Inhibition of AKT induces p53/SIRT6/PARP1-dependent parthanatos to suppress tumor growth.

BACKGROUND: Targeting AKT suppresses tumor growth through inducing apoptosis, however, during which whether other forms of cell death occurring is poorly understood. METHODS: The effects of increasing PARP1 dependent cell death (parthanatos) induced by inhibiting AKT on cell proliferation were determined by CCK-8 assay, colony formation assay, Hoechst 33,258 staining and analysis of apoptotic cells by flow cytometry. For the detailed mechanisms during this process, Western blot analysis, qRT-PCR analysis, immunofluorescence and co-immunoprecipitation were performed. Moreover, the inhibition of tumor growth by inducing p53/SIRT6/PARP1-dependent parthanatos was further verified in the xenograft mouse model. RESULTS: For the first time, we identified that inhibiting AKT triggered parthanatos, a new form of regulated cell death, leading to colon cancer growth suppression. For the mechanism investigation, we found that after pharmacological or genetic AKT inhibition, p53 interacted with SIRT6 and PARP1 directly to activate it, and promoted the formation of PAR polymer. Subsequently, PAR polymer transported to outer membrane of mitochondria and resulted in AIF releasing and translocating to nucleus thus promoting cell death. While, blocking PARP1 activity significantly rescued colon cancer from death. Furthermore, p53 deletion or mutation eliminated PAR polymer formation, AIF translocation, and PARP1 dependent cell death, which was promoted by overexpression of SIRT6. Meanwhile, reactive oxygen species production was elevated after inhibition of AKT, which might also play a role in the occurrence of parthanatos. In addition, inhibiting AKT initiated protective autophagy simultaneously, which advanced tumor survival and growth. CONCLUSION: Our findings demonstrated that AKT inhibition induced p53-SIRT6-PARP1 complex formation and the activation of parthanatos, which can be recognized as a novel potential therapeutic strategy for cancer. Video Abstract.

Biomimetic synthesis of a novel O2-regeneration nanosystem for enhanced starvation/chemo-therapy.

Glucose oxidase-mediated starvation therapy that effectively cuts off energy supply holds great promise in cancer treatment. However, high glutathione (GSH) contents and anoxic conditions severely reduce therapy efficiency and cannot fully kill cancer cells. Herein, to resolve the above problem, this study constructed a biomimetic nanosystem based on nanreproo-MnO2with porous craspedia globose-like structure and high specific surface area, and it was further modified with dopamine and folic acid to guarantee good biocompatibility and selectivity toward cancer cells. This nanosystem responsively degraded and reacted with GSH and acid to regenerate O2, which significantly increased intracellular O2levels, accelerated glucose consumption, and improved starvation therapy efficiency. Moreover, anticancer drug of camptothecin was further loaded, and notably enhanced cancer growth inhibition was obtained at very low drug concentrations. Most importantly, this novel therapy could unprecedentedly inhibit cancer cell migration to a very low ratio of 19%, and detailed cell apoptosis analyses revealed late stage apoptosis contributed most to the good therapeutic effect. This work reported a new train of thought to improve starvation therapy in biomedicine, and provided a new strategy to design targeted nanocarrier to delivery mixed drugs to overcome the restriction of starvation therapy and develop new therapy patterns.

Optimization and scale-up of the production of rhamnolipid by Pseudomonas aeruginosa in solid-state fermentation using high-density polyurethane foam as an inert support.

To be competitive with common synthetic surfactants, the cost of production of rhamnolipid must be minimized by the fermentation process of non-foaming and low impurities. Herein, a novel solid-state fermentation process was developed for production of rhamnolipid by Pseudomonas aeruginosa SKY. The results were shown that high-density polyurethane foam is a satisfactory alternative to agro-industrial by-products for SSF of rhamnolipid. Palm oil and NaNO3 were promising carbon source and nitrogen source, respectively. Response surface methodology was employed to enhance the production of rhamnolipid. Palm oil, NaNO3 and liquid-to-solid ratios were significant factors. The optimal medium was developed as: 73.6 g/l palm oil; 3.0 g/l g NaNO3; 1.1 g NaCl; 1.1 g KCl; 3.4 g KH2PO4; 4.4 g K2HPO4; 0.5 g MgSO4·7H2O and 37.2 liquid-to-solid ratios. An overall 1.39-fold increase in rhamnolipid production was achieved in the optimized medium as compared with the unoptimized basal medium. Air pressure pulsation solid-state fermentation (APP-SSF) was applied to the experiment of scale-up for improving transfer efficiency of heat and mass. The yield of rhamnolipid reached 39.8 g/l in a 30 l APP-SSF fermenter. The crude extract of rhamnolipid lowered the surface tension of water to 28 mN/m and kept the critical micelle concentration at 50 mg/l. The work revealed the SSF with HPUF as an inert support was a promising fermentation system that could effectively produce rhamnolipid with low impurities, high productivity and low cost of production at a large scale.

Settling behaviors of microplastic disks in acceleration fall.

The settling of microplastics (MPs) in the initial acceleration fall stage, i.e., before reaching the terminal settling velocity, has not been investigated, which is however important for understanding MP transport and fate. MP disks sized 3-5 mm, of three shapes and made of three polymers (1.038-1.343 g/cm3) were examined. Five release ways and three release angles (0°, 45°, 90°) were used. MP disks with the release angle of 0° start to zigzag immediately after the release, while the MP disks with the release angles of 45° and 90° first adjust to a horizontal position and then zigzag. The adjustment distances in the vertical and horizontal directions, as well as the maximum vertical settling velocity, are influenced by MP density, size, release angle and release way. The detailed settling trajectory and velocity were also analyzed. Finally, the time-changing drag coefficient of MP disks was examined and discussed.

Tribbles pseudokinase 3 promotes enterovirus A71 infection via dual mechanisms.

Enterovirus A71 (EV-A71) is the main pathogen causing hand, foot and mouth disease (HFMD) in children and occasionally associated with neurological diseases such as aseptic meningitis, brainstem encephalitis (BE) and acute flaccid paralysis. We report here that cellular pseudokinase tribbles 3 (TRIB3) facilitates the infection of EV-A71 via dual mechanisms. In one hand, TRIB3 maintains the metabolic stability of scavenger receptor class B member 2 (SCARB2), the bona fide receptor of EV-A71, to enhance the infectious entry and spreading of the virus. On the other hand, TRIB3 facilitates the replication of EV-A71 RNA in a SCARB2-independent manner. The critical role of TRIB3 in EV-A71 infection and pathogenesis was further demonstrated in vivo in mice. In comparison to wild-type C57BL/6 mice, EV-A71 infection in TRIB3 knockdown mice (Trib3+/-) resulted in significantly lower viral loads in muscular tissues and reduced lethality and severity of clinical scores and tissue pathology. In addition, TRIB3 also promoted the replication of coxsackievirus B3 (CVB3) and coxsackievirus A16 (CVA16) in vitro. In conclusion, our results suggest that TRIB3 is one of key host cellular proteins required for the infection and pathogenesis of EV-A71 and some other human enteroviruses and may thus be a potential therapeutic target for combating the infection of those viruses.

Settling behaviors of microplastic disks in water.

Microplastic (MP) disks have not been studied for settling behaviors in aquatic environments, which affects the transport and fate of MPs. Therefore, settling experiments were conducted on MP disks of three shapes and four common-seen materials. Lighter MP disks (with density ρs = 1.038 g/cm3 and length l ≤ 5 mm) followed rectilinear vertical trajectories, while heavier MP disks (ρs = 1.161-1.343 g/cm3 and l = 5 mm) followed zigzag trajectories with oscillations and rotations. The mean terminal settling velocities of MP disks were 19.6-48.8 mm/s. Instantaneous settling velocities of heavier MP disks fluctuated. Existing formulas could not accurately predict the settling velocity of MP disks; thus, a new model was proposed with an error of 15.5 %. Finally, the Red - I* diagram (Red is the disk Reynolds number and I* is the dimensionless moment of inertia) was extended for MP disks to predict settling trajectories.

Heavy Ion Radiation Directly Induced the Shift of Oral Microbiota and Increased the Cariogenicity of Streptococcus mutans.

Radiation caries is one of the most common complications of head and neck radiotherapy. A shift in the oral microbiota is the main factor of radiation caries. A new form of biosafe radiation, heavy ion radiation, is increasingly being applied in clinical treatment due to its superior depth-dose distribution and biological effects. However, how heavy ion radiation directly impacts the oral microbiota and the progress of radiation caries are unknown. Here, unstimulated saliva samples from both healthy and caries volunteers and caries-related bacteria were directly exposed to therapeutic doses of heavy ion radiation to determine the effects of radiation on oral microbiota composition and bacterial cariogenicity. Heavy ion radiation significantly decreased the richness and diversity of oral microbiota from both healthy and caries volunteers, and a higher percentage of Streptococcus was detected in radiation groups. In addition, heavy ion radiation significantly enhanced the cariogenicity of saliva-derived biofilms, including the ratios of the genus Streptococcus and biofilm formation. In the Streptococcus mutans-Streptococcus sanguinis dual-species biofilms, heavy ion radiation increased the ratio of S. mutans. Next, S. mutans was directly exposed to heavy ions, and the radiation significantly upregulated the gtfC and gtfD cariogenic virulence genes to enhance the biofilm formation and exopolysaccharides synthesis of S. mutans. Our study demonstrated, for the first time, that direct exposure to heavy ion radiation can disrupt the oral microbial diversity and balance of dual-species biofilms by increasing the virulence of S. mutans, increasing its cariogenicity, indicating a potential correlation between heavy ions and radiation caries. IMPORTANCE The oral microbiome is crucial to understanding the pathogenesis of radiation caries. Although heavy ion radiation has been used to treat head and neck cancers in some proton therapy centers, its correlation with dental caries, especially its direct effects on the oral microbiome and cariogenic pathogens, has not been reported previously. Here, we showed that the heavy ion radiation directly shifted the oral microbiota from a balanced state to a caries-associated state by increasing the cariogenic virulence of S. mutans. Our study highlighted the direct effect of heavy ion radiation on oral microbiota and the cariogenicity of oral microbes for the first time.

A new model for the terminal settling velocity of microplastics.

Microplastic (MP) settling process is important for the transport of microplastic particles (MPs, <5 mm) in water bodies. However, for the control parameter of the drag coefficient (Cd), no generalized formula has been proposed for MPs of different shapes and materials. In this study, a total of 1343 MP settling data were collected from the literature. It was found that the drag law for perfect spheres cannot reasonably predict Cd for MPs with particle Reynolds number of 1-103. A new formula for Cd was developed by introducing the dimensionless particle diameter (d⁎) and two shape descriptors. The absolute error of the new formula is 15.2%, smaller than those (42.5-72.8%) of other existing formulas. Moreover, an explicit model was developed for MP settling velocity by correlating Cd, d⁎, and shape descriptors, with lower absolute error (8.8%) than those (15.4-77.2%) of existing models.

Beclin1 Binds to Enterovirus 71 3D Protein to Promote the Virus Replication.

Enterovirus 71 (EV71) is the main pathogen causing hand-foot-mouth disease (HFMD) in infants and children, which can also lead to severe neurological diseases and even death. Therefore, understanding the replication mechanism of EV71 is of great significance for the prevention and control of EV71-induced diseases. Beclin1 (BECN1, a mammalian homologue of ATG6 in yeast) is an important core protein for the initiation and the normal process of autophagy in cells. In addition to its involvement in autophagy, Beclin1 has also been reported to play an important role in cancer and innate immune signaling pathways. However, the role of Beclin1 in EV71 replication remains elusive. Here, we primarily found that Beclin1 facilitates EV71 replication in human rhabdomyosarcoma (RD) cells and the autophagy was actually induced, but Beclin1 was not significantly affected at either mRNA level or protein level during early EV71 infection. Further studies discovered that Beclin1 could interacts with EV71 non-structural protein 3D mainly through its evolutionary conserved domain (ECD) and coiled-coiled domain (CCD), thus promoting the replication of EV71 in human rhabdomyosarcoma (RD) cells and human astroglioma (U251) cells. Collectively, we reveal a novel regulatory mechanism associated with Beclin1 to promote EV71 replication, thus providing a potential therapeutic target for the prevention and control of EV71-associated diseases.

Vertebral Augmentation can Induce Early Signs of Degeneration in the Adjacent Intervertebral Disc: Evidence from a Rabbit Model.

STUDY DESIGN: An experimental study. OBJECTIVE: The aim of this study was to determine the effect of polymethylmethacrylate (PMMA) augmentation on the adjacent disc. SUMMARY OF BACKGROUND DATA: Vertebral augmentation with PMMA reportedly may predispose the adjacent vertebra to fracture. The influence of PMMA augmentation on the adjacent disc, however, remains unclear. METHODS: Using a retroperitoneal approach, PMMA augmentation was performed for 23 rabbits. For each animal, at least one vertebra was augmented with 0.2 to 0.3 mL PMMA. The disc adjacent to the augmented vertebra and a proximal control disc were studied using magnetic resonance (MR) imaging, histological and molecular level evaluation at 1, 3, and 6 months postoperatively. Marrow contact channels in the endplate were quantified in histological slices and number of invalid channels (those without erythrocytes inside) was rated. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was performed to determine disc cell apoptosis. RESULTS: On MR images, the signal and height of the adjacent disc did not change 6 months after vertebral augmentation. Histological scores of the adjacent disc increased over time, particularly for the nucleus pulposus. The adjacent disc had greater nucleus degeneration score than the control disc at 3 months (5.7 vs. 4.5, P < 0.01) and 6 months (6.9 vs. 4.4, P < 0.001). There were more invalid marrow contact channels in the endplate of augmented vertebra than the control (43.3% vs. 11.1%, P < 0.01). mRNA of ADAMTS-5, MMP-13, HIF-1α, and caspase-3 were significantly upregulated in the adjacent disc at 3 and 6 months (P < 0.05 for all). In addition, there were more TUNEL-positive cells in the adjacent disc than in the control disc (43.4% vs. 24.0%, P < 0.05) at 6 months postoperatively. CONCLUSION: Vertebral augmentation can induce early degenerative signs in the adjacent disc, which may be due to impaired nutrient supply to the disc. LEVEL OF EVIDENCE: N/A.

Deep eutectic solvent and inorganic salt pretreatment of lignocellulosic biomass for improving xylose recovery.

Deep eutectic solvents (DESs) have received considerable attention in recent years due to their low cost, low toxicity, and biodegradable properties. In this study, a sequential pretreatment comprising of a DES (choline chloride:urea in a ratio of 1:2) and divalent inorganic salt (CuCl2) was evaluated, with the aim of recovering xylose from oil palm fronds (OPF). At a solid-to-liquid ratio of 1:10 (w/v), DES alone was ineffective in promoting xylose extraction from OPF. However, a combination of DES (120°C, 4h) and 0.4mol/L of CuCl2 (120°C, 30min) resulted in a pretreatment hydrolysate containing 14.76g/L of xylose, remarkably yielding 25% more xylose than the CuCl2-only pretreatment (11.87g/L). Characterization studies such as FE-SEM, BET, XRD, and FTIR confirmed the delignification of OPF when DES was implemented. Thus, the use of this integrated pretreatment system enabled xylose recoveries which were comparable with other traditional pretreatments.

The anti-caries effects of dental adhesive resin influenced by the position of functional groups in quaternary ammonium monomers.

OBJECTIVES: A new quaternary ammonium monomer (QAM), triethylaminododecyl acrylate (TEADDA) was synthesized, in which the position of the functional groups was different from that of dimethylaminododecyl methacrylate (DMADDM). The objectives were to: (1) investigate the effect of the changed position of the functional groups on the mechanical properties, anti-biofilm activity and biocompatibility of adhesive resin, and (2) study the anti-bacterial mechanism of QAM to improve the performance of the adhesive system modified by QAM. METHODS: TEADDA and DMADDM were added into adhesives. Microtensile bond strength and surface charge density were measured. Multi-species biofilms were incubated on specimens for 16h, 48h and 72h and analyzed via MTT assay, lactic acid measurement and confocal laser scanning microscopy. The ratio of different species of bacteria was measured by real-time polymerase chain reaction. Cytotoxicity and biocompatibility were analyzed by eluents cytotoxicity test and histological images of H&E staining via an animal study in rats. RESULTS: The mass fraction of TEDDA allowed to be added into adhesive was higher than that of DMADDM. However, even 10% TEADDA did not yield a strong anti-biofilm effect on biofilm growth, lactic acid production and bacteria compositions. TEADDA added into adhesives showed better mechanical properties but weaker anti-bacterial effect. There was no significant difference on cytotoxicity and biocompatibility between DMADDM and TEADDA. SIGNIFICANCE: The study could be helpful for the investigation of the anti-caries mechanism of QAMs, the design of new QAMs and the improvement of the anti-caries activity of the modified dental materials.

Bacillus sp. strain P38: an efficient producer of L-lactate from cellulosic hydrolysate, with high tolerance for 2-furfural.

In this study, efficient polymer-grade L-lactic acid production was achieved with the strain Bacillus sp. P38 by using cellulosic hydrolysate as the sole carbon source. In fed-batch fermentation, 180 g L(-1)L-lactic acid was obtained with a volumetric productivity of 2.4 g L(-1)h(-1) and a yield of 0.96 g g(-1) total reducing sugars. No D-isomer of lactic acid was detected in the broth. Strain P38 tolerated up to 10 g L(-1) 2-furfural, and lactate production was sharply inhibited only when the 2-furfural concentration was higher than 6 g L(-1). Moreover, strain P38 also tolerated high concentrations (>6 g L(-1)) of other fermentation inhibitors in cellulosic hydrolysate, such as vanillin and acetic acid, although it was slightly sensitive to formic acid. The efficient L-lactic acid production, combined with high inhibitor tolerance and efficient pentose utilization, indicate that Bacillus sp. P38 is a promising producer of polymer-grade L-lactic acid from cellulosic biomass.

[Clinical manifestation and diagnosis of non-Hodgkin's lymphoma (NHL) in head and neck : analysis of 138 cases].

PURPOSE: To analyze the clinical features, pathological classification, causes of misdiagnosis, treatment and prognosis of non-Hodgkin's lymphoma (NHL) originated in head and neck and to afford experience in early diagnosis and treatment. METHODS: The clinical manifestation , treatment and prognosis of 138 cases of primary head and neck NHL were studied retrospectively. RESULTS: The incidence in male was higher than that in female (male:female=1.26:1). Among non-Hodgkin's lymphomas, 86.2% were neoplasms originating from B lymphocytes,which was remarkably higher than those originating from T lymphocyte(13.8%). The early symptom was mainly painless mass(71.7%). The primary location was in the following order: cervical region, submandibular region , parotid gland, skin of face, maxilla and mandible, tongue and palate. The main treatment was multimodal therapy(chemotherapy and operations).The five- year survival rate was 59.4%. CONCLUSIONS: The clinical manifestation of head and neck NHL is not specific and has a high misdiagnosis rate,so we must pay more attention to improve the early diagnosis and treatment of NHL.