Human Brain In Vitro Model for Pathogen Infection-Related Neurodegeneration Study.

Recent advancements in stem cell biology and tissue engineering have revolutionized the field of neurodegeneration research by enabling the development of sophisticated in vitro human brain models. These models, including 2D monolayer cultures, 3D organoids, organ-on-chips, and bioengineered 3D tissue models, aim to recapitulate the cellular diversity, structural organization, and functional properties of the native human brain. This review highlights how these in vitro brain models have been used to investigate the effects of various pathogens, including viruses, bacteria, fungi, and parasites infection, particularly in the human brain cand their subsequent impacts on neurodegenerative diseases. Traditional studies have demonstrated the susceptibility of different 2D brain cell types to infection, elucidated the mechanisms underlying pathogen-induced neuroinflammation, and identified potential therapeutic targets. Therefore, current methodological improvement brought the technology of 3D models to overcome the challenges of 2D cells, such as the limited cellular diversity, incomplete microenvironment, and lack of morphological structures by highlighting the need for further technological advancements. This review underscored the significance of in vitro human brain cell from 2D monolayer to bioengineered 3D tissue model for elucidating the intricate dynamics for pathogen infection modeling. These in vitro human brain cell enabled researchers to unravel human specific mechanisms underlying various pathogen infections such as SARS-CoV-2 to alter blood-brain-barrier function and Toxoplasma gondii impacting neural cell morphology and its function. Ultimately, these in vitro human brain models hold promise as personalized platforms for development of drug compound, gene therapy, and vaccine. Overall, we discussed the recent progress in in vitro human brain models, their applications in studying pathogen infection-related neurodegeneration, and future directions.

Probing Catalytic Sites and Adsorbate Spillover on Ultrathin FeO2-x Film on Ir(111) during CO Oxidation.

The spatially resolved identification of active sites on the heterogeneous catalyst surface is an essential step toward directly visualizing a catalytic reaction with atomic scale. To date, ferrous centers on platinum group metals have shown promising potential for low-temperature CO catalytic oxidation, but the temporal and spatial distribution of active sites during the reaction and how molecular-scale structures develop at the interface are not fully understood. Here, we studied the catalytic CO oxidation and the effect of co-adsorbed hydrogen on the FeO2-x/Ir(111) surface. Combining scanning tunneling microscopy (STM), isotope-labeled pulse reaction measurements, and DFT calculations, we identified both FeO2/Ir and FeO2/FeO sites as active sites with different reactivity. The trilayer O-Fe-O structure with its Moiré pattern can be fully recovered after O2 exposure, where molecular O2 dissociates at the FeO/Ir interface. Additionally, as a competitor, dissociated hydrogen migrates onto the oxide film with the formation of surface hydroxyl and water clusters down to 150 K.

Aging behavior of microplastics accelerated by mechanical fragmentation: alteration of intrinsic and extrinsic properties.

Microplastics (MPs) inevitably undergo multiple aging processes during their life cycle in the environment. However, the information regarding the mechanical fragmentation behavior of MPs remained unclear, including the changes in the intrinsic properties of aged MPs, the measurement of aging degree, the underlying mechanism, and the interaction with heavy metals. Here, MPs (PS, PP, PET) were aged by crushing (-CR) and ball-milling (-BM) to simulate mild and severe mechanical fragmentation, respectively. Our results indicated that mechanical fragmentation significantly affected the morphology of MPs. The aging degree of MP-BM was deeper compared to MP-CR owing to smaller particle size, larger specific surface area, poorer heat resistance, better hydrophilicity, and richer oxygen-containing functional groups. The carbonyl index (CI) and O/C ratio were used to measure the aging degree of the two mechanical aging treatments. Besides, the mechanism was proposed and the discrepancy between the two treatments was elaborated from three aspects including the excitation energy source, reaction interface, and reaction dynamics. Furthermore, the extrinsic properties of MPs altered with the increase of aging degree; specifically, the adsorption capacities of heavy metals were enhanced. Meanwhile, it was unveiled that the CI value and O/C ratio played a vital role in estimating the adsorption ability of heavy metals. The findings not only reveal the mechanical fragmentation behavior of MPs but also provide new insights into the assessment of the potential risks of the aged MPs via chemical indexes.

Study on the mechanism of NOx reduction by NH3-SCR over a ZnXCu1-XFe2O4 catalyst.

Experimental evidence shows that CuFe2O4 exhibits excellent catalytic performance in the SCR reaction. However, there is a lack of in-depth research on its specific reaction mechanism. Our study begins by computing the adsorption model of molecules like NH3 and then goes on to examine the SCR reaction mechanism of NH3 on CuFe2O4 before and after Zn doping. The results indicate that NH3 is chemically adsorbed (-1.26 eV) on the surface and has a strong interaction with the substrate. Importantly, Zn doping provides more favorable reactive sites for NH3 molecules. Subsequent investigation into the NH3 dehydrogenation and SCR reaction processes showed that incorporating Zn can greatly decrease the energy barrier of the most critical step in the reaction (0.58 eV). Additionally, the study also assesses the feasibility of the reaction of adsorbed NO with surface active O atoms to form NO2 (barrier 0.86 eV). Lastly, the sulfur resistance of the catalyst before and after doping is calculated and analyzed, and it is found that Zn doping effectively improves the sulfur resistance. Our study provides valuable theoretical guidance for the development of ferrite spinel and doping modification.

CircTmeff1 Promotes Muscle Atrophy by Interacting with TDP-43 and Encoding A Novel TMEFF1-339aa Protein.

Skeletal muscle atrophy is a common clinical feature of many acute and chronic conditions. Circular RNAs (circRNAs) are covalently closed RNA transcripts that are involved in various physiological and pathological processes, but their role in muscle atrophy remains unknown. Global circRNA expression profiling indicated that circRNAs are involved in the pathophysiological processes of muscle atrophy. circTmeff1 is identified as a potential circRNA candidate that influences muscle atrophy. It is further identified that circTmeff1 is highly expressed in multiple types of muscle atrophy in vivo and in vitro. Moreover, the overexpression of circTmeff1 triggers muscle atrophy in vitro and in vivo, while the knockdown of circTmeff1 expression rescues muscle atrophy in vitro and in vivo. In particular, the knockdown of circTmeff1 expression partially rescues muscle mass in mice during established atrophic settings. Mechanistically, circTmeff1 directly interacts with TAR DNA-binding protein 43 (TDP-43) and promotes aggregation of TDP-43 in mitochondria, which triggers the release of mitochondrial DNA (mtDNA) into cytosol and activation of the cyclic GMP-AMP synthase (cGAS)/ stimulator of interferon genes (STING) pathway. Unexpectedly, TMEFF1-339aa is identified as a novel protein encoded by circTmeff1 that mediates its pro-atrophic effects. Collectively, the inhibition of circTmeff1 represents a novel therapeutic approach for multiple types of skeletal muscle atrophy.

Identification of a novel small-molecule inhibitor of miR-29b attenuates muscle atrophy.

Muscle atrophy is debilitating and can be induced by several stressors. Unfortunately, there are no effective pharmacological treatment until now. MicroRNA (miR)-29b is an important target that we identified to be commonly involved in multiple types of muscle atrophy. Although sequence-specific inhibition of miR-29b has been developed, in this study, we report a novel small-molecule miR-29b inhibitor that targets miR-29b hairpin precursor (pre-miR-29b) (Targapremir-29b-066 [TGP-29b-066]) considering both its three-dimensional structure and the thermodynamics of interaction between pre-miR-29b and the small molecule. This novel small-molecule inhibitor has been demonstrated to attenuate muscle atrophy induced by angiotensin II (Ang II), dexamethasone (Dex), and tumor necrosis factor α (TNF-α) in C2C12 myotubes, as evidenced by increase in the diameter of myotube and decrease in the expression of Atrogin-1 and MuRF-1. Moreover, it can also attenuate Ang II-induced muscle atrophy in mice, as evidenced by a similar increase in the diameter of myotube, reduced Atrogin-1 and MuRF-1 expression, AKT-FOXO3A-mTOR signaling activation, and decreased apoptosis and autophagy. In summary, we experimentally identified and demonstrated a novel small-molecule inhibitor of miR-29b that could act as a potential therapeutic agent for muscle atrophy.

High Photocatalytic Hydrogen Production of Ag@TiO2 with Different Sizes by Simple Chemical Synthesis.

The photocatalytic enhancement of sliver-based metals compounded with semiconductor materials has been demonstrated. However, there are relatively few studies on the effect of particle size in the system on photocatalytic performance. In this paper, silver nanoparticles of two different sizes, 25 and 50 nm, were prepared by a wet chemical method and subsequently sintered to obtain a photocatalyst with a core-shell structure. The photocatalyst Ag@TiO2-50/150 prepared in this study has a hydrogen evolution rate as high as 4538.90 µmol·g-1·h-1. It is interesting to find that when the ratio of silver core size to composite size is 1:3, the hydrogen yield is almost not affected by the silver core diameter, and the hydrogen production rate is basically the same. In addition, the rate of hydrogen precipitation in air for 9 months was still more than 9 times those of previous studies. This provides a new idea for the study of the oxidation resistance and stability of photocatalysts.

Accuracy of 2 direct digital scanning techniques-intraoral scanning and stereophotogrammetry-for complete arch implant-supported fixed prostheses: A prospective study.

STATEMENT OF PROBLEM: Conventional impression techniques for complete arch implant-supported prostheses are technique-sensitive. Stereophotogrammetry (SPG) and intraoral scanning (IOS) may offer an alternative to conventional impression making. PURPOSE: The purpose of this prospective study was to compare the accuracy of IOS and SPG for complete arch implant scans and to evaluate the passive fit of frameworks fabricated with SPG. MATERIAL AND METHODS: Laboratory scanning of gypsum casts, SPG, and IOS were performed for all participants. The data regarding the abutment platform were superimposed to calculate the 3D deviation of SPG and IOS compared with that of laboratory scanning as an evaluation of accuracy. The effect of implant position and number on accuracy was analyzed. The more accurate technique between SPG and IOS was used to fabricate the titanium frameworks, as was laboratory scanning. The passive fit of the frameworks was assessed by clinical examination, the Sheffield test, and panoramic radiography. RESULTS: Seventeen participants (21 arches, 120 implants) were included. The accuracy of SPG ranged from 2.70 µm to 92.80 µm, with a median (Q1, Q3) of 17.00 (11.68, 22.50) µm, which was significantly more accurate than that of IOS, ranging from 21.30 µm to 815.60 µm, with a median (Q1, Q3) of 48.95 (34.78, 75.88) µm. No significant correlation was found between position or number of implants and 3D deviation in the SPG group. A weak positive correlation was found between implant number and 3D deviation in the IOS group. SPG and laboratory scanning were used to fabricate titanium frameworks. The passive fit between the frameworks and abutment platforms was confirmed. CONCLUSIONS: SPG, which was not affected by position or number of implants, was more accurate than IOS and comparable with laboratory scanning. The frameworks fabricated based on SPG and laboratory scanning were comparable in their passive fit. The SPG technique may be an alternative to laboratory scanning for complete arch implant scans.

A completely digital workflow aided by cone beam computed tomography scanning to maintain jaw relationships for implant-supported fixed complete dentures: A clinical study.

STATEMENT OF PROBLEM: The conventional workflow for the fabrication of implant-supported fixed complete dentures (IFCDs) is complex and makes it impossible to maintain jaw relationships. A fully digital workflow might solve this problem. PURPOSE: The purpose of this clinical study was to develop a completely digital workflow aided by a cone beam computed tomography (CBCT) scan for the fabrication of IFCDs and to evaluate the accuracy of this workflow with regard to the maintenance of jaw relationships. MATERIAL AND METHODS: All participants received a preoperative CBCT scan while wearing radiographic diagnosis dentures and occluding in the maximum intercuspal position. After the implant surgery, CBCT scanning, intraoral scanning, and stereophotogrammetry were performed to identify jaw anatomy, soft tissue, and the 3-dimensional (3D) locations of the implants, respectively. Then, all data were merged to transfer jaw relationships and generate digital casts to fabricate interim prostheses. A posttreatment CBCT scan was performed while the participants were wearing the interim prostheses and occluding in the maximum intercuspal position. The preoperative and postoperative jaw relationships were compared by CBCT cephalometric analysis. A meaningful and unacceptable difference was defined as 0.8 degrees and 2.4 degrees, respectively. RESULTS: Six participants (6 jaw relationships, 9 arches, and 58 implants) were included. All interim prostheses were stable and achieved symmetric occlusion after only minimal adjustment. A total of 18 angles were measured. Three angles revealed a meaningful minimal difference, and 1 angle revealed an unacceptable minimal difference. No prosthodontic complications were reported during the study. CONCLUSIONS: A completely digital workflow for fabricating IFCDs achieved sufficient accuracy for the maintenance of jaw relationships throughout the treatment.

Correlation study of biomechanical changes between diabetic eye disease and glaucoma using finite element model of human eye with different iris-lens channel distances.

PURPOSE: To find out the effect on the biomechanical response of the eye in the setting of diabetes combined with glaucoma. METHOD: Five finite element models of the human eyes with various iris-lens channel (ILC) distances (2 µm-20 µm) were constructed, respectively. The human eye model used for finite element analysis contain all the ocular contents and the optic nerve head. All these models with different ILC distances were used to simulate the effect of pupillary block and abnormal aqueous flow induced by diabetes. And those models were also used for the biomechanical properties study of ocular tissues under the elevated intraocular pressure (IOP), using unidirectional fluid-solid coupling numerical simulation method. RESULTS: For the most severe cases of pupil block (2 µm), a significant difference in chamber pressure caused the iris to move forward and had posterior adhesion to the lens. And the strain, stress, and displacement of the whole eyeball were significantly higher than those of the other four cases, while the Optic Nerve Head (ONH) region was the opposite. The promotion of IOP to biomechanical response at both global eye and ONH region was close to the normal eye conditions, or even ease for ILC = 2 µm. But in the cases of glaucoma with pupil block and high aqueous flow, the biomechanical properties of the whole eyeball were remarkably enhanced for all IOP conditions. Less influence was observed in the ONH region. CONCLUSION: The promotion of diabetes for glaucoma is not directly on the optic nerve, instead, it indirectly affects the optic nerve by affecting the global eye. Glaucoma combined with diabetes complications may increase the biomechanical damage of IOP to the whole eye.

Effectiveness and Mechanisms of Low-Intensity Pulsed Ultrasound on Osseointegration of Dental Implants and Biological Functions of Bone Marrow Mesenchymal Stem Cells.

Dental implant restoration is the preferred choice for patients with dentition defects or edentulous patients, and obtaining stable osseointegration is the determining factor for successful implant healing. The risk of implant failure during the healing stage is still an urgent problem in clinical practice due to differences in bone quality at different implant sites and the impact of some systemic diseases on bone tissue metabolism. Low-intensity pulsed ultrasound (LIPUS) is a noninvasive physical intervention method widely recognized in the treatment of bone fracture and joint damage repair. Moreover, many studies indicated that LIPUS could effectively promote the osseointegration of dental implants and improve the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). This review is aimed at investigating the research progress on the use of LIPUS in dental implant medicine from three aspects: (1) discuss the promoting effects of LIPUS on osseointegration and peri-implant bone regeneration, (2) summarize the effects and associated mechanisms of LIPUS on the biological functions of BMSCs, and (3) introduce the application and prospects of LIPUS in the clinical work of dental implantation. Although many challenges need to be overcome in the future, LIPUS is bound to be an efficient and convenient therapeutic method to improve the dental implantation success rate and expand clinical implant indications.

Two transporters mobilize magnesium from vacuolar stores to enable plant acclimation to magnesium deficiency.

Magnesium (Mg) is an essential metal for chlorophyll biosynthesis and other metabolic processes in plant cells. Mg is largely stored in the vacuole of various cell types and remobilized to meet cytoplasmic demand. However, the transport proteins responsible for mobilizing vacuolar Mg2+ remain unknown. Here, we identified two Arabidopsis (Arabidopsis thaliana) Mg2+ transporters (MAGNESIUM TRANSPORTER 1 and 2; MGT1 and MGT2) that facilitate Mg2+ mobilization from the vacuole, especially when external Mg supply is limited. In addition to a high degree of sequence similarity, MGT1 and MGT2 exhibited overlapping expression patterns in Arabidopsis tissues, implying functional redundancy. Indeed, the mgt1 mgt2 double mutant, but not mgt1 and mgt2 single mutants, showed exaggerated growth defects as compared to the wild type under low-Mg conditions, in accord with higher expression levels of Mg-starvation gene markers in the double mutant. However, overall Mg level was also higher in mgt1 mgt2, suggesting a defect in Mg2+ remobilization in response to Mg deficiency. Consistently, MGT1 and MGT2 localized to the tonoplast and rescued the yeast (Saccharomyces cerevisiae) mnr2Δ (manganese resistance 2) mutant strain lacking the vacuolar Mg2+ efflux transporter. In addition, disruption of MGT1 and MGT2 suppressed high-Mg sensitivity of calcineurin B-like 2 and 3 (cbl2 cbl3), a mutant defective in vacuolar Mg2+ sequestration, suggesting that vacuolar Mg2+ influx and efflux processes are antagonistic in a physiological context. We further crossed mgt1 mgt2 with mgt6, which lacks a plasma membrane MGT member involved in Mg2+ uptake, and found that the triple mutant was more sensitive to low-Mg conditions than either mgt1 mgt2 or mgt6. Hence, Mg2+ uptake (via MGT6) and vacuolar remobilization (through MGT1 and MGT2) work synergistically to achieve Mg2+ homeostasis in plants, especially under low-Mg supply in the environment.

Biomechanical analysis of ocular diseases and its in vitro study methods.

Ocular diseases are closely related to the physiological changes in the eye sphere and its contents. Using biomechanical methods to explore the relationship between the structure and function of ocular tissue is beneficial to reveal the pathological processes. Studying the pathogenesis of various ocular diseases will be helpful for the diagnosis and treatment of ocular diseases. We provide a critical review of recent biomechanical analysis of ocular diseases including glaucoma, high myopia, and diabetes. And try to summarize the research about the biomechanical changes in ocular tissues (e.g., optic nerve head, sclera, cornea, etc.) associated with those diseases. The methods of ocular biomechanics research in vitro in recent years are also reviewed, including the measurement of biomechanics by ophthalmic equipment, finite element modeling, and biomechanical analysis methods. And the preparation and application of microfluidic eye chips that emerged in recent years were summarized. It provides new inspiration and opportunity for the pathogenesis of eye diseases and personalized and precise treatment.

Delivery of engineered extracellular vesicles with miR-29b editing system for muscle atrophy therapy.

Muscle atrophy is a frequently observed complication, characterized by the loss of muscle mass and strength, which diminishes the quality of life and survival. No effective therapy except exercise is currently available. In our previous study, repressing miR-29b has been shown to reduce muscle atrophy. In our current study, we have constructed artificially engineered extracellular vesicles for the delivery of CRISPR/Cas9 to target miR-29b (EVs-Cas9-29b). EVs-Cas9-29b has shown a favorable functional effect with respect to miR-29b repression in a specific and rapid manner by gene editing. In in vitro conditions, EVs-Cas9-29b could protect against muscle atrophy induced by dexamethasone (Dex), angiotensin II (AngII), and tumor necrosis factor-alpha (TNF-α). And EVs-Cas9-29b introduced in vivo preserved muscle function in the well-established immobilization and denervation-induced muscle atrophy mice model. Our work demonstrates an engineered extracellular vesicles delivery of the miR-29b editing system, which could be potentially used for muscle atrophy therapy.

Targeting miR-30d reverses pathological cardiac hypertrophy.

BACKGROUND: Pathological cardiac hypertrophy occurs in response to numerous stimuli and precedes heart failure (HF). Therapies that ameliorate pathological cardiac hypertrophy are highly needed. METHODS: The expression level of miR-30d was analyzed in hypertrophy models and serum of patients with chronic heart failure by qRT-PCR. Gain and loss-of-function experiments of miR-30d were performed in vitro. miR-30d gain of function were performed in vivo. Bioinformatics, western blot, luciferase assay, qRT-PCR, and immunofluorescence were performed to examine the molecular mechanisms of miR-30d. FINDINGS: miR-30d was decreased in both murine and neonatal rat cardiomyocytes (NRCMs) models of hypertrophy. miR-30d overexpression ameliorated phenylephrine (PE) and angiotensin II (Ang II) induced hypertrophy in NRCMs, whereas the opposite phenotype was observed when miR-30d was downregulated. Consistently, the miR-30d transgenic rat was found to protect against isoproterenol (ISO)-induced pathological hypertrophy. Mechanistically, methyltransferase EZH2 could promote H3K27me3 methylation in the promotor region of miR-30d and suppress its expression during the pathological cardiac hypertrophy. miR-30d prevented pathological cardiac hypertrophy via negatively regulating its target genes MAP4K4 and GRP78 and inhibiting pro-hypertrophic nuclear factor of activated T cells (NFAT). Adeno-associated virus (AAV) serotype 9 mediated-miR-30d overexpression exhibited beneficial effects in murine hypertrophic model. Notably, miR-30d was reduced in serum of patients with chronic heart failure and miR-30d overexpression could significantly ameliorate pathological hypertrophy in human embryonic stem cell-derived cardiomyocytes. INTERPRETATION: Overexpression of miR-30d may be a potential approach to treat pathological cardiac hypertrophy. FUNDING: This work was supported by the grants from National Key Research and Development Project (2018YFE0113500 to J Xiao), National Natural Science Foundation of China (82020108002 to J Xiao, 81900359 to J Li), the grant from Science and Technology Commission of Shanghai Municipality (20DZ2255400 and 21XD1421300 to J Xiao, 22010500200 to J Li), Shanghai Sailing Program (19YF1416400 to J Li), the "Dawn" Program of Shanghai Education Commission (19SG34 to J Xiao), the "Chen Guang" project supported by the Shanghai Municipal Education Commission and Shanghai Education Development Foundation (19CG45 to J Li).

DATA-driven shock impact of COVID-19 on the market financial system.

The Corona Virus Disease 2019(COVID-19) has a dramatic effect on my country's market and financial system. Although China has controlled the deterioration of the epidemic, this global epidemic will inevitably have an impact on the global economy including China. In order to study the shock effect of the COVID-19 on the market financial system, this paper builds a data model processing system based on the event analysis method, and analyzes the shock effect from three aspects of supply chain finance, financial securities, and corporate financial systems. Moreover, this paper uses crawler technology to obtain valid data from major websites, analyzes model data with mathematical statistics combined with event models, and outputs the results and compares them with the actual situation. Through data analysis, it can be seen that the model constructed in this paper can effectively reflect the shock effect of the COVID-19 on the market financial system. Finally, the comparison method is used to compare the research results with the actual situation. The results show that the two are basically the same. Therefore, it can be seen that the proposed research method has significant effects and has certain reference value for studying the shock effect of the epidemic on the financial system.

Formation of trihalomethanes and haloacetic acids from 2,6-dichloro-1,4-benzoquinone during chlorination: Decomposition kinetics, conversion rates, and pathways.

As a typical aromatic disinfection byproduct (DBP), 2,6-dichloro-1,4-benzoquinone (2,6-DCBQ) attracts much concern due to the potential toxicity. To further evaluate the role of 2,6-DCBQ as an intermediate DBP in water with or without chlorine, their decomposition characteristics and transformation potential to the regulated DBPs (i.e., trihalomethanes (THMs) and haloacetic acids (HAAs)) were investigated under different chlorine doses, pH values, temperatures, contact times, and bromide levels. The decomposition of 2,6-DCBQ under different conditions all fit apparent first-order kinetics. The hydrolysis rate constants of 2,6-DCBQ significantly increased with pH. The half-live values of 2,6-DCBQ were 108.3-568.7 h at pH 6.0-6.5, and 1.8-31.1 h at pH 7.0-8.5. During the hydrolysis of 2,6-DCBQ, there was no THMs and HAAs generated. During chlorination, 2,6-DCBQ decayed rapidly accompanied by the fast formation of trichloromethane (TCM) and the gradual generation of dichloroacetic acid and trichloroacetic acid. The molar conversion rates of 2,6-DCBQ-to-THMs (i.e., TCM) and 2,6-DCBQ-to-HAAs were 2.9-10.0% and 0.1-2.2% under different conditions. The presence of bromide increased the conversion rates of 2,6-DCBQ-to-THMs and caused the generation of brominated THMs and HAAs. According to the decomposition characteristics of 2,6-DCBQ and the formation trends of THMs and HAAs under different conditions, multiple formation pathways from 2,6-DCBQ to THMs and HAAs were proposed.

LncRNA SAMMSON Mediates Adaptive Resistance to RAF Inhibition in BRAF-Mutant Melanoma Cells.

The long noncoding RNA (lncRNA) SAMMSON is required for human melanoma cell growth and survival. However, whether SAMMSON regulates the response of mutant BRAF melanoma cells to RAF inhibitors remains unknown. In this work, we showed that SAMMSON is rapidly induced upon inhibition of ERK signaling, and SAMMSON overexpression conferred resistance to vemurafenib-induced cytotoxicity in melanoma cells. SOX10 mediated transcriptional induction of SAMMSON by vemurafenib, and SOX10 sumoylation at K55 was essential for this function. In addition, depletion of SAMMSON activated p53 signaling, which is dependent on the SAMMSON-interacting protein CARF. Depletion of SAMMSON sensitized mutant BRAF melanoma cells to RAF inhibitors in vitro and in vivo, while CARF knockdown reversed the enhanced sensitivity. In summary, these findings suggest that SAMMSON may function as a new mediator of adaptive resistance to RAF inhibitors in melanoma by modulating CARF-p53 signaling. SIGNIFICANCE: This study highlights the role of a SAMMSON/CARF/p53 signaling axis in modulating the adaptive resistance of mutant BRAF melanoma to RAF inhibitors.

Rose bengal-mediated photodynamic inactivation against periodontopathogens in vitro.

BACKGROUND: The main goal of periodontal therapy is to eliminate the spread of infection in the periodontium. Antimicrobial photodynamic therapy (aPDT) is a bactericidal method that has been recently introduced for controlling periodontal infection. The aim of this in vitro study was to evaluate the effect of aPDT using a combination of medium-power blue light-emitting diodes (LEDs) and rose bengal (RB) on selected key periodontopathogens. METHODS: Porphyromonas gingivalis ATCC33277, Aggregatibacter actinomycetemcomitans ATCC29523 and Fusobacterium nucleatum ATCC10953 were used in the experiments. Each bacterial suspension was irradiated with a blue LED (BL) (450-470 nm, output power density of 1.2 W/cm2) for 20-60 s (6-18 J/cm2), treated with RB (1 min), or subjected to a combination of RB treatment and BL irradiation (40 s, 12 J/cm2). All bacterial suspensions were serially diluted, plated and incubated anaerobically or microaerobically, and the numbers of colony-forming units (CFUs) were counted on day 7. One-way analysis of variance (ANOVA) and Tukey's HSD tests were used for statistical analysis. RESULTS: Treatment with BL irradiation from 6 to 18 J/cm2 did not significantly reduce the number of CFUs, whereas treatment with RB alone induced a low-to-high reduction in the bacterial CFUs in a dye concentration-dependent manner. Furthermore, the difference in the effects obtained with 16 µg/mL and 160 µg/mL RB was not statistically significant. Treatment with the BL at 12 J/cm2 combined with 160 µg/mL RB yielded maximal log reductions of 3.03, 4.2 and 2.23 in P. gingivalis, A. actinomycetemcomitans and F. nucleatum CFUs, respectively. CONCLUSION: Within the limits of this study, the three periodontal pathogens, especially A. actinomycetemcomitans, were susceptible to photodynamic inactivation by the combination of the BL and RB. RB-mediated aPDT may offer a viable alternative tool for periodontal pathogen treatment, especially for A. actinomycetemcomitans eradication. aPDT may be a valuable tool for the treatment of periodontal diseases, particularly those in which A. actinomycetemcomitans is a dominating pathogen.

CIC Is a Mediator of the ERK1/2-DUSP6 Negative Feedback Loop.

DUSP6 functions as an important negative feedback component of the MAPK/ERK signaling pathway. Although DUSP6 expression is tightly regulated by ERK1/2 signaling, the molecular mechanism of this regulation remains partially understood. In this work, we show that the transcriptional repressor CIC functions downstream of the ERK1/2 signaling to negatively regulate DUSP6 expression. CIC directly represses DUSP6 transcription by binding to three cis-regulatory elements (CREs) in DUSP6 promoter. p90RSK, a downstream target of ERK1/2, phosphorylates CIC at S173 and S301 sites, which creates a 14-3-3 recognition motif, resulting in 14-3-3-mediated nuclear export of CIC and derepression of DUSP6. Finally, we demonstrate that the oncogenic CIC-DUX4 fusion protein acts as a transcriptional activator of DUSP6 and its nuclear/cytoplasmic distribution remains regulated by ERK1/2 signaling. These results complete an ERK1/2/p90RSK/CIC/DUSP6 negative feedback circuit and elucidate the molecular mechanism of how RTK/MAPK signaling harnesses the transcriptional repressor activity of CIC in mammalian cells.