Exploring Aeromonas dhakensis in Aldabra giant tortoises: a debut report and genetic characterization.

Aeromonas dhakensis (A. dhakensis) is becoming an emerging pathogen worldwide, with an increasingly significant role in animals and human health. It is a ubiquitous bacteria found in terrestrial and aquatic milieus. However, there have been few reports of reptile infections. In this study, a bacterial strain isolated from a dead Aldabra giant tortoise was identified as A. dhakensis HN-1 through clinical observation, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS), and gene sequencing analysis. Subsequently, to evaluate its pathogenicity, the detection of virulence genes and mice infection experiments were performed. A. dhakensis HN-1 was found to contain seven virulence genes, including alt, ela, lip, act, aerA, fla, and hlyA. Mice infected with A. dhakensis HN-1 exhibited hemorrhage of varying degrees in multiple organs. The half-maximal lethal dose (LD50) value of A. dhakensis HN-1 for mice was estimated to be 2.05 × 107 colony forming units (CFU)/mL. The antimicrobial susceptibility test revealed that A. dhakensis HN-1 was resistant to amoxicillin, penicillin, ampicillin and erythromycin. This is the first report of A. dhakensis in Aldabra giant tortoises, expanding the currently known host spectrum. Our findings emphasize the need for One Health surveillance and extensive research to reduce the spread of A. dhakensis across the environment, humans, and animals.

α-Ketoglutarate alleviates osteoarthritis by inhibiting ferroptosis via the ETV4/SLC7A11/GPX4 signaling pathway.

Osteoarthritis (OA) is the most common degenerative joint disorder that causes disability in aged individuals, caused by functional and structural alterations of the knee joint. To investigate whether metabolic drivers might be harnessed to promote cartilage repair, a liquid chromatography-mass spectrometry (LC-MS) untargeted metabolomics approach was carried out to screen serum biomarkers in osteoarthritic rats. Based on the correlation analyses, α-ketoglutarate (α-KG) has been demonstrated to have antioxidant and anti-inflammatory properties in various diseases. These properties make α-KG a prime candidate for further investigation of OA. Experimental results indicate that α-KG significantly inhibited H2O2-induced cartilage cell matrix degradation and apoptosis, reduced levels of reactive oxygen species (ROS) and malondialdehyde (MDA), increased superoxide dismutase (SOD) and glutathione (GSH)/glutathione disulfide (GSSG) levels, and upregulated the expression of ETV4, SLC7A11 and GPX4. Further mechanistic studies observed that α-KG, like Ferrostatin-1 (Fer-1), effectively alleviated Erastin-induced apoptosis and ECM degradation. α-KG and Fer-1 upregulated ETV4, SLC7A11, and GPX4 at the mRNA and protein levels, decreased ferrous ion (Fe2+) accumulation, and preserved mitochondrial membrane potential (MMP) in ATDC5 cells. In vivo, α-KG treatment inhibited ferroptosis in OA rats by activating the ETV4/SLC7A11/GPX4 pathway. Thus, these findings indicate that α-KG inhibits ferroptosis via the ETV4/SLC7A11/GPX4 signaling pathway, thereby alleviating OA. These observations suggest that α-KG exhibits potential therapeutic properties for the treatment and prevention of OA, thereby having potential clinical applications in the future.

Disappearing Cerebral Communicating Vessels in Embryonic Development-With a Case of Carotid-Vertebrobasilar Artery Anastomosis.

In early embryonic development, anastomoses between carotid and basilar arteries occur in the primitive trigeminal, primitive otic, primitive hypoglossal, and primitive proatlantal arteries, and these anastomoses gradually disappear as the embryo continues to develop. This article reported a case of vascular anastomosis occurring between the external carotid artery and vertebral artery.

Improved FAST algorithm for non-uniform rotational distortion correction in OCT endoscopic imaging.

Optical Coherence Tomography (OCT) is widely used for endoscopic imaging in endoluminal organs because of its high imaging accuracy and resolution. However, OCT endoscopic imaging suffers from Non-Uniform Rotational Distortion (NURD), which can be caused by many factors, such as irregular motor rotation and changes in friction between the probe and the sheath. Correcting this distortion is essential to obtaining high-quality Optical Coherence Tomography Angiography (OCTA) images. There are two main approaches for correcting NURD: hardware-based methods and algorithm-based methods. Hardware-based methods can be costly, challenging to implement, and may not eliminate NURD. Algorithm-based methods, such as image registration, can be effective for correcting NURD but can also be prone to the problem of NURD propagation. To address this issue, we process frames by coarse and fine registration, respectively. The new reference frame is generated by filtering out the A-scan that may have the NURD problem by coarse registration. And the fine registration uses this frame to achieve the final NURD correction. In addition, we have improved the Features from Accelerated Segment Test (FAST) algorithm and put it into coarse and fine registration process. Four evaluation functions were used for the experimental results, including signal-to-noise ratio (SNR), peak signal-to-noise ratio (PSNR), mean squared error (MSE), and structural similarity index measure (SSIM). By comparing with Scale-invariant feature transform (SIFT), Speeded up robust features (SURF), Oriented FAST and Rotated BRIEF (ORB), intensity-based (Cross-correlation), and Optical Flow algorithms, our algorithm has a higher similarity between the corrected frames. Moreover, the noise in the OCTA data is better suppressed, and the vascular information is well preserved. Our image registration-based algorithm reduces the problem of NURD propagation between B-scan frames and improves the imaging quality of OCT endoscopic images.

Misdiagnosis of Chronic Subdural Hematoma in Children.

Chronic subdural hematoma is a common intracranial disease in elderly individuals, but in very rare cases, it can also develop in children. Some subdural hematomas are prone to misdiagnosis as epidural hematomas, which can affect the surgical approach and patient prognosis. We report a case of a 12-year-old male child with a misdiagnosed subdural hematoma as an epidural hematoma. We reviewed the data of this misdiagnosed patient with chronic subdural hematoma, analyzed relevant imaging information, identified the reasons for misdiagnosis, and summarized our experience to enhance our understanding of this condition.

Correction: Selection and characterization of ultrahigh potency designed ankyrin repeat protein inhibitors of C. difficile toxin B.

[This corrects the article DOI: 10.1371/journal.pbio.3000311.].

Selection and characterization of ultrahigh potency designed ankyrin repeat protein inhibitors of C. difficile toxin B.

Clostridium difficile infection (CDI) is a major nosocomial disease associated with significant morbidity and mortality. The pathology of CDI stems primarily from the 2 C. difficile-secreted exotoxins-toxin A (TcdA) and toxin B (TcdB)-that disrupt the tight junctions between epithelial cells leading to the loss of colonic epithelial barrier function. Here, we report the engineering of a series of monomeric and dimeric designed ankyrin repeat proteins (DARPins) for the neutralization of TcdB. The best dimeric DARPin, DLD-4, inhibited TcdB with a half maximal effective concentration (EC50) of 4 pM in vitro, representing an approximately 330-fold higher potency than the Food and Drug Administration (FDA)-approved anti-TcdB monoclonal antibody bezlotoxumab in the same assay. DLD-4 also protected mice from a toxin challenge in vivo. Cryo-electron microscopy (cryo-EM) studies revealed that the 2 constituent DARPins of DLD-4-1.4E and U3-bind the central and C-terminal regions of the delivery domain of TcdB. Competitive enzyme-linked immunosorbent assay (ELISA) studies showed that the DARPins 1.4E and U3 interfere with the interaction between TcdB and its receptors chondroitin sulfate proteoglycan 4 (CSPG4) and frizzled class receptor 2 (FZD2), respectively. Our cryo-EM studies revealed a new conformation of TcdB (both apo- and DARPin-bound at pH 7.4) in which the combined repetitive oligopeptides (CROPS) domain points away from the delivery domain. This conformation of the CROPS domain is in stark contrast to that seen in the negative-stain electron microscopy (EM) structure of TcdA and TcdB at the same pH, in which the CROPS domain bends toward and "kisses" the delivery domain. The ultrapotent anti-TcdB molecules from this study serve as candidate starting points for CDI drug development and provide new biological tools for studying the pathogenicity of C. difficile. The structural insights regarding both the "native" conformation of TcdB and the putative sites of TcdB interaction with the FZD2 receptor, in particular, should help accelerate the development of next-generation anti-C. difficile toxin therapeutics.

Sesame allergy: mechanisms, prevalence, allergens, residue detection, effects of processing and cross-reactivity.

Sesame allergy is a serious public health problem and is mainly induced by IgE-mediated reactions, whose prevalence is distributed all over the world. Sesame has been included on the priority allergic food list in many countries. This review summarizes the mechanism and prevalence of sesame allergy. The characteristics, structures and epitopes of sesame allergens (Ses i 1 to Ses i 7) are included. Moreover, the detection methods for sesame allergens are evaluated, including nucleic-acid, immunoassays, mass spectrometry, and biosensors. Various processing techniques for reducing sesame allergenicity are discussed. Additionally, the potential cross-reactivity of sesame with other plant foods is assessed. It is found that the allergenicity of sesame is related to the structures and epitopes of sesame allergens. Immunoassays and mass spectrometry are the major analytical tools for detecting and quantifying sesame allergens in food. Limited technologies have been successfully used to reduce the antigenicity of sesame, involving microwave heating, high hydrostatic pressure, salt and pH treatment. More technologies for reducing the allergenicity of sesame should be widely investigated in future studies. The reduction of allergenicity in processed sesames should be ultimately confirmed by clinical studies. What's more, sesame may exhibit cross-reactivity with peanut and tree nuts.

High Tensile Strength of Engineered ß-Solenoid Fibrils via Sonication and Pulling.

We present estimates of ultimate tensile strength (UTS) for two engineered ß-solenoid protein mutant fibril structures (spruce budworm and Rhagium inquisitor antifreeze proteins) derived from sonication-based measurements and from force pulling molecular dynamics simulations, both in water. Sonication experiments generate limiting scissioned fibrils with a well-defined length-to-width correlation for the mutant spruce budworm protein and the resultant UTS estimate is 0.66 ± 0.08 GPa. For fibrils formed from engineered R. inquisitor antifreeze protein, depending upon geometry, we estimate UTSs of 3.5 ± 3.2-5.5 ± 5.1 GPa for proteins with interfacial disulfide bonds, and 1.6 ± 1.5-2.5 ± 2.3 GPa for the reduced form. The large error bars for the R. inquisitor structures are intrinsic to the broad distribution of limiting scission lengths. Simulations provide pulling velocity-dependent UTSs increasing from 0.2 to 1 GPa in the available speed range, and 1.5 GPa extrapolated to the speeds expected in the sonication experiments. Simulations yield low-velocity values for the Young's modulus of 6.0 GPa. Without protein optimization, these mechanical parameters are similar to those of spider silk and Kevlar, but in contrast to spider silk, these proteins have a precisely known sequence-structure relationship.

Extraordinarily Stable Amyloid Fibrils Engineered from Structurally Defined ß-Solenoid Proteins.

The self-assembly of biological molecules into ordered nanostructures is an attractive method for fabricating novel nanomaterials. Nucleic acid-based nanostructures suffer from limitations to functionalization and stability. Alternatively, protein-based nanostructures have advantageous chemical properties, but design facility lags behind that of nucleic acids. Structurally defined fibrils engineered from ß-solenoid proteins (BSPs) form under mild conditions [Peralta, M. D. R., et al. (2015) ACS Nano 9, 449-463] and are good candidates for novel nanomaterials because of the defined sequence-to-structure relationship and tunable properties. Here, the stability of two types of engineered fibrils was examined using circular dichroism spectroscopy, transmission electron microscopy, and electrophoresis. Both are stable to at least 90 °C, and one survives autoclaving. They are stable toward organic solvents, urea, and pH extremes. One is even stable in 2% sodium dodecyl sulfate with heating. The fibrils show variable resistance to proteolytic digestion: one is resistant to trypsin, but chymotrypsin and proteinase K degrade both. These results show that BSPs have excellent potential for bottom-up design of rugged, functional, amyloid-based nanomaterials.

Self-attention CNN for retinal layer segmentation in OCT.

The structure of the retinal layers provides valuable diagnostic information for many ophthalmic diseases. Optical coherence tomography (OCT) obtains cross-sectional images of the retina, which reveals information about the retinal layers. The U-net based approaches are prominent in retinal layering methods, which are usually beneficial to local characteristics but not good at obtaining long-distance dependence for contextual information. Furthermore, the morphology of retinal layers with the disease is more complex, which brings more significant challenges to the task of retinal layer segmentation. We propose a U-shaped network combining an encoder-decoder architecture and self-attention mechanisms. In response to the characteristics of retinal OCT cross-sectional images, a self-attentive module in the vertical direction is added to the bottom of the U-shaped network, and an attention mechanism is also added in skip connection and up-sampling to enhance essential features. In this method, the transformer's self-attentive mechanism obtains the global field of perception, thus providing the missing context information for convolutions, and the convolutional neural network also efficiently extracts local features, compensating the local details the transformer ignores. The experiment results showed that our method is accurate and better than other methods for segmentation of the retinal layers, with the average Dice scores of 0.871 and 0.820, respectively, on two public retinal OCT image datasets. To perform the layer segmentation of retinal OCT image better, the proposed method incorporates the transformer's self-attention mechanism in a U-shaped network, which is helpful for ophthalmic disease diagnosis.

pH-induced physiochemical and structural changes of milk proteins mixtures and its effect on foaming behavior.

Milk proteins are well known to produce aerated food due to the amphiphilicity. However, milk proteins are commonly added in blends for the desirable properties in food industry. In this study, the foaming properties of milk protein mixtures (MPM), a mixtures of whey protein isolated (WPI) and milk protein concentrate (MPC), was studied through foaming capacity (FC), foam stability (FS), and foam morphology at pH 3.0-9.0. Physiochemical, structural, surface properties, and Pearson correlation analysis were measured to gain insight into foaming behavior. Results indicated that MPM showed excellent FC (113.0-114.3 %) and FS (90.7-93.0 %) at pH 6.0-9.0, and foam displayed a smaller size and uniform distribution. MPM solutions showed smaller particles, higher solubility, and lower apparent viscosity at pH 6.0-9.0, which resulted in an increase in surface pressure and adsorption rate (Kdiff), facilitating more protein absorbed to interface. To further investigate structural changes, various spectral methods were used, in which the structure of MPM was changed with pH. Correlation analysis further suggests that Kdiff and solubility positively affect the formation of foam, while free sulfhydryl and ß-sheet contributed to stabilizing foams. These findings provide valuable information on MPM as ingredients for aerated foods under acidic, neutral, and alkaline conditions.

The properties of the rice resistant starch processing and its application in skimmed yogurt.

Extrusion is typically employed to prepare resistant starch (RS). However, the process is complicated. In this study, the effects of twin-screw extrusion on the crystallinity, thermal properties, and functional properties of starch formed in different extrusion zones were investigated. The effects of this process on the rheological properties and microstructure of RS-added skimmed yogurt were also studied. According to the results, the RS content increased from 7.40 % in the raw material to 33.79 % in the extrudate. The A-type crystal structure of the starch was not observed. The dissociation temperature of the extruded starch ranged from 87.76 °C to 100.94 °C. The glycemic index (GI) of skimmed yogurt fortified with 0.4 % RS was 48.7, and the viscosity was also improved. The microstructure exhibited a uniform network of the starch-protein structure. The findings may serve as a theoretical basis for the application of RS in the food industry.

Effect of pH on the soybean whey protein-gum arabic emulsion delivery systems for curcumin: Emulsifying, stability, and digestive properties.

A novel curcumin (CUR) delivery system was developed using soybean whey protein (SWP)-based emulsions, enhanced by pH-adjustment and gum arabic (GA) modification. Modulating electrostatic interactions between SWP and GA at oil/water interface, pH provides favorable charging conditions for stable distribution between droplets. GA facilitated the SWP form a stable interfacial layer that significantly enhanced the emulsifying properties and CUR encapsulation efficiency of the system at pH 6.0, which were 90.15 ± 0.67%, 870.53 ± 3.22 m2/g and 2157.62 ± 115.31%, respectively. Duncan's test revealed significant improvements in thermal, UV, oxidative, and storage stabilities of CUR (P < 0.05). At pH 6.0, GA effectively protected CUR by inhibiting SWP degradation during gastric digestion and promoting the release of CUR by decreasing steric hindrance with oil droplets during intestinal digestion, achieving the highest CUR bioaccessibility (69.12% ± 0.28%) based on Duncan's test. The SWP-GA-CUR emulsion delivery system would be a novel carrier for nutrients.

Integrated Network Pharmacology and Experimental Validation Approach to Investigate the Mechanisms of Radix Rehmanniae Praeparata - Angelica Sinensis - Radix Achyranthis Bidentatae in Treating Knee Osteoarthritis.

Background: Knee osteoarthritis (KOA) is a persistent degenerative condition characterized by the deterioration of cartilage. The Chinese herbal formula Radix Rehmanniae Praeparata- Angelica Sinensis-Radix Achyranthis Bidentatae (RAR) has often been used in effective prescriptions for KOA as the main functional drug, but its underlying mechanism remains unclear. Therefore, network pharmacology and verification experiments were employed to investigate the impact and mode of action of RAR in the treatment of KOA. Methods: The destabilization of the medial meniscus model (DMM) was utilized to assess the anti-KOA effect of RAR by using gait analysis, micro-computed tomography (Micro-CT), and histology. Primary chondrocytes were extracted from the rib cartilage of a newborn mouse. The protective effects of RAR on OA cells were evaluated using a CCK-8 assay. The antioxidative effect of RAR was determined by measuring reactive oxygen species (ROS), superoxide dismutase (SOD), and glutathione (GSH) production. Furthermore, network pharmacology and molecular docking were utilized to propose possible RAR targets for KOA, which were further verified through experiments. Results: In vivo, RAR significantly ameliorated DMM-induced KOA characteristics, such as subchondral bone sclerosis, cartilage deterioration, gait abnormalities, and the degree of knee swelling. In vitro, RAR stimulated chondrocyte proliferation and the expression of Col2a1, Comp, and Acan. Moreover, RAR treatment significantly reduced ROS accumulation in an OA cell model induced by IL-1ß and increased the activity of antioxidant enzymes (SOD and GSH). Network pharmacology analysis combined with molecular docking showed that Mapk1 might be a key therapeutic target. Subsequent research showed that RAR could downregulate Mapk1 mRNA levels in IL-1ß-induced chondrocytes and DMM-induced rats. Conclusion: RAR inhibited extracellular matrix (ECM) degradation and oxidative stress response via the MAPK signaling pathway in KOA, and Mapk1 may be a core target.

Gubi Zhitong formula alleviates osteoarthritis in vitro and in vivo via regulating BNIP3L-mediated mitophagy.

BACKGROUND: Osteoarthritis (OA) is characterized by degeneration of articular cartilage, leading to joint pain and dysfunction. Gubi Zhitong formula (GBZTF), a traditional Chinese medicine formula, has been used in the clinical treatment of OA for decades, demonstrating definite efficacy. However, its mechanism of action remains unclear, hindering its further application. METHODS: The ingredients of GBZTF were analyzed and performed with liquid chromatography-mass spectrometry (LC-MS). 6 weeks old SD rats were underwent running exercise (25 m/min, 80 min, 0°) to construct OA model with cartilage wear and tear. It was estimated by Micro-CT, Gait Analysis, Histological Stain. RNA-seq technology was performed with OA Rats' cartilage, and primary chondrocytes induced by IL-1ß (mimics OA chondrocytes) were utilized to evaluated and investigated the mechanism of how GBZTF protected OA cartilage from being damaged with some functional experiments. RESULTS: A total of 1006 compounds were identified under positive and negative ion modes by LC-MS. Then, we assessed the function of GBZTF through in vitro and vivo. It was found GBZTF could significantly up-regulate OA rats' limb coordination and weight-bearing capacity, and reduce the surface and sub-chondral bone erosions of OA joints, and protect cartilage from being destroyed by inflammatory factors (iNOS, IL-6, IL-1ß, TNF- α, MMP13, ADAMTS5), and promote OA chondrocytes proliferation and increase the S phage of cell cycle. In terms of mechanism, RNA-seq analysis of cartilage tissues revealed 1,778 and 3,824 differentially expressed genes (DEGs) in model vs control group and GBZTF vs model group, respectively. The mitophagy pathway was most significantly enriched in these DEGs. Further results of subunits of OA chondrocytes confirmed that GBZTF could alleviate OA-associated inflammation and cartilage damage through modulation BCL2 interacting protein 3-like (BNIP3L)-mediated mitophagy. CONCLUSION: The therapeutic effectiveness of GBZTF on OA were first time verified in vivo and vitro through functional experiments and RNA-seq, which provides convincing evidence to support the molecular mechanisms of GBZTF as a promising therapeutic decoction for OA.

TRPC4 aggravates hypoxic pulmonary hypertension by promoting pulmonary endothelial cell apoptosis.

Pulmonary hypertension (PH) is a devastating disease that lacks effective treatment options and is characterized by severe pulmonary vascular remodeling. Pulmonary arterial endothelial cell (PAEC) dysfunction drives the initiation and pathogenesis of pulmonary arterial hypertension. Canonical transient receptor potential (TRPC) channels, a family of Ca2+-permeable channels, play an important role in various diseases. However, the effect and mechanism of TRPCs on PH development have not been fully elucidated. Among the TRPC family members, TRPC4 expression was markedly upregulated in PAECs from hypoxia combined with SU5416 (HySu)-induced PH mice and monocrotaline (MCT)-treated PH rats, as well as in hypoxia-exposed PAECs, suggesting that TRPC4 in PAECs may participate in the occurrence and development of PH. In this study, we aimed to investigate whether TRPC4 in PAECs has an aggravating effect on PH and elucidate the molecular mechanisms. We observed that hypoxia treatment promoted PAEC apoptosis through a caspase-12/endoplasmic reticulum stress (ERS)-dependent pathway. Knockdown of TRPC4 attenuated hypoxia-induced apoptosis and caspase-3/caspase-12 activity in PAECs. Accordingly, adeno-associated virus (AAV) serotype 6-mediated pulmonary endothelial TRPC4 silencing (AAV6-Tie-shRNA-TRPC4) or TRPC4 antagonist suppressed PH progression as evidenced by reduced right ventricular systolic pressure (RVSP), pulmonary vascular remodeling, PAEC apoptosis and reactive oxygen species (ROS) production. Mechanistically, unbiased RNA sequencing (RNA-seq) suggested that TRPC4 deficiency suppressed the expression of the proapoptotic protein sushi domain containing 2 (Susd2) in hypoxia-exposed mouse PAECs. Moreover, TRPC4 activated hypoxia-induced PAEC apoptosis by promoting Susd2 expression. Therefore, inhibiting TRPC4 ameliorated PAEC apoptosis and hypoxic PH in animals by repressing Susd2 signaling, which may serve as a therapeutic target for the management of PH.

Comparison of proanthocyanidins A2 and B2 on IgE-reactivity and epitopes in Gly m 6 using multispectral, LC/MS-MS and molecular docking.

This study comparatively analyzed the changes in IgE-reactivity and epitopes in proanthocyanidins A2- (PA-Gly m 6) and B2-Gly m 6 (PB-Gly m 6) conjugates prepared by alkali treatment at 80 °C for 20 min. Similar to the western blot, ELISA also showed a higher reduced IgE-reactivity in PA-Gly m 6 (70.12 %) than PB-Gly m 6 (63.17 %). SDS-PAGE demonstrated that proanthocyanidins A2 caused more formation of >180 kDa polymers than proanthocyanidins B2. Multispectral analyses revealed that PA-Gly m 6 exhibited more structural alteration (e.g., a decrease of α-helical content and ANS fluorescence intensity) to unfold protein structure than proanthocyanidins B2, improving the accessibility to modify Gly m 6 for shielding or destroying conformational epitopes. LC/MS-MS revealed that PA-Gly m 6 conjugates had a lower abundance of allergens, peptides and linear epitopes than PB-Gly m 6 conjugates. Molecular docking showed that proanthocyanidins A2 and B2 reacted with Gln-317 and Asn-94 of epitopes, respectively. Overall, proanthocyanidins A2 is more effective than proanthocyanidins B2 to decrease the IgE-reactivity of Gly m 6 due to more shielding or destruction of conformational epitopes and lower content allergens and linear epitopes, which was attributed to more protein-crosslinks formation and structural changes in PA-Gly m 6 conjugates.

Multi-spectral and proteomic insights into the impact of proanthocyanidins on IgE binding capacity and functionality in soy 11S protein during alkali-heating treatment.

This study evaluated the impact of proanthocyanidins on immunoglobulin E (IgE) binding capacity, antioxidant, foaming and emulsifying properties in soy 11S protein following alkali treatment at 80 °C for 20 min. The formation of >180 kDa polymer was observed in the combined heating and proanthocyanidins-conjugation treatment sample (11S-80PC) rather than in the heating treated sample (11S-80) using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The structural analyzes demonstrated that 11S-80PC exhibited more protein unfolding than 11S-80. Heatmap analysis revealed that 11S-80PC had more alteration of peptide and epitope profiles in 11S than in 11S-80. Molecular docking showed that PC could well react with soy protein 11S. Liquid chromatography tandem MS analysis (LC/MS-MS) demonstrated that there was a 35.6 % increase in 11S-80, but a 14.5 % decrease in 11S-80PC for the abundance of total linear epitopes. As a result, 11S-80PC exhibited more reduction in IgE binding capacities than 11S-80 owing to more obscuring and disruption of linear and conformational epitopes induced by structural changes. Moreover, 11S-80PC exhibited higher antioxidant capacities, foaming properties and emulsifying activity than 11S-80. Therefore, the addition of proanthocyanidins could decrease allergenic activity and enhance the functional properties of the heated soy 11S protein.

The alteration of composition, conformation, IgE-reactivity and functional attributes in proanthocyanidins-soy protein 7S conjugates formed by alkali-heating treatment: Multi-spectroscopic and proteomic analyses.

This study assessed the alteration of IgE-reactivity and functional attribute in soy protein 7S-proanthocyanidins conjugates (7S-80PC) formed by alkali-heating treatment (pH 9.0, 80 °C, 20 min). SDS-PAGE demonstrated that 7S-80PC exhibited the formation of >180 kDa polymers, although the heated 7S (7S-80) had no changes. Multispectral experiments revealed more protein unfolding in 7S-80PC than in 7S-80. Heatmap analysis showed that 7S-80PC showed more alteration of protein, peptide and epitope profiles than 7S-80. LC/MS-MS demonstrated that the content of total dominant linear epitopes was increased by 11.4 % in 7S-80, but decreased by 47.4 % in 7S-80PC. As a result, Western-blot and ELISA showed that 7S-80PC exhibited lower IgE-reactivity than 7S-80, probably because 7S-80PC exhibited more protein-unfolding to increase the accessibility of proanthocyanidins to mask and destroy the exposed conformational epitopes and dominant linear epitopes induced by heating treatment. Furthermore, the successful attachment of PC to soy 7S protein significantly increased antioxidant activity in 7S-80PC. 7S-80PC also showed higher emulsion activity than 7S-80 owing to its high protein flexibility and protein unfolding. However, 7S-80PC exhibited lower foaming properties than 7S-80. Therefore, the addition of proanthocyanidins could decrease IgE-reactivity and alter the functional attribute of the heated soy 7S protein.