Effects of Dityrosine on Lactic Acid Metabolism in Mice Gastrocnemius Muscle During Endurance Exercise via the Oxidative Stress-Induced Mitochondria Damage.

Dityrosine (Dityr) has been detected in commercial food as a product of protein oxidation and has been shown to pose a threat to human health. This study aims to investigate whether Dityr causes a decrease in lactic acid metabolism in the gastrocnemius muscle during endurance exercise. C57BL/6 mice were administered Dityr or saline by gavage for 13 weeks and underwent an endurance exercise test on a treadmill. Dityr caused a severe reduction in motion displacement and endurance time, along with a significant increase in lactic acid accumulation in the blood and gastrocnemius muscle in mice after exercise. Dityr induced significant mitochondrial defects in the gastrocnemius muscle of mice. Additionally, Dityr induced serious oxidative stress in the gastrocnemius muscle, accompanied by inflammation, which might be one of the causes of mitochondrial dysfunction. Moreover, significant apoptosis in the gastrocnemius muscle increased after exposure to Dityr. This study confirmed that Dityr induced oxidative stress in the gastrocnemius muscle, which further caused significant mitochondrial damage in the gastrocnemius muscle cell, resulting in decreased capacity of lactic acid metabolism and finally affected performance in endurance exercise. This may be one of the possible mechanisms by which highly oxidized foods cause a decreased muscle energy metabolism.

The expression, immune infiltration, prognosis, and experimental validation of OSBPL family genes in liver cancer.

BACKGROUND: Liver cancer is the third most deadly malignant tumor in the world with poor prognosis and lacks early diagnostic markers. It is urgent need to explore new biomarkers and prognostic factors. The oxysterol-binding protein-like family proteins (OSBPLs) are essential mediators of lipid transportation and cholesterol balancing which has been reported to participate in cancer progression. So far, the expression, immune infiltration, and prognosis of OSBPLs have not been elucidated in liver cancer. METHODS: The differential expressions of OSBPLs between liver tumor and normal tissues were assessed by analyzing RNA-seq data from TCGA and protein data from CPTAC, respectively. Subsequently, genetic variations, potential functional enrichment analysis, and immune cell infiltration were analyzed. Further, the prognostic effects of OSBPLs were identified via constructing lasso models and performing receiver operating characteristic curve (ROC) analysis. Moreover, 10 local liver cancer specimens were involved to validate the expression of OSBPL3 via immunohistochemistry (IHC) assay. Finally, CCK-8, cell cycle, apoptosis, transwell assays, real time qPCR (RT-qPCR), and western blot assays were conducted to explore the function of OSBPL3 in liver cancer cells. RESULTS: The mRNA of OSBPL2, OSBPL3, and OSBPL8 were highly expressed while OSBPL6 was lowly expressed in liver cancer samples compared with normal samples. As to the protein expression, OSBPL2 and OSBPL3 were significantly elevated and OSBPL5, OSBPL6, OSBPL9, OSBPL10, OSBPL11 were downregulated in tumor samples. A positive correlation was found between copy number variations (CNV) and the expression of OSBPL2, OSBPL8, OSBPL9, OSBPL11, while DNA methylation was negatively associated with the expressions of OSBPLs. Of these, CNV amplification mainly contributed to the overexpression of OSBPL2 and DNA methylation may be responsible for the high expression of OSBPL3. Interestingly, OSBPL3, OSBPL5, SOBPL7, and OSBPL10 were significantly positively correlated with immune infiltration. Notably, OSBPL3 was identified correlated to overall survival (OS) and disease specific survival (DSS) in liver cancer. Functionally, knocking down OSBPL3 reduced liver cancer cell viability, induced a G2/M cell cycle arrest, promoted apoptosis, and restrained cell migration. CONCLUSION: In aggregate, we reported a heretofore undescribed role of OSBPLs in liver cancer by analyzing multi-omics data. Importantly, we identified OSBPL3 was overexpressed in liver tumor compared with normal and its high expression was correlated with poor OS and DSS. Inhibition of OSBPL3 resulted in a pronounced decrease in cell proliferation and migration.

Degradation of poly(butylene adipate-co-terephthalate) films by Thermobifida fusca FXJ-1 isolated from compost.

In recent years, Poly(butylene adipate-co-terephthalate) (PBAT) films were wildly used due to its biodegradable properties. However, there are few reports of strains that can high efficiently degrade PBAT. Thermobifida fusca FXJ-1, a thermophilic actinomycete, was screened and identified from compost. FXJ-1 can efficiently degrade PBAT at 55 °C in MSM medium. The degradation rates of the pure PBAT film (PF), PBAT film used for mulching on agricultural fields (PAF), and PBAT-PLA-ST film (PPSF) were 82.87 ± 1.01%, 87.83 ± 2.00% and 52.53 ± 0.54%, respectively, after nine days of incubation in MSM medium. Cracking areas were monitored uniformly distributed on the surfaces of three kinds of PBAT-based films after treatment with FXJ-1 using scanning electron microscopy. The LC-MS results showed that PBAT might be degraded into adipic acid, terephthalic acid, butylene adipate, butylene terephthalate and butylene adipate-co-terephthalate, and these products are involved in the cleavage of ester bonds. We also found that amylase produced by FXJ-1 played an important role in the degradation of PPSF. FXJ-1 also showed an efficient PBAT-based films degradation ability in simulating compost environment, which implied its potential application in PBAT and starch-based film degradation by industrial composting.

Identification of Essential Genes Associated With Prodigiosin Production in Serratia marcescens FZSF02.

Prodigiosin is a promising secondary metabolite produced mainly by Serratia strains. To study the global regulatory mechanism of prodigiosin biosynthesis, a mutagenesis library containing 23,000 mutant clones was constructed with the EZ-Tn5 transposon, and 114 clones in the library showed altered prodigiosin production ability. For 37 of the 114 clones, transposon insertion occurred on the prodigiosin biosynthetic cluster genes; transposon inserted genes of the 77 clones belonged to 33 different outside prodigiosin biosynthetic cluster genes. These 33 genes can be divided into transcription-regulating genes, membrane protein-encoding genes, and metabolism enzyme-encoding genes. Most of the genes were newly reported to be involved in prodigiosin production. Transcriptional levels of the pigA gene were significantly downregulated in 22 mutants with different inserted genes, which was in accordance with the phenotype of decreased prodigiosin production. Functional confirmation of the mutant genes involved in the pyrimidine nucleotide biosynthesis pathway was carried out by adding orotate and uridylate (UMP) into the medium. Gene complementation confirmed the regulatory function of the EnvZ/OmpR two-component regulatory system genes envZ and ompR in prodigiosin production.

Immunoregulatory function of Dictyophora echinovolvata spore polysaccharides in immunocompromised mice induced by cyclophosphamide.

The purpose of this study was to investigate whether the Dictyophora echinovolvata spore polysaccharides (DESP) affect the immunity in immunocompromised mice induced by cyclophosphamide (CTX). The healthy female Kunming mice were randomly divided into six groups, including a normal control (NC) group, a positive control group, a model control (MC) group, and three groups treated with low-, intermediate-, and high-dose polysaccharide, respectively. A series of immunoregulatory properties were determined, including humoral and cellular immunity, immune function, and immune factors of mononuclear macrophages. Compared with NC and MC groups, treatment with DESP significantly increased the spleen index and decreased the thymus index; increased the serum concentrations of immunoglobulin (Ig)A, IgG, IgM, hemolysin, IL-1ß, and IL-2; delayed the allergic reaction; and improved the splenic lymphocyte transformation ability; and enhanced the phagocytosis of macrophages and the ability to secrete IL-6, TNF-α, caspase-1, and NO with DESP supplementation. These results indicated that DESP might have a good regulatory effect on CTX-induced immunodeficiency in mice, adjust the body's immune imbalance, and improve the symptoms of low immunity.

An Evolutionary Game Theory Study for Construction and Demolition Waste Recycling Considering Green Development Performance under the Chinese Government's Reward-Penalty Mechanism.

The low efficiency of the closed-loop supply chain in construction and demolition waste (CDW) recycling has restricted the green development of China's construction industry. Additionally, the government's reward-penalty mechanism has a huge influence on green development. This study aimed to investigate the effect of green development performance (GDP) and the government's reward-penalty mechanism on the decision-making process of production and recycling units, as well as to reveal the optimal strategies under different conditions. Therefore, the strategies' evolutionary paths of production and recycling units were investigated by using evolutionary game theory. Firstly, an evolutionary game model between production units and recycling units was proposed under the government's reward-penalty mechanism. Then, the evolutionary stability strategies in different scenarios were discussed. Finally, the effects of the relevant parameters on the evolutionary paths of the game model were analyzed using numerical simulations. The main conclusions are as follows. (1) When the range of GDP changes, the evolutionary stable strategy changes accordingly. GDP plays a positive role in promoting the high-quality development of the CDW recycling supply chain, but an increase in GDP can easily lead to the simultaneous motivation of free-riding. (2) The government's reward-penalty mechanism effectively regulates the decision-making process of production and recycling units. An increase in the subsidy rate and supervision probability helps to reduce free-riding behavior. Moreover, the incentive effect of the subsidy probability on recycling units is more obvious, while the effect of the supervision probability on improving the motivation of active participation for production units is more remarkable. This paper not only provides a decision-making basis to ensure production and recycling units to make optimal strategy choices under different conditions but also provides a reference for the government to formulate a reasonable reward-penalty mechanism that is conducive to a macro-control market.

High temperatures can effectively degrade residual tetracyclines in chicken manure through composting.

Tetracyclines (TCs) residues in livestock manure are the main origin of environmental tetracyclines contamination and have serious environmental and health risks. This work aimed to examine the effect of temperature on the degradation patterns of TCs antibiotics. Tetracycline (TC), doxycycline (DC), oxytetracycline (OTC) and chlortetracycline (CTC) were all degraded much more quickly in sterilized double distilled H2O (ddH2O) at higher temperatures and lost antibacterial activity after being incubated at 70 °C for 72 h. High pH value enhanced the degradation process of TCs solutions. Degradation products of the TCs were identified with LC/MS. The TCs from simulated composting with sterilized chicken manure and nonsterilized manure all showed temperature-dependent thermal degradation. The degradation pattern fitted the availability-adjusted loss model well. The fitted equations showed that the half-lifes of degradation of the TCs were 1.66-7.62 h at 70 °C, 3.29-21.39 h at 60 °C and 9.25-57.19 h at 50 °C. Compared with those of nonsterilized manures, we concluded that high temperature can effectively degrade the residual TCs in chicken manure by thermal degradation during high temperature composting. Temperatures that are elevated moderately, for example, up to 70 °C, during the composting process can make the degradation process more effective and rapid.

Enhanced production of prodigiosin by Serratia marcescens FZSF02 in the form of pigment pellets

Background: Prodigiosin has been demonstrated to be an important candidate in investigating anticancer drugs and in many other applications in recent years. However, industrial production of prodigiosin has not been achieved. In this study, we found a prodigiosin-producing strain, Serratia marcescens FZSF02, and its fermentation strategies were studied to achieve the maximum yield of prodigiosin. Results: When the culture medium consisted of 16.97 g/L of peanut powder, 16.02 g/L of beef extract, and 11.29 mL/L of olive oil, prodigiosin reached a yield of 13.622 ± 236 mg/L after culturing at 26 °C for 72 h. Furthermore, when 10 mL/L olive oil was added to the fermentation broth at the 24th hour of fermentation, the maximum prodigiosin production of 15,420.9 mg/L was obtained, which was 9.3-fold higher than the initial level before medium optimization. More than 60% of the prodigiosin produced with this optimized fermentation strategy was in the form of pigment pellets. To the best of our knowledge, this is the first report on this phenomenon of pigment pellet formation, which made it much easier to extract prodigiosin at low cost. Prodigiosin was then purified and identified by absorption spectroscopy, HPLC, and LCMS. Purified prodigiosin obtained in this study showed anticancer activity in separate experiments on several human cell cultures: A549, K562, HL60, HepG2, and HCT116. Conclusions: This is a promising strain for producing prodigiosin. The prodigiosin has potential in anticancer medicine studies.

Effect of thermotolerant bacterial inoculation on the microbial community during sludge composting.

A thermophilic bacterium (Geobacillus stearothermophilus CHB1) was inoculated in a sludge compost, and the effects of the inoculation on the abundance and structure of the bacterial community in the sludge compost were investigated using quantitative PCR and Illumina MiSeq sequencing. The results showed that the high-temperature stage (>50 °C) of the CHB1 and CK (control without inoculum) piles started on days 5 and 8, respectively, and lasted for 7 and 2 days, respectively, indicating the extension of the thermophilic phase by CHB1 inoculation in the sludge compost. At the end of composting, the CHB1 piles showed a higher loss of total organic carbon, lower C/N ratio, and lower moisture content. The abundance of bacteria in the CHB1 piles was significantly higher in the heating and thermophilic phase of composting but were lower than those of the CK in the cooling phase. The richness and diversity of the bacterial community in the thermophilic phase increased after inoculation with CHB1. After inoculation of CHB1, there were higher relative abundances of Firmicutes, Thermopolyspora, Thermobacillus, Thermomonas, Thermomonospora, and Thermovum, which can grow in a high-temperature environment. Furthermore, redundancy analysis indicated that total organic carbon, total nitrogen, C/N ratio, pH, temperature, and moisture were the significant parameters that affected the bacterial community structure during sludge composting. Our findings suggested that inoculation with CHB1 would enhance the quality and efficiency of composting.

Corrosion Behaviors of Q345R Steel at the Initial Stage in an Oxygen-Containing Aqueous Environment: Experiment and Modeling.

The ingress of oxygen into pressure vessels used in oil & gas production and transportation could easily result in serious corrosion. In this work, the corrosion behaviors of Q345R steel at the initial stage in 1 wt.% NaCl solution were investigated using electrochemical techniques. The effects of oxygen concentration, temperature and pH on corrosion behaviors were discussed. Simultaneously, a numerical model based on the mixed potential theory was proposed. The results show that the proposed model accords well with the experimental data in the pH range from 9.0 to 5.0. In this pH range, the oxygen reduction reaction, H⁺ reduction, water reduction, and iron oxidation can be quantitatively analyzed using this model. However, this model shows a disagreement with the experimental data at lower pH. This can be attributed to the fact that actual area of reaction on the electrode is much smaller than the preset area due to the block effect resulted from hydrogen bubbles adsorbed on the electrode surface.

Fusion of a family 20 carbohydrate-binding module (CBM20) with cyclodextrin glycosyltransferase of Geobacillus sp. CHB1 improves catalytic efficiency.

Cyclodextrin glycosyltransferase (CGTase) is an important industrial enzyme for production of cyclodextrins (CDs) from starch by intramolecular transglycosylation. CGTase consists of five domains labeled A to E. For optimizing catalytic activity of CGTase, CGTase of Geobacillus sp. was fused with the family 20 carbohydrate-binding module (CBM) of the Bacillus circulans strain 251 CGTase. The CBMbc251 that has a low binding free energy with maltohexaose, was selected by in silico design. Then the fusion enzyme, CGTΔE-CBMbc251, was constructed by fusing the CBMbc251 to the C-terminal region of CGTΔE. The fusion enzyme displayed an even greater enhancement of total α-cyclization activity (40.2%) and γ-cyclization activity (181.58%). Optimal reaction pH range was wilder and the thermal stability was better under 50 and 60 °C. Compared to the wild-type CGTase, the fusion enzyme showed a remarkable decrease in Km and a slight alteration in Vmax. The enhancement of soluble starch catalytic efficiency might be due to the changes of substrate binding ability in the critical substrate binding sites between the CBM and starch granule.

[Clinical outcome analysis on using extensor digitorum communis splitting approach for the treatment of radial head fractures].

OBJECTIVE: To explore the clinical outcomes of internal fixation or replacement for the treatment of radial head fractures through the extensor digitorum communis splitting approach. METHODS: From July 2012 to May 2015, 25 patients with radial head fractures were reviewed. There were 17 males and 8 females, ranging in age from 20 to 67 years old, with a mean age of 39 years old. Twenty-one patients were treated with reconstruction of plate internal fixation, and 4 patients were treated with radial head replacement. According to Mason classification, 19 cases were type II and 6 cases were type III. All the patients underwent internal fixation or replacement through the extensor digitorum communis splitting approach. The patients were followed up clinically and radiographically until the beginning of fracture union and the entrance of function recovery of elbow motion into a plateau. The functional status of the elbow was evaluated using the Mayo Elbow Performance Index(MEPI). Radiographic signs of post-traumatic arthritis were rated according to the Broberg and Morrey system. RESULTS: All the patients were followed up, and the average duration was 29 months (ranged, 12 to 56 months). The average range of flexion and extension was 120°, the extension was limited by 10°, and the flexion was 135°. The average forearm rotation range was 142°, pronation was 75°, supination was 67°. The mean MEPI was 93±7(ranged, 80 to 100 scores); according to the MEPI scoring criceria, 19 patients got an excellent functional result, 6 good. According to the Broberg and Morrey systems of traumatic arthritis, 19 patients were in grade 0, 6 in grade 1, and no patients in grade 2 or 3. No patients with nonunion of the radial head and failure of internal fixation were found. There were no complications of nerve or vascular injuries, and obvious limitation of elbow rotation. Heterotopic ossification around the elbow occurred in 4 cases, but the motion was not significantly limited. There were no complications such as prosthesis loosening and infection in 4 cases after radial head prosthesis replacement. CONCLUSIONS: The extensor digitorum communis splitting approach is an effective exposure method for internal fixation or replacement in the treatment of radial head fractures.

[Improving soluble expression of Geobacillus sp. B1 CGTase by errorprone PCR].

Objective: This study was aimed to enhance the extracellular enzymes activities and soluble expression of CGTase from Geobacillus sp. B1 by directed evolution. Methods: A library of CGTase mutants was constructed by introducing random mutagenesis using error-prone PCR to screen mutant enzymes with improved extracellular enzyme activities and soluble expression. After induction, expression and purification, the mutant enzyme was characterized. Results: After screening, two optimum mutants ds-6 and ep-9 with extracellular alpha-cyclization activity are respectively 1.72 times and 2.18 times of the original enzyme. The sequence of ep-9 cgt gene showed that three nucleotides substitution, G2005A, A2037G and T2081G were observed, and two of them caused amino acid changes. According to the 3D structure of Geobacillus sp. B1 cyclodextrin glucosetransferase mimicked by SWISS-MODEL Repository, two amino acid mutations were in rotating angle between beta angle and the random coil. The wild-type CGTase or ep-9 genes was ligated with pET-28 (a)-OmpA vector, and expressed in E. coli BL21 (DE3). After induced by lactose, CGTases were purified and characterized. The results showed that the specific ß-cyclization activity of the evolved CGTase was 1.31-fold than that of the wild-type CGTase, and the Km decreased from 4.3 to 3.74 g/L. The pH stability of the evolved CGTase was better than wild-type CGTase. Site-directed mutagenesis demonstrated the key to improve the soluble expression level and extracellular enzyme activity was G2005A. Conclusion: Directed evolution by error-prone PCR of Geobacillus sp. B1 CGTase gene is effective to improve extracellular enzyme activities and soluble expression, in particular mutation occurred in the G2005A.

[Genetic diversity and genetic relationships of spacies containing extremely aromatic compounds in leaves of Chimonanthus].

OBJECTIVE: Species containing extremely aromatic compounds in leaves of Chimonanthus was analyzed to evaluate its genetic diversity and genetic relationships. METHOD: AFLP molecular marker technique was used in the study, UPGMA cluster analysis was conducted with the software of POPGENE32. RESULTS: Five hundred and fifty-nine bands were amplified by 10 pairs of primers screened, of which 226 bands were polymorphic, and the percentage of polymorphic bands was 36.8%. Observed number of alleles, effective number of alleles, Nei's genetic diversity index and Shannon's information index were 1.992 6, 1.306 5, 0.199 2 and 0.325 1, respectively. Genetic distances of the 21 populations were ranged from 0.039 2 to 0.289 4. CONCLUSION: Species containing extremely aromatic compounds in leaves of Chimonanthus with low genetic diversity play an important role in enhancing the protection of species and germplasm resources. Form the molecular level, the studies demonstrated the correctness of the result by Zhang Ruohui that species containing extremely aromatic compounds in leaves of Chimonanthus were divided into Ch. salicifolius, Ch. Zhejiangensis, Ch. nitens and Ch. grammatus.