Search

The liver is the central organ for digestion and detoxification and has unique metabolic and regenerative capacities. The hepatobiliary system originates from the foregut endoderm, in which cells undergo multiple events of cell proliferation, migration, and differentiation to form the liver parenchyma and ductal system under the hierarchical regulation of transcription factors. Studies on liver development and diseases have revealed that SRY-related high-mobility group box 9 (SOX9) plays an important role in liver embryogenesis and the progression of hepatobiliary diseases. SOX9 is not only a master regulator of cell fate determination and tissue morphogenesis, but also regulates various biological features of cancer, including cancer stemness, invasion, and drug resistance, making SOX9 a potential biomarker for tumor prognosis and progression. This review systematically summarizes the latest findings of SOX9 in hepatobiliary development, homeostasis, and disease. We also highlight the value of SOX9 as a novel biomarker and potential target for the clinical treatment of major liver diseases.

Abstract Corn cob is a major waste mass-produced in corn agriculture. Corn cob hydrolysate containing xylose, arabinose, and glucose is the hydrolysis product of corn cob. Herein, a recombinant Escherichia coli strain BT-10 was constructed to transform corn cob hydrolysate into 1,2,4-butanetriol, a platform substance with diversified applications. To eliminate catabolite repression and enhance NADPH supply for alcohol dehydrogenase YqhD catalyzed 1,2,4-butanetriol generation, ptsG encoding glucose transporter EIICBGlc and pgi encoding phosphoglucose isomerase were deleted. With four heterologous enzymes including xylose dehydrogenase, xylonolactonase, xylonate dehydratase, α-ketoacid decarboxylase and endogenous YqhD, E. coli BT-10 can produce 36.63 g/L 1,2,4-butanetriol with a productivity of 1.14 g/[L·h] using xylose as substrate. When corn cob hydrolysate was used as the substrate, 43.4 g/L 1,2,4-butanetriol was generated with a productivity of 1.09 g/[L·h] and a yield of 0.9 mol/mol. With its desirable characteristics, E. coli BT-10 is a promising strain for commercial 1,2,4-butanetriol production.

In order to clarify the relationships in Rhododendron subg. Tsutsusi, micromorphological characters of leaf epidermis were examined for 37 species in Rhododendron subg. Tsutsusi using scanning electron microscopy (SEM), among which the relevant features of 29 species are reported for the first time. The results are as follows: (1) The stomatal apparatus are anomocytic and all on abaxial surface. (2) According to the micromorphological features of glands, trichome types and stomatal apparatus of leaf epidermis, subg. Tsutsusi can be divided into five types, i.e., R. simsii-type, R. mariae-type, R. indicum-type, R. saxatile-type and R. farrerae-type. (3) The plants in R. simsii-type have the similar morphological features, have no or discontinuous ringed striates around stomatal apparatus. (4) The plants in R. mariae-type do not have distinct T-pieces at the polar regions of guard cells but have multilayer ringed striates. (5) It is implied that the plants in R. indicum-type have the close affinity with subg. Therorhodion in having distinct T-pieces at the polar regions of guard cells. (6) R. saxatile-type differs from other types in its simple trichomes being virgate but without glands. (7) The leaf epidermis of R. farrerae-type has glands only. (8) The relationships between the closely related species are also discussed on the basis of leaf epidermal micromorphological features. For example, R. hypoblematosum and R. polyraphidoideum tend to be treated as separate species. Support the preservation of R. naamkwanense var. cryptonerve as a variety of R. naamkwanense, and do not support the R. adenanthum into R. tenue as a synonym. The results of this study show that micromorphological features of leaf epidermis are of significant value for species delimitation in subg. Tsutsusi.

Abstract The clinical benefit of tyrosine kinase inhibitors (TKIs)-based systemic therapy for advanced hepatocellular carcinoma (HCC) is limited due to drug resistance. Here, we uncover that lipid metabolism reprogramming mediated by unconventional prefoldin RPB5 interactor (URI) endows HCC with resistance to TKIs-induced ferroptosis. Mechanistically, URI directly interacts with TRIM28 and promotes p53 ubiquitination and degradation in a TRIM28-MDM2 dependent manner. Importantly, p53 binds to the promoter of stearoyl-CoA desaturase 1 (SCD1) and represses its transcription. High expression of URI is correlated with high level of SCD1 and their synergetic expression predicts poor prognosis and TKIs resistance in HCC. The combination of SCD1 inhibitor aramchol and deuterated sorafenib derivative donafenib displays promising anti-tumor effects in p53-wild type HCC patient-derived organoids and xenografted tumors. This combination therapy has potential clinical benefits for the patients with advanced HCC who have wild-type p53 and high levels of URI/SCD1.

Hollow microspheres as the filler material of syntactic foams have been adopted in extensive practical applications, where the physical parameters and their homogeneity have been proven to be critical factors during the design process, especially for high-specification scenarios. Based on double-emulsion droplet templates, hollow microspheres derived from microfluidics-enabled soft manufacturing have been validated to possess well-controlled morphology and composition with a much narrower size distribution and fewer defects compared to traditional production methods. However, for more stringent requirements, the innate density difference between the core–shell solution of the double-emulsion droplet template shall result in the wall thickness heterogeneity of the hollow microsphere, which will lead to unfavorable mechanical performance deviations. To clarify the specific mechanical response of microfluidics-derived hollow silica microspheres with varying eccentricities, a hybrid method combining experimental nanoindentation and a finite element method (FEM) simulation was proposed. The difference in eccentricity can determine the specific mechanical response of hollow microspheres during nanoindentation, including crack initiation and the evolution process, detailed fracture modes, load-bearing capacity, and energy dissipation capability, which should shed light on the necessity of optimizing the concentricity of double-emulsion droplets to improve the wall thickness homogeneity of hollow microspheres for better mechanical performance.

Context Ellagic acid (EA) is a phenolic constituent in certain fruits and has largely been recognized for its role as an antioxidant compound.Objective To evaluate the effect of EA on beryllium sulphate-induced splenic toxicity in rats.Materials and methods Male Sprague-Dawley rats were divided into four groups. The first group was used as control. Group 2 was exposed to BeSO4 (12 mg/kg, b.w.). Groups 3 and 4 were treated with EA (100 and 300 mg/kg, b.w.) daily for 6 weeks after exposing to BeSO4 (12 mg/kg, b.w.). Various biochemical and molecular biomarkers were assessed in blood and spleen.Results BeSO4-intoxicated rats showed significant higher WBC (6.74 ± 0.20 × 109/L vs. 11.02 ± 1.31 × 109/L, p

Abstract Cholangiocarcinoma (CCA) is an epithelial malignancy with a dismal prognosis owing to limited treatment options. Here, we identified several compound candidates against CCA using a high-throughput drug screen with approved or emerging oncology drugs, among which kinesin spindle protein (KSP) inhibitors showed potent cytotoxic effects on CCA cells. Treatment with KSP inhibitors SB743921 and ARRY520 caused significant tumor suppression in CCA xenograft models in vivo. Mechanistically, KSP inhibitors led to the formation of abnormal monopolar spindles, which further resulted in the mitotic arrest and cell death of CCA cells both in vivo and in vitro. KEGG pathway analysis of transcriptional data confirmed this finding. Moreover, our clinical data as well as the TCGA database showed KIF11 expression was abundant in most CCA tumor specimens and associated with poor outcomes of CCA patients. Our results demonstrate that the therapeutic regimen of KSP inhibitors could be a promising treatment strategy in CCA.

IntroductionCholangiocarcinoma (CCA) is a malignant tumor of the biliary epithelium with a poor prognosis. The lack of biomarkers to predict therapeutic response and prognosis is one of the major challenges for CCA treatment. Tertiary lymphoid structures (TLS) provide a local and pivotal microenvironment for tumor immune responses. The prognostic value and clinical relevance of TLS in CCA remain unclear. We aimed to explore the characteristics and clinical significance of TLS in CCA.MethodsWe investigated the prognostic value and clinical relevance of TLS in CCA using a surgery cohort containing 471 CCA patients (cohort 1) and an immunotherapy cohort containing 100 CCA patients (cohort 2). Hematoxylin and eosin (H&E) and immunohistochemical (IHC) staining were used to evaluate the maturity of TLS. Multiplex IHC (mIHC) was employed to characterize the composition of TLS.ResultsDifferent maturity of TLS were observed in CCA tissue sections. Strong staining of the four-gene signature including PAX5, TCL1A, TNFRSF13C, and CD79A were found in TLS regions. A high density of intra-tumoral TLS (T-score high) were significantly correlated with longer overall survival (OS) both in CCA cohort 1 (p = 0.002) and cohort 2 (p = 0.01), whereas a high density of peri-tumoral TLS (P-score high) were associated with shorter OS in these two cohorts (p = 0.003 and p = 0.03, respectively).ConclusionThe established four-gene signature efficiently identified the TLS in CCA tissues. The abundance and spatial distribution of TLS were significantly correlated with the prognosis and immune checkpoint inhibitors (ICIs) immunotherapy response of CCA patients. The presence of intra-tumoral TLS are positive prognostic factors for CCA, which provide a theoretical basis for the future diagnosis and treatment of CCA.

The effect of stand density on the soil bacterial community and diversity remains unclear. Spectrophotometry and full-length 16S rRNA sequences were used to determine the effects of planting density on soil physicochemical attributes and the diversity of soil bacterial and understory vegetation in a young Phoebe zhennan plantation at five densities. The findings showed that stand density had significant effects on the total nitrogen, ammonium nitrogen (NH4+-N), nitrate-nitrogen (NO3−-N), organic carbon, and the dominance and evenness of shrubs. Candidatus Udaeobacter and Candidatus Soilbacter were the two most common genera across the five stand densities. The density D5 (850 stems/hm2) demarcated from the others with a lower diversity of soil bacteria. Overall, the relatively low- and middle-density plantations were more conducive to complex and stable understory vegetation, bacterial communities, and soil nutrient cycles. The functional categories of the bacterial communities revealed a high proportion associated with chemoheterotrophy, aerobic chemoheterotrophy, and nitrogen fixation. Bacterial diversity and function were significantly influenced by soil pH, NH4+-N, NO3−-N, total phosphorus, and available phosphorus. However, there were no significant correlations between soil physicochemical attributes, understory vegetation, and bacterial diversity. Therefore, we speculated that the key drivers of the soil bacterial community were the soil physicochemical attributes and that stand density affected the soil bacterial community diversity by changing the soil physicochemical attributes. Overall, P. zhennan plantations with densities below 600 stems/hm2 were conducive to complex and stable soil bacterial communities and nutrient cycles.

Abstract NRF2 is the master transcriptional activator of cytoprotective genes and Kelch-like ECH-associated protein 1 (Keap1), a biosensor for electrophiles and oxidation, promotes NRF2 degradation in unstressed conditions. SQSTM1/p62, an oncogenic protein aberrantly accumulated in hepatocellular carcinoma (HCC), binds and sequestrates Keap1, leading to the prevention of NRF2 degradation. Here, we show that p15INK4b-related sequence/regulation of nuclear pre-mRNA domain-containing protein 1A (RPRD1A) is highly expressed in HCC tumors and correlated with aggressive clinicopathological features. RPRD1A competitively interacts with TRIM21, an E3 ubiquitin ligase of p62, resulting in the decrease of p62 ubiquitination and the increased sequestration for Keap1. Therefore, RPRD1A enhances the nuclear translocation of NRF2, which induces gene expression for counteracting oxidative stress, maintaining cancer cells survival, and promoting HCC development. Moreover, disturbing the redox homeostasis of cancer cells by genetic knockdown of RPRD1A sensitizes cancer cells to platinum-induced cell death. Our study reveals RPRD1A is involved in the oxidative stress defense program and highlights the therapeutic benefits of targeting pathways that support antioxidation.

Fluoride anion (F−) affects environmental, biological, and chemical processes significantly. Therefore, its detection has received increasing attention, and sensitive, effective, and convenient probes for F− detection need to be developed urgently. In this work, two perylene tetra-(alkoxycarbonyl) (PTAC) based colorimetric and ratiometric probes, P1 and P2, were developed for the detection of F−. The interactions between F− and these two probes were investigated by absorption, electrochemistry, 1H NMR, and density functional methods. Both the two probes were complexed with F− with a ratio of 1:1. The detection limits of P1 and P2 were 0.22 μM and 0.87 μM, respectively. It was worth noting that the absorption peak of P1 showed a 190 nm red shift when sensing F−, and P1 is the largest red shift value reported in F− probes based on PTAC derivatives. This phenomenon was resulted from the unique configuration and deprotonation of P1 that can promote the intramolecular charge transfer (ICT). This strategy provides an example for the development of other ion probes based on D-A type ICT mechanism.

Abstract 1-aminoperylene diimide/TiO2/MoS2 composite (NH2-PDI/TiO2/MoS2) with ordered structure was prepared by hydrothermal synthesis method. The composite was characterized by XRD, SEM, FTIR, XPS, BET, DRS, PL, EIS, Raman, photocurrent, and Mott-Schottky plots spectroscopy. The potential positions of the conduction and valence bands, and the band gap energy of the semiconductors were estimated. The composite exhibited higher photocatalytic activity compared with the mono-component systems. The apparent rate constants (k) were determined as 0.00616, 0.00352, 0.00738, 0.00517, 0.00752, and 0.00806 min−1 for TiO2, NH2-PDI, NH2-PDI/TiO2, MoS2, MoS2/TiO2, and NH2-PDI/TiO2/MoS2, respectively. The detection of radical scavengers confirmed that superoxide radicals, photogenerated holes, and photogenerated electrons were the main active substances for MB degradation. Between type II- heterojunction mechanism and Z-scheme mechanism, the latter could explain the enhanced photocatalytic activity of the composite better. The Z-scheme mechanism accumulates more electrons at CB level of NH2-PDI and hence generates more super oxide radicals.

The pH detection in the alkaline range is particularly important in many fields such as leather processing, waste water treatment, paper industry, and metal mining and finishing. Compared with traditional analysis methods such as colorimetric sensors and electrochemical sensors, the fluorescence and colorimetric probes for pH measurements have attracted much more attention due to their advantages of high sensitivity, excellent selectivity, noninvasiveness, low cost, fast response time, the possibility of continuously measurement, etc. However, there are few fluorescent probes fiting for alkaline pH monitoring. Acturally, the design and synthesis of them were more significant for new probes producing. In this study, the design, synthesis, and practical application of two novel fluorescent pH probes for alkaline pH assay were discussed. Both of the two probes were derived from perylene tetra-(alkoxycarbonyl). The red or blue shift of the absorption/fluorescence spectrum was caused by the introduction of electron donor amino or oxygen ring in the bay region. Due to electronic separation of the OH group from the electron-withdrawing core, the probes have high pKa values and cover the pH range from 8 to 12. They exist in either fluorescent acid form or non-fluorescent basic form. It was investigated that the amino substituent at bay region had a higher pKa value than O-heterocyclic annulated perylene, which showed that the adjustable pKa value could be achieved by the modification of electron withdrawing groups. The probes would have a wide use for testing strips measurements and monitoring pH changes in concrete.

The increase in food allergies and diabetes leads to the assumption that they are related. This study aimed to (1) verify the interaction between food allergy and diabetes and (2) explore the potential mechanisms by which food allergy promotes diabetes. Female BALB/c mice were grouped into a control group (CK), an ovalbumin-sensitized group (OVA), a diabetes group (STZ), and a diabetic allergic group (STZ + OVA) (Mice were modeled diabetes with STZ first, then were given OVA to model food allergies), and an allergic diabetic group (OVA + STZ) (Mice were modeled food allergies with OVA first, then were given STZ to model diabetes). The results showed that OVA + STZ mice exhibited a more serious Th2 humoral response, and they were more susceptible to diabetes. Furthermore, when the OVA + STZ mice were in the sensitized state, the intestinal barrier function was severely impaired, and mast cell activation was promoted. Moreover, we found that the effect of food allergy on diabetes is related to the inhibition of GLP-1 secretion and the up-regulation of the PI3K/Akt/mTOR/NF-κB P65 signaling pathway in the jejunum. Overall, our results suggest that food allergies have interactions with diabetes, which sheds new light on the importance of food allergies in diabetes.

The ketogenic diet with extremely high fat and very low carbohydrate levels is very popular in society today. Although it has beneficial effects on epilepsy and neurodegenerative diseases, how ketogenic diets impact host glucose and lipid metabolism and gut microbiota still needs further investigation.

Oxidative stress, which could be evoked by numerous inducements including mycotoxins like deoxynivalenol (DON), cause severe damages to organisms. Antioxidants are promising protectants against oxidative stress that could be applied in pharmaceutical, cosmetic, and food and feed industries. In this study, a thermostable and acidophilic superoxide dismutase (AaSOD) was used to develop an antioxidant product that can potentially protect organisms from oxidative stress related damages. The enzyme was successfully expressed as an extracelluar protein in Bacillus subtilis with a high yield. To obtain a feasible protocol for industrial production of AaSOD, the fermentation mediums that are commonly used for culturing B. subtilis were screened, the feasibility of expressing AaSOD without antibiotic as selection pressure was confirmed, and the effect of using lactose as an inducer instead of isopropyl-β-d-thiogalactoside (IPTG) was investigated. Batch fermentation was conducted to validate the optimized conditions for AaSOD production, and 6530 U mL-1 of SOD activity was obtained in the fermentation broth. The dry powder product of AaSOD with an activity of 22202 U g-1 was prepared by spray-drying and was administrated on zebrafish to test its function as a protectant against DON, and thus gained a significant redress of the reactive oxygen species (ROS) accumulation induced by DON. Taken together, this study provides a feasible protocol to prepare the AaSOD-based antioxidant product that is potentially applied in livestock industry.

Hepatocarcinogenesis is frequently accompanied by substantial metabolic reprogramming to maximize the growth and proliferation of cancer cells. In this study, we carried out a comprehensive study of metabolomics and lipidomics profiles combined with gene expression analysis to characterize the metabolic reprogramming in hepatocellular carcinoma (HCC). Compared with adjacent noncancerous liver tissue, the enhanced aerobic glycolysis and de novo lipogenesis (DNL) and the repressed urea cycle were underscored in HCC tissue. Furthermore, multiscale embedded correlation analysis was performed to construct differential correlation networks and reveal pathologically relevant molecule modules. The obtained hub nodes were further screened according to the maximum biochemical diversity and the least intraclass correlation. Finally, a panel of ornithine, FFA 18:1, PC O-32:1 and TG (18:1_17:1_18:2) was generated to achieve the prognostic risk stratification of HCC patients (p < 0.001 by log-rank test). Altogether, our findings suggest that the metabolic dysfunctions of HCC detected via metabolomics and lipidomics would contribute to a better understanding of clinical relevance of hepatic metabolic reprogramming and provide potential sources for the identification of therapeutic targets and the discovery of biomarkers.

Hepatocellular carcinoma (HCC) displays a high degree of metabolic and phenotypic heterogeneity and has dismal prognosis in most patients. Here, a gas chromatography–mass spectrometry (GC-MS)-based nontargeted metabolomics method was applied to analyze the metabolic profiling of 130 pairs of hepatocellular tumor tissues and matched adjacent noncancerous tissues from HCC patients. A total of 81 differential metabolites were identified by paired nonparametric test with false discovery rate correction to compare tumor tissues with adjacent noncancerous tissues. Results demonstrated that the metabolic reprogramming of HCC was mainly characterized by highly active glycolysis, enhanced fatty acid metabolism and inhibited tricarboxylic acid cycle, which satisfied the energy and biomass demands for tumor initiation and progression, meanwhile reducing apoptosis by counteracting oxidative stress. Risk stratification was performed based on the differential metabolites between tumor and adjacent noncancerous tissues by using nonnegative matrix factorization clustering. Three metabolic clusters displaying different characteristics were identified, and the cluster with higher levels of free fatty acids (FFAs) in tumors showed a worse prognosis. Finally, a metabolite classifier composed of six FFAs was further verified in a dependent sample set to have potential to define the patients with poor prognosis. Together, our results offered insights into the molecular pathological characteristics of HCC.

Xanthan gum is prone to thermal oxidative degradation, which limits its applications. However, conformational changes in xanthan gum and appropriate stabilizers may improve its thermal stability. Therefore, in this study, we aimed to establish a strategy to maintain the viscosity of xanthan gum during long-term storage at high temperatures. We modified the original strain used for xanthan gum production by genetic engineering and added stabilizers during the production process. The structure and thermal stability of the resulting xanthan gum samples were then determined. Pyruvyl deficiency, combined with the addition of sodium sulfite and glyoxal during the production process, was found to significantly improve the maintenance of viscosity. The apparent viscosity of the new xanthan gum solution remained above 100 mPa·s after being stored at 90 °C for 48 days. Fourier-transform infrared spectra and scanning electron microscopy images showed that pyruvate-free xanthan gum with added stabilizers had more extensive cross-linking than natural xanthan gum. In conclusion, these findings may contribute to the use of xanthan gum in applications that require high temperatures for a long period of time.

Metal rubber (MR) is an entangled fibrous functional material, and its mechanical properties are crucial for its applications; however, numerical constitutive models of MR for prediction and calculation are currently undeveloped. In this work, we provide a numerical constitutive model to express the mechanics of MR materials and develop an efficient finite elements method (FEM) to calculate the performance of MR components. We analyze the nonlinearity and anisotropy characteristics of MR during the deformation process. The elasticity matrix is adopted to express the nonlinearity and anisotropy of MR. An artificial neural network (ANN) model is built, trained, and tested to output the current elastic moduli for the elasticity matrix. Then, we combine the constitutive ANN model with the finite element method simulation to calculate the mechanics of the MR component. Finally, we perform a series of static and shock experiments and finite element simulations of an MR isolator. The results demonstrate the feasibility and accuracy of the numerical constitutive MR model. This work provides an efficient and convenient method for the design and analysis of MR components.

Chinese jujube has been valued for its medical properties. To examine the beneficial role of jujube in regulating the immune response, we investigated the anti-allergic effects of jujube extract and cyclic adenosine monophosphate (cAMP), a functional compound of jujube extract, using a mouse allergy model and rat basophilic leukemia (RBL)-2H3 mast cells. The investigations of the immunomodulatory effects of jujube extract on RBL-2H3 mast cell degranulation showed that the release of β-hexosaminidase (β-hex) was inhibited. In mice with ovalbumin (OVA)-induced allergic rhinitis, cAMP reduced the secretion of IgE and histamine in the plasma. cAMP regulated both Th1 and Th2 cytokines levels in the plasma. In addition, cAMP inhibited β-hex release and blocked extracellular Ca2+ influxes in RBL-2H3 cells. Our results demonstrate that jujube, as well as cAMP, suppressed cytokine production in the allergic response pathway, which resulted in prevention or alleviation of allergy symptoms.

Microfluidic technologies have enabled generation of exquisite multiple emulsion droplets, which have been used in many fields, including single-cell assays, micro-sized chemical reactions, and material syntheses. Electrical controlling is an important technique for droplet manipulation in microfluidic systems, but the dielectrophoretic behaviors of multiple emulsion droplets in electrical fields are rarely studied. Here, we report on the dielectrophoresis response of double emulsion droplets in AC electric fields in microfluidic channel. A core-shell model is utilized for analyzing the polarization of droplet interfaces and the overall dielectrophoresis (DEP) force. The water-in-oil-in-water droplets, generated by glass capillary devices, experience negative DEP at low field frequency. At high frequency, however, the polarity of DEP is tunable by adjusting droplet shell thickness or core conductivity. Then, the behavior of droplets with two inner cores is investigated, where the droplets undergo rotation before being repelled or attracted by the strong field area. This work should benefit a wide range of applications that require manipulation of double emulsion droplets by electric fields.

Supplementation of probiotics is a promising gut microbiota-targeted therapeutic method for hyperlipidemia and atherosclerosis. However, the selection of probiotic candidate strains is still empirical. Here, we obtained a human-derived strain, Lactobacillus mucosae A1, which was shown by metagenomic analysis to be promoted by a high-fiber diet and associated with the amelioration of host hyperlipidemia, and validated its effect on treating hyperlipidemia and atherosclerosis as well as changing structure of gut microbiota in ApoE-/- mice on a Western diet. L. mucosae A1 attenuated the severe lipid accumulation in serum, liver and aortic sinus of ApoE-/- mice on a Western diet, while it also reduced the serum lipopolysaccharide-binding protein content of mice, reflecting the improved metabolic endotoxemia. In addition, L. mucosae A1 shifted the gut microbiota structure of ApoE-/- mice on a Western diet, including recovering a few members of gut microbiota enhanced by the Western diet. This study not only suggests the potential of L. mucosae A1 to be a probiotic in the treatment of hyperlipidemia and atherosclerosis, but also highlights the advantage of such function-based rather than taxonomy-based strategies for the selection of candidate strains for the next generation probiotics.

The gallium-based eutectic liquid metal alloys exhibit unique properties of deformability, excellent electrical conductivity and low vapour pressure. The liquid metal-based circuits’ element or actuator have drawn considerable attention in stretchable electronics and microelectromechanical (MEMS) actuators. Yet, the motion of the liquid metal within the electrolyte needs to be precisely regulated to satisfy application requirements. Herein, we investigated the locomotion of liquid metal within the alkaline aqueous solution under electrostatic actuation. The relationship between the travelling speed of the liquid metal slug and the relative influential parameters, such as the voltage amplitude and frequencies of the applied electric field, electrolyte concentration, electrodes distance and the liquid metal volume, were experimentally characterized. A travelling speed up to 20.33 mm/s was obtained at the applied voltage of 4 Vpp at 150 Hz at 6 V DC offset. Finally, the frequency-dependent liquid metal marble movements were demonstrated, namely oscillation and forward locomotion while oscillating. The oscillation frequency was determined by the frequency of the applied alternate current (AC) signal. The remarkable transportation and oscillating characteristic of the liquid metal marble under the electrostatic actuation may present potentials towards the development of flexible electronics and reconfigurable structures.

A two-phase flow axisymmetric numerical model was proposed to understand liquid metal droplet formation in a co-flowing capillary microfluidics device based on a phase field model. The droplet detachment processes were observed in the experiment and are in good agreement with the simulation method. The effects of the viscosities and flowrates of the continuous phase fluid, interfacial tension as well as the wetting property of the metallic needle against the bulk liquid metal on the droplet formation and production rate were numerically investigated. It was found that the droplet diameter decreased with the increment of the viscosities and flowrates of the outer phase carrier fluid. The dispersed phase fluid with high interfacial tension tended to prolong the time for equilibrium between the viscous drag force and interfacial tension on the liquid–liquid fluid surface, delaying the droplet to be pinched off from the capillary orifice and causing large droplet diameter. Finally, the wetting performance of the metallic needle against the liquid metal was explored. The result indicate that the droplet diameter became less dependent on the contact angle while the size distribution of the liquid metal droplet was affected by their wetting performance. A more hydrophilic wetting performance were expected to prepare liquid metal droplet with more monodispersity. The numerical model and simulation results provide the feasibility of predicting the droplet formation with a high surface tension in a glass capillary microfluidic device.

In this paper we present a novel microfluidic approach for continuous, rapid and switchable particle concentration, using induced-charge electroosmosis (ICEO) in 3D electrode layouts. Field-effect control on non-linear electroosmosis in the transverse direction greatly facilitates a selective concentration of biological yeast cells from a straight main microchannel into one of the three downstream branch channels in our microfluidic device. For the geometry configuration of 3D driving electrode plates on sidewalls and a 2D planar gate electrode strip on the channel bottom surface, we briefly describe the underlying physics of an ICEO-based particle flow-focusing method, and provide relevant simulation results to show how gate voltage amplitude can be used to guide the motion trajectory of the concentrated particle stream. With a relatively simple geometrical configuration, the proposed microfluidic device provides new possibilities to controllably concentrate micro/nanoparticles in continuous flow by using ICEO, and is suitable for a high-throughput front-end cell concentrator interfacing with various downstream biosensors.

As one of the most commonly used eukaryotic recombinant protein expression systems, P. pastoris relies heavily on the AOX1 promoter (PAOX1), which is strongly induced by methanol but strictly repressed by glycerol and glucose. However, the complicated signaling pathways involved in PAOX1 regulation when supplemented with different carbon sources are poorly understood. Here we constructed a kinase deletion library in P. pastoris and identified 27 mutants which showed peculiar phenotypes in cell growth or PAOX1 regulation. We analyzed both annotations and possible functions of these 27 targets, and then focused on the MAP kinase Hog1. In order to locate its potential downstream components, we performed the phosphoproteome analysis on glycerol cultured WT and Δhog1 strains and identified 157 differentially phosphorylated proteins. Our results identified important kinases involved in P. pastoris cell growth and PAOX1 regulation, which could serve as valuable targets for further mechanistic studies.

In this study, we make use of the AC field-effect flow control on induced-charge electroosmosis (ICEO), to develop an electrokinetic micromixer with 3D electrode layouts, greatly enhancing the device performance compared to its 2D counterpart of coplanar metal strips. A biased AC voltage wave applied to the central gate terminal, i.e., AC field-effect control, endows flow field-effect-transistor of ICEO the capability to produce arbitrary symmetry breaking in the transverse electrokinetic vortex flow pattern, which makes it fascinating for microfluidic mixing. Using the Debye-Huckel approximation, a mathematical model is established to test the feasibility of the new device design in stirring nanoparticle samples carried by co-flowing laminar streams. The effect of various experimental parameters on constructing a viable micromixer is investigated, and an integrated microdevice with a series of gate electrode bars disposed along the centerline of the channel bottom surface is proposed for realizing high-flux mixing. Our physical demonstration on field-effect nonlinear electroosmosis control in 3D electrode configurations provides useful guidelines for electroconvective manipulation of nanoscale objects in modern microfluidic systems.

We present herein a novel method of bipolar field-effect control on DC electroosmosis (DCEO) from a physical point of view, in the context of an intelligent and robust operation tool for stratified laminar streams in microscale systems. In this unique design of the DC flow field-effect-transistor (DC-FFET), a pair of face-to-face external gate terminals are imposed with opposite gate-voltage polarities. Diffuse-charge dynamics induces heteropolar Debye screening charge within the diffuse double layer adjacent to the face-to-face oppositely-polarized gates, respectively. A background electric field is applied across the source-drain terminal and forces the face-to-face counterionic charge of reversed polarities into induced-charge electroosmotic (ICEO) vortex flow in the lateral direction. The chaotic turbulence of the transverse ICEO whirlpool interacts actively with the conventional plug flow of DCEO, giving rise to twisted streamlines for simultaneous DCEO pumping and ICEO mixing of fluid samples along the channel length direction. A mathematical model in thin-layer approximation and the low-voltage limit is subsequently established to test the feasibility of the bipolar DC-FFET configuration in electrokinetic manipulation of fluids at the micrometer dimension. According to our simulation analysis, an integrated device design with two sets of side-by-side, but upside-down gate electrode pair exhibits outstanding performance in electroconvective pumping and mixing even without any externally-applied pressure difference. Moreover, a paradigm of a microdevice for fully electrokinetics-driven analyte treatment is established with an array of reversed bipolar gate-terminal pairs arranged on top of the dielectric membrane along the channel length direction, from which we can obtain almost a perfect liquid mixture by using a smaller magnitude of gate voltages for causing less detrimental effects at a small Dukhin number. Sustained by theoretical analysis, our physical demonstration on bipolar field-effect flow control for the microfluidic device of dual functionalities in simultaneous electroconvective pumping and mixing holds great potential in the development of fully-automated liquid-phase actuators in modern microfluidic systems.

Improvement of phosphorus circulation in the soil is necessary to enhance phosphorus availability to plants. Phosphorus circulation activity is an index of soil’s ability to supply soluble phosphorus from organic phosphorus in the soil solution. To understand the relationship among phosphorus circulation activity; bacterial biomass; pH; and Fe, Al, and Ca concentrations (described as mineral concentration in this paper) in agricultural soil, 232 soil samples from various agricultural fields were collected and analyzed. A weak relationship between phosphorus circulation activity and bacterial biomass was observed in all soil samples (R2 = 0.25), and this relationship became significantly stronger at near-neutral pH (6.0–7.3; R2 = 0.67). No relationship between phosphorus circulation activity and bacterial biomass was observed at acidic (pH < 6.0) or alkaline (pH > 7.3) pH. A negative correlation between Fe and Al concentrations and phosphorus circulation activity was observed at acidic pH (R2 = 0.72 and 0.73, respectively), as well as for Ca at alkaline pH (R2 = 0.64). Therefore, bacterial biomass, pH, and mineral concentration should be considered together for activation of phosphorus circulation activity in the soil. A relationship model was proposed based on the effects of bacterial biomass and mineral concentration on phosphorus circulation activity. The suitable conditions of bacterial biomass, pH, and mineral concentration for phosphorus circulation activity could be estimated from the relationship model.

High-fat diet (HFD)-induced obesity is associated with insulin resistance, which may affect brain synaptic plasticity through impairment of insulin-sensitive processes underlying neuronal survival, learning, and memory. The experimental model consisted of 3 month-old C57BL/6J mice fed either a normal chow diet (control group) or a HFD (60% of calorie from fat; HFD group) for 12 weeks. This model was characterized as a function of time in terms of body weight, fasting blood glucose and insulin levels, HOMA-IR values, and plasma triglycerides. IRS-1/Akt pathway was assessed in primary hepatocytes and brain homogenates. The effect of HFD in brain was assessed by electrophysiology, input/output responses and long-term potentiation. HFD-fed mice exhibited a significant increase in body weight, higher fasting glucose- and insulin levels in plasma, lower glucose tolerance, and higher HOMA-IR values. In liver, HFD elicited (a) a significant decrease of insulin receptor substrate (IRS-1) phosphorylation on Tyr608 and increase of Ser307 phosphorylation, indicative of IRS-1 inactivation; (b) these changes were accompanied by inflammatory responses in terms of increases in the expression of NFκB and iNOS and activation of the MAP kinases p38 and JNK; (c) primary hepatocytes from mice fed a HFD showed decreased cellular oxygen consumption rates (indicative of mitochondrial functional impairment); this can be ascribed partly to a decreased expression of PGC1α and mitochondrial biogenesis. In brain, HFD feeding elicited (a) an inactivation of the IRS-1 and, consequentially, (b) a decreased expression and plasma membrane localization of the insulin-sensitive neuronal glucose transporters GLUT3/GLUT4; (c) a suppression of the ERK/CREB pathway, and (d) a substantial decrease in long-term potentiation in the CA1 region of hippocampus (indicative of impaired synaptic plasticity). It may be surmised that 12 weeks fed with HFD induce a systemic insulin resistance that impacts profoundly on brain activity, i.e., synaptic plasticity.

BACKGROUND: Various Pseudomonas strains can use L-lactate as their sole carbon source for growth. However, the L-lactate-utilizing enzymes in Pseudomonas have never been identified and further studied. METHODOLOGY/PRINCIPAL FINDINGS: An NAD-independent L-lactate dehydrogenase (L-iLDH) was purified from the membrane fraction of Pseudomonas stutzeri SDM. The enzyme catalyzes the oxidation of L-lactate to pyruvate by using FMN as cofactor. After cloning its encoding gene (lldD), L-iLDH was successfully expressed, purified from a recombinant Escherichia coli strain, and characterized. An lldD mutant of P. stutzeri SDM was constructed by gene knockout technology. This mutant was unable to grow on L-lactate, but retained the ability to grow on pyruvate. CONCLUSIONS/SIGNIFICANCE: It is proposed that L-iLDH plays an indispensable function in Pseudomonas L-lactate utilization by catalyzing the conversion of L-lactate into pyruvate.

BACKGROUND: Bacterial GreA is an indispensable factor in the RNA polymerase elongation complex. It plays multiple roles in transcriptional elongation, and may be implicated in resistance to various stresses. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we show that Escherichia coli GreA inhibits aggregation of several substrate proteins under heat shock condition. GreA can also effectively promote the refolding of denatured proteins. These facts reveal that GreA has chaperone activity. Distinct from many molecular chaperones, GreA does not form stable complexes with unfolded substrates. GreA overexpression confers the host cells with enhanced resistance to heat shock and oxidative stress. Moreover, GreA expression in the greA/greB double mutant could suppress the temperature-sensitive phenotype, and dramatically alleviate the in vivo protein aggregation. The results suggest that bacterial GreA may act as chaperone in vivo. CONCLUSIONS/SIGNIFICANCE: These results suggest that GreA, in addition to its function as a transcription factor, is involved in protection of cellular proteins against aggregation.