarthritis (OA) patients has shown positive outcomes in analgesic effect and functional recovery. However, few studies applied such therapy on large area of quadriceps muscle in these patients. The aim of this study was to evaluate immediate effect of multi-focal LLLT on quadriceps of knee OA patients in pain and functional performance. Fifty-one participants with knee OA were enrolled and evaluated before (T1) and immediately after intervention (T2) by knee joint pain in numeric rating scale (NRS), walking speed, timed five-chair stands, and quadriceps strength by isokinetic dynamometer. Intervention with two multi-focal Gallium-Aluminum-Arsenide laser devices, each device with 36 laser diodes (wavelength 808 ± 10 nm, continuous, mean power 50 mW, 30 minutes), applied simultaneously over bilateral quadriceps with a total dose of 180 J for each thigh. The multi-focal LLLT significantly improved knee joint pain as measured by the NRS (54% reduction), timed five-chair stands, and walking speed (P < .05). Knee extensor strength also increased in terms of peak torque and force of concentric and eccentric contraction as measured by isokinetic dynamometer (P < .05). In conclusion, single-session multi-focal LLLT on quadriceps in knee OA patients has immediate beneficial effect on knee pain reduction, quadriceps strengthening and functional performance recovery. Long-term effect requires further investigation. Multi-focal LLLT on quadriceps might serve as an alternative non-invasive treatment option in these patients.

Porphyrin-Containing Polymer as a Superior Blue Light-Absorbing Additive To Afford High- Jsc Ternary Solar Cells.

Integrating an additional component featuring complementary light absorption into binary polymer solar cells is a superior tactic to ameliorate solar cell efficiency and stability. An appropriate additive not only extends the absorption range but may also facilitate charge separation and transport processes. In this work, we elucidate the effects of incorporating a porphyrin-containing conjugated polymer (PPor-1), which displays absorption in 350-500 nm, into binary PTB7-Th:4TIC and PTB7-Th:ITIC blends, affording devices with an average power conversion efficiency approaching 9%. We successfully demonstrate that PPor-1 can be incorporated as an additive to impart improved Jsc (up to 19.1 mA cm-2).

The conversion and deglycosylation of isoflavones and anthocyanins in black soymilk process.

The conversion of isoflavones and anthocyanins during black soymilk processing including soaking and heating was investigated. During soaking procedure, the ß-glucosidase activity, content of bioactive compounds including daidzein, genistein, delphinidin and cyanidin were significantly increased by deglycosylation reaction. After heating treatment at 90 °C for 1 h, the content of isoflavones ß-glucosides including daidzin, glycitin and genistin were increased through a de-esterification reaction from malonyl-glucosides at high temperature. On the contrary, aglycone content including daidzein and genistein were no significant changed, this result indicated aglycones displayed well thermal stability. Moreover, anthocyanins including delphinidin-3-O-glucoside (D3G) and cyanidin-3-O-glucoside (C3G) were converted into delphinidin and cyanidin by glucose removal. HPLC analysis suggested that daidzein, genistein, D3G and C3G were bound with ß-conglycinin and glycinin. Our results establish the conversion of bioactive components including daidzein, genistein, delphinidin and cyanidin by deglycosylation during soaking and heating progress and benefit to the production of soymilk.

Comparison of different dosing strategies of intratracheally instilled budesonide on meconium injured piglet lungs.

BACKGROUND: Severe inflammation plays a vital role in the pathogenesis of meconium aspiration syndrome (MAS). Intratracheal (IT) instillation of corticosteroids may be beneficial for MAS in optimizing local effect and reducing systemic adverse effects, but the optimum dosing course remains open to question. METHODS: Thirty meconium-injured newborn piglets were enrolled into six study groups. The first four groups consisted of the IT instillation of 0.25/0.5 mg/kg using either one (IT-B251/IT-B501) or two (IT-B252/IT-B502) doses of budesonide, while the other two groups were the intravenous (IV) dexamethasone (0.5 mg/kg) (IV-Dex) group and the control group (Ctrl). Vital signs and cardiopulmonary functions were monitored throughout the experiments. Pulmonary histology was examined after completing the experiments. RESULTS: Both the IV-Dex and IT-B501 groups got significant improvement in oxygenation (P < 0.05). Lung compliance became worse after one dose of 0.25 mg/kg of IT budesonide. Pulmonary histology revealed that there were significantly lower lung injury scores for all treatment groups compared to control group, especially at the non-dependent sites of both the IT-B501 and IT-B502 groups. There was no significant difference between double- and single-dose groups, no matter whether 0.25 or 0.5 mg/kg of budesonide was used. CONCLUSIONS: IT instillation of one dose of 0.5 mg/kg budesonide is beneficial in treating meconium-injured piglet lungs during the first 8 h of injury, but a second dose at an interval of 4 h does not have a superior beneficial effect compared to one dose.

Coacervation of ß-conglycinin, glycinin and isoflavones induced by propylene glycol alginate in heated soymilk.

This study investigated the propylene glycol alginate (PGA)-induced coacervation of ß-conglycinin (7S), glycinin (11S) and isoflavones in heated soymilk. The addition of 0.9% PGA caused 7S, 11S, daidzein and genistein to coacervate following a 1h incubation period. SDS-PAGE showed that the protein bands corresponding to the 7S α', 7S α, 7S ß, 11S A3, and 11S acidic subunits and the 11S basic proteins in the soymilk supernatant fraction (SSF) decreased to 37.7 ± 12.7%, 24.7 ± 3.9%, 4.9 ± 1.8%, 8.5 ± 2.7%, 18.1 ± 1.8% and 6.0 ± 1.6%, respectively. In addition, isoflavones including daidzein and genistein were also coacervated from the SSF into the soymilk pellet fraction (SPF) following incubation with 0.9% PGA for 1h. The amounts of daidzein and genistein in the SSF decreased to 8.6 ± 1.6% and 2.0 ± 1.0%, respectively. HPLC analysis suggested that daidzein and genistein were bound to the 7S and 11S proteins. These results suggested that daidzein and genistein were co-precipitated with the 7S and 11S proteins into the SPF by 0.9% PGA. Our results demonstrated that PGA is a potent coagulant for the coacervation of 7S, 11S, daidzein and genistein.

Aggregation of soy protein-isoflavone complexes and gel formation induced by glucono-δ-lactone in soymilk.

This study investigated the glucono-δ-lactone (GDL)-induced aggregation of isoflavones and soy proteins in soymilk. High-performance liquid chromatography (HPLC) analysis indicated that isoflavones mixed with ß-conglycinin (7S) and glycinin (11S) proteins formed 7S-isoflavone and 11S-isoflavone complexes in soymilk supernatant fraction (SSF). Most of the soy protein-isoflavone complexes then precipitated into the soymilk pellet fraction (SPF) following the addition of 4 mM GDL, whereupon the pH value of the soymilk dropped from 6.6 to 5.9. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and HPLC analysis suggest that the addition of 4 mM GDL induced the aggregation of most 7S (α', α and ß subunits), 11S acidic and 11S basic proteins as well as isoflavones, including most aglycones, including daidzein, glycitein, genistein and a portion of glucosides, including daidzin, glycitin, genistin, malonyldaidzin and malonylgenistin. These results provide an important reference pertaining to the effects of GDL on the aggregation of soy protein-isoflavone complexes and could benefit future research regarding the production of tofu from soymilk.