The Potential of Soluble Proteins in High-Moisture Soy Protein-Gluten Extrudates Preparation.

In this study, the effects of different soluble proteins, including collagen peptides (CP), soy protein hydrolysate (HSPI), whey protein isolate (WPI), sodium caseinate (SC), and egg white protein (EWP), on the structural and mechanical properties of blends containing soy protein isolate (SPI) and wheat gluten (WG) were investigated using high-moisture extrusion. The addition of CP and HSPI resulted in a more pronounced fibrous structure with increased voids, attributing to their plasticizing effect that enhanced polymer chain mobility and reduced viscosity. WPI, SC, and EWP acted as crosslinking agents, causing early crosslink formation and decreased polymer chain mobility. These structural variations directly influenced the tensile properties of the extrudates, with CP displaying the highest anisotropic index. Moreover, the presence of soluble proteins impacts the permeability of the extrudates. These insights shed light on how soluble proteins can be used to modify the properties of SPI-WG blends, making them suitable for meat analogue production.

Isotope dilution-HPLC-MS/MS to investigate the production patterns and possible pathways of free and protein-bound AGEs and 4-MI in cookies.

A qualitative and quantitative method for detecting free and protein-bound advanced glycation end products (AGEs) and 4-methylimidazole (4-MI) was established using isotope dilution-HPLC-MS/MS, and successfully applied in cookies and model systems. The effects of different temperatures (160-220 °C) on the formation of free and protein-bound AGEs and 4-MI in cookies were discussed, and the possible model systems (Maillard reaction pathway 1 using wheat gluten protein + glucose + sucrose; direct addition pathway 1 using wheat gluten protein + CML/CEL/4-MI) of protein-bound AGEs and 4-MI were verified. The results showed that the contents of protein-bound CML, CEL, and 4-MI were higher than free content with a tendency of increasing first and subsequently decreasing with temperature, reaching a maximum at 200 °C in cookies. In the model systems, the levels of protein-bound CML, CEL, and 4-MI are higher than those of free CML, CEL, and 4-MI. The protein-bound CML, CEL, and 4-MI accounted for 90.73, 87.64, and 97.56% of the total amount in the model system 1, while accounting for 68.19, 59.00, and 50.96% in the model system 2, respectively. In comparison, protein-bound CML, CEL, and 4-MI could be easily generated directly by Maillard reaction.

Performance of atrazine adsorption behavior and microbial community structure in Mollisol aggregate fraction.

Owing to complex pore systems and chemical substances, soil aggregates provide a spatially heterogeneous microenvironment for adsorption capacity and microbial survival. As the widely used pesticide in farmlands, atrazine environmental behavior is not well known at the aggregate scale. In this study, Mollisol soil samples were sieved into four aggregate-size classes: large macroaggregates (>2 mm, LMa), small macroaggregates (1-2 mm, SMa), microaggregates (0.25-1 mm, Mia) and primary particles (<0.25 mm, P). The pore characteristics of each aggregate fraction was visualized by non-invasive X-ray three-dimensional microscopic computed tomography (3D-CT) combined with pore network extraction. The adsorption kinetics of atrazine in each aggregate-size fraction can be described well by a pseudo-second-order kinetic model. The adsorption isothermal process of atrazine can be better fitted by the Langmuir isotherm model than Freundlich isotherm model. There was an obvious linear correlation between the maximum atrazine adsorption capacity and aggregate SOC content as well as TN. In addition, the abundance of bacteria, actinomycetes and anaerobic bacteria in P was totally higher than those in SMa and Mia. Although pH is strongly linked to the bacterial community in the aggregate fraction, aggregate particle size explained 18 % for shaping the microbial community. Therefore, chemical properties and pore characteristics of each soil aggregate fraction both contributed to performance of atrazine adsorption behavior and microbial community.

Effects of AMF Compound Inoculants on Growth, Ion Homeostasis, and Salt Tolerance-Related Gene Expression in Oryza sativa L. Under Salt Treatments.

Increased soil salinization is among the main factors that limits safe rice production. Arbuscular mycorrhizal fungi (AMF) have been shown to alleviate the toxic effects of salt stress in plants. However, more studies on AMF combined with other functional microorganisms are needed to further improve salt tolerance in rice. Therefore, the compound inoculum Funneliformis mosseae (Fm) together with two functional microorganisms, Piriformospora indica (Pi) and Agrobacterium rhizogenes (Ar) was evaluated for their effect on the rice growth, photosynthetic gas exchange parameters, ion homeostasis, and the expression of salt tolerance-related genes under 0, 80, 120 and 160 mM salt stress conditions. The results showed that: (1) the rice seedling biomass of the AMF compound inoculant treatment group was significantly higher than that of the non-inoculation treatment group (P < 0.05); (2) under NaCl stress, inoculation with AMF compound inoculants can activate the rice antioxidant enzyme system and improve osmoregulation ability; (3) AMF compound inoculants can increase the concentration of K+ in the plant and inhibit the transfer of Na+ to rice leaves, maintaining a high K+/Na+; and (4) AMF compound inoculants could induce and regulate the overexpression of genes related to salt tolerance, photosynthesis and ion homeostasis in rice, and improve the tolerance of rice under salt stress. Our study showed that AMF compound inoculants could improve the adaptability of rice under NaCl stress and promote plant growth by regulating the photosynthetic gas exchange parameter, reactive oxygen species (ROS) scavenging ability, and ion homeostasis of plants. These results suggest that AMF compound inoculants may play an important role in improving rice productivity in salinized soil.

Soy Protein Hydrolysates Affect the Structural and Mechanical Properties of Soy Protein-Wheat Gluten Extrudates Using High Moisture Extrusion.

This study aimed to investigate the effect of hydrolyzed soy protein isolate (HSPI) as a plasticizer in the soy protein mixture-wheat gluten (SP-WG) extrudates on its structural and mechanical properties during high moisture extrusion. Those SP were prepared by mixing soy protein isolate (SPI) and HSPI with different ratios. HSPI primarily consisted of small molecular weight peptides measured with size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The elastic modulus of SP-WG blends decreased with increased HSPI contents through the closed cavity rheometer. Adding HSPI at low concentrations (≤30 wt% of SP) enhanced a fibrous appearance and higher mechanical anisotropy while adding more HSPI resulted in a compact and brittle structure and tended to be isotropic. It can be concluded that the partial addition of HSPI as a plasticizer can promote the formation of a fibrous structure with enhanced mechanical anisotropy.

Selection of Reliable Reference Genes for RT-qPCR Analysis of Bursaphelenchus mucronatus Gene Expression From Different Habitats and Developmental Stages.

Quantitative reverse transcription polymerase chain reaction (RT-qPCR), a sensitive technique for gene expression analysis, depends on the stability of the reference genes used for data normalization under different experimental conditions. Bursaphelenchus mucronatus, a pine-parasitic nematode varying in virulence, is widely distributed in natural pine forests throughout the northern hemisphere, but has not been investigated with respect to the identification of reference genes suitable for the normalization of RT-qPCR data. In the present study, eight candidate reference genes were analyzed in B. mucronatus under different habitat conditions and at different developmental stages. The expression stability of these genes was assessed by geNorm, NormFinder, BestKeeper, delta Cq, and RefFinder algorithms. In general, our results identified encoding beta-tubulin as the most stable gene. Moreover, pairwise analysis showed that three reference genes were sufficient to normalize the gene expression data under each set of conditions, with genes encoding beta-tubulin, 18S ribosomal RNA and ubiquitin-conjugating enzyme being the most suitable reference genes for different habitat conditions, whereas genes encoding beta-tubulin, histone, and 18S ribosomal RNA exhibited the most stable expression at different developmental stages. Validation of the selected reference genes was performed by profiling the expression of the fatty acid- and retinol-binding protein gene in different habitats, and by profiling the expression of the arginine kinase gene at different developmental stages. This first systematic analysis for the selection of suitable reference genes for RT-qPCR in B. mucronatus will facilitate future functional analyses and deep mining of genetic resources in this nematode.

Wilms' tumor 1-associating protein plays an aggressive role in diffuse large B-cell lymphoma and forms a complex with BCL6 via Hsp90.

BACKGROUND: Wilms' tumor 1-associating protein (WTAP) is a nuclear protein, which is ubiquitously expressed in many tissues. Furthermore, in various types of malignancies WTAP is overexpressed and plays a role as an oncogene. The function of WTAP in diffuse large B-cell lymphoma (DLBCL), however, remains unclear. METHODS: Immunohistochemistry was applied to evaluate the levels of WTAP expression in DLBCL tissues and normal lymphoid tissues. Overexpression and knock-down of WTAP in DLBCL cell lines, verified on mRNA and protein level served to analyze cell proliferation and apoptosis in DLBCL cell lines by flow cytometry. Finally, co-immunoprecipitation (Co-IP), IP, and GST-pull down assessed the interaction of WTAP with Heat shock protein 90 (Hsp90) and B-cell lymphoma 6 (BCL6) as well as determined the extend of its ubiquitinylation. RESULTS: WTAP protein levels were consistently upregulated in DLBCL tissues. WTAP promoted DLBCL cell proliferation and improved the ability to confront apoptosis, while knockdown of WTAP in DLBCL cell lines allowed a significant higher apoptosis rate after treatment with Etoposide, an anti-tumor drug. The stable expression of WTAP was depended on Hsp90. In line, we demonstrated that WTAP could form a complex with BCL6 via Hsp90 in vivo and in vitro. CONCLUSION: WTAP is highly expressed in DLBCL, promoting growth and anti-apoptosis in DLBCL cell lines. WTAP is a client protein of Hsp90 and can appear in a complex with BCL6 and Hsp90 in DLBCL. Down-regulation of WTAP could improve the chemotherapeutic treatments in DLBCL.

LITAF is a potential tumor suppressor in pancreatic cancer.

Early diagnosis of pancreatic cancer, one of the most deadly cancers with low survival rates, is difficult, and effective biomarkers are urgently needed. Lipopolysaccharide-induced tumor necrosis factor-α factor (LITAF) has been recently proposed as a potential tumor suppressor gene in several types of cancer. Here, we analyzed the biological function of LITAF in pancreatic cancer. The LITAF gene and protein levels were decreased in pancreatic tumor tissues compared with their paired adjacent non-cancerous tissues. In addition, patients with the lower LITAF protein expression had lower disease-free survival rates. The decreased LITAF expression correlated with LITAF promoter hypermethylation in pancreatic cancer cells and tissues. Moreover, promoter demethylation dose-dependently increased the LITAF transcription. Importantly, LITAF demethylation suppressed proliferation and cell cycle progression, and enhanced apoptosis of pancreatic cancer cells. Together, our results indicate that LITAF functions as a tumor suppressor gene in pancreatic cancer cells, and might serve as a novel biomarker for early diagnosis of pancreatic cancer.

Oxyresveratrol and ascorbic acid O/W microemulsion: Preparation, characterization, anti-isomerization and potential application as antibrowning agent on fresh-cut lotus root slices.

The purpose of this study is to prepare an oxyresveratrol (Oxy) microemulsion (ME) with improved Oxy's solubility and stability and to investigate its antibrowning effects on fresh-cut lotus root slices. The formula of OxyME consisted of ethyl butyrate, Tween 80, PEG400, and water with w/w of 4%, 10.67%, 5.33%, and 80%, respectively. Encapsulating Oxy into OxyME greatly increased its solubility and stability compared with that of in water. Strong antibrowning effects were observed on fresh-cut lotus root slices treated with OxyME, even better than 4-hexylresorcinol. The addition of ascorbic acid (VC) into OxyME greatly improved the Oxy stability in long-term storage and antibrowning effects on fresh-cut lotus root slices. However, the simultaneous addition of calcium chloride and VC did not obviously improve the antibrowning effects compared with the addition of VC alone. These results indicated that Oxy+VCME may be suitable as an antibrowning agent for fresh-cut vegetables.

The feedback loop of LITAF and BCL6 is involved in regulating apoptosis in B cell non-Hodgkin's-lymphoma.

Dysregulation of the apoptotic pathway is widely recognized as a key step in lymphomagenesis. Notably, LITAF was initially identified as a p53-inducible gene, subsequently implicated as a tumor suppressor. Our previous study also showed LITAF to be methylated in 89.5% B-NHL samples. Conversely, deregulated expression of BCL6 is a pathogenic event in many lymphomas. Interestingly, our study found an oppositional expression of LITAF and BCL6 in B-NHL. In addition, LITAF was recently identified as a novel target gene of BCL6. Therefore, we sought to explore the feedback loop between LITAF and BCL6 in B-NHL. Here, our data for the first time show that LITAF can repress expression of BCL6 by binding to Region A (-87 to +65) containing a putative LITAF-binding motif (CTCCC) within the BCL6 promoter. Furthermore, the regulation of BCL6 targets ( PRDM1 or c-Myc) by LITAF may be associated with B-cell differentiation. Results also demonstrate that ectopic expression of LITAF induces cell apoptosis, activated by releasing cytochrome c, cleaving PARP and caspase 3 in B-NHL cells whereas knockdown of LITAF robustly protected cells from apoptosis. Interestingly, BCL6, in turn, could reverse cell apoptosis mediated by LITAF. Collectively, our findings provide a novel apoptotic regulatory pathway in which LITAF, as a transcription factor, inhibits the expression of BCL6, which leads to activation of the intrinsic mitochondrial pathway and tumor apoptosis. Our study is expected to provide a possible biomarker as well as a target for clinical therapies to promote tumor cell apoptosis.

Identifying Virulence-Associated Genes Using Transcriptomic and Proteomic Association Analyses of the Plant Parasitic Nematode Bursaphelenchus mucronatus.

Bursaphelenchus mucronatus (B. mucronatus) isolates that originate from different regions may vary in their virulence, but their virulence-associated genes and proteins are poorly understood. Thus, we conducted an integrated study coupling RNA-Seq and isobaric tags for relative and absolute quantitation (iTRAQ) to analyse transcriptomic and proteomic data of highly and weakly virulent B. mucronatus isolates during the pathogenic processes. Approximately 40,000 annotated unigenes and 5000 proteins were gained from the isolates. When we matched all of the proteins with their detected transcripts, a low correlation coefficient of r = 0.138 was found, indicating probable post-transcriptional gene regulation involved in the pathogenic processes. A functional analysis showed that five differentially expressed proteins which were all highly expressed in the highly virulent isolate were involved in the pathogenic processes of nematodes. Peroxiredoxin, fatty acid- and retinol-binding protein, and glutathione peroxidase relate to resistance against plant defence responses, while ß-1,4-endoglucanase and expansin are associated with the breakdown of plant cell walls. Thus, the pathogenesis of B. mucronatus depends on its successful survival in host plants. Our work adds to the understanding of B. mucronatus' pathogenesis, and will aid in controlling B. mucronatus and other pinewood nematode species complexes in the future.

Encapsulation artocarpanone and ascorbic acid in O/W microemulsions: Preparation, characterization, and antibrowning effects in apple juice.

The aim of this study is to improve artocarpanone solubility by developing an O/W microemulsion with the evaluation of its antibrowning effects. The chemical and physical stabilities as well as antibrowning effects in apple juice were also evaluated. The formulation of artocarpanone microemulsion consisted of 4% w/w of ethyl butyrate, 10.67% w/w of Tween 80, 5.33% w/w of polyethylene glycol 400, and 80% w/w of water, with a maximum solubility of artocarpanone up to 10.54 ± 0.01 mg/mL, at least 3000-folds increase in solubility compared that in water. Encapsulating artocarpanone and ascorbic acid (VC) into microemulsion simultaneously decreased modest artocarpanone solubility whereas improving its stability in long-term storage. Blank, artocarpanone and artocarpanone-Vc-loaded microemulsions demonstrated steadily during accelerated and long-term storage. Artocarpanone-Vc-loaded microemulsion showed strong antibrowning effects in apple juice at room temperature in 24h, suggesting that artocarpanone-Vc-loaded microemulsion is a good antibrowning agent for apple juices.

Impact of phase separation of soy protein isolate/sodium alginate co-blending mixtures on gelation dynamics and gels properties.

The influence of sodium alginate (SA) on soy protein isolate (SPI)-based co-blending system gelling properties was studied under thermodynamic compatibility and incompatibility conditions using a direct addition (SPI/SA) or co-drying (SPI/SA-CO) process. For an SPI/SA (30:1) or SPI/SA-CO (30:1) system, the addition of too little SA did not significantly modify the SPI, and the gelation temperature (Tgel) and storage modulus (G') were similar to an SPI solution alone. For SPI/SA (20:1) and SPI/SA-CO (10:1), the Tgel and G' were between the values for solutions of SPI or SA alone; however, SPI/SA-CO (20:1) and SPI/SA-CO (10:1) gels could nearly double the equilibrium value of G' (Geq'), thus improving the barrier and mechanical properties of the final formed films. The cryo-transmission electron microscope morphology of the SPI/SA-CO (20:1) and SPI/SA-CO (10:1) systems after heating was of the core-shell type in which the core comprised SPI gel.

The truncate mutation of Notch2 enhances cell proliferation through activating the NF-κB signal pathway in the diffuse large B-cell lymphomas.

The Notch2 is a critical membrane receptor for B-cell functions, and also displays various biological roles in lymphoma pathogenesis. In this article, we reported that 3 of 69 (4.3%) diffuse large B-cell lymphomas (DLBCLs) exhibited a truncate NOTCH2 mutation at the nucleotide 7605 (G/A) in the cDNA sequence, which led to partial deletion of the C-terminal of PEST (proline-, glutamic acid-, serine- and threonine-rich) domain. The truncate Notch2 activated both the Notch2 and the NF-κB signals and promoted the proliferation of B-cell lymphoma cell lines, including DLBCL and Burkitt's lymphoma cell lines. Moreover, the ectopic proliferation was completely inhibited by ammonium pyrrolidinedithiocarbamate (PDTC), an NF-κB inhibitor. Simultaneously, PDTC also reduced the expression level of Notch2. Based on these results, we conclude that the Notch2 receptor with PEST domain truncation enhances cell proliferation which may be associated with the activation of the Notch2 and the NF-κB signaling. Our results are expected to provide a possible target for new DLBCL therapies by suppressing the Notch2 and the NF-κB signaling.

Critical role of SHP2 (PTPN11) signaling in germinal center-derived lymphoma.

Germinal center lymphoma is a heterogeneous human lymphoma entity. Here we report that constitutive activity of SHP2 (PTPN11) and its downstream kinase ERK is essential for the viability of germinal center lymphoma cells and disease progression. Mechanistically, SHP2/ERK inhibition impedes c-Myc transcriptional activity, which results in the repression of proliferative phenotype signatures of germinal center lymphoma. Furthermore, SHP2/ERK signaling is required to maintain the CD19/c-Myc loop, which preferentially promotes survival of a distinct subtype of germinal center lymphoma cells carrying the MYC/IGH translocation. These findings demonstrate a critical function for SHP2/ERK signaling upstream of c-Myc in germinal center lymphoma cells and provide a rationale for targeting SHP2 in the therapy of germinal center lymphoma.

Assessment of the physical, mechanical, and moisture-retention properties of pullulan-based ternary co-blended films.

Multi-component substances made through direct blending or blending with co-drying can form films on the surfaces of intermediate moisture foods (IMFs), which help retain moisture and protect food texture and flavor. An IMF film system based on pullulan, with glycerol serving as the plasticizer, was studied using alginate and four different types of polysaccharides (propyleneglycol alginate, pectin, carrageenan, and aloe polysaccharide) as the blend-modified substances. The physical, mechanical, color, transparency, and moisture-retention properties of the co-blended films with the polysaccharides were assessed. A new formula was established for the average moisture retention property, water barrier, tensile strength, elongation at break, and oxygen barrier property of the ternary co-blended films using the Design Expert software. The new model established for moisture content measurement used an indirect method of film formation on food surfaces by humectants, which should expedite model validation and allow a better comprehension of moisture transfer through edible films.

Evaluation of specific marker CK13 and CK10/13 combined with APM staining for the diagnosis of amniotic fluid embolism and aspiration.

OBJECTIVE: To explore the value of CK13 (Rab) and CK10/13(Mab) as objective and specific biomarkers combined with Alcian-Phloxine-Martius yellow (APM) staining for the diagnosis of amniotic fluid embolism (AFE) and amniotic fluid aspiration (AFA). METHODS: A retrospective study of forensic autopsy cases of 148 neonatal deaths and 19 maternal deaths in the perinatal stage was conducted at the Institute of Pathology and Forensic Medicine, Zhejiang University. Medical records were reviewed and monoclonal antibody for CK13 (Rab) and CK10/13 (Mab) as immunostaining of amniotic fluid squamous cells, APM staining, and Hematoxylin and Eosin (HE) staining were used to diagnose the AFE and AFA. Descriptive statistics of the patient population were analyzed using SPSS 20.0 software. RESULTS: Immunoreactivity of CK13 and CK10/13 specifically identified squamous cells of all the AFE and AFA cases. The amniotic fluid squamous cells were stained positive in a deep brown color with the monoclonal antibody to CK 13 and CK10/13 whereas the endothelial cells and alveolar epithelial cells were negative. There was no CK13 or CK10/13 expression in the non-AFE and non-AFA cases. With APM staining keratinized squamous cells were pink and mucus was blue-green in marked contrast with the surrounding tissue, which improved the detection rates of both keratinized squamous cells and mucus. CONCLUSIONS: CK13 (Rab) and CK10/13 (Mab) are valuable and reliable biomarkers of amniotic fluid squamous cells. APM reveals enriched mucus and keratinized squamous cells of amniotic fluid. Immunohistochemical detection of CK13 and CK10/13 combined with APM staining can improve the accuracy and reduce the difficulty in the diagnosis of AFE and AFA.

Blend-modification of soy protein/lauric acid edible films using polysaccharides.

Different types of polysaccharides (propyleneglycol alginate (PGA), pectin, carrageenan and aloe polysaccharide) were incorporated into soy protein isolate (SPI)/lauric acid (La) films using a co-drying process or by direct addition to form biodegradable composite films with modified water vapour permeability (WVP) and mechanical properties. The WVP of SPI/La/polysaccharide films decreased when polysaccharides were added using the co-drying process, regardless of the type of polysaccharide. The tensile strength of SPI/La film was increased by the addition of polysaccharides, and the percentage elongation at break was increased by incorporating PGA using the co-drying process. Regarding oxygen-barrier performance, no notable differences were observed between the SPI/La and SPI/La/polysaccharide films. The most significant improvement was observed by blending PGA, with the co-dried preparation exhibiting better properties than the direct-addition preparation. Scanning electron microscopy (SEM) revealed that the microstructures of the films are the basis for the differences in the barrier and mechanical properties of the modified blends of SPI, polysaccharides and La.

Role of human-mediated dispersal in the spread of the pinewood nematode in China.

BACKGROUND: Intensification of world trade is responsible for an increase in the number of alien species introductions. Human-mediated dispersal promotes not only introductions but also expansion of the species distribution via long-distance dispersal. Thus, understanding the role of anthropogenic pathways in the spread of invading species has become one of the most important challenges nowadays. METHODOLOGY/PRINCIPAL FINDINGS: We analysed the invasion pattern of the pinewood nematode in China based on invasion data from 1982 to 2005 and monitoring data on 7 locations over 15 years. Short distance spread mediated by long-horned beetles was estimated at 7.5 km per year. Infested sites located further away represented more than 90% of observations and the mean long distance spread was estimated at 111-339 km. Railways, river ports, and lakes had significant effects on the spread pattern. Human population density levels explained 87% of the variation in the invasion probability (P<0.05). Since 2001, the number of new records of the nematode was multiplied by a factor of 5 and the spread distance by a factor of 2. We combined a diffusion model to describe the short distance spread with a stochastic, individual based model to describe the long distance jumps. This combined model generated an error of only 13% when used to predict the presence of the nematode. Under two climate scenarios (stable climate or moderate warming), projections of the invasion probability suggest that this pest could expand its distribution 40-55% by 2025. CONCLUSIONS/SIGNIFICANCE: This study provides evidence that human-induced dispersal plays a fundamental role in the spread of the pinewood nematode, and appropriate control measures should be taken to stop or slow its expansion. This model can be applied to Europe, where the nematode had been introduced later, and is currently expanding its distribution. Similar models could also be derived for other species that could be accidentally transported by humans.