Organic phosphorus levels change the hyphosphere phoD-harboring bacterial community of Funneliformis mosseae.

The phoD-harboring bacterial community is responsible for organic phosphorus (P) mineralization in soil and is important for understanding the interactions between arbuscular mycorrhizal (AM) fungi and phosphate-solubilizing bacteria (PSB) at the community level for organic P turnover. However, current understanding of the phoD-harboring bacterial community associated with AM fungal hyphae responses to organic P levels remains incomplete. Here, two-compartment microcosms were used to explore the response of the phoD-harboring bacterial community in the hyphosphere to organic P levels by high-throughput sequencing. Extraradical hyphae of Funneliformis mosseae enriched the phoD-harboring bacterial community and organic P levels significantly altered the composition of the phoD-harboring bacterial community in the Funneliformis mosseae hyphosphere. The relative abundance of dominant families Pseudomonadaceae and Burkholderiaceae was significantly different among organic P treatments and were positively correlated with alkaline phosphatase activity and available P concentration in the hyphosphere. Furthermore, phytin addition significantly decreased the abundance of the phoD gene, and the latter was significantly and negatively correlated with available P concentration. These findings not only improve the understanding of how organic P influences the phoD-harboring bacterial community but also provide a new insight into AM fungus-PSB interactions at the community level to drive organic P turnover in soil.

MiR-222-3p suppresses C2C12 myoblast proliferation and differentiation via the inhibition of IRS-1/PI3K/Akt pathway.

Numerous studies have revealed the profound impact of microRNAs on regulating skeletal muscle development and regeneration. However, the biological function and regulation mechanism of miR-222-3p in skeletal muscle remains largely unknown. In this study, miR-222-3p was found to be abundantly expressed in the impaired skeletal muscles, indicating that it might have function in the development and regeneration process of the skeletal muscle. MiR-222-3p overexpression impeded C2C12 myoblast proliferation and myogenic differentiation, whereas inhibition of miR-222-3p got the opposite results. The dual-luciferase reporter assay showed that insulin receptor substrate-1 (IRS-1) was the target gene of miR-222-3p. We next found that knockdown of IRS-1 could obviously suppress C2C12 myoblast proliferation and differentiation. Additionally, miR-222-3p-induced repression of myoblast proliferation and differentiation was verified to be associated with a decrease in phosphoinositide 3-kinase (PI3K)-Akt signaling. Overall, we demonstrated that miR-222-3p inhibited C2C12 cells myogenesis via IRS-1/PI3K/Akt pathway. Therefore, miR-222-3p may be used as a therapeutic target for alleviating muscle loss caused by inherited and nonhereditary diseases.

Phase Target-Based Calibration of Projector Radial Chromatic Aberration for Color Fringe 3D Measurement Systems.

The camera and projector are indispensable hardware parts of a color fringe projection 3D measurement system. Chromatic aberration between different color channels of the projector and camera has an impact on the measurement accuracy of the color fringe projection 3D profile measurement. There are many studies on camera calibration, but the chromatic aberration of the projector remains a question deserving of further investigation. In view of the complex system architecture and theoretical derivation of the traditional projector radial chromatic aberration method, a phase target based on projector radial chromatic aberration measurement and the correction method are proposed in this paper. This method uses a liquid crystal display with a holographic projection film as the phase target. The liquid crystal display sequentially displays red, green, and blue horizontal and vertical sinusoidal fringe images. The projector projects red, green, and blue horizontal and vertical sinusoidal fringe images to the phase target in turn, and calculates the absolute phases of the display fringes and reflection fringes, respectively. Taking the green channel as the reference channel, a phase coordinate system is established based on the phases of the vertical and horizontal directions displayed on the display screen, using the phase of the reflection fringes on the display screen as the ideal phase value of the phase point. Then, the phase coordinate system of the red and blue channels is transferred to the green phase coordinate system to calculate the chromatic aberration of the red-green channels and the blue-green channels, and pre-compensation is conducted. Experimental results prove that this method can measure and calibrate the radial chromatic aberration of the projector without being affected by the image quality of the camera. The correction effect of this method is that the maximum chromatic aberration of the red-green channel decreases from 1.9591/pixel to 0.5759/pixel, and the average chromatic aberration decreases from 0.2555/pixel to 0.1865/pixel. In addition, blue-green channel maximum chromatic aberration decreased from 1.8906/pixel to 0.5938/pixel, and the average chromatic aberration decreased from 0.2347/pixel to 0.1907/pixel. This method can improve the projection quality for fringe projection 3D profile measurement technology.

An aptly industrialized bioprocess for lactic acid production from corn stover using thermotolerant microbial consortia.

Chemical pretreatment of lignocellulosic biomass is a critical step in the conversion of lignocellulose to biofuels and biochemical. The main drawback of this pretreatment process is the formation of inhibitors which exhibit combined toxicity to microorganisms and result to low product concentrations and yields. In this study, the selection of microbial consortia by enrichment on hydrolysate of H2SO4-pretreated corn stover (pre-CS) without detoxification has been investigated as an efficient way to develop new strategies for lignocellulose utilization. The analysis of cattle stomach-dervied microbial consortia domesticated to degrade hydrolysate of pre-CS to produce lactic acid (LA) at different temperatures was investigated. Bacterial 16S rRNA gene amplicon sequencing analyses indicated that the three microbial consortia were taxonomically distinct and Enterococcus became dominant at high temperature. The highest glucose consumption rate was observed at 45 °C, while the three microbial consortia showed similar consumption rates of xylose and arabinose. The selected microbial consortia DUT37, DUT45 and DUT47 showed preferable resistances to inhibitors in hydrolysate of pre-CS and abilities of xylose utilization. A batch simultaneous saccharification and fermentation (SSF) process was developed by microbial consortium DUT47 at 47 °C to produce LA from pre-CS under non-detoxified and non-sterile conditions. The LA concentration and yield were 43.73 g/L and 0.50 g/g-corn stover (CS), respectively. Microbial consortium DUT47 has been shown to be suitable for LA production from H2SO4-pretreated corn stover without detoxification due to its thermophilic growth characteristics, robust tolerance of inhibitors, and the simultaneous utilization of glucose and xylose.

High-efficient L-lactic acid production from inedible starchy biomass by one-step open fermentation using thermotolerant Lactobacillus rhamnosus DUT1908.

The purpose of this study was to establish a simplified operational process for lactic acid (LA) production from inedible starchy biomass by open fermentation using thermotolerant Lactobacillus rhamnosus DUT1908. One step simultaneous liquefaction, saccharification and fermentation (SLSF) was proposed to produce LA using aging paddy rice with hull (APRH) as feedstock. First, a robust microbial strain was obtained by adaptive laboratory evolution under high temperature stress. As a result, L. rhamnosus DUT1908 showed high thermotolerance up to 50 °C and high efficiency of substrate utilization. Then, the performance of this thermotolerant L-lactic acid producing strain was demonstrated. Finally, various fermentation strategies were compared for LA production from APRH, including simultaneous saccharification and fermentation (SSF) and SLSF. In one-step open SLSF process, 107.8 g/L lactic acid was obtained with a productivity of 3.4 g/(L.h) and a yield to theoretical glucose of 0.89 g/g. This is the highest yield and productivity of lactic acid reported on starchy residues, and provides an efficient route for the development of high value-added products.

Sequential fed-batch fermentation of 1,3-propanediol from glycerol by Clostridium butyricum DL07.

The demand for 1,3-propanediol (1,3-PDO) has increased sharply due to its role as a monomer for the synthesis of polytrimethylene terephthalate (PTT). Although Clostridium butyricum is considered to be one of the most promising bioproducers for 1,3-PDO, its low productivity hinders its application on industrial scale because of the longer time needed for anaerobic cultivation. In this study, an excellent C. butyricum (DL07) strain was obtained with high-level titer and productivity of 1,3-PDO, i.e., 104.8 g/L and 3.38 g/(L•h) vs. 94.2 g/L and 3.04 g/(L•h) using pure or crude glycerol as substrate in fed-batch fermentation, respectively. Furthermore, a novel sequential fed-batch fermentation was investigated, in which the next bioreactor was inoculated by C. butyricum DL07 cells growing at exponential phase in the prior bioreactor. It could run steadily for at least eight cycles. The average concentration of 1,3-PDO in eight cycles was 85 g/L with the average productivity of 3.1 g/(L•h). The sequential fed-batch fermentation could achieve semi-continuous production of 1,3-PDO with higher productivity than repeated fed-batch fermentation and would greatly contribute to the industrial production of 1,3-PDO by C. butyricum. KEY POINTS: • A novel C. butyricum strain was screened to produce 104.8 g/L 1,3-PDO from glycerol. • Corn steep liquor powder was used as a cheap nitrogen source for 1,3-PDO production. • A sequential fed-batch fermentation process was established for 1,3-PDO production. • An automatic glycerol feeding strategy was applied in the production of 1,3-PDO.

Simultaneous liquefaction, saccharification, and fermentation of L-lactic acid using aging paddy rice with hull by an isolated thermotolerant Enterococcus faecalis DUT1805.

Simultaneous liquefaction, saccharification, and fermentation (SLSF) has attracted much attention for the production of bio-based chemicals, including L-lactic acid, due to its high efficiency and low cost. In this study, a lactic acid-producing bacterium with high tolerance of temperature up to 55 °C was isolated and characterized as Enterococcus faecalis DUT1805. Various strategies of stepwise controlled temperature were proposed and investigated for glucose utilization. The results indicated that E. faecalis DUT 1805 exhibited an optimal temperature at 50 °C, which could achieve temperature compatibility of enzyme, saccharification, and fermentation, and decrease the possibility of contamination by the other microorganisms during the large-scale fermentation. To reduce the cost of raw material and operation for lactic acid production, aging paddy rice with hull (APRH) was used in L-lactic acid production by simultaneous liquefaction, saccharification, and fermentation (SLSF). An open SLSF operation at 50 °C and pH 6.5, and 17% (w/v) solid loading in 5 L bioreactors was demonstrated with the lactic acid titer, yield, and productivity of 73.75 g/L, 87% to initial starch, and 2.17 g/(L h), respectively.

Stability and oscillatory behavior of microbial consortium in continuous conversion of crude glycerol to 1,3-propanediol.

Microbial consortium is an alternative for bioconversion of crude glycerol to value-added products whereas concerns about the process stability in long-term operation existed. The aim of this study is to evaluate the feasibility of using an anaerobic microbial consortium as inoculum for continuous conversion of crude glycerol to 1,3-propanediol (1,3-PDO). Performances of continuous fermentations with the consortium inoculum were evaluated under different dilution rates and glycerol feed concentrations. The highest 1,3-PDO production of 57.86 g/L was achieved with a productivity of 5.55 g/(L·h). Analyses of kinetic data showed that the consortium maintained a consistent pattern for 1,3-PDO production under different operating conditions despite changes in community composition. The continuous fermentation by the consortium was able to operate for a longer period of time (31 volume changes) than that using pure culture (24 volume changes) with the average 1,3-PDO concentration of 53.52 g/L and productivity of 6.69 g/(L·h) under glycerol-excess condition, which could be contributed to the intraspecies diversity among Clostridium butyricum in the consortium. Under glycerol-limited conditions, however, a spontaneous oscillation of the consortium was observed after continuous operation for about 120 h, along with severe fluctuations of the microbial community. The oscillatory behavior could be reduced by increasing the dilution rates and was likely the metabolic feature of C. butyricum.

The effects of ionic liquid 1-ethyl-3-methylimidazolium trifluoromethanesulfonate on the production of 1,3-propanediol from crude glycerol by microbial consortium.

Ionic liquids (ILs) as "green" solvents have been widely used owing to their excellent properties, e.g., for biodiesel production. Crude glycerol as a by-product in biodiesel production is an ideal feedstock for the microbial production of 1,3-propanediol (PDO), which is a versatile bulk chemical. PDO can be produced by microbial consortium with the advantages of high substrate tolerance and narrow by-product profile. In the present study, the effect of IL 1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([Emim][TfO]) was evaluated on the capacity of PDO production from crude glycerol by microbial consortium DL38-BH. In the batch fermentation at 60 g/L crude glycerol and 10 g/L [Emim][TfO], the concentration and yield of PDO from glycerol increased from 23.14 g/L and 0.45 mol/mol to 31.17 g/L and 0.60 mol/mol, respectively. Our results showed that [Emim][TfO] decreased the ratio of intracellular NADH to NAD+ and increased the concentration of 3-HPA during batch fermentation. The activities of three key enzymes in glycerol metabolism were stimulated by [Emim][TfO] during the batch fermentation by microbial consortium DL38-BH. Compared to the control, the proportion of Klebsiella genus which could convert glycerol to PDO increased significantly from 79.19% to 89.49% and the other genera that did not produce PDO were dramatically decreased (P < 0.05) at the end of batch fermentation. This work demonstrated that [Emim][TfO] significantly improved the concentration and yield of PDO from crude glycerol by adjusting microbial community during batch fermentation by microbial consortium.

Selection and characterization of an anaerobic microbial consortium with high adaptation to crude glycerol for 1,3-propanediol production.

Crude glycerol is an ideal feedstock for bioproduction of 1,3-propanediol (1,3-PDO) while pure culture always shows low substrate tolerance and limited productivity. In this study, an anaerobic microbial consortium for conversion of crude glycerol was selected and its 1,3-PDO production capacity was evaluated. The consortium was obtained from anaerobic activated sludge by 19 serial transfers and mainly consisted of 94.64% Clostridiaceae and 4.47% Peptostreptococcaceae. The consortium adapted well with high glycerol concentration of 120 g/L as well as wide substrate concentration fluctuation from 15 to 80 g/L, producing 60.61 and 82.66 g/L 1,3-PDO in the batch and fed-batch fermentation, with the productivity of 3.79 and 3.06 g/(L∙h), respectively, which are among the best results published so far. Furthermore, mini consortia isolated by serial dilution exhibited similar microbial composition but gradually decreasing tolerance to crude glycerol. Four randomly selected Clostridium butyricum displayed different substrate tolerance and insufficient 1,3-PDO production capacity. This work demonstrated that the high adaptation to crude glycerol of the consortium was the collaborative effort of different individuals.

Value of Multimode Sonography for Assessment of Pelvic Lipomatosis Compared With Computed Tomography.

OBJECTIVES: To assess imaging features of pelvic lipomatosis with a multimode sonographic technique, including 2-dimensional (2D) grayscale sonography, 3-dimensional (3D) sonography, and transrectal sonography, and compare its diagnostic features with those of computed tomography (CT). METHODS: In this study, 7 patients with different clinical manifestations were incidentally discovered by 2D sonography and followed by 3D and transrectal sonography before CT was performed for comparison. The urinary tract morphologic characteristics, bladder shape, and amount and distribution of perivesical or perirectal fatty tissue in all 7 patients were evaluated by both imaging modalities. Ten healthy participants were recruited and imaged as a control group. RESULTS: The following sonographic features were shown in all 7 patients: (1) bilateral hydroureters and hydronephrosis on 2D sonography; (2) bladder shifting anteriorly and superiorly on 2D sonography; (3) nonvisualization of the prostate through the bladder window on transabdominal scanning and nonvisualization of the bladder when the prostate was scanned by transrectal sonography; (4) bladder elongation in the craniocaudal dimension, which appeared as a gourd or pear shape on 3D sonography; and (5) excessive fat accumulation between the prostate and rectum as well as in perivesical or perirectal regions on 2D and transrectal sonography. Computed tomography in these patients confirmed the sonographic findings, with evidence of hydronephrosis, bladder location shift and elongation, and excessive fitty tissue with extrinsic compression. CONCLUSIONS: This preliminary study has shown that the application of a multimode sonographic technique (ie, 2D, 3D, and transrectal sonography) can provide unique evidence and imaging features of pelvic lipomatosis, which are comparable with CT for making a suggestive diagnosis of pelvic lipomatosis. Thus, multimode sonography may be the modality of choice for assessment of patients with a suspicion of lipomatosis.

[Tolerance of hyperosmolar Candida krusei].

The utilization of industrial microorganisms for the conversion of lignocellulose into high value-added chemicals is an essential pathway towards achieving carbon neutrality and promoting sustainable bioeconomy. However, the pretreated lignocellulase hydrolysate often contains various sugars, salts, phenols/aldehydes and other substances, which requires microorganisms to possess strong tolerance for direct fermentation. This study aims to investigate the tolerance of Candida krusei to substrate, salt, and high temperature shock, in order to validate its potential for utilizing the enzymatic hydrolysate of Pennisetum giganteum in seawater for fermentation. The experimental results showed that the adaptively domesticated C. krusei exhibited tolerance to glucose at a concentration of 200 g/L and became a hypertonic strain. When seawater was used instead of freshwater without sterilization, the yield of glycerol in fermentation was 109% higher than that in freshwater with sterilization. Moreover, the combined thermal shock at 32 hours of fermentation and addition of 10 Na2SO3 at 48 hours resulted in a yield of glycerol to glucose 0.37 g/g, which was 225% higher than the control group. By fermenting the enzymatic hydrolysate of P. giganteum pretreated in seawater, the total conversion rate of glucose into glycerol and ethanol reached 0.45 g/g. This study indicates that hypertonic C. krusei exhibits remarkable adaptability to substrate, salt, and temperature. It not only can directly utilize complex lignocellulosic hydrolysates, but also exhibits strong tolerance to them. Therefore, it provides a potential candidate strain for the production of bio-based chemicals using lignocellulosic processes.

Kinetics-based development of two-stage continuous fermentation of 1,3-propanediol from crude glycerol by Clostridium butyricum.

BACKGROUND: Glycerol, as a by-product, mainly derives from the conversion of many crops to biodiesel, ethanol, and fatty ester. Its bioconversion to 1,3-propanediol (1,3-PDO) is an environmentally friendly method. Continuous fermentation has many striking merits over fed-batch and batch fermentation, such as high product concentration with easy feeding operation, long-term high productivity without frequent seed culture, and energy-intensive sterilization. However, it is usually difficult to harvest high product concentrations. RESULTS: In this study, a three-stage continuous fermentation was firstly designed to produce 1,3-PDO from crude glycerol by Clostridium butyricum, in which the first stage fermentation was responsible for providing the excellent cells in a robust growth state, the second stage focused on promoting 1,3-PDO production, and the third stage aimed to further boost the 1,3-PDO concentration and reduce the residual glycerol concentration as much as possible. Through the three-stage continuous fermentation, 80.05 g/L 1,3-PDO as the maximum concentration was produced while maintaining residual glycerol of 5.87 g/L, achieving a yield of 0.48 g/g and a productivity of 3.67 g/(L·h). Based on the 14 sets of experimental data from the first stage, a kinetic model was developed to describe the intricate relationships among the concentrations of 1,3-PDO, substrate, biomass, and butyrate. Subsequently, this kinetic model was used to optimize and predict the highest 1,3-PDO productivity of 11.26 g/(L·h) in the first stage fermentation, while the glycerol feeding concentration and dilution rate were determined to be 92 g/L and 0.341 h-1, separately. Additionally, to achieve a target 1,3-PDO production of 80 g/L without the third stage fermentation, the predicted minimum volume ratio of the second fermenter to the first one was 11.9. The kinetics-based two-stage continuous fermentation was experimentally verified well with the predicted results. CONCLUSION: A novel three-stage continuous fermentation and a kinetic model were reported. Then a simpler two-stage continuous fermentation was developed based on the optimization of the kinetic model. This kinetics-based development of two-stage continuous fermentation could achieve high-level production of 1,3-PDO. Meanwhile, it provides a reference for other bio-chemicals production by applying kinetics to optimize multi-stage continuous fermentation.

Dynamic flux balance analysis of 1,3-propanediol production by clostridium butyricum fermentation.

To study the relationship between the yield of 1,3-propanediol (1,3-PDO) and the flux change of the Clostridium butyricum metabolic pathway, an optimized calculation method based on dynamic flux balance analysis was used by combining genome-scale flux balance analysis with a kinetic model. A more comprehensive and extensive metabolic pathway was obtained by optimization calculations. The primary extended branches include: the dihydroxyacetone node, which enters the pentose phosphate pathway; the α-oxoglutarate node, which has synthetic metabolic pathways for glutamic acid and amino acids; and the serine and homocysteine nodes, which produce cystathionine before homocysteine enters the methionine cycle pathway. According to the expanded metabolic network, the flux distribution of key nodes in the metabolic pathway and the relationship between the flux distribution ratio of nodes and the yield of 1,3-PDO were analyzed. At the dihydroxyacetone node, the flux of dihydroxyacetone converted to dihydroxyacetone phosphate was positively correlated with the yield of 1,3-PDO. As an important intermediate product, the flux change in the metabolic pathway of α-oxoglutarate reacting with amino acids to produce glutamic acid is positively correlated with the yield. When pyruvate was used as the central node to convert into lactic acid and α-oxoglutarate, the proportion of branch flux was negatively correlated with the yield of 1,3-PDO. These studies provide a theoretical basis for the optimization and further study of the metabolic pathway of C. butyricum.

Dynamic immobilization of bacterial cells on biofilm in a polyester nonwoven chemostat.

Cell immobilization plays an important role in biocatalysis for high-value products. It is necessary to maintain the viability of immobilized cells for bioconversion using viable cells as biocatalysts. In this study, a novel polyester nonwoven chemostat was designed for cell immobilization to investigate biofilm formation and the dynamic balance between adsorption and desorption of cells on polyester nonwoven. The polyester nonwoven was suitable for cell immobilization, and the cell numbers on the polyester nonwoven can reach 6.5 ± 0.38 log CFU/mL. After adding the polyester nonwoven to the chemostat, the fluctuation phenomenon of free bacterial cells occurred. The reason for this phenomenon was the balance between adsorption and desorption of bacterial cells on the polyester nonwoven. Bacterial cells could adhere to the surface of polyester nonwoven via secreting extracellular polymeric substances (EPS) to form biofilms. As the maturation of biofilms, some dead cells inside the biofilms can cause the detachment of biofilms. This process of continuous adsorption and desorption of cells can ensure that the polyester nonwoven chemostat has lasting biological activity.

Co-production of 1,3-propanediol and phage phiKpS2 from the glycerol fermentation by Klebsiella pneumoniae.

As an alternative to antibiotics in response to antimicrobial-resistant infections, bacteriophages (phages) are garnering renewed interest in recent years. However, the massive preparation of phage is restricted using traditional pathogens as host cells, which incurs additional costs and contamination. In this study, an opportunistic pathogen, Klebsiella pneumoniae used to convert glycerol to 1,3-propanediol (1,3-PDO), was reused to prepare phage after fermentation. The phage infection showed that the fed-batch fermentation broth containing 71.6 g/L 1,3-PDO can be directly used for preparation of phage with a titer of 1 × 108 pfu/mL. Then, the two-step salting-out extraction was adopted to remove most impurities, e.g. acetic acid (93.5%), ethanol (91.5%) and cells (99.4%) at the first step, and obtain 1,3-PDO (56.6%) in the top phase as well as phage (97.4%) in the middle phase at the second step. This integrated process provides a cheap and environment-friendly manner for coproduction of 1,3-PDO and phage.

Effects of thermophilic and acidophilic microbial consortia on maize wet-milling steeping.

To understand the ecology of species and promote biotechnology through beneficial strain selection for improving starch yield in maize wet-milling steeping, bacterial diversity and community structure during the counter-current steeping process in a commercial steeping system were characterized and investigated. The microbial diversity in the steeping liquor, which consisted of 16 phyla, 131 families, and 290 genera, was more abundant compared to those present on the surface of unsteeped maize. As the counter-current steeping progressed, exposing newer maize to the older steepwater, Lactobacillus dominated, replacing Rahnella, Pseudomonas, Pantoea, and Serratia. The thermophilic and acidophilic microbial consortia were enriched through adaptive evolution engineering and employed to improve starch yield. Several steeping strategies were evaluated, including water alone, SO2 alone, mono-culture of B. coagulans, microbial consortia, and a combination of consortium and SO2. Combining the microbial consortium with SO2 significantly increased the starch yield to, about 66.4 ± 0.5%, a 22% and 46% increase over SO2 alone and the consortium alone, respectively. Scanning electron microscope (SEM) of steeped maize structure indicated that the combination of consortium and SO2 disrupted the protein matrix and widened gaps between starch granules in maize endosperm. This released proteins into the steepwater and left starch granules in the aleurone layer. The steeping strategy of using thermophilic and acidophilic microbial consortium as additives shows potential application as an environmentally friendly alternative to conventional maize steeping procedures.

Hsc70 promotes anti-tumor immunity by targeting PD-L1 for lysosomal degradation.

Immune checkpoint inhibition targeting the PD-1/PD-L1 pathway has become a powerful clinical strategy for treating cancer, but its efficacy is complicated by various resistance mechanisms. One of the reasons for the resistance is the internalization and recycling of PD-L1 itself upon antibody binding. The inhibition of lysosome-mediated degradation of PD-L1 is critical for preserving the amount of PD-L1 recycling back to the cell membrane. In this study, we find that Hsc70 promotes PD-L1 degradation through the endosome-lysosome pathway and reduces PD-L1 recycling to the cell membrane. This effect is dependent on Hsc70-PD-L1 binding which inhibits the CMTM6-PD-L1 interaction. We further identify an Hsp90α/ß inhibitor, AUY-922, which induces Hsc70 expression and PD-L1 lysosomal degradation. Either Hsc70 overexpression or AUY-922 treatment can reduce PD-L1 expression, inhibit tumor growth and promote anti-tumor immunity in female mice; AUY-922 can further enhance the anti-tumor efficacy of anti-PD-L1 and anti-CTLA4 treatment. Our study elucidates a molecular mechanism of Hsc70-mediated PD-L1 lysosomal degradation and provides a target and therapeutic strategies for tumor immunotherapy.

Oleanolic acid alleviates obesity-induced skeletal muscle atrophy via the PI3K/Akt signaling pathway.

Oleanolic acid (OA) is a pentacyclic triterpene with reported protective effects against various diseases, including diabetes, hepatitis, and different cancers. However, the effects of OA on obesity-induced muscle atrophy remain largely unknown. This study investigated the effects of OA on skeletal muscle production and proliferation of C2C12 cells. We report that OA significantly increased skeletal muscle mass and improved glucose intolerance and insulin resistance. OA inhibited dexamethasone (Dex)-induced muscle atrophy in C2C12 myoblasts by regulating the PI3K/Akt signaling pathway. In addition, it also inhibited expression of MuRF1 and Atrogin1 genes in skeletal muscle of obese mice suffering from muscle atrophy, and increased the activation of PI3K and Akt, thereby promoting protein synthesis, and eventually alleviating muscle atrophy. Taken together, these findings suggest OA may have potential for the prevention and treatment of muscle atrophy.

Exercise-inducible circulating extracellular vesicle irisin promotes browning and the thermogenic program in white adipose tissue.

AIM: Exercise can reduce body weight and promote white fat browning, but the underlying mechanisms remain largely unknown. This study investigated the role of fibronectin type III domain-containing protein 5 (FNDC5)/Irisin, a hormone released from exercising muscle, in the browning of white fat in circulating extracellular vesicles (EVs). METHODS: Mice were subjected to a 4 weeks of running table exercise, and fat browning was analyzed via histology, protein blotting and qPCR. Circulating EVs were extracted by ultrahigh-speed centrifugation, and ELISA was used to measure the irisin concentration in the circulating EVs. Circulating EVs that differentially expressed irisin were applied to adipocytes, and the effect of EV-irisin on adipocyte energy metabolism was analyzed by immunofluorescence, protein blotting, and cellular oxygen consumption rate analysis. RESULTS: During sustained exercise, the mice lost weight and developed fat browning. FNDC5 was induced, cleaved, and secreted into irisin, and irisin levels subsequently increased in the plasma during exercise. Interestingly, irisin was highly expressed in circulating EVs that effectively promoted adipose browning. Mechanistically, the circulating EV-irisin complex is transported intracellularly by the adipocyte membrane receptor integrin αV, which in turn activates the AMPK signaling pathway, which is dependent on mitochondrial uncoupling protein 1 to cause mitochondrial plasmonic leakage and promote heat production. After inhibition of the AMPK signaling pathway, the effects of the EV-irisin on promoting fat browning were minimal. CONCLUSION: Exercise leads to the accumulation of circulating EV-irisin, which enhances adipose energy metabolism and thermogenesis and promotes white fat browning in mice, leading to weight loss.