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Aim Emperipolesis, the cell-in-cell structure, is one of the pathological diagnostic standard of AIH. We have already identified that CD8+ T cells participated in the emperipolesis process in liver tissue of AIH patients. In this study, we aimed to clarify the characteristics and molecular mechanisms of emperipolesis in the patients with AIH in vitro and in mice with α-GalCer-induced acute hepatitis. Methods Peripheral blood mononuclear cell (PBMC) of patients with variant chronic liver diseases and healthy controls were co-cultured with hepatic cell line to induce emperipolesis in vitro. Confocal staining was performed to illustrate the cellular types and potential mechanisms of emperipolesis in AIH. In addition, the mouse model of α-GalCer-induced acute hepatitis which mimics human AIH in several features, was used to confirm the role of CD44/p-ERM/F-actin in the emperipolesis process in vivo. Results In co-culture system with PBMC and hepatic cell line, emperipolesis was observed most common in the patients with AIH. CD8+ T cells were the major cells participated in the emperipolesis process, and penetrated into hepatocytes actively via CD44/p-ERM/F-actin pathway. As a result, most CD8+ T cells engulfed in hepatocytes undergo self-apoptosis. In α-GalCer-induced acute hepatitis model, emperipolesis was observed around the inflammatory foci, and was inhibited by Ezrin phosphorylation inhibitor NSC668394. Similarly, activated murine CD8+ T cells penetrated into primary hepatocytes via CD44/p-ERM/F-actin pathway in vitro. Conclusions CD8+ T cells penetrated into hepatocytes actively via CD44/p-ERM/F-actin signaling pathway and undergo apoptosis, may be a compensative mechanism to attenuate overwhelming immune attack in AIH. This article is protected by copyright. All rights reserved.

Objectives Collagen triple helix repeat containing-1 (CTHRC1) is a highly conserved extracellular matrix glycoprotein that is overexpressed in two murine models of cholestatic liver fibrosis. Elevated CTHRC1 has been found to attenuate liver fibrosis in these murine models, thus we aimed to study the expression of CTHRC1 in patients with cholestatic liver diseases and its correlation with hepatic conditions. Methods Ninety patients with chronic liver disease, including 48 had primary biliary cholangitis (PBC), 18 had primary sclerosing cholangitis (PSC) and 24 had chronic hepatitis B (CHB), together with five healthy controls (HC), were recruited to this study. Participants' liver sections were analyzed using immunohistochemistry. Serum CTHRC1 levels in another cohort of 59 patients with PBC and 10 age-matched HC were detected by enzyme-linked immunosorbent assay. Results CTHRC1 protein was primarily expressed in activated hepatic stellate cells (HSC). CTHRC1 expression was significantly increased in the PBC and PSC groups, compared with the HC and CHB groups. Importantly, the hepatic fibrosis stage of the PBC group was positively correlated with hepatic CTHRC1 expression (r = 0.425, P = 0.003). Meanwhile, there were significant correlations between serum CTHRC1 levels and both the degrees of hepatic inflammation and fibrosis stage in the PBC group (r = 0.300, P = 0.022; r = 0.321, P = 0.012). Conclusion CTHRC1 may play a role in hepatic fibrogenesis in PBC and that serum CTHRC1 may be a potential novel noninvasive biomarker in the assessment of liver fibrosis and inflammation.

Rutin is a common dietary flavonoid with important antioxidant and pharmacological activities. However, its application in the food industry is limited mainly because of its poor water solubility. The subcritical water (SW) treatment provides an efficient technique to solubilize and achieve the enrichment of rutin in soy protein isolate (SPI) by inducing their complexation. The physicochemical, interfacial, and emulsifying properties of the complex were investigated and compared to the mixtures. SW treatment had much enhanced rutin-combined capacity of SPI than that of conventional method, ascribing to the well-contacted for higher water solubility of rutin with stronger collision-induced hydrophobic interactions. Compared to the mixtures of rutin with proteins, the complex exhibited an excellent surface activity and improved the physical and oxidative stability of its stabilized emulsions. This improving effect could be attributed to the targeted accumulation of rutin at the oil-water interface accompanied by the adsorption of SPI resulting in the thicker interfacial layer, as evidenced by higher interfacial protein and rutin concentrations. This study provides a novel strategy for the design and enrichment of nanovehicle providing water-insoluble hydrophobic polyphenols for interfacial delivery in food emulsified systems.

BACKGROUND: Several methods have been reported for the conjugation of proteins with polysaccharides. Protein–polysaccharide conjugates can be formed by traditional dry heating, but this process is not attractive from an industrial viewpoint, and no commercial conjugates have been manufactured in this way. In the present study, in order to develop a more practical reaction method, macromolecular crowding was used to attach polysaccharides to proteins. RESULTS: Soy protein isolate–dextran conjugates (SDCs) were prepared via the initial stage of the Maillard reaction in macromolecular crowding conditions. The impact of various processing conditions on the formation of SDCs was investigated. The optimal conditions chosen from the experiments were a soy protein isolate/dextran ratio of 1:1 (w/w), a pH of 6.5, a reaction temperature of 60 °C and a reaction time of 30 h. Circular dichroism spectroscopy showed that the secondary and tertiary structures of the conjugates were changed significantly. Structural flexibility increased, allowing better display of their functional characteristics. The conjugates had a composition with various sizes, especially macromolecules, according to gel permeation chromatography. Thermal analysis showed that the thermal stability of the conjugates was improved. CONCLUSION: The production of SDCs under macromolecular crowding conditions appears to be an effective and promising technique, representing an advance over classic protein glycosylation methods. Copyright © 2012 Society of Chemical Industry

Summary The aim of this investigation was to explore the pH-induced conformational changes of phaseolin using differential scanning calorimetry (DSC), fluorescence and circular dichroism (CD) spectra. The surface charges at the varying pH and ionic strength were also determined to probe the possible relationship between the surface charges of phaseolin and its conformation. The ζ-potential of phaseolin decreased from 27.9 mV at pH 3.0 to –44 mV at pH 9.4, and the isoelectric point (iep) occurred at pH 4.21, which were comparable with the theoretical estimations derived from amino acid composition and pK values. The iep of phaseolin was independent of ionic strength, while the magnitude of ζ-potential decreased as the ionic strength increased. The near-UV CD, DSC and fluorescence spectra analyses indicated highly acidic and alkaline pH conditions induced the loss of tertiary conformation of phaseolin. The far-UV CD spectra analysis confirmed pH-shift led to the decreases in ordered secondary structure compositions. The present results suggested the loss of secondary and/or tertiary conformation of phaseolin as a result of the varying pattern in electrostatic interactions as pH shifted from 7.0 to acidic or alkalic pH.

Summary Cold-set soybean glycinin gel was prepared by cross-linking of soybean glycinin (non-preheated and preheated) at pH 8.0 and 50 °C using transglutaminase (TG), followed by cold-set acidification using glucono-δ-lactone (GDL). Dynamic oscillatory rheological measurements (both under non-preheat and preheat conditions) showed that there was a substantial increase in the elastic (G′) and viscous (G″) moduli of enzyme treated gels. The moduli also showed similar frequency dependence. The yield strains of enzyme treated gels were clearly higher than that of the gels without enzyme treatment. Texture profile analysis indicated that preheat and enzyme treatments have beneficial effect on hardness, fracturability and gumminess of gels. Thermal and enzyme treatments have synergetic effect to improve the strength of gels, which is also consistent with the results of rheological measurements. These results demonstrated that heat treatment followed by TG treatment and GDL acidification improved the functional properties of soybean glycinin cold-set gel.

Summary The conjugation reaction between soybean acid-precipitated protein (SAPP) and dextran in liquid systems via the initial stage of the Maillard reaction was studied. Functional SAPP–dextran conjugates were prepared in 80% ethanol-reacting system at 50 °C for 6 h, along with 95% ethanol-reacting system at 60 °C for 24 h. The covalent attachment of dextran to SAPP was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and gel filtration chromatography.Compared to the classical dry-heating, the reaction time of glycosylation in the two ethanol systems was largely shortened. Emulsifying activity of SAPP–dextran conjugates obtained by dry-heating incubation and in ethanol was similar at pH 7.0 and10.0, significantly higher than that of SAPP–dextran mixture or SAPP alone. In addition, SAPP–dextran conjugates obtained in 80% ethanol-reacting system for 6 h were completely soluble after heating at 90 °C for 20 min. The impact of various processing conditions on the formation of SAPP–dextran conjugates was investigated. This study provides important guidance to create protein–polysaccharide conjugates at mild temperatures in liquid systems.

This study aimed at establishing an effective preparative isolation method of soy β-conglycinin constituent subunits and characterizing some of their physicochemical properties and their heat-induced aggregation. These subunits were isolated in relatively large amounts and in high purity by dissociating β-conglycinin in 6 M urea and using a combination of DEAE-Sepharose fast flow column chromatography and immobilized metal ion affinity chromatography (IMAC). At a pH deviating from isoelectric point (pI), zeta potentials of α′ and α subunits were much larger than that of β subunit, while in the latter case, the hydrophobic groups were more buried within the proteins. Dynamic light scattering analysis indicated that the extent of heat-induced aggregation of β subunit was much higher than that of α′ and α subunits, and the aggregation was also more affected by the increase in ionic strength. Atomic force microscopy analysis indicated that more ordered and stranded aggregates were formed for α′ and α subunits. These results confirm a close relationship between physicochemical properties and heat-induced aggregation of β-conglycinin subunits.

Summary The purpose of this study was to characterise the Maillard reaction in a pressurised liquid system containing β-conglycinin and dextran. The rate of browning and the free amino content in this liquid system, with an initial pH of 7.2, were measured after reaction under atmospheric pressure and at 10 MPa. The secondary and tertiary structures of the conjugates were analysed by circular dichroism, showing that the increased pressure accelerated the reaction rate of the free amino groups and reduced the rate of browning. Significant structural changes occurred in the tertiary structure of the conjugates. Additionally, the emulsifying properties of the β-conglycinin–dextran conjugates produced were studied. The emulsifying properties of the conjugates produced at 10 MPa were higher than those made at atmospheric pressure.

A simple two step precipitation method was used to investigate the effect of MgCl 2 instead of CaCl 2 on fractionation of soybean glycinin and β-conglycinin. Compositional and physicochemical properties of the resulting protein fractions were characterised. The optimised procedure, in terms of protein yield, purity, phytate content and physicochemical properties, was obtained when the addition of 5 mM MgCl 2 was used. After application of 5 mM MgCl 2 , the phytate content of the glycinin-rich and β-conglycinin-rich fractions was about 0.4% and 1.3%, respectively, but the addition of 5 mM CaCl 2 increased the phytate content of the glycinin-rich fraction to 1.25% and decreased that of β-conglycinin-rich fraction to 0.67%. Low phytate protein product was suitable for use in infant formula and acidic food. The solubility of the glycinin-rich fractions with MgCl 2 was significantly higher than that with CaCl 2 at pH < 4.5. Application of MgCl 2 improved thermal stability of the β-conglycinin-rich fraction.

Summary The functional acid-precipitated soy protein (SAPP)–dextran conjugate was prepared by dry-heated storage at 60 °C under 79% relative humidity (RH) for 5 days through Maillard reaction between the e-amino of lysine in soy proteins and the reducing-end carbonyl residue in the dextran. The covalent attachment of dextran to SAPP was confirmed by SDS-polyacrylamide gel electrophoresis and gel filtration chromatography. Functional properties of soy protein depend on the structural and aggregation characteristics of their major components (storage globulins 7S and 11S). The conjugate seemed to be predominantly formed by 7S, and the acidic subunits of 11S in soy protein. The emulsifying properties of the SAPP–dextran conjugate were about four times higher than those of SAPP. The solubility of the protein was not enhanced as a result of preheating, but rather it was not decreased when the conjugated protein was heated at 90 °C for 20 min due to the presence of the polysaccharide. The excellent emulsifying properties of SAPP–dextran conjugate were maintained even at pH 3.0 and were further improved at pH 10.0. The object of Maillard reaction is to guarantee the suitable reaction degree, and the resulting soluble conjugate can have excellent emulsifying properties.

Two novel fulvene-type organic ligands L1 and L2 have been synthesized by aroylation reaction of substituted cyclopentadienyl anions. L1 crystallized in the monoclinic space group P21, a = 6.7498(4), b = 20.5769(11), c = 8.9532(5) A, β = 109.5060(10)°, and Z = 2. L2 crystallized in the triclinic space group P , a = 11.4126(10), b = 11.4763(10), c = 11.7719(10) A, α = 67.050(2), β = 74.509(2), γ = 61.148(2)°, and Z = 2. The coordination chemistry of L1 and L2 has been investigated. One novel AgI-containing organometallic coordination polymer 3, based on both AgI−C and AgI−N interactions, has been synthesized by combination of L1 and AgClO4 in a benzene/dichloromethane mixed-solvent system. Two new MII-containing [M = CuII (4) and CoII (5)] supramolecular complexes with seven-membered metalla-ring unit have been prepared by combination of L2 and the corresponding M(OAc)2·2H2O (M = CuII and CoII) salts in ethanol. The isostructural compounds 4 and 5 crystallized in the triclinic space group P [4: a = 8.8808(5), b = 10.9928(7), c = 14.2196(9) A, α = 99.9750(10), β = 95.8190(10), γ = 93.5610(10)°, and Z = 1; 5: a = 8.7964(10), b = 10.8103(12), c = 14.3765(15) A, α = 99.681(2), β = 95.930(2), γ = 93.429(2)°, and Z = 1]. In the solid state, molecular complexes 4 and 5 are bound by strong intermolecular N···O−H hydrogen bonds in a hand-in-hand fashion to give a novel one-dimensional chain motif. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)

Scrotal elephantiasis can be physically disabling and physchologically distressing to the victim. Ablative procedure has been used in its treatment and has achieved limited success. The authors developed a microlymphaticovenous procedure to treat elephantiasis of the scrotum and applied it clinically in three patients. The immediate and long-term (13–24 months) results have been very satisfactory. The scrotum size was dramatically reduced to a nearly normal level, and subjective symptoms and objective signs were improved. The operative techniques are described, the three case histories are illustrated, and the advantages of microlymphaticovenous anastomosis, the selection of patients, and the factors required for success of the surgery are discussed.