Regulation of Fluorescence and Self-assembly of a Salicylaldehyde Azine-Containing Amphiphile by Pillararene.

Regulation of fluorescence and self-assembly of a salicylaldehyde azine-containing amphiphile by a water-soluble pillar[5]arene via host-guest recognition in water was realized. The fluorescence and the self-assembled aggregates of the bola-type amphiphile G can be tailored by adding different amounts of water-soluble pillar[5]arene (WP5). In addition, the emission property and self-assembly behavior of G and WP5 are responsive to pH conditions. Furthermore, the fluorescence emission property of G and the regulation by WP5 or pH conditions was applied as information encryption material, rewritable paper, and erasable ink. We believe that this fluorescence regulation strategy is promising for the construction of advanced fluorescent organic materials.

BioCaster in 2021: automatic disease outbreaks detection from global news media.

SUMMARY: BioCaster was launched in 2008 to provide an ontology-based text mining system for early disease detection from open news sources. Following a 6-year break, we have re-launched the system in 2021. Our goal is to systematically upgrade the methodology using state-of-the-art neural network language models, whilst retaining the original benefits that the system provided in terms of logical reasoning and automated early detection of infectious disease outbreaks. Here, we present recent extensions such as neural machine translation in 10 languages, neural classification of disease outbreak reports and a new cloud-based visualization dashboard. Furthermore, we discuss our vision for further improvements, including combining risk assessment with event semantics and assessing the risk of outbreaks with multi-granularity. We hope that these efforts will benefit the global public health community. AVAILABILITY AND IMPLEMENTATION: BioCaster web-portal is freely accessible at http://biocaster.org.

Self-Luminescent Drug Delivery Vehicle: Synthesis, Self-Assembly Behavior, Cysteine-Responsive Property, and Application in the Visualization of Drug Release.

Targeted drug delivery systems have gained great attention from the chemistry and biomedical fields in recent years due to the minimized harm to normal cells. When designing targeted drug delivery systems, the property of harmlessness to normal cells and the tracking ability of the whole process are quite crucial. These two characters can be brought into the related systems by applying a drug carrier that is self-luminescent and its drug release can be induced by the microenvironment of cancer cells. Therefore, the design and synthesis of drug delivery vehicles are significant for the fabrication of target drug delivery systems. Herein, we have synthesized a cysteine-responsive and fluorescent molecule, maleic acid-modified tetraphenylethylene derivative (MATPE), by a facile method. In addition, a drug delivery system with self-luminescence and cysteine-responsiveness based on the self-assembly of MATPE was fabricated. In this system, MATPE and cysteine both played dual roles as cysteine probe/drug carrier and emission-enhanced inducement/drug-release stimulus. The drug-release process was successfully realized in cancer cells and can be visualized, exhibiting great potential in the field of theranostics.

Palladium-Catalyzed Double N-Arylation to Access Unsymmetric N,N'-Bicarbazole Scaffolds.

Herein, the palladium-catalyzed double C-N coupling of 9H-carbazol-9-amines and 2,2'-dibromo-1,1'-biphenyl is reported. This protocol offers access to N,N'-bicarbazole scaffolds, which have frequently been used as linkers in the construction of functional covalent organic frameworks (COFs). A variety of substituted N,N'-bicarbazoles were synthesized in moderate to high yields based on this chemistry, and the potential application of this method was showcased by the synthesis of COF monomers like tetrabromide 4 and tetraalkynylate 5.

Golgi-localized calcium/manganese transporters FgGdt1 and FgPmr1 regulate fungal development and virulence by maintaining Ca2+ and Mn2+ homeostasis in Fusarium graminearum.

Calcium and manganese transporters play important roles in regulating Ca2+ and Mn2+ homeostasis in cells, which is necessary for the normal physiological activities of eukaryotes. Gdt1 and Pmr1 function as calcium/manganese transporters in the Golgi apparatus. However, the functions of Gdt1 and Pmr1 have not been previously characterized in the plant pathogenic fungus Fusarium graminearum. Here, we identified and characterized the biological functions of FgGdt1 and FgPmr1 in F. graminearum. Our study shows that FgGdt1 and FgPmr1 are both localized to the cis- and medial-Golgi. Disruption of FgGdt1 or FgPmr1 in F. graminearum caused serious defects in vegetative growth, conidiation, sexual development and significantly decreased virulence in wheat but increased deoxynivalenol (DON) production. Importantly, FgGdt1 is involved in Ca2+ and Mn2+ homeostasis and the severe phenotypic defects of the ΔFggdt1 mutant were largely due to loss of FgGdt1 function in Mn2+ transportation. FgGdt1-mCherry colocalizes with FgPmr1-GFP at the Golgi, and FgGDT1 exerts its biological function upstream of FgPMR1. Taken together, our results collectively demonstrate that the cis- and medial-Golgi-localized proteins FgGdt1 and FgPmr1 regulate Ca2+ and Mn2+ homeostasis of the Golgi apparatus, and this function is important in modulating the growth, development, DON biosynthesis and pathogenicity of F. graminearum.

The Small GTPase FgRab1 Plays Indispensable Roles in the Vegetative Growth, Vesicle Fusion, Autophagy and Pathogenicity of Fusarium graminearum.

Rab GTPases are key regulators of membrane and intracellular vesicle transports. However, the biological functions of FgRab1 are still unclear in the devastating wheat pathogen Fusarium graminearum. In this study, we generated constitutively active (CA) and dominant-negative (DN) forms of FgRAB1 from the wild-type PH-1 background for functional analyses. Phenotypic analyses of these mutants showed that FgRab1 is important for vegetative growth, cell wall integrity and hyphal branching. Compared to the PH-1 strain, the number of spores produced by the Fgrab1DN strain was significantly reduced, with obviously abnormal conidial morphology. The number of septa in the conidia of the Fgrab1DN mutant was fewer than that observed in the PH-1 conidia. Fgrab1DN was dramatically reduced in its ability to cause Fusarium head blight symptoms on wheat heads. GFP-FgRab1 was observed to partly localize to the Golgi apparatus, endoplasmic reticulum and Spitzenkörper. Furthermore, we found that FgRab1 inactivation blocks not only the transport of the v-SNARE protein FgSnc1 from the Golgi to the plasma membrane but also the fusion of endocytic vesicles with their target membranes and general autophagy. In summary, our results indicate that FgRab1 plays vital roles in vegetative growth, conidiogenesis, pathogenicity, autophagy, vesicle fusion and trafficking in F. graminearum.

Gender- and Age-Specific Relationships Between Phthalate Exposures and Obesity in Shanghai Adults.

Phthalate exposure has been reported to be associated with obesity (measured by body mass index [BMI]) and central obesity (measured by waist circumference [WC]). Yet, reported associations and the potential gender and age differences are inconsistent. We conducted a cross-sectional study involving 2330 participants in the fall of 2012. Urinary metabolites of ten phthalates were measured. Height, body weight, and waist circumference (WC) were measured using standardized methods. We performed logistic regression analyses to estimate the association between each urine phthalate metabolite (categorized into quartiles) and obesity and central obesity and conducted an additional, stratified analysis to explore the gender and age differences. In the overall study population, higher urinary levels of MMP, MEHHP, and MECPP were associated with increased ratios of central obesity. When stratifying by gender and central obesity, higher urinary levels of MMP, MEHHP, and MEOHP were associated with increased odds of central obesity in females, whereas MBzP was significantly associated inversely with central obesity in females. In males, it showed no significant P value for trend (P trend). When stratifying by age in females, higher urinary levels of MEHP, MEOHP, MEHHP, and MECPP were associated with increased odds of central obesity in women aged ≤45 years. In females aged >45 years, it showed no significant P trend. In conclusion, we found that association between phthalates and central obesity was stronger than between phthalates and obesity; association between phthalates and central obesity was stronger in females than in males and was stronger in younger females (aged ≤45 years) than in older females (aged >45 years).

[Cumulative risk assessment of phthalates exposure among infants and their mothers in Shanghai].

OBJECTIVE: To estimate the levels of phthalates exposure and cumulative risk assessment among infants and their mothers in Shanghai by phthalate monoesters inurine specimens. METHODS: To collect urine specimens and questionnaires from 152 pairs of infants and mothers in Shanghai and conduct a cross-sectional study. Ten phthalate monoesters were determined by ultra performance liquid chromatography coupled with tandem mass spectroscopy. To estimate the internal exposure of 6 phthalates by creatininemodel and performe cumulative risk assessment by hazard index( HI). RESULTS: The top five of the concentration level of phthalate monoesters in mothers' urine specimens were mono-isobutyl phthalate( Mi BP), mono-n-butyl phthalate( Mn BP), mono-ethyl phthalate( MEP), mono-methyl phthalate( MMP) and mono-2-ethylhexyl phthalate( MEHP), while in infants ', they were Mi BP, Mn BP, MEP, MEHP and mono-2-ethyl-5-carboxypentyl phthalate( MECPP). The estimated daily intake( EDI) was calculated according to the concentration of urinary phthalates monoesters. And the DEHP was at the highest level both in mothers and infants, the geometric means( GMs) reached to 6. 64 and 14. 02 µg/(kg·d). While others from high to low were Di BP, Dn BP, DEP, DMP and BBP respectively. HI was used to perform cumulative risk assessment. The proportion of HI over 1 among mothers was 5. 92%, and among infants was 32. 24%. CONCLUSION: There are some health risks of the phthalates exposure among mothers and infants in communities in Shanghai, and the cumulative risks of infants are higher than mothers.

Two acetyl-CoA synthetase isoenzymes are encoded by distinct genes in marine yeast Rhodosporidium diobovatum.

OBJECTIVES: Two genes encoding two acetyl-CoA synthetase (ACS) isoenzymes have been identified in the marine yeast Rhodosporidium diobovatum MCCC 2A00023. RESULTS: ACS1 encoded a polypeptide with a sequence of 578 amino acid residues, a predicted molecular weight of 63.73 kDa, and pI of 8.14, while the ACS2 encoded a polypeptide containing 676 amino acid residues with a deduced molecular mass of 75.61 kDa and a pI of 5.95. Biological activity of Acs1p and Acs2p was confirmed by heterologous expression in Escherichia coli. A 1.5-kb DNA fragment of the ACS1 gene and a 2.7-kb DNA fragment of the ACS2 gene were deleted using the RNA guide CRISPR-Cas9 system. The strain lacking ACS1 was unable to grow on acetate and ethanol media, while the ACS2 deletant was unable to grow on glucose medium. ACS1-ACS2 double mutants of R. diobovatum were non-viable. CONCLUSIONS: ACS isoenzymes are essential to the yeast metabolism, and other sources of ACSs cannot compensate for the lack of ACSs encoded by the two genes.

Phthalate monoesters in association with uterine leiomyomata in Shanghai.

Phthalates are ubiquitous environmental pollutants because of the broad use of plastics. We conducted a case-control study to determine whether uterine leiomyomata were related to exposure to phthalates. Urine specimens and questionnaires were collected from 61 cases and 61 age-matched controls. Nine phthalate monoesters were determined by ultra performance liquid chromatography coupled with tandem mass spectroscopy. Cases had significantly higher levels of creatinine-adjusted mono-iso-butyl phthalate (MiBP), mono-n-butyl phthalate (MnBP), mono-2-ethylhexyl phthalate (MEHP), mono-2-ethyl-5-oxohexyl phthalate, mono-2-ethyl-5-hydroxyhexyl phthalate (MEHHP), mono(2-ethyl-5-carboxypentyl) phthalate (MECPP), total di(2-ethylhexyl) phthalate metabolites (∑DEHPmet), and total dibutyl phthalate metabolites (∑DBP(met)) than controls. After adjusting for potential confounders, logistic regression analyses demonstrated that leiomyomata were positively associated with MiBP, MnBP, MEHP, MEHHP, MECPP, ∑DEHP(met), and ∑DBP(met). In summary, our data support the hypothesis that uterine leiomyomata are related to phthalate exposure.

Diselenide-Bridged Mesoporous Silica-Based Nanoplatform with a Triple ROS-Scavenging Effect for Intracerebral Hemorrhage Treatment.

Oxidative stress (OS) is a major mediator of secondary brain injury following intracerebral hemorrhage (ICH). Thus, antioxidant therapy is emerging as an attractive strategy to combat ICH. To achieve both reactive oxygen species (ROS) scavenging ability and on-demand drug release ability, we constructed a novel polydopamine (PDA)-coated diselenide-bridged mesoporous silica nanoparticle (DSeMSN) drug delivery system (PDA-DSeMSN). Edaravone (Eda) was blocked in the pores of DSeMSN by covering the pores with PDA as a gatekeeper. The drug maintained nearly "zero release" before reaching the lesion site, while in the ROS-enriched circumstances, the PDA shell went through degradation and the doped diselenide bonds broke up, triggering the disintegration of nanoparticles and leading to Eda release. Interestingly, the ROS-degradable property of the PDA shell and diselenide bond endowed the system with enhanced ROS-eliminating capacity. The synergistic effect of ROS-responsive drug delivery and ROS-scavenging PDA-DSeMSN showed efficient antioxidative and mitochondria protective performance without apparent toxicity in vitro. Importantly, PDA-DSeMSN@Eda through intravenous administration specifically accumulated in perihematomal sites and demonstrated robust neuroprotection in an ICH mouse model through antioxidative and antiapoptotic effects with high biological safety. Thus, the PDA-DSeMSN platform holds tremendous potential as an excellent carrier for on-demand delivery of drugs and provides a new and effective strategy for the clinical treatment of ICH.

Fenton-like nanoparticles capable of H2O2 self-supply and glutathione consumption for chemodynamic and chemotherapy of cancer.

Chemodynamic therapy (CDT) utilizing the Fenton reaction to convert hydrogen peroxide (H2O2) into cytotoxic hydroxyl radicals (˙OH) has recently drawn extensive interest in tumor treatment. However, the therapeutic efficiency of CDT often suffers from high concentrations of glutathione (GSH), insufficient endogenous H2O2 and inefficient Fenton activity. Herein, a GSH-depleting and H2O2 self-providing nanosystem that can efficiently load copper ions and doxorubicin (DOX) (MSN-Cu2+-DOX) to induce enhanced CDT and chemotherapy is proposed. The results show that MSN-Cu2+-DOX could release Cu2+ and DOX under acidic conditions. Particularly, both the released Cu2+ and Cu2+ in MSN-Cu2+-DOX are available for ˙OH production via a Fenton-like reaction for CDT. Meanwhile, Cu2+ undergoes a reduction to Cu+ by depleting overexpressed GSH, thereby enhancing CDT. Moreover, the released DOX could not only be used for chemotherapy, but also promote the generation of endogenous H2O2 to improve the efficiency of a Cu-based Fenton-like reaction. Resultantly, this nanosystem featuring Fenton-like activity, GSH consumption, H2O2 self-sufficiency and chemotherapy exhibits a great antitumor effect with a tumor inhibition ratio of 93.05%. Overall, this study provides a promising strategy to enhance CDT for effective tumor therapy.

Flexible Sono-Piezo Patch for Functional Sweat Gland Repair through Endogenous Microenvironmental Remodeling.

Remodeling the endogenous regenerative microenvironment in wounds is crucial for achieving scarless, functional tissue regeneration, especially the functional recovery of skin appendages such as sweat glands in burn patients. However, current approaches mostly rely on the use of exogenous materials or chemicals to stimulate cell proliferation and migration, while the remodeling of a pro-regenerative microenvironment remains challenging. Herein, we developed a flexible sono-piezo patch (fSPP) that aims to create an endogenous regenerative microenvironment to promote the repair of sweat glands in burn wounds. This patch, composed of multifunctional fibers with embedded piezoelectric nanoparticles, utilized low-intensity pulsed ultrasound (LIPUS) to activate electrical stimulation of the target tissue, resulting in enhanced pro-regenerative behaviors of niche tissues and cells, including peripheral nerves, fibroblasts, and vasculatures. We further demonstrated the effective wound healing and regeneration of functional sweat glands in burn injuries solely through such physical stimulation. This noninvasive and drug-free therapeutic approach holds significant potential for the clinical treatment of burn injuries.

FgAP1σ Is Critical for Vegetative Growth, Conidiation, Virulence, and DON Biosynthesis in Fusarium graminearum.

The AP1 complex is a highly conserved clathrin adaptor that plays important roles in regulating cargo protein sorting and intracellular vesicle trafficking in eukaryotes. However, the functions of the AP1 complex in the plant pathogenic fungi including the devastating wheat pathogen Fusarium graminearum are still unclear. In this study, we investigated the biological functions of FgAP1σ, a subunit of the AP1 complex in F. graminearum. Disruption of FgAP1σ causes seriously impaired fungal vegetative growth, conidiogenesis, sexual development, pathogenesis, and deoxynivalenol (DON) production. The ΔFgap1σ mutants were found to be less sensitive to KCl- and sorbitol-induced osmotic stresses but more sensitive to SDS-induced stress than the wild-type PH-1. Although the growth inhibition rate of the ΔFgap1σ mutants was not significantly changed under calcofluor white (CFW) and Congo red (CR) stresses, the protoplasts released from ΔFgap1σ hyphae were decreased compared with the wild-type PH-1, suggesting that FgAP1σ is necessary for cell wall integrity and osmotic stresses in F. graminearum. Subcellular localization assays showed that FgAP1σ was predominantly localized to endosomes and the Golgi apparatus. In addition, FgAP1ß-GFP, FgAP1γ-GFP, and FgAP1µ-GFP also localize to the Golgi apparatus. FgAP1ß interacts with FgAP1σ, FgAP1γ, and FgAP1µ, while FgAP1σ regulates the expression of FgAP1ß, FgAP1γ, and FgAP1µ in F. graminearum. Furthermore, the loss of FgAP1σ blocks the transportation of the v-SNARE protein FgSnc1 from the Golgi to the plasma membrane and delays the internalization of FM4-64 dye into the vacuole. Taken together, our results demonstrate that FgAP1σ plays vital roles in vegetative growth, conidiogenesis, sexual reproduction, DON production, pathogenicity, cell wall integrity, osmotic stress, exocytosis, and endocytosis in F. graminearum. These findings unveil the functions of the AP1 complex in filamentous fungi, most notably in F. graminearum, and lay solid foundations for effective prevention and control of Fusarium head blight (FHB).

Cell mechanical responses to subcellular perturbations generated by ultrasound and targeted microbubbles.

The inherently dynamic and anisotropic microenvironment of cells imposes not only global and slow physical stimulations on cells but also acute and local perturbations. However, cell mechanical responses to transient subcellular physical signals remain unclear. In this study, acoustically activated targeted microbubbles were used to exert mechanical perturbations to single cells. The cellular contractile force was sensed by elastic micropillar arrays, while the pillar deformations were imaged using brightfield high-speed video microscopy at a frame rate of 1k frames per second for the first 10s and then confocal fluorescence microscopy. Cell mechanical responses are accompanied by cell membrane integrity changes. Both processes are determined by the perturbation strength generated by microbubble volumetric oscillations. The instantaneous cellular traction force relaxation exhibits two distinct patterns, correlated with two cell fates (survival or permanent damage). The mathematical modeling unveils that force-induced actomyosin disassembly leads to gradual traction force relaxation in the first few seconds. The perturbation may also influence the far end subcellular regions from the microbubbles and may propagate into connected cells with attenuations and delays. This study carefully characterizes the cell mechanical responses to local perturbations induced by ultrasound and microbubbles, advancing our understanding of the fundamentals of cell mechano-sensing, -responsiveness, and -transduction. STATEMENT OF SIGNIFICANCE: Subcellular physical perturbations commonly exist but haven't been fully explored yet. The subcellular perturbation generated by ultrasound and targeted microbubbles covers a wide range of strength, from mild, intermediate to intense, providing a broad biomedical relevance. With µm2 spatial sensing ability and up to 1ms temporal resolution, we present spatiotemporal details of the instantaneous cellular contractile force changes followed by attenuated and delayed global responses. The correlation between the cell mechanical responses and cell fates highlights the important role of the instantaneous mechanical responses in the entire cellular reactive processes. Supported by mathematical modeling, our work provides new insights into the dynamics and mechanisms of cell mechanics.

Drainage Type Classification and Key Controlling Factors of Productivity for CBM Wells in the Zheng Zhuang Area, Southern Qinshui Basin, North China.

The production of coalbed methane (CBM) wells varies greatly in the Qinshui Basin, North China. Analyzing the primary factors controlling the CBM well productivity is essential to improve their development efficiency. Based on the geological conditions and production data of CBM wells in the Zheng zhuang area, the principal component analysis (PCA) method was used to classify the drainage types and screen the key factors influencing the production of gas and water. The drainage types of the CBM wells in the study area can be divided into four categories. The gas production shows an increasing trend with the increase of the comprehensive score of the PCA. The key controlling factors of productivity for CBM wells can be summarized by the gas-bearing property, permeability, groundwater fluid potential, and burial depth. The impact of burial depth on CBM well productivity is manifested in its control of gas content and permeability. The groundwater flows to a low fluid potential area, which leads to a high water production and a small pressure drop. The gas production shows a positive correlation with post-fracturing permeability. The gas content is a key factor for controlling the critical desorption pressure, critical gas production pressure, and pressure drop at the gas breakthrough point. High gas content is a prerequisite for the high productivity of CBM wells.

Phthalate exposure in association with the use of personal care products among general population from Shanghai.

Phthalates are used in a wide variety of personal care products (PCPs) as solubilizer, denaturant or color fixative. However, there are few studies referring the correlation between phthalate exposure and PCPs use among general population in China. In this study, ten metabolites of phthalates in spot urine samples (N = 500) were quantified using ultra-performance liquid chromatography tandem mass spectrometry. The frequency and duration of 12 types of PCPs were collected by questionnaire survey. The associations of phthalate metabolites and PCPs use were assessed by multivariable liner regression models. Median concentration of mono-benzylphthalate (MBzP) was significantly higher among frequent users of almost all PCPs. Low molecular weight phthalates (monomethyl phthalate (MMP), mono-n-butylphthalate (MnBP) and monoisobutylphthalate (MiBP)) were significantly lower among frequent user of some rinse-off PCPs (e.g., shampoo, facial cleanser, and body wash). Meanwhile, monoethylphthalate (MEP) was positive correlated with frequently use of facial moisturizer. Mono-2-ethylhexylphthalate (MEHP) and MBzP showed a significant positive association with frequently use of body lotion. Univariate linear analyses indicated a significant decreasing trend between urinary concentrations of MMP, MnBP, and the number of rinse-off PCPs being frequently used, and a significant increasing trend between urinary MBzP and the number of leave-on products being frequently used. These findings indicated that the use of some PCPs, especially leave-on PCPs, could be a potential source of exposure to some phthalates.

Trehalose Phosphate Synthase Complex-Mediated Regulation of Trehalose 6-Phosphate Homeostasis Is Critical for Development and Pathogenesis in Magnaporthe oryzae.

Trehalose biosynthesis pathway is a potential target for antifungal drug development, and trehalose 6-phosphate (T6P) accumulation is widely known to have toxic effects on cells. However, how organisms maintain a safe T6P level and cope with its cytotoxicity effects when accumulated have not been reported. Herein, we unveil the mechanism by which the rice blast fungus Magnaporthe oryzae avoids T6P accumulation and the genetic and physiological adjustments it undergoes to self-adjust the metabolite level when it is unavoidably accumulated. We found that T6P accumulation leads to defects in fugal development and pathogenicity. The accumulated T6P impairs cell wall assembly by disrupting actin organization. The disorganization of actin impairs the distribution of chitin synthases, thereby disrupting cell wall polymer distribution. Additionally, accumulation of T6P compromise energy metabolism. M. oryzae was able to overcome the effects of T6P accumulation by self-mutation of its MoTPS3 gene at two different mutation sites. We further show that mutation of MoTPS3 suppresses MoTps1 activity to reduce the intracellular level of T6P and partially restore ΔMotps2 defects. Overall, our results provide insights into the cytotoxicity effects of T6P accumulation and uncover a spontaneous mutation strategy to rebalance accumulated T6P in M. oryzae. IMPORTANCE M. oryzae, the causative agent of the rice blast disease, threatens rice production worldwide. Our results revealed that T6P accumulation, caused by the disruption of MoTPS2, has toxic effects on fugal development and pathogenesis in M. oryzae. The accumulated T6P impairs the distribution of cell wall polymers via actin organization and therefore disrupts cell wall structure. M. oryzae uses a spontaneous mutation to restore T6P cytotoxicity. Seven spontaneous mutation sites were found, and a mutation in MoTPS3 was further identified. The spontaneous mutation in MoTPS3 can partially rescue ΔMotps2 defects by suppressing MoTps1 activity to alleviate T6P cytotoxicity. This study provides clear evidence for better understanding of T6P cytotoxicity and how the fungus protects itself from T6P's toxic effects when it has accumulated to severely high levels.

Effects of extracellular matrix rigidity on sonoporation facilitated by targeted microbubbles: Bubble attachment, bubble dynamics, and cell membrane permeabilization.

In this study, we investigated the effects of extracellular matrix rigidity, an important physical property of microenvironments regulating cell morphology and functions, on sonoporation facilitated by targeted microbubbles, highlighting the role of microbubbles. We conducted mechanistic studies at the cellular level on physiologically relevant soft and rigid substrates. By developing a unique imaging strategy, we first resolved details of the 3D attachment configurations between targeted microbubbles and cell membrane. High-speed video microscopy then unveiled bubble dynamics driven by a single ultrasound pulse. Finally, we evaluated the cell membrane permeabilization using a small molecule model drug. Our results demonstrate that: (1) stronger targeted microbubble attachment was formed for cells cultured on the rigid substrate, while six different attachment configurations were revealed in total; (2) more violent bubble oscillation was observed for cells cultured on the rigid substrate, while one third of bubbles attached to cells on the soft substrate exhibited deformation shortly after ultrasound was turned off; (3) higher acoustic pressure was needed to permeabilize the cell membrane for cells on the soft substrate, while under the same ultrasound condition, acoustically-activated microbubbles generated larger pores as compared to cells cultured on the soft substrate. The current findings provide new insights to understand the underlying mechanisms of sonoporation in a physiologically relevant context and may be useful for the clinical translation of sonoporation.

Rosmarinic acid alleviates ethanol-induced lipid accumulation by repressing fatty acid biosynthesis.

Recent studies have demonstrated that rosmarinic acid is a valuable natural product for treatment of alcoholic liver disease. However, the mechanisms whereby rosmarinic acid improves alcoholic liver disease remain unclear. Here we performed experiments using a non-transformed mouse hepatocyte cell line (AML12). Oil-red O staining demonstrated that rosmarinic acid reduced ethanol-induced lipid accumulation. It was shown that rosmarinic acid prevented ethanol-induced elevation of the malondialdehyde level. We also found that rosmarinic acid inhibited ethanol-induced mRNA expression of tumor necrosis factor-α and interleukin 6. Metabolomics analysis revealed that rosmarinic acid ameliorated ethanol-induced fatty acid biosynthesis in the cytoplasm. In addition, palmitic acid was a candidate biomarker in cells exposed to ethanol or ethanol plus rosmarinic acid. Rosmarinic acid prevented the ethanol-induced increase in sorbitol that is a component of the polyol pathway. Moreover, we confirmed that rosmarinic acid attenuated ethanol-induced mRNA expression of fatty acid synthase, probably by modulating the AMPK/SREBP-1c pathway. Furthermore, rosmarinic acid prevented the ethanol-induced decrease in eight metabolites that are involved in mitochondrial metabolism, including glycine and succinic acid which are the components of carnitine synthesis. These results provide a crucial insight into the molecular mechanism of rosmarinic acid in alleviating ethanol-induced injury.