Cation transport regulator homolog-1 (CHAC1), a crucial component of the glutathione (GSH) metabolic pathway, plays a central role in intracellular redox homeostasis. Programmed cell death (PCD) is an active cellular death process initiated by gene regulation in response to internal and external stressors. Previous research has demonstrated that CHAC1 participates in various cell death processes by modulating glutathione metabolism, endoplasmic reticulum stress, and oxidative stress. In this comprehensive review, we first provide an overview of the involvement of CHAC1 in diverse cell death processes, including the apoptosis, ferroptosis, and paraptosis pathways. Emphasizing its molecular complexity, we explore the potential of CHAC1 as a therapeutic target across different PCD modes. Subsequently, this review summarizes the current status of the usage of various drugs targeting CHAC1 and explores the significance of CHAC1 regulation in diverse diseases, including cancer, acute injury, and metabolic disorders, with particular emphasis on its prognostic value and biomarker candidates for cancer. Next, we have critically evaluated CHAC1-centered therapeutic approaches that primarily focused on tumor interventions, while discussing their implications for drug resistance and tumor suppression. Overall, interference with CHAC1 can induce programmed cell death across a spectrum of diseases, thereby providing novel opportunities for targeted therapy.

The etiology of moyamoya disease (MMD) remains unknown. The main pathological finding is fibrocellular thickening of the intima, irregular undulation of the internal elastic lamina affecting the distal portions of the internal carotid artery and A1 and M1 segments. Our aim is to describe the histological and electron microscope ultrastructural characteristics of cortical MMD vessels (middle cerebral artery) in hemorrhagic and ischemic presentation along different Suzuki stages. From January 2022 to December 2022, we collected clinical and radiological data of 310 patients with MMD, among them we identified 52 patients that underwent superficial temporal artery-middle cerebral artery (STA-MCA) bypass. We collected arterial walls (excisional arteriotomy) of the recipient arteries specifically, M3 or M4 segments of the MCA. Observations and micrographs were captured utilizing an HT7700 transmission electron microscope. MMD patients exhibit severe internal elastic lamina (IEL) changes as compared to patients with intracranial atherosclerosis. Hemorrhagic MMD presentation showed a higher score of IEL ruptured when comparing to ischemic presentation. Endothelial cells in hemorrhagic MMD showed more significant contraction compared to those in ischemic moyamoya disease. Hemorrhagic and ischemic MMD patients showed no statistically significant differences when correlated to Suzuki stages and cerebral perfusion. MMD patients exhibit IEL changes and endothelial cells contraction extending into the distal segments of the middle cerebral artery. Hemorrhagic MMD presentation has higher IEL rupture score making these patients probably more susceptible for hemorrhage. This study provides an inside of the extension of MMD into the brain surface.

Coenzyme Q10 (CoQ10) is a powerful antioxidant. However, the poor water solubility and low bioavailability still remain challenges for its application. An embedded delivery system of CoQ10 based on whey protein concentrate (WPC) and polymerized whey protein concentrate (PWPC) was prepared, and the physicochemical properties, antioxidant capacity and bioavailability were characterized in this study. Both groups of nanoparticles showed a particle size distribution from 241 to 331 nm in the protein-to-CoQ10 mass ratio range of 100:1 to 20:1. In addition, the minimum polydispersity index value was observed at the mass ratio of 20:1. Differential scanning calorimetry and Fourier transform infrared spectra analysis revealed that the CoQ10 was successfully dispersed in the WPC and PWPC particles through hydrophobic interaction in both groups in addition to the hydrogen bond present in the WPC group. All nanoparticles exhibited irregular spherical or aggregate structure in the transmission electron microscopy diagram. The PWPC-based nanoparticles showed a slightly higher antioxidant capacity than that of the WPC, and both values were significantly higher than that of its corresponding physical mixture and free CoQ10 (p < 0.05). The results of the simulated gastrointestinal digestion experiments denoted that these two nanoparticles could protect CoQ10 from gastric digestion and then deliver it to the intestine. Compared with its free state, the bioavailability of CoQ10 embedded in WPC and PWPC increased by nearly 7.58 times and 7.48 times, respectively. The data indicated that WPC and PWPC could be effective delivery carriers to enhance the bioavailability of active substances like CoQ10.

The composition of milk lipids varies across different ethnic sources. The lipidome profiles of Chinese Han human milk (HHM) and Chinese Korean human milk (KHM) were investigated in this study. A total of 741 lipids were identified in HHM and KHM. Twenty-eight differentially expressed lipids (DEL) were screened between the 2 milk groups; among these, 6 triacylglycerols (TG), 13 diacylglycerols (DG), 7 free fatty acids (FFA), and 1 monoglyceride (MG) were upregulated in KHM. Carnitine (CAR) was upregulated in HHM. Most DEL showed a single peak distribution in both groups. The correlations, related pathways and diseases of these DEL were further analyzed. The results demonstrated that DG, MG, and FFA showed highly positive correlations with each other (r > 0.8). The most enriched Kyoto Encyclopedia of Genes and Genomes (https://www.kegg.jp/kegg/) and Human Metabolome Database (http://www.hmdb.ca) pathways were inositol phosphate metabolism, and α-linolenic acid and linolenic acid metabolism, respectively. Major depressive disorder-related FFA (20:5) and FFA (22:6) were more abundant in KHM, whereas HHM showed more obesity-related CAR. These data potentially provide lipidome information regarding human milk from different ethnicities in China.

BackgroundIn some MMD patients, the digital subtraction angiography (DSA) examination found, occlusion in the ipsilateral internal carotid artery or middle cerebral artery, accompanied by the formation of numerous moyamoya vessels. Conversely, the contralateral internal carotid artery or middle cerebral artery shows signs of stenosis without the presence of moyamoya vessels. Notably, cerebral perfusion studies reveal a similar or even more severe reduction in perfusion on the occluded side compared to the stenotic side. Importantly, clinical symptoms in these patients are typically attributed to ischemia caused by the stenotic side. This condition is referred to as unstable moyamoya disease (uMMD).ObjectiveThis clinical research focuses on evaluating risk factors related to MMD and developing strategies to minimize postoperative complications. The study aims to analyze vascular characteristics and identify potential risk factors in patients with uMMD.MethodsThe authors reviewed consecutive cases with complete clinical and radiological documentation of patients who underwent surgery between January 2018 and June 2023. Univariate analysis and multivariate logistic regression analysis were employed to understand the risk factors and prognosis of postoperative complications in uMMD.ResultsPostoperative complications were retrospectively analyzed in 1481 patients (aged 14 to 65). Among them, 1,429 patients were assigned to the conventional treatment group, while 52 were in the unstable moyamoya disease group. The uMMD treatment group showed a significantly higher incidence of early postoperative complications such as RIND, cerebral infarction, and cerebral hemorrhage (p < 0.05). Univariate and multivariate logistic regression analyses were conducted on the postoperative complications of 52 uMMD patients. Initial symptoms of stenosis ≤50% (univariate: p = 0.008, multivariate: p = 0.015; OR [95% CI] =23.149 [1.853–289.217]) and choosing occluded side surgery (univariate: p = 0.043, multivariate: p = 0.018; OR [95% CI] =0.059 [0.006–0.617]) were identified as significant risk factors for postoperative neurological complications.ConclusionCompared to the conventional treatment group, uMMD has higher complication rates, with vascular stenosis degree and surgical side selection identified as significant risk factors. A comprehensive understanding of preoperative clinical symptoms and vascular characteristics in moyamoya disease patients, coupled with the formulation of rational surgical plans, contributes positively to decreasing postoperative mortality and disability rates in uMMD.

Impacts of co-cold extrusion (<= 50 degrees C) of whey protein isolate (WPI) and cysteine (Cys, 0, 20, 40, 60, 80 and 100 mmol/L) on its physicochemical, in vitro digestion and rheological properties were investigated. As Cys concentration increased, the emulsifying properties and in vitro digestibility of co-extruded WPI-Cys products showed an increasing trend. Specifically, when Cys reached 100 mmol/L, surface hydrophobicity, emulsification activity index (EAI), emulsification stability index (ESI) and in vitro stomach digestibility of the co-extruded WPI-Cys products increased by 205.07%, 77.51%, 193.95% and 71.81% compared with WPI, respectively. Principal component analysis (PCA) results further indicated that co-extruded WPI-Cys at a concentration of 100 mmol/L had the best functional properties. In addition, co-extruded WPI-Cys exhibited the strongest P & eacute;clet number (Pe) value and apparent viscosity at a Cys concentration of 100 mmol/L among all samples. Therefore, co-extrusion would be an effective method for modifying WPI, providing whey protein-based ingredients with excellent functional properties for food processing.

Endometriosis is a disease characterized by the ectopic growth of endometrial tissue (glands and stroma) outside the confines of the uterus and often involves vital organs such as the intestines and urinary system. Endometriosis is considered a refractory disease owing to its enigmatic etiology, propensity for recurrence following conservative or surgical interventions, and the absence of radical treatment and long-term management. In recent years, the incidence of endometriosis has gradually increased, rendering it a pressing concern among women of childbearing age. A more profound understanding of its pathogenesis can significantly improve prognosis. Recent research endeavors have spotlighted the molecular mechanisms by which microRNAs (miRNAs) regulate the occurrence and progression of endometriosis. Many miRNAs have been reported to be aberrantly expressed in the affected tissues of both patients and animal models. These miRNAs actively participate in the regulation of inflammatory reactions, cellular proliferation, angiogenesis, and tissue remodeling. Their capacity to modulate crucial signaling pathways, such as the Wnt/β-catenin signaling pathway, reinforces their potential utility as diagnostic markers or therapeutic agents for endometriosis. In this review, we provide the latest insights into the role of miRNAs that interact with the Wnt/β-catenin pathway to regulate the biological behaviors of endometriosis cells and disease-related symptoms, such as pain and infertility. We hope that this review will provide novel insights and promising targets for innovative therapies addressing endometriosis.

This paper investigated the antibacterial mechanism of spermidine-capped carbon dots (S-PCDs) against Staphylococcus aureus. The results showed that there were a large number of amino groups on the surface of S-PCDs and they had a high positive charge (+47.06 mV), which could be adsorbed on the negatively charged bacterial surface through electrostatic interaction and changed the permeability of the bacterial cell membrane. The extracellular protein and nucleic acid contents of S. aureus treated with S-PCDs were 5.4 and 1.2 times higher than those of the control group, respectively. The surface folds and defects of the bacterial cell membrane, and the leakage of cell contents were observed using SEM and TEM. The expression of metabolic oxidation regulatory genes dmpI, narJ and narK was upregulated and the intracellular ROS generation was induced, causing bacterial oxidative stress and eventually bacterial death. S-PCDs can effectively inhibit biofilm formation and had low cytotoxicity. The S-PCD treatment successfully inhibited microbial reproduction when pasteurized milk was stored at 25 °C and 4 °C. These results provide important insights into the antimicrobial mechanism of S-PCDs and lay the foundation for their application in the food field as a potentially novel bacteriostatic nanomaterial.

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. Physi-ochemical, 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 fl-sheet contributed to stabilizing foams. These findings provide valuable information on MPM as ingredients for aerated foods under acidic, neutral, and alkaline conditions.

This paper investigated antibacterial mechanism against Staphylococcus aureus of spermidine-capped carbon dots (S-PCDs) synthesized by hydrothermal synthesis. Results showed that the surface of S-PCDs with a diameter of 6.87 nm contained a large number of amino groups and high positive charge (+47.06 mV), which made it adsorbed on the surface of negatively charged bacteria through electrostatic interaction, and changed the permeability of bacterial cell membrane. After S-PCDs treatment of S. aureus, the absorption fluorescence intensity of propyl iodide was 2.1 times that of the control group, and extracellular protein and nucleic acid contents were increased 5.4 and 1.2 times, respectively. The surface folds and defects of bacterial cell membrane and the leakage of cell contents were observed by SEM and TEM. After S-PCDs treatment, the expression of metabolic oxidation regulatory genes dmpI, narJ and narK was up-regulated, and more oxidative metabolic enzymes were encoded, leading to metabolic imbalance. S-PCDs treatment can induce intracellular ROS generation, causing bacterial oxidative stress and eventually bacterial death. S-PCDs can effectively inhibit biofilm formation and had low cytotoxicity. In addition, the S-PCDs treatment successfully inhibited microbial reproduction when pasteurized milk was stored at 25 °C and 4 °C. These results provide important insights into the antimicrobial mechanism of S-PCDs and lay the foundation for its application in food field as a potentially novel bacteriostatic nanomaterial.

ObjectivesWe aimed to explore the results of OA-PICA-protected bypass grafting in patients with severe stenosis of the vertebral artery combined with PICA.MethodsThree patients with vertebral artery stenosis involving the posterior inferior cerebellar artery, treated by the Department of Neurosurgery of Henan Provincial People's Hospital from January 2018 to December 2021, were retrospectively analyzed. All the patients underwent Occipital Artery–Posterior Inferior Cerebellar Artery (OA-PICA) bypass surgery followed by elective vertebral artery stenting. Intraoperative indocyanine green fluorescence angiography (ICGA) showed patency of the bridge-vessel anastomosis. Postoperatively, the ANSYS software was used to assess the flow pressure changes and vascular shear in combination with the reviewed DSA angiogram. CTA or DSA was reviewed 1–2 years postoperatively, and the prognosis was evaluated by the modified Rankin Scale (mRS) one year postoperatively.ResultsOA-PICA bypass surgery was completed in all patients, with intraoperative ICGA showing a patent bridge anastomosis, followed by stenting of the vertebral artery, and a review of the DSA angiogram. We also employed ANSYS software evaluation of the bypass vessel, which showed stable pressure and low turnover angle, suggesting a low rate of long-term occlusion of the vessel. All patients had no procedure-related complications during their hospitalization, and were followed up for a mean of 24 months postoperatively, with a good prognosis (mRS score of 1) at 1 year postoperatively.ConclusionOA-PICA-protected bypass grafting is an effective treatment for patients with severe stenosis of the vertebral artery combined with PICA.

乳清蛋白作为主要蛋白成分,与婴儿配方奶粉的起泡特性息息相关.脱盐乳清粉D70、D90和浓缩乳清蛋白(whey protein concentrate,WPC)是婴儿配方奶粉的3种主要乳清蛋白原料.该研究表征并比较了热处理工艺下3种乳清蛋白原料起泡特性、理化与结构特性变化,探究起泡性与理化及结构变化影响规律.结果表明,D70的起泡能力(95.62％)和泡沫稳定性(65.02％)最高,其次为D90(73.97％/24.47％),最小是WPC(60.39％/13.91％).不同加热处理均提高了乳清蛋白的起泡能力,而其中以80 ℃,20 s影响最大.通过粒径、电位、溶解度表征蛋白的理化特性,借助光谱技术测定蛋白的结构特性.皮尔逊相关性分析表明,起泡能力及泡沫稳定性与Zeta电位(0.78/0.69)、β-转角含量(0.82/0.67)、无规则卷曲含量(0.74/0.60)、表面疏水性(0.67/0.46)呈正相关,与α-螺旋呈负相关(-0.81/-0.84).研究结果对于改善婴儿配方奶粉冲调起泡性具有指导意义.

The modification, structure, functionality and IgE binding capacity of soybean protein (SPI) upon covalent conjugation with gallic acid (GA), caffeic acid (CA), and tannic acid (TA) under alkali treatment were assessed. SDS-PAGE showed the formation of SPI-polyphenol conjugates and the cross-linking of SPI. Protein unfolding in the conjugates was observed, characterized by a reduction in α-helix and an increase in UV ultraviolet absorption, surface hydrophobicity and free sulfhydryl groups. LC/MS-MS demonstrated that the modification of protein and major allergens varied with the types of polyphenols. Western-blot and ELISA demonstrated that SPI-polyphenol conjugates exhibited a significant reduced IgE binding capacity due to the masking or destruction of epitopes among Gly m 4, Gly m 5, Gly m 6 and P28, resulting from structural changes. Additionally, antioxidant capacity and emulsifying properties were increased in SPI-polyphenol conjugates. Therefore, polyphenol treatment may be a promising method to prepare hypoallergenic soybean products with desired functionality.

This study was performed to determine the crosslinking formed by proanthocyanidins (PC) with respect to IgE binding capacities, functionality, structure and composition of soybean protein (SPI) following the alkali treatment at 60-100 degrees C. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed the formation of >180 kDa polymers, resulting from the formation of SPI-PC conjugates and protein cross-links. Structural analyses demonstrated that SPI-PC conjugates exhibited structural changes to unfold proteins and increase molecular flexibility. Liquid chromatography coupled to tandem mass spectrometry (LC/MS-MS) showed a decrease in unique protein and peptide numbers as well as major allergen and dominant epitopes abundance. When SPI was treated with PC under the alkali treatment at 80 degrees C, it exhibited a maximum reduction (68.8 %) in the immunoglobulin E (IgE) binding capacity and a maximum increase in DPPH' radical scavenging activities (6.11-fold), ABTS '+ radical scavenging activities (4.80-fold), foaming stability (6.1 %) and emulsifying activity (27.3 %), compared to the control SPI. Overall, this study demonstrates that alkali treatment at 60-100 degrees C to form SPI-PC conjugates has potential applications for producing hypoallergenic soybean products with the desired functionality, most especially from alkali treatment at 80 degrees C. Moreover, the addition of PC pronouncedly alleviates the undesirable functional properties in heated SPI.

This study investigated the changes in functionality, structure, and protein composition in soybean protein-proanthocyanidins (SPI-PC) conjugates prepared by the alkali method with dose-ranging PC molecular graft-ing (0.5, 1, 2 mg/mL). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that more >180 kDa polymers were formed in SPI-PC conjugates with the increase of PC levels. The structural analysis showed that conjugation to PC altered the molecular structure of SPI, appearing the protein-unfolding with PC level-dependent. Liquid chromatography coupled to tandem mass spectrometry (LC/MS-MS) analysis demonstrated that the composition of protein and peptides, as well as allergen and epitopes, were changed after SPI was conjugated with PC. The antioxidant activity of SPI-PC conjugates was increased with the increase of PC levels because high PC contents caused more hydroxyl groups in polyphenols to introduce into the SPI. Emul-sifying properties were increased with the increase of PC levels, probably resulting from the unfolding of pro-teins, increase of random coil and decrease of surface hydrophobicity. Western-bolt and enzyme-linked immunosorbent assay (ELISA) showed a reduced IgE binding capacity in SPI-PC conjugates with PC level -dependent, which was attributed to the reduction of major soybean allergens and the masking or destruction of epitopes induced by structural changes. However, the foaming properties varied from PC levels. Overall, this study demonstrated that SPI-PC conjugates formed using the alkali method have a potential application as hy-poallergenic soybean products or functional ingredients in foods. In future studies, suitable reactions (e.g., PC levels) are required to be investigated in SPI-PC conjugates for balancing various functional properties.

This multiscale study aimed to evaluate the effect of different salts (NaCl, KCl, MgCl2, and CaCl2) on foaming capacity (FC) and foam stability (FS) of model protein system (MPS) for infant formula via the changes of surface and structural properties. Results showed that the FC and FS of MPS were increased at NaCl, KCl, and MgCl2 condition, whereas CaCl2 significantly (P < 0.05) decreased FC (79.5 ± 10.6%) and increased FS (93.2 ± 2.2%). The H0 was increased, while the net charge and surface tension was reduced after addition of salts. Structural analysis revealed the reduction of intensity of intrinsic fluorescence spectroscopy and UV (UV) absorption, and the conversion of α-helix into β-strand, which was attributed to protein agglomeration. Additionally, MgCl2 and CaCl2 exhibited larger size and lower net charge compared with NaCl and KCl, indicating a greater ability to bind to charged amino acids and formed larger aggregates. Correlation analysis indicated that FC was positively related to adsorbed protein and β-turn, while negatively correlated with particle size. FS showed a positive correlation with β-strand, apparent viscosity, and zeta potential. However, it exhibited a negative correlation with β-turn, α-helix, and SH content. These results provide a theoretical reference for further understanding of the effect of salts on the foaming properties of MPS.

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.

In this study, the differences in effects of (-)-epigallocatechin gallate (EGCG) and proanthocyanidins (PC) on the functionality and allergenicity of soybean protein isolate (SPI) were studied. SDS-PAGE demonstrated that SPI-PC conjugates exhibited more high-molecular-weight polymers (>180 kDa) than SPI-EGCG conjugates. Structural analysis showed that SPI-PC conjugates exhibited more disordered structures and protein-unfolding, improving the accessibility of PC to modify SPI, compared to SPI-EGCG conjugates. LC/MS-MS demonstrated that PC caused more modification of SPI and major soybean allergens than EGCG, resulting in a lower abundance of epitopes. The successful attachment of EGCG and PC to SPI significantly increased antioxidant capacity in conjugates. Furthermore, SPI-PC conjugates exhibited greater emulsifying activity and lower immunoglobulin E (IgE) binding capacity than SPI-EGCG conjugates, which was attributed to more disordered structure and protein-unfolding in SPI-PC conjugates. It is implied that proanthocyanidins may be promising compounds to interact with soybean proteins to produce functional and hypoallergenic foods.

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 Mesp bHLH genes play a conserved role during segmental patterning of the mesoderm in the vertebrate embryo by specifying segmental boundaries and anteroposterior (A–P) segmental polarity. Here we use a xenotransgenic approach to compare the transcriptional enhancers that drive expression of the Mesp genes within segments of the presomitic mesoderm (PSM) of different vertebrate species. We find that the genomic sequences upstream of the mespb gene in the pufferfish Takifugu rubripes (Tr-mespb) are able to drive segmental expression in transgenic Xenopus embryos while those from the Xenopus laevis mespb (Xl-mespb) gene drive segmental expression in transgenic zebrafish. In both cases, the anterior segmental boundary of transgene expression closely matches the expression of the endogenous Mesp genes, indicating that many inputs into segmental gene expression are highly conserved. By contrast, we find that direct retinoic acid (RA) regulation of endogenous Mesp gene expression is variable among vertebrate species. Both Tr-mespb and Xl-mespb are directly upregulated by RA, through a complex, distal element. By contrast, RA represses the zebrafish Mesp genes. We show that this repression is mediated, in part, by RA-mediated activation of the Ripply genes, which together with Mesp genes form an RA-responsive negative feedback loop. These observations suggest that variations in a direct response to RA input may allow for changes in A–P patterning of the segments in different vertebrate species.