Mechanical overload induces TMJ disc degeneration via TRPV4 activation.

OBJECTIVE: The temporomandibular joint (TMJ) disc cushions intraarticular stress during mandibular movements. While mechanical overloading is related to cartilage degeneration, the pathogenesis of TMJ disc degeneration is unclear. Here, we determined the regulatory role of mechanoinductive transient receptor potential vanilloid 4 (TRPV4) in mechanical overload-induced TMJ disc degeneration. METHODS: We explored the effect of mechanical overload on the TMJ discs in a rat occlusal interference model in vivo, and by applying sustained compressive force in vitro. TRPV4 inhibition was delivered by small interfering RNA or GSK2193874; TRPV4 activation was delivered by GSK1016790A. The protective effect of TRPV4 inhibition was validated in the rat occlusal interference model. RESULTS: Occlusal interference induced TMJ disc degeneration with enhanced extracellular matrix degradation in vivo and mechanical overload promoted inflammatory responses in the TMJ disc cells via Ca2+ influx with significantly upregulated TRPV4. TRPV4 inhibition reversed mechanical overload-induced inflammatory responses; TRPV4 activation simulated mechanical overload-induced inflammatory responses. Moreover, TRPV4 inhibition alleviated TMJ disc degeneration in the rat occlusal interference model. CONCLUSION: Our findings suggest TRPV4 plays a pivotal role in the pathogenesis of mechanical overload-induced TMJ disc degeneration and may be a promising target for the treatment of degenerative changes of the TMJ disc.

Investigation on the quality regulating mechanism of antifreeze peptides on frozen surimi: From macro to micro.

Freeze denaturation of protein caused by ice crystals is the main motivation for the quality deterioration of surimi during circulation and storage. This investigation aimed to cryoprotect surimi by adding antifreeze peptides from Takifugu obscurus skin (TsAFP) which can inhibit ice recrystallization, and to elucidate regulating mechanism. The comprehensive results showed that 4% TsAFP, half dosage of commercial cryoprotectant, had good cryoprotection on surimi by reducing the moisture variation and maintaining protein solubility of surimi at macro level, as well as inhibiting the degeneration and structure changes of myofibrillar proteins at micro level. Meanwhile, TsAFP could directly bind to the structural cavity of myosin, inhibit protein freezing-induced oxidation, maintain the spatial structure of myosin and water retention ability to preserve the surimi quality. This study helped better comprehend the protective mechanisms of antifreeze peptides in frozen surimi and was expected to provide a promising cryoprotectant for low-sweetness and low-calorie surimi.

New insight into the mechanism by which antifreeze peptides regulate the physiological function of Streptococcus thermophilus subjected to freezing stress.

INTRODUCTION: Antifreeze peptides regulate the physiological functions of frozen cells and even their apoptosis; however, the mechanisms by which antifreeze peptides regulate these processes remain unclear, although the interactions between cell membranes and ice are well known to be important in this process. OBJECTIVES: Our study aims to investigate how antifreeze peptides regulate cell physiological functions during the freezing process. METHODS: We investigated the cryoprotective effect of rsfAFP on the physiological functions of S. thermophilus under freezing stress by measuring cellular metabolism activity, intracellular enzyme activity, cell membrane characterization, and cell apoptosis. The mechanism by which rsfAFP impacts S. thermophilus physiological functions under freezing stress was investigated using multispectral techniques and cryo-TEM. RESULTS: We show that a recombinant antifreeze peptide (rsfAFP) interacts with the extracellular capsular polysaccharides and peptidoglycan of Streptococcus thermophilus and ice to cover the outer layer of the membrane, forming a dense protective layer that regulates the molecular structure of extracellular ice crystals, which results in reduced extracellular membrane damage, depressed apoptosis and increased intracellular metabolic activity. This interaction mechanism was indicated by the fact that S. thermophilus better maintained its permeability barrier, membrane fluidity, membrane structural integrity, and cytoplasmic membrane potential during freezing stress with rsfAFP treatment. CONCLUSION: These results provide new insights into the mechanism by which rsfAFP regulates frozen cellphysiological functionsand apoptosis under freezing stress.

Identification of Novel Antifreeze Peptides from Takifugu obscurus Skin and Molecular Mechanism in Inhibiting Ice Crystal Growth.

Foodborne hydrolyzed antifreeze peptides have been widely used in the food industry and the biomedical field. However, the components of hydrolyzed peptides are complex and the molecular mechanism remains unclear. This study focused on identification and mechanism analysis of novel antifreeze peptides from Takifugu obscurus skin by traditional methods and computer-assisted techniques. Results showed that three peptides (EGPRAGGAPG, GDAGPSGPAGPTG, and GEAGPAGPAG) possessed cryoprotection via reducing the freezing point and inhibiting ice crystal growth. Molecular docking confirmed that the cryoprotective property was related to peptide structure, especially α-helix, and hydrogen bond sites. Moreover, the antifreeze peptides were double-faces, which controlled ice crystals while affecting the arrangement of surrounding water molecules, thus exhibiting a strong antifreeze activity. This investigation deepens the comprehension of the mechanism of antifreeze peptides at molecular scale, and the novel efficient antifreeze peptides can be developed in antifreeze materials design and applied in food industry.

Effects and mechanism of antifreeze peptides from silver carp scales on the freeze-thaw stability of frozen surimi.

The objective of this work was to investigate the cryoprotective effects of antifreeze peptides obtained from silver carp scales (ScAFPs) on the freeze-thaw stability of surimi, and to explore the action mechanisms of ScAFPs on frozen surimi. The comprehensive analysis of ice crystal size, myofibril protein oxidation, water retention, surimi gel properties, and rheological properties of surimi after different freeze-thaw cycles were investigated. Results showed that frozen surimi treated with ScAFPs exhibited a higher Ca2+-ATPase activity, salt-soluble protein concentration and sulfhydryl group content, while lower surface hydrophobicity, carbonyl content and disulfide bond content. Moreover, the gel properties and water holding capacity of surimi and surimi gel were improved significantly by regulating the size of ice crystals during freeze-thaw process. These findings indicate that ScAFPs could serviced as a new food ingredient with anti-freezing function for frozen products.

New insights into the pollutant composition of stormwater treating wetlands.

With intensified climate change and urbanisation, constructed wetland (CW) serves as an alternative to conventional wastewater treatment plants. In Australia, the primary function of CW is to reduce sediments, nutrients from runoffs and attenuate floods. However, water quality analysis after construction is limited, hence, pollutant composition in established CWs and target pollutants in many guidelines remain outdated. To refresh the understanding of pollutants in urban discharges, this study reviewed two CWs in industrialised regions of Victoria, Australia. A total number of 26 pollutants were analysed in the collected water and sediment samples from both CW. The findings highlighted excessive concentrations of Zinc, Aluminium, Iron and Copper in one wetland and less commonly found pollutants like Barium, Titanium and Strontium are also detected. While Arsenic, Zinc, Copper, Nickel and hydrocarbons' accumulations are particularly significant in the other wetland. This study also reviews the pollutants discovered in 136 stormwater wetlands and covers the sources and impacts of various metal pollutants in stormwater runoffs. Overall, it is found that the concentrations of Zinc, Aluminium and Iron are particularly high in the CWs reviewed. This study brings attention to the pollutants profile of established CWs and the impact of heavy metals on the aquatic environment. The findings from this research revealed that the existing design and management guidelines for constructed wetlands in urban catchments are lacking in reduction targets for metal pollutants, thus improvements are essential to safeguard the water quality and performance of CWs.

Snow flea antifreeze peptide for cryopreservation of lactic acid bacteria.

Cryogenic machining is one of the most commonly used techniques for processing and preserving in food industry, and traditional antifreeze agents cannot regulate the mechanical stress damage caused by ice crystals formed during recrystallization or thawing. In this study, we successfully developed an express system of a novel recombinant snow flea antifreeze peptide (rsfAFP), which has significant ice recrystallization inhibition ability, thermal hysteresis activity and alters ice nucleation, thus regulating extracellular ice crystal morphology and recrystallization. We showed that rsfAFP improved the survival rate, acid-producing ability, freezing stability, and cellular metabolism activity of Streptococcus thermophilus. We further showed that rsfAFP interacts with the membrane and ice crystals to cover the outer layer of cells, forming a dense protective layer that maintains the physiological functions of S. thermophilus under freezing stress. These findings provide the scientific basis for using rsfAFP as an effective antifreeze agent for lactic acid bacteria cryopreservation or other frozen food.

Investigation of the cryoprotective mechanism and effect on quality characteristics of surimi during freezing storage by antifreeze peptides.

Freezing technology is important for storage of animal products such as surimi. However, mechanical damage caused by ice crystals would lead to quality deterioration. This study aims to investigate the protective effect of antifreeze peptides (AFPs) on the quality of surimi during freezing storage and its possible mechanism. We found that AFPs exhibited a strong inhibition of ice crystal recrystallization, and the molecular weight ranged from 180 to 3000 Da. AFPs can prevent the degeneration of myofibrillar protein by reducing the loss of Ca2+-ATPase activity, slowing oxidation of sulfhydryl groups to disulfide bonds, and maintaining surface hydrophobicity and solubility of myofibrillar protein. Moreover, AFPs can reduce the influence of freezing stress on water mobility, thereby protecting the surimi from losing immobilized water and bound water during frozen storage. These findings indicate that AFPs could potentially serve as a food ingredient with antifreeze functional for the storage of surimi products.

Extension of Drosophila Lifespan by Astragalus polysaccharide through a Mechanism Dependent on Antioxidant and Insulin/IGF-1 Signaling.

Historical literature and pharmacological studies demonstrate that Astragalus polysaccharide (APS) has anti-inflammatory and antioxidative effects. Studies into the longevity effects of APS are limited, and the molecular mechanism of lifespan extension by APS is not elucidated yet. Here, the longevity effect of APS was investigated in Drosophila melanogaster by feeding dose-dependent APS. APS significantly extended the lifespan and improved the reproduction. Meanwhile, APS increased locomotion, TAG level, and starvation resistance and reduced the mortality rate induced by hydrogen peroxide. The activities of superoxide dismutase (SOD) and catalase (CAT) were increased in flies treated with APS diet. Moreover, APS significantly enhanced expressions of antioxidant genes (Sod1, Sod2, and Cat), dFoxO, and 4E - BP, decreased the expressions of insulin-like peptides (dilp2, dilp3, and dilp5), and longevity gene MTH. Together, these results indicate that APS can prolong the lifespan by regulating antioxidant ability and insulin/IGF-1 signaling and also enhance the reproduction ability in Drosophila. APS may be explored as a novel agent for slowing the aging process and improving reproduction.

Ice-binding proteins: a remarkable ice crystal regulator for frozen foods.

Ice crystal growth during cold storage presents a quality problem in frozen foods. The development of appropriate technical conditions and ingredient formulations is an effective method for frozen food manufacturers to inhibit ice crystals generated during storage and distribution. Ice-binding proteins (IBPs) have great application potential as ice crystal growth inhibitors. The ability of IBPs to retard the growth of ice crystals suggests that IBPs can be used as a natural ice conditioner for a variety of frozen products. In this review, we first discussed the damage caused by ice crystals in frozen foods during freezing and frozen storage. Next, the methods and technologies for production, purification and evaluation of IBPs were summarized. Importantly, the present review focused on the characteristics, structural diversity and mechanisms of IBPs, and the application advances of IBPs in food industry. Finally, the challenges and future perspectives of IBPs are also discussed. This review may provide a better understanding of IBPs and their applications in frozen products, providing some valuable information for further research and application of IBPs.

Interaction among protein, daidzein and surfactants in the WPI-based daidzein self-microemulsifying delivery system.

The self-microemulsifying delivery system was fabricated by whey protein isolate (WPI), daidzein (Dai) and surfactants, the interaction of WPI, Dai and D-α-Tocopherol polyethylene glycol succinate (TPGS) was hereby studied in the absence or presence of Tween 20. The increase of surfactant concentration led to the decrease of the modulus and changes of protein interfacial conformation, which allowed the formation of a strong intermolecular network. The environment and structure of WPI and daidzein could be changed by TPGS, and the addition of Tween 20 could further enhance the interaction between the components by changing TPGS structure. With the increase of surfactants and oil phase, Ksv and Ka values of WPI-Dai increased first and then decreased. Therefore, the interaction between the components was also dependent on the WPI-surfactant ration. These findings provide a potential strategy for designing microemulsion food system based on the understanding of the interactions among individual composition of microemulsions.

Effects of gelatin-based antifreeze peptides on cell viability and oxidant stress of Streptococcus thermophilus during cold stage.

Cold stage adversely affects cell proliferation and cell viability of probiotics such as Streptococcus thermophilus in food industry, new type of cryoprotectants continues to be needed. Gelatin-based antifreeze peptide becomes a popular topic because of its cryoprotective effects on cold-stressed probiotics. In this study the effects of tilapia scales antifreeze peptides (TSAPP) on cell viability and oxidant stress of S. thermophilus during cold stage were investigated. The results showed that the percentage of viable cells was increased 10.85 folds compared with control groups. Addition of TSAPP activated the activities of ATPases, relieved the hyperpolarization of cell membrane potential and regulated the intracellular Ca2+ concentration. Furthermore, TSAPP significantly inhibited reactive oxygen species level and malonaldehyde content in cells. Under cryopreservation with TSAPP, cells of S. thermophilus maintained higher activities of antioxidant enzymes including catalase, peroxidase and total antioxidant capacity. These findings indicate that TSAPP likely offered its cellular protection by maintaining membrane integrity and alleviation of oxidative stress.

Long-term protection against dengue viruses in mice conferred by a tetravalent DNA vaccine candidate.

The development of an effective tetravalent vaccine against dengue viruses (DENVs) has become a world priority. We previously showed that four monovalent dengue DNA vaccines expressing premembrane (prM) and envelope (E) proteins displayed effective protection against corresponding challenges in mice. Thus, to elucidate the overall immunity and persistence of the tetravalent formulation (TetraME), we evaluated the humoral and cellular immune responses as well as the long-term protection in the current study. TetraME-immunized mice displayed increased production of Th1/Th2-typed cytokines upon stimulation with heterologous DENV antigens. Moreover, high levels of tetravalent DENV antibodies and sterilized immunity were detected long-term (30 weeks after immunization). These findings provide feasible validation for the potential utility of this vaccine formulation.

Preparation and Characterization of Chitosan-Based Ternary Blend Edible Films with Efficient Antimicrobial Activities for Food Packaging Applications.

To improve the mechanical and antibacterial properties of chitosan (CS) films, a ternary blend edible film was prepared by incorporating CS, gelatin (GE), and natural cinnamon essential oil (CEo). Scanning electron microscopy (SEM), Atomic force microscopy (AFM), Fourier transform infrared spectroscopy, and X-ray diffraction were performed to evaluate the films. The mechanical properties, light transmission, thermal stability, hydrophilicity, and antibacterial activity of the films were also determined. The results confirmed all of the films exhibited excellent UV protection with low transparency at 600 nm. Compared with the CS films, the ternary composite film (CSGEo film, containing CS, GE, and CEo) had a higher elongation at break but a lower tensile strength. SEM images revealed that all films had smooth surfaces, although some obvious differences between CS and CSGEo films were observed by AFM. Additionally, the incorporation of GE and CEo to the films enhanced their thermal stability and contact angle, but decreased their crystallinity and wettability. The antimicrobial activity results showed that CSGEo films had excellent antimicrobial activity against Escherichia coli and Staphylococcus aureus, for which the antibacterial rate exceeded 98%. The minimum inhibitory concentrations of the CSGEo solution against E. coli and S. aureus were both 52.06 µg/mL, and the minimal bactericidal concentrations were 104.12 and 52.06 µg/mL, respectively. These results suggest that CSGEo films possess good mechanical and antibacterial properties, and therefore, their application in the food packaging industry is promising. PRACTICAL APPLICATION: The main raw materials of the edible films developed in this study are aquatic by-products, so the films are edible and biodegradable. The addition of gelatin and CEo improved the UV barrier and thermal properties but decreased the crystallinity and hydrophilicity of the films, making them suitable for use as packaging materials. CEo-incorporated films exhibited excellent mechanical properties and antibacterial activity and can, therefore, be used in the food packaging industry.

Analysis of the spectrum and antibiotic resistance of uropathogens in outpatients at a tertiary hospital.

The objective of this study was to analyse the distribution and antimicrobial resistance of bacterial uropathogens isolated from outpatients at Henan Provincial People's Hospital. A total of 1419 samples from 823 newly diagnosed and 596 recurrent UTI outpatients culture positive. Escherichia coli was the most common uropathogen. Compared with the recurrent group, the newly diagnosed group had a higher isolation rate of E. coli and Enterobacter cloacae but a lower isolation rate of Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter spp. Except for P. aeruginosa, the resistance of Gram-negative bacteria to most antibiotics was less than 30%. All Enterococcus and Staphylococcus spp. were sensitive to linezolid, vancomycin and teicoplanin. Both Gram-negative and -positive bacteria exhibited high susceptibility to fosfomycin. Uropathogens isolated from recurrent outpatients had higher resistance rates than did those isolated from newly diagnosed outpatients. Our study indicated that fosfomycin might be an excellent treatment option for outpatients with UTIs.