Synthesis and anti-liver fibrosis activity of imidazole and thiazole compounds containing amino acids.

Four series of imidazoles (15a-g, 20c, and 20d) and thiazoles (18a-g, 22a, and 22b) possessing various amino acids were synthesized and evaluated for activin receptor-like kinase 5 (ALK5) inhibitory activities in an enzymatic assay. Among them, compounds 15g and 18c showed the highest inhibitory activity against ALK5, with IC50 values of 0.017 and 0.025 µM, respectively. Compounds 15g and 18c efficiently inhibited extracellular matrix (ECM) deposition in TGF-ß-induced hepatic stellate cells (HSCs), and eventually suppressed HSC activation. Moreover, compound 15g showed a good pharmacokinetic (PK) profile with a favorable half-life (t1/2 = 9.14 h). The results indicated that these compounds exhibited activity targeting ALK5 and may have potential in the treatment of liver fibrosis; thus they are worthy of further study.

The Bbotf1 Zn(â ¡)2Cys6 transcription factor contributes to antioxidant response, fatty acid assimilation, peroxisome proliferation and infection cycles in insect pathogenic fungus Beauveria bassiana.

The abilities to withstand oxidation and assimilate fatty acids are critical for successful infection by many pathogenic fungi. Here, we characterized a Zn(II)2Cys6 transcription factor Bbotf1 in the insect pathogenic fungus Beauveria bassiana, which links oxidative response and fatty acid assimilation via regulating peroxisome proliferation. The null mutant ΔBbotf1 showed impaired resistance to oxidants, accompanied by decreased activities of antioxidant enzymes including CATs, PODs and SODs, and down-regulated expression of many antioxidation-associated genes under oxidative stress condition. Meanwhile, Bbotf1 acts as an activator to regulate fatty acid assimilation, lipid and iron homeostasis as well as peroxisome proliferation and localization, and the expressions of some critical genes related to glyoxylate cycle and peroxins were down-regulated in ΔBbotf1 in presence of oleic acid. In addition, ΔBbotf1 was more sensitive to osmotic stressors, CFW, SDS and LDS. Insect bioassays revealed that insignificant changes in virulence were seen between the null mutant and parent strain when conidia produced on CZP plates were used for topical application. However, propagules recovered from cadavers killed by ΔBbotf1 exhibited impaired virulence as compared with counterparts of the parent strain. These data offer a novel insight into fine-tuned aspects of Bbotf1 concerning multi-stress responses, lipid catabolism and infection cycles.

Properties of co-gel between Tenebrio Molitor larvae protein and myofibrillar protein induced by transglutaminase.

This study explored the effect of adding transglutaminase (TGase) to a co-gel of Tenebrio Molitor larvae protein (TMLP) and myofibrillar protein (MP). Different concentrations of TGase (0-90 U/g) were added to the co-gel. The results showed that 60 U/g TGase treatment significantly improved the gel strength and water holding capacity (WHC) by 26.51 g and 9.2 %, respectively. TGase promoted the rheological properties and accelerated the three-dimensional network structure of the co-gel. Moreover, TGase significantly increased (P < 0.05) the tyrosine residues, tryptophan residues content and hydrophobic interactions of the aliphatic groups. The chemical forces between the protein molecules changed. TGase promoted the transition of α-helix to ß-sheet and free water to immobilized water, thereby improving the WHC of co-gel. The principal component analysis reflected the links among indicators. This study illustrated that TGase might be an effective strategy to improve the co-gel of TMLP and MP and emulsified meat products with insects.

Ameliorating the stability of native/thermally denatured chicken-derived myofibrillar proteins particles in an aqueous system: The synergistic effect of acidification combined with inulin and inulin/sodium alginate.

The effect of acidification through hydrochloric acid combined with inulin (In), and inulin/sodium alginate (In/SA) on the stability of native/thermally denatured myofibrillar proteins (MPs/TMPs) particles in an aqueous system was investigated. At the same pH, MPs-In and TMPs-In particles were smaller and had higher absolute potentials than MPs-In/SA and TMPs-In/SA particles. Additionally, the size of MPs-In particles reached 1 µm, and the solubility increased from 21.73 ± 0.57 % to 76.26 ± 1.27 % when the pH was reduced from 5.0 to 3.0. The absolute potential of TMPs 3-In particles increased from 15.77 ± 0.72 to 28.20 ± 0.30 mV, and the solubility increased from 18.65 ± 0.72 % to 74.53 ± 0.74 %. Confocal laser microscopy revealed that, compared with pH 5.0 or 4.0, MPs-In/TMPs-In particles dispersed more evenly at pH 3.0 compared with pH 5.0 or 4.0. This further confirmed that electrostatic repulsion between particles maximally contributed to particle stability. Furthermore, the α-helix content in TMPs-In particles at pH 3.0 decreased from 41.51 ± 1.09 % (TMPs control) to 16.61 ± 1.87 %. This decrement of an up to 60 % led to decreased intramolecular hydrogen bonds and improved surface hydrophobicity. Therefore, a single polysaccharide (In) combined with MPs/TMPs particles exhibited higher dispersion and stability at pH 3.0. These findings could provide new insights into chicken-derived protein beverage processing.

Effects of protein aggregation induced by NaCl and temperature on gelation of silkworm (Antheraea pernyi) pupa raw powder.

Edible insects have great potential for producing protein-rich ingredients. This study aimed to investigate the effects of protein aggregation induced by NaCl (0-1 M) and temperature (65-95 °C) on gelation of Antheraea pernyi (A. pernyi) pupa raw powder. No thermal aggregates were observed at low temperature (65 °C), on the basis of there being no significant enhancement in turbidity and particle size (P > 0.05), regardless of NaCl concentrations. At elevated temperatures (75-95 °C), protein solutions exhibited significantly higher turbidity and particle size (P < 0.05), accompanied by an initial rise in surface hydrophobicity followed by a decline, alongside declining sulfhydryl. This marks the beginning of massive thermal aggregation driven by molecular forces. In addition, covalent (disulfide bonds) and non-covalent (hydrogen bonding, electrostatic interactions, and hydrophobicity) forces were influenced by NaCl, leading to variability in the protein aggregation and gelation. Correlation analysis indicates that the higher protein aggregation induced by ions was beneficial to the construction of more compact three-dimensional structures, as well as to the rheology, texture, and water-holding capacity of A. pernyi pupa gels. However, excessive salt ions destroyed the gel structure. Our findings will aid the use of A. pernyi pupae as textural ingredients in formula foods.

Improved gelling and emulsifying properties of chicken wooden breast myofibrillar protein by high-intensity ultrasound combination with pH-shifting.

The functional properties of chicken wooden breast myofibrillar protein (WBMP) are impaired. The protein structure and functional properties of WBMP are investigated using high-intensity ultrasound (HIU, 20 kHz, 200, 400, 600, and 800 W) combined with pH-shifting. HIU promoted the unfolding of WBMP, reduced the particle size of WBMP, and enhanced electrostatic repulsion. Medium-power (200 and 400 W) HIU promoted the α-helix to ß-sheet transformation, while high-power (600 and 800 W) HIU significantly (P < 0.05) increased the content of the random coil. The microstructure and images after storage further showed that 400 W HIU in combination with pH-shifting made the WBMP emulsion more uniform. In addition, gel performance analysis showed that the gel strength and water-holding capacity of the protein gel increased gradually after 400 W. Scanning electron microscope images also showed the formation of a stable network structure in the protein gel. This work could help promote the utilization of inferior proteins similar to WBMP, but the utilization rate still needs to be further improved.

Regulation mechanism of curcumin-loaded oil on the emulsification and gelation properties of myofibrillar protein: Emphasizing the dose-response of curcumin.

The regulation mechanism of curcumin (CUR) in the oil phase on the emulsification and gelation properties of myofibrillar protein (MP) was investigated. CUR enhanced the emulsifying activity index (EAI) of MP but decreased its turbiscan stability index (TSI) and surface hydrophobicity, which exacerbated oil droplet aggregation. Medium amounts (200 mg/L) of CUR changed the 3D network architectures of emulsion gels from lamellar to reticular, improving the gels' water-holding capacity (WHC), storage modulus, springiness, and cohesiveness. Besides, the LF-NMR revealed that CUR had limited effects on the mobility of immobilized and free water. The α-helix of MP in gels with medium amounts of CUR decreased from 51% to 45%, but the ß-sheet increased from 23% to 27% compared to those without CUR. Overall, CUR has the potential to become a novel structural modifier in emulsified meat products due to its dose-response.

Synthesis and antimicrobial activity evaluation of pyrazole derivatives containing the imidazo[2,1-b][1,3,4]thiadiazole moiety.

Four series of novel pyrazole derivatives (compounds 17a-m, 18a-m, 19a-g, and 20a-g) were synthesized, and their antibacterial and antifungal activities were evaluated. Most of the target compounds (17a-m, 18k-m, and 19b-g) showed strong antifungal activity and high selectivity relative to both Gram-positive and Gram-negative bacteria. Among them, compounds 17l (minimum inhibitory concentration [MIC] = 0.25 µg/mL) and 17m (MIC = 0.25 µg/mL) showed the strongest antifungal activity, being 2- and 4-fold more active than the positive controls gatifloxacin and fluconazole, respectively. In particular, compound 17l showed little cytotoxicity against human LO2 cells and did not exhibit hemolysis at ultrahigh concentrations, as did the positive control compounds gatifloxacin and fluconazole. These results indicate that these compounds are valuable for further development as antifungal agents.

The physicochemical properties and stability of myofibrillar protein oil-in-water emulsions as affected by the structure of sugar.

Different sugars (glucose, GL; fructose, FR; hyaluronic acid, HA; cellulose, CE) were added to a myofibrillar protein (MP) emulsion (MP: 1.2 w/v%, sugar: 0.1% w/v) to study the effect of sugar structure on the physicochemical properties and stability of the MP emulsions. The emulsifying properties of MP-HA were significantly (P < 0.05) higher than those of the other groups. The monosaccharide (GL/FR) exerted negligible effects on the emulsifying performance of the MP emulsions. The ζ-potential and particle size implied that HA introduced stronger negative charges, significantly reducing the final particle size (190-396 nm). Rheological examinations indicated that the introduction of polysaccharides considerably increased the viscosity and network entanglement; confocal laser scanning microscopy and creaming index revealed that MP-HA was stable during storage, whereas MP-GL/FR/CE exhibited severe delamination after long-term storage. HA, a heteropolysaccharide, is most suitable for improving MP emulsion quality.

Synthesis and Antibacterial Activity Evaluation of Imidazole Derivatives Containing 6-Methylpyridine Moiety.

A series of 2-cyclopropyl-5-(5-(6-methylpyridin-2-yl)-2-substituted-1H-imidazol-4-yl)-6-phenylimidazo[2,1-b][1,3,4]thiadiazoles (15a-t and 16a-f) were synthesized and their antibacterial activities were evaluated. More than half of the compounds showed moderate or strong antibacterial activity. Among them, compounds 15t (MIC=1-2 µg/mL) and 16d (MIC=0.5 µg/mL) showed the strongest antibacterial activities. Notably, compound 16d did not exhibit cytotoxicity in HepG2 cells and did not show hemolysis like the positive control compound Gatifloxacin. The results suggest that compound 16d should be further investigated as a candidate antibacterial agent.

"Water-Carbon" redistribution caused by China's interprovincial grain transportation.

Water resource and carbon emission involved in grain production in mainland China are redistributed among provinces as the grain was transported in recent years. This study first calculated the water consumption and carbon emission during the growth of grain crops based on the water-carbon footprint theory, and then used the social-equity method to calculate the inter-regional grain virtual water and virtual carbon flow. Finally, the regional and national trends in water saving and carbon emission reduction were calculated based on the spatial and temporal differences in grain planting among provinces. In terms of virtual water-carbon, from 2000 to 2017, the amount of the inter-provincial grain virtual water flow increased from 717.4 × 108 m3 to 1472.6 × 108 m3. Heilongjiang and Guangdong are the provinces with the largest amount of grain virtual water outflow (670.9 × 108 m3) and inflow (402.8 × 108 m3) in 2017, respectively. And the total inter-provincial grain virtual carbon flow increased from 2362.7 × 104 t CO2e to 12,680.6 × 104 t CO2e. Grain transport leads to water saving and carbon emission reduction, the amount of water saving increased from 25.6 × 108 m3 to 77.0 × 108 m3 and the carbon emission reduction increased from 2.4 × 104 t CO2e to 847.4 × 104 t CO2e from 2000 to 2017. Based on research results and from the perspective of socio-hydrology combined with water saving and carbon emission reduction, the regions could optimize the integration of water saving, carbon emission reduction, and sustainable development based on coordinating the grain planting structure according to their own climatic condition, soil and water resource condition, and socioeconomic condition.

Effects of ultrasound-assisted slightly acidic electrolyzed water thawing on myofibrillar protein conformation and gel properties of chicken breasts.

The effects of air thawing, water thawing, slightly acidic electrolyzed water thawing, ultrasound-assisted water thawing (WUT) and ultrasound-assisted slightly acidic electrolyzed water thawing (EUT) on the myofibrillar protein conformation and gel quality of chicken breasts were investigated. In the EUT group, protein solubility was higher (52.43%) than other thawing groups, and particle size was 24.57% smaller than the control group. Gel of EUT and WUT groups had stronger elasticities than the control group. Gel whiteness level was not significantly different between the EUT and control group. Gel water holding capacity of the EUT group was 9.07% greater than the control group. Gel water distributions and mobilities of the EUT and WUT groups were closer to the control group. The gel of the EUT group had a compact and homogeneous network. In conclusion, EUT treatment effectively reduced conformation disruption and improved gel properties, which was conducive to producing gel products.

Preheating and high-intensity ultrasound synergistically affect the physicochemical, structural, and gelling properties of chicken wooden breast myofibrillar protein.

The effect of different preheating temperatures (PT, 30, 40, 50, and 60 °C) cooperate with high-intensity ultrasound (HIU) treatment on the physicochemical, structural, and gelling properties of wooden breast myofibrillar protein (WBMP) was studied. The results show that G' of WBMP was the highest with preheating to 50 °C (PT50) (P < 0.05). Notably, the PT50 gel had the best hardness (9.38 ± 0.31 N) and springiness (0.51 ± 0.11), and water-holding capacity (89.95 ± 4.03 %). An analysis of the Raman spectra showed that the ß-fold content of PT50 was the highest (33.85 ± 0.03 %, P < 0.05). Low-field NMR data showed that the immobilized-water content of PT50 was also the highest (91.04 ± 1.36 %, P < 0.05). Scanning electron microscopy images showed that the PT50 gel was uniform and dense. Therefore, preheating to 50 °C enhances the HIU effect to produce an improved WBMP gel.

Quality changes of duck meat during thermal sterilization processing caused by microwave, stepwise retort, and general retort heating.

The quality changes of duck meat during thermal sterilization using microwave, stepwise retort and general retort heating were evaluated. Results showed that compared with stepwise retort and general retort, duck meat subjected to microwave showed significantly higher gumminess, chewiness, cohesiveness and resilience as well as glutamic acid, lysine and total amino acids. Low-field NMR revealed that the relative content of immobilized water after microwave and stepwise retort treatment was significantly higher than that after general retort treatment. The relative content of 1-octen-3-ol with characteristic mushroom aroma was significantly higher with microwave and stepwise retort heating than with general retort heating, while 2-pentyl-furan with poor taste was only detected with general retort heating. The muscle bundles subjected to microwave were neatly arranged, similar to those with no thermal sterilization. Overall, the meat quality after three thermal sterilization treatment was microwave > stepwise retort > general retort.

Fabrication and Characterization of Chicken- and Bovine-Derived Chondroitin Sulfate/Sodium Alginate Hybrid Hydrogels.

The physicochemical properties and microstructure of hybrid hydrogels prepared using sodium alginate (SA) and chondroitin sulfate (CS) extracted from two animal sources were investigated. SA-based hybrid hydrogels were prepared by mixing chicken- and bovine-derived CS (CCS and BCS, respectively) with SA at 1/3 and 2/3 (w/w) ratios. The results indicated that the evaporation water loss rate of the hybrid hydrogels increased significantly upon the addition of CS, whereas CCS/SA (2/3) easily absorbed moisture from the environment. The thermal stability of the BCS/SA (1/3) hybrid hydrogel was higher than that of CCS/SA (1/3) hybrid hydrogel, whereas the hardness and adhesiveness of the CCS/SA (1/3) hybrid hydrogel were lower and higher, respectively, than those of the BCS/SA (1/3) hybrid hydrogel. Low-field nuclear magnetic resonance experiments demonstrated that the immobilized water content of the CCS/SA (1/3) hybrid hydrogel was higher than that of the BCS/SA (1/3) hybrid hydrogel. FTIR showed that S=O characteristic absorption peak intensity of BCS/SA (2/3) was obviously higher, suggesting that BCS possessed more sulfuric acid groups than CCS. SEM showed that the hybrid hydrogels containing CCS have more compact porous microstructure and better interfacial compatibility compared to BCS.

Synergistic effect of preheating and different power output high-intensity ultrasound on the physicochemical, structural, and gelling properties of myofibrillar protein from chicken wooden breast.

The effects of preheating to 50 ℃ and the subsequent application of high-intensity ultrasound (HIU, 20 kHz) at 200, 400, 600, and 800 W on the physicochemical, structural, and gelling properties of wooden breast myofibrillar protein (WBMP) were studied. Results suggested that the WBMP structure expanded to the balanced state at 600 W, and rheological properties exhibit that 600 W HIU (P < 0.05) significantly improved the storage modulus (G') of WBMP. Notably, the WBMP gel (600 W) had the best hardness (65.428 ± 0.33 g), springiness (0.582 ± 0.01), and water-holding capacity (86.11 ± 0.83%). Raman spectra and low-field NMR indicated that 600 W HIU increased the ß-fold content (37.94 ± 0.04%) and enlarged the immobilized-water proportion (93.87 ± 0.46%). Scanning electron micrographs confirmed that the gel was uniform and dense at 600 W. Therefore, preheating to 50 ℃ followed by HIU (600 W) helped form a superior WBMP gel.

Effect of High-Voltage Electrostatic Field Heating on the Oxidative Stability of Duck Oils Containing Diacylglycerol.

High-voltage electrostatic field (HVEF) as an emerging green technology is just at the beginning of its use in meat products and by-products processing. In this study, we employed duck oil to produce duck-oil-based diacylglycerol (DAG), termed DDAG. Three different DDAG volume concentrations (0, 20%, and 100%) of hybrid duck oils, named 0%DDAG, 20%DDAG, and 100%DDAG, respectively, were used to investigate their thermal oxidation stability in high-voltage electrostatic field heating and ordinary heating at 180 ± 1 ℃. The results show that the content of saturated fatty acids and trans fatty acids of the three kinds of duck oils increased (p < 0.05), while that of polyunsaturated fatty acids decreased (p < 0.05) from 0 h to 8 h. After heating for 8 h, the low-field nuclear magnetic resonance showed that the transverse relaxation time (T21) of the three oils decreased (p < 0.05), while the peak area ratio (S21) was increased significantly (p < 0.05). The above results indicate that more oxidation products were generated with heating time. The peroxide value, the content of saturated fatty acids, and the S21 increased with more DAG in the duck oil, which suggested that the oxidation stability was likely negatively correlated with the DAG content. Moreover, the peroxide value, the content of saturated fatty acids and trans fatty acids, and the S21 of the three concentrations of duck oils were higher (p < 0.05) under ordinary heating than HVEF heating. It was concluded that HVEF could restrain the speed of the thermal oxidation reaction occurring in the duck oil heating and be applied in heating conditions.

Improving myofibrillar proteins solubility and thermostability in low-ionic strength solution: A review.

The development of myofibrillar proteins drinks (MPDs) can provide meat protein nutrition to specific groups of people. However, one major challenge is that myofibrillar proteins (MPs) are insoluble in solutions with a low ionic strength. Another functional constraint is the susceptibility of MPs to heat-induced aggregation. Currently, the primary approach used to improve the water solubility of MPs is to inhibit the assembly of myofilaments. Increasing the thermostability of MPs primarily inhibits the aggregation of myosin or oxidizes myosin to soluble substances. This review focuses on the description of several chemical and physical strategies, with an emphasis on the advantages, disadvantages, and recent progress. Under the myosin filament assembly process and the cross-linking aggregation mechanism, this summary helps improve our understanding of the solution and thermostability of MPs in low-ionic-strength solutions, thus providing new ideas to the development of MPDs.

Effects of heat treatment on physicochemical and microstructure properties of myofibrillar proteins combined with glucose and cellulose nanofibers.

The effects of unheated, 80 °C, 100 °C, and 121 °C on the physicochemical and microstructural properties of myofibrillary protein (MP) supplemented with glucose (Glc) and cellulose nanofibers (CNFs) were studied. The results showed that Glc and CNFs cross-linked with MP through non-covalent bonds when unheated compared with the unheated MP group, which increased the particle size and surface hydrophobicity of MP, and changed the secondary structure. At 80 °C and 100 °C, the particle size and hydrophobicity of MP significantly increased. At 121 °C, the contents of leucine and lysine in MP decreased by 16.2% and 19.9%, respectively, however, both Glc and CNFs could prevent this decrease. Meanwhile, microscopic images showed that the dispersion of Glc-MP and CNFs-MP were more uniform than the MP groups. Compared with Glc-MP, CNFs-MP had more uniform particle size distribution and better thermal stability at high temperatures (121 °C).

A secretory phospholipase A2 of a fungal pathogen contributes to lipid droplet homeostasis, assimilation of insect-derived lipids, and repression of host immune responses.

Secretory phospholipase A2s (sPLA2s) are found in a wide range of organisms from bacteria to higher plants and animals and are involved in varied and cellular processes. However, roles of these enzymes in microbial pathogens remain unclear. Here, an sPLA2 (BbPLA2) was characterized in the filamentous insect pathogenic fungus, Beauveria bassiana. BbPLA2 was exclusively expressed in insect hemolymph-derived cells (hyphal bodies), and its expression was induced by insect-derived nutrients and lipids, and nutrient starvation. High levels of secretion of BbPLA2 were observed as well as its distribution in hyphal body lipid drops (LDs). Overexpression of BbPLA2 increased the ability of B. bassiana to utilize insect-derived nutrients and lipids, and promoted LD accumulation, indicating functions for BbPLA2 in mediating LD homeostasis and assimilation of insect-derived lipids. Strains overexpressing BbPLA2 showed moderately increased virulence, including more efficient penetration of the insect cuticle and evasion of host immune responses as compared to the wild type strain. In addition, B. bassiana-activated host immune genes were downregulated in the BbPLA2 overexpression strain, but upregulated by infections with a ΔBbPLA2 strain. These data demonstrate that BbPLA2 contributes to LD homeostasis, assimilation of insect-derived lipids, and repression of host immune responses.