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Piezo-driven rotator is convenient for tilted magnetic field experiments due to its precise angle control. However, the rotator itself and the sample mounted on it are difficult to be cooled down because of extra heat leaks and presumably bad thermal contacts from the piezo. Here, we report a piezo-driven sample rotation system designed for ultra-low temperature environment. The sample, as well as the rotating sample holder, can be cooled to as low as 25 mK by customized thermal links and thermal contacts. More importantly, the electron temperature in the electrical transport measurements can also be cooled down to 25 mK with the help of home-made filters. To demonstrate the application of our rotation system at ultra-low electron temperature, a measurement revealing tilt-induced localization and delocalization in the second Landau level of two-dimensional electron gas is provided., Comment: 22 pages, 6 figures

Fiber-reinforced composite laminates (FRCL) is susceptible to the external impacting. Understanding the crack propagation and structural mechanical properties of the damaged FRCL under low-velocity oblique impact is of great value in practical application. A new analytical dynamic model is developed in this work to research the dynamic response and damage property of FRCL under oblique impacting. The displacement field and strain–displacement relations of the FRCL are established by utilizing higher-order shear plate theory. The matrix damage and fiber rupture in FRCL under oblique impacting are captured by an internal variable-based continuum damage constitutive relation. To accurately predict the oblique impacting force, an analytical dynamic impacting model is proposed basing on a developed contact model, where normal and tangential contact is coupled and solved simultaneously. The whole initial boundary value problem is iteratively solved by synthetically using finite differential method and Newmark- $$\beta $$ method. The solving convergence and accuracy of the model is demonstrated and validated. Simulations show that the matrix damage is more easily to appear in FRCL under shear force due to oblique contact when under oblique impacting, and the damage profile is different from normal impacting. The dynamic responses of the FRCL plate under oblique impacting differ also greatly from normal impacting. The current research provides a theoretical basis for FRCL design and its engineering application when under low-velocity impacting.

Numerous studies showed that mesenchymal stem cells derived exosome (MSC-Ex) markedly enhanced tissue regeneration, however, the issue of whether MSC-Ex could control stem cells expansion after a regenerative response to prevent tissue from overcrowding and dysplasia remains to be established. Herein, we found that human umbilical cord MSC (hucMSC)-exosomal14-3-3ζ mediated the binding of YAP and p-LATS by forming a complex to promote the phosphorylation of YAP, which orchestrate exosomal Wnt4 signal in cutaneous regeneration. First, we assessed deep second-degree burn rats treated with hucMSC-Ex and discovered that hucMSC-Ex promoting self-regulation of Wnt/β-catenin signaling at the remodeling phase of cutaneous regeneration. HucMSC-Ex restricted excessive skin cell expansion and collagen deposition at 4 weeks. Under high cell density conditions, hucMSC-Ex inhibited Wnt/β-catenin signaling through induction of YAP phosphorylation. Second, hucMSC-Ex proteomic analysis revealed that 14-3-3 proteins could be transported by exosome. Using gain- and loss-of-function studies, our results showed that hucMSC-exosomal 14-3-3ζ controlled YAP activities and phosphorylation at Ser127 site, and were required for the binding of YAP and p-LATS. Further studies revealed that 14-3-3ζ recruited YAP and p-LATS to form a complex under high cells density status and 14-3-3ζ other than YAP or p-LATS was the key regulatory molecule of this complex. These findings collectively indicate that hucMSC-Ex functions not only as an “accelerator” of the Wnt/β-catenin signal to repair damaged skin tissue but also as a “brake” of the signal by modulating YAP to orchestrate controlled cutaneous regeneration.

Novel and non-invasive biomarkers are urgently needed for early detection of cancer. Exosomes are nano-sized particles released by cells and contain various bioactive molecules including proteins, DNA, mRNAs, and non-coding RNAs. Increasing evidence suggests that exosomes play critical roles in tumorigenesis, tumor growth, metastasis, and therapy resistance. Exosomes could be readily accessible in nearly all the body fluids. The altered production of exosomes and aberrant expression of exosomal contents could reflect the pathological state of the body, indicating that exosomes and exosomal contents can be utilized as novel cancer biomarkers. Herein, we review the basic properties of exosomes, the functional roles of exosomes in cancer, and the methods of detecting exosomes and exosomal contents. In particular, we highlight the clinical values of exosomal non-coding RNAs in cancer diagnosis and prognosis.

This review article summarizes the requirement of low temperature conditions in existing experimental approaches to quantum computation and quantum simulation., 23 pages, 4 figures, 2 tables

The $\ensuremath{\nu}=0$ quantum Hall state of bilayer graphene is a fertile playground to realize many-body ground states with various broken symmetries. Here we report the experimental observations of a previously unreported metallic phase. The metallic phase resides in the phase space between the previously identified layer polarized state at large transverse electric field and the canted antiferromagnetic state at small transverse electric field. We also report temperature dependence studies of the quantum spin Hall state of $\ensuremath{\nu}=0$. Complex nonmonotonic behavior reveals concomitant bulk and edge conductions and excitations. These results provide a timely experimental update to understand the rich physics of the $\ensuremath{\nu}=0$ state.

A novel adsorbent is described for magnetic solid-phase extraction (MSPE) of the aflatoxins AFB1 and AFB2 (AFBs). Magnetic agarose microspheres (MAMs) were functionalized with an aptamer to bind the AFBs which then were quantified by HPLC and on-line post-column photochemical derivatization with fluorescence detection. Streptavidin-conjugated MAMs were synthesized first by a highly reproducible strategy. They possess strong magnetism and high surface area. The MAMs were characterized by transmission electron microscopy, scanning electron microscopy, optical microscopy, laser diffraction particle size analyzer, Fourier transform infrared spectrometry, vibrating sample magnetometry and laser scanning confocal microscopy. Then, the AFB-aptamers were immobilized on MAMs through biotin–streptavidin interaction. Finally, the MSPE is performed by suspending the aptamer-modified MAMs in the sample. They are then collected by an external magnetic field and the AFBs are eluted with methanol/buffer (20:80). Several parameters affecting the coupling, capturing and eluting efficiency were optimized. Under the optimized conditions, the method is fast, has good linearity, high selectivity, and sensitivity. The LODs are 25 pg·mL−1 for AFB1 and 10 pg·mL−1 for AFB2. The binding capacity is 350 ± 8 ng·g−1 for AFB1 and 384 ± 8 ng·g−1 for AFB2, and the precision of the assay is

A novel adsorbent is described for magnetic solid-phase extraction (MSPE) of the aflatoxins AFB

Gastric cancer tissue-derived MSC-like cells (GC-MSC) share similar characteristics to bone marrow MSC (BM-MSC); however, the phenotypical and functional differences and the molecular mechanism of transition between the two cell types remain unclear. Compared to BM-MSC, GC-MSC exhibited the classic phenotype of reactive stroma cells, a stronger gastric cancer promoting capacity and lower expression of miR-155-5p. Inhibition of miR-155-5p by transfecting miRNA inhibitor induced a phenotypical and functional transition of BM-MSC into GC-MSC-like cells, and the reverse experiment deprived GC-MSC of tumor-promoting phenotype and function. NF-kappa B p65 (NF-κB p65) and inhibitor of NF-kappa B kinase subunit epsilon (IKBKE/IKKε) were identified as targets of miR-155-5p and important for miRNA inhibitor activating NF-κB p65 in the transition. Inactivation of NF-κB by pyrrolidine dithiocarbamic acid (PDTC) significantly blocked the effect of miR-155-5p inhibitor on BM-MSC. IKBKE, NF-κB p65 and phospho-NF-κB p65 proteins were highly enriched in MSC-like cells of gastric cancer tissues, and the latter two were correlated with the pathological progression of gastric cancer. In GC-MSC, the expression of miR-155-5p was downregulated and NF-κB p65 protein was increased and activated. NF-κB inactivation by PDTC or knockdown of its downstream cytokines reversed the phenotype and function of GC-MSC. Taken together, our findings revealed that miR-155-5p downregulation induces BM-MSC to acquire a GC-MSC-like phenotype and function depending on NF-κB p65 activation, which suggests a novel mechanism underlying the cancer associated MSC remodeling in the tumor microenvironment and offers an effective target and approach for gastric cancer therapy.

As a natural health supplement, 3,3′-diindolylmethane (DIM) is proposed as a preventive and chemotherapeutic agent for cancer by inhibiting cell proliferation and inducing cell apoptosis. However, we found that in contrary to high level of DIM (30 μM), low level of DIM (1 μM and 10 μM) obviously promoted gastric cancer cell growth and migration. In addition, we found that low level of DIM increased the expression of stemness factors and enhanced the pluripotency of gastric cancer cells. Low level of DIM promoted gastric cancer progression by inducing the PORCN-dependent secretion of Wnt4 and the activation of β-catenin signaling. Wnt4 knockdown reversed the effects of low level of DIM on gastric cancer cells. The results of in vivo studies showed that gastric cancer cells treated with low level of DIM (1 μM) grew faster and expressed higher level of Wnt4 than control cells. Taken together, our findings indicate that low level of DIM activates autocrine Wnt4 signaling to enhance the progression of gastric cancer, which may suggest an adverse aspect of DIM in cancer therapy. Our findings will provide a new aspect for the safety of DIM in its clinical application.

Cooling to see the effects of disorder In sufficiently strong external magnetic fields, thin superconducting films typically become insulating. The presence of disorder can affect this phase transition. Theorists have proposed that disorder can cause the so-called Griffiths singularity, where the behavior of the system is determined by a small number of superconducting islands that form above the critical magnetic field. Xing et al. observed a signature of such a singularity in thin films of gallium by analyzing transport data taken at very low temperatures (see the Perspective by Markovic). In this regime, thermal fluctuations were not strong enough to homogenize the system, which allowed the rare islands to form. Science , this issue p. 542 ; see also p. 509

Table S3. Sequences of miR-20a mimics and inhibitor. (DOCX 18 kb)

Among the various geometries to study the fractional quantum Hall effect in two dimensional electron gas, the Corbino disc owns the advantage to probe the bulk properties directly. In this work we explore the influence of in-plane electric fields on the stability of the 5/2 fractional quantum Hall state realized in Corbino geometry. The effect of weak electric fields is investigated at ultra-low temperatures in order to compare with a theoretical proposal of enhanced Pfaffian state under weak electric fields.

Table S2. Primer sequences and amplified fragment products. (DOCX 19 kb)

Table S1. Sequences of TRIM3-siRNA. (DOCX 18 kb)

Two-dimensional (2D) transition metal dichalcogenides (TMDs) have a range of unique physics properties and could be used in the development of electronics, photonics, spintronics, and quantum computing devices. The mechanical exfoliation technique of microsize TMD flakes has attracted particular interest due to its simplicity and cost effectiveness. However, for most applications, large-area and high-quality films are preferred. Furthermore, when the thickness of crystalline films is down to the 2D limit (monolayer), exotic properties can be expected due to the quantum confinement and symmetry breaking. In this paper, we have successfully prepared macro-size atomically flat monolayer NbSe

Exosomes are critically involved in tumor growth, metastasis, and therapy resistance. Exosomes have the potential to be utilized as cancer biomarkers. In this study, we aimed to explore the roles and clinical values of exosomal miRNAs in gastric cancer. We found that the concentration of exosomes was significantly higher in the serum of gastric cancer patients and the culture supernatants of gastric cancer cells than that in healthy volunteers and gastric mucosa epithelial cells. In particular, miR-423-5p was elevated in the serum exosomes of gastric cancer patients, and the level of exosomal miR-423-5p was remarkably correlated with lymph node metastasis. High level of exosomal miR-423-5p was associated with poor outcome in gastric cancer patients. MiR-423-5p enriched exosomes could be internalized into gastric cancer cells, which enhanced cell proliferation and migration both in vitro and in vivo. Mechanistically, miR-423-5p inhibited the expression of suppressor of fused protein (SUFU) to enhance the proliferation and migration of gastric cancer cells. The expression levels of SUFU were significantly decreased in gastric cancer cells and the tumor tissues of gastric cancer patients. Taken together, our findings indicate that exosomes could deliver miR-423-5p to promote cancer growth and metastasis and serum exosomal miR-423-5p may serve as a potential marker for gastric cancer diagnosis and prognosis.

Double horse-head pumping unit, being one of the most classical mechanical equipment, has high efficiency and good balance ability during the oil extraction owing to its horse-head structure connecting with the rod by the steel wire rope. But its characteristic of energy consumption reduction is limited because of the motor torque fluctuation and negative torque appearing while the pumping unit is working in the upstroke and downstroke. The compound balance design is applied to the double horse-head pumping unit by the crank balance and walking beam balance, which is completed by the equal energy principle during the up and down circulation of the oil suction unit. The finite element model of the whole equipment is built, and the simulation analysis is completed by the software ADAMS, under the conditions of the compound balance and that of the crank balance. The output torque of the crank, the forces from the back horse-head rope, and the connection pin are calculated. From the viewpoint of system design to compare with the traditional crank balance pumping unit, the compound balance design can reduce the torque fluctuation greatly, decrease the forces of steel wire rope connecting with the back horse-head, and get rid of the structure problems from the traditional pumping unit. The stress test of the double horse-head pumping unit designed by the compound balance method is completed in the oilfields. It has proved the correctness and reasonability of the compound balance design. The methodology of the compound balance design is helpful in improving the work efficiency and reliability and bringing about better abilities of energy consumption reduction for the pumping unit during its work circulation.

Background & Aims The mechanisms of glycogen synthase kinase-3 (GSK-3)-mediated cytoprotection during liver ischemia/reperfusion (I/R) remain controversial, particularly in older organs. This study explores the role and potential mechanisms of GSK-3 in young and aging livers. Methods A rodent partial warm I/R model was used to evaluate the therapeutic potential of GSK-3 modulation during hepatic I/R in young and aging Sprague-Dawley rats. Results GSK-3 inhibition through IPC or SB216763 (SB21) preconditioning protected young rats from I/R-induced liver injury. This protection was absent in old animals but could be restored by glucose infusion prior to the I/R insult. The protection conferred by GSK-3 inhibition depended on mitochondrial metabolism regulation. Indeed, the inhibition of GSK-3 suppressed mitochondrial permeability transition pore (MPTP) opening, triggering mitohormesis in young animals, whereas insufficient fuel suppressed mitochondrial metabolism and inactivated the GSK-3-related protection in old animals. SB21 and glucose reactivated the mitochondrial F 0 F 1 -ATPase and subsequent protective cascades in the senescent liver. These effects were antagonized by an ATPase inhibitor and by an MPTP opener. Conclusions The protection conferred by GSK-3 inhibition during hepatic I/R insult is energy dependent, particularly in senescent livers. These findings demonstrate a key role for GSK-3-related mitochondrial energy homeostasis, which may shed new light on the clinical use of GSK-3 inhibitors to protect liver function in surgical settings, particularly for older patients.

Liver regeneration after partial hepatectomy (PH) is a synchronized process that is precisely controlled by system-wide transcriptional regulatory networks. To clarify the transcriptional changes and regulatory networks that involve transcription factors (TFs) and their target genes during the priming phase, an advanced mouse oligonucleotide array-based transcription factor assay (MOUSE OATFA), mRNA microarray analysis, bioinformatic analysis and ChIP-on-chip experiments were used. A total of 774 genes were upregulated or downregulated in PH liver samples compared with the sham operation (SH) group. Seventeen TFs showed significant changes in activity in the regenerating livers, some of which have not been extensively studied in previous reports, including upstream stimulatory transcription factor 1 (USF1). The TF signatures from MOUSE OATFA were combined with mRNA expression profiles and ChIP-on-chip analyses to construct experimental transcriptional regulatory networks in regenerating livers. USF1-centered regulatory networks were further confirmed by ChIP assays, revealing some of its target genes and novel coregulatory networks. The combination of MOUSE OATFA with transcriptome profiling and bioinformatic analysis represents a novel paradigm for the comprehensive prediction of transcriptional coregulatory networks during the early phase of liver regeneration.

Our goal was to gain a better understanding of the contribution of the burial of polar groups and their hydrogen bonds to the conformational stability of proteins. We measured the change in stability, Δ(ΔG), for a series of hydrogen bonding mutants in four proteins: villin headpiece subdomain (VHP) containing 36 residues, a surface protein from Borrelia burgdorferi (VlsE) containing 341 residues, and two proteins previously studied in our laboratory, ribonucleases Sa (RNase Sa) and T1 (RNase T1). Crystal structures were determined for three of the hydrogen bonding mutants of RNase Sa: S24A, Y51F, and T95A. The structures are very similar to wild type RNase Sa and the hydrogen bonding partners form intermolecular hydrogen bonds to water in all three mutants. We compare our results with previous studies of similar mutants in other proteins and reach the following conclusions. (1) Hydrogen bonds contribute favorably to protein stability. (2) The contribution of hydrogen bonds to protein stability is strongly context dependent. (3) Hydrogen bonds by side chains and peptide groups make similar contributions to protein stability. (4) Polar group burial can make a favorable contribution to protein stability even if the polar groups are not hydrogen bonded. (5) The contribution of hydrogen bonds to protein stability is similar for VHP, a small protein, and VlsE, a large protein.

Propofol pretreatment before reperfusion, or propofol conditioning, has been shown to be cardioprotective, while its mechanism is unclear. The current study investigated the roles of endocannabinoid signaling in propofol cardioprotection in an in vivo model of myocardial ischemia/reperfusion (I/R) injury and in in vitro primary cardiomyocyte hypoxia/reoxygenation (H/R) injury. The results showed that propofol conditioning increased both serum and cell culture media concentrations of endocannabinoids including anandamide (AEA) and 2-arachidonoylglycerol (2-AG) detected by LC-MS/MS. The reductions of myocardial infarct size in vivo and cardiomyocyte apoptosis and death in vitro were accompanied with attenuations of oxidative injuries manifested as decreased reactive oxygen species (ROS), malonaldehyde (MDA), and MPO (myeloperoxidase) and increased superoxide dismutase (SOD) production. These effects were mimicked by either URB597, a selective endocannabinoids degradation inhibitor, or VDM11, a selective endocannabinoids reuptake inhibitor. In vivo study further validated that the cardioprotective and antioxidative effects of propofol were reversed by selective CB2 receptor antagonist AM630 but not CB1 receptor antagonist AM251. We concluded that enhancing endogenous endocannabinoid release and subsequent activation of CB2 receptor signaling represent a major mechanism whereby propofol conditioning confers antioxidative and cardioprotective effects against myocardial I/R injury.

Some theories predict that the filling factor 5/2 fractional quantum Hall state can exhibit non-Abelian statistics, which makes it a candidate for fault-tolerant topological quantum computation. Although the non-Abelian Pfaffian state and its particle-hole conjugate, the anti-Pfaffian state, are the most plausible wave functions for the 5/2 state, there are a number of alternatives with either Abelian or non-Abelian statistics. Recent experiments suggest that the tunneling exponents are more consistent with an Abelian state rather than a non-Abelian state. Here, we present edge-current-tunneling experiments in geometrically confined quantum point contacts, which indicate that Abelian and non-Abelian states compete at filling factor 5/2. Our results are consistent with a transition from an Abelian state to a non-Abelian state in a single quantum point contact when the confinement is tuned. Our observation suggests that there is an intrinsic non-Abelian 5/2 ground state, but that the appropriate confinement is necessary to maintain it. This observation is important not only for understanding the physics of the 5/2 state, but also for the design of future topological quantum computation devices.

MicroRNAs (miRNAs) are short non-coding RNAs that are involved in post-transcriptional regulation of gene expression. Hsa-miR-26b is an intronic miRNA located in the intron of CTDSP1 (carboxy-terminal domain, RNA polymerase II, polypeptide A, small phosphatase 1). In the present study, the expression of hsa-miR-26b was examined during human preadipocyte differentiation. 15 days after induction of differentiation, mature human adipocytes were treated with adipokines. Hsa-miR-26b was differentially expressed during human preadipocyte differentiation. Tumor necrosis factor-α (TNF-α), leptin and resistin, but not interleukin-6, caused downregulation of hsa-miR-26b expression in adipocytes. These results suggest that the expression of hsa-miR-26b is affected by TNF-α, leptin and resistin and that hsa-miR-26b may be an important mediator in regulating the obesity-related insulin sensitivity and inflammatory responses.

NYGGF4, also known as phosphotyrosine interaction domain containing 1(PID1), is a recently discovered gene which is involved in obesity-related insulin resistance (IR) and mitochondrial dysfunction. We aimed to further elucidate the effects and mechanisms underlying NYGGF4-induced IR by investigating the effect of overexpressing mitochondrial transcription factor A (TFAM), which is essential for mitochondrial DNA transcription and replication, on NYGGF4-induced IR and mitochondrial abnormalities in 3T3-L1 adipocytes. Overexpression of TFAM increased the mitochondrial copy number and ATP content in both control 3T3-L1 adipocytes and NYGGF4-overexpressing adipocytes. Reactive oxygen species (ROS) production was enhanced in NYGGF4-overexpressing adipocytes and reduced in TFAM-overexpressing adipocytes; co-overexpression of TFAM significantly attenuated ROS production in NYGGF4-overexpressing adipocytes. However, overexpression of TFAM did not affect the mitochondrial transmembrane potential (ΔΨm) in control 3T3-L1 adipocytes or NYGGF4-overexpressing adipocytes. In addition, co-overexpression of TFAM-enhanced insulin-stimulated glucose uptake by increasing Glucose transporter type 4 (GLUT4) translocation to the PM in NYGGF4-overexpressing adipocytes. Overexpression of NYGGF4 significantly inhibited tyrosine phosphorylation of Insulin receptor substrate 1 (IRS-1) and serine phosphorylation of Akt, whereas overexpression of TFAM strongly induced phosphorylation of IRS-1 and Akt in NYGGF4-overexpressing adipocytes. This study demonstrates that NYGGF4 plays a role in IR by impairing mitochondrial function, and that overexpression of TFAM can restore mitochondrial function to normal levels in NYGGF4-overexpressing adipocytes via activation of the IRS-1/PI3K/Akt signaling pathway.

The continuous synthesis of bis(indolyl)phenylmethane has been successfully accomplished for the first time by Friedel-Crafts alkylation of indole with benzaldehyde in a fixed-bed reactor over different zeolites. A citric acid modified Hβ (CA-Hβ) zeolite showed better catalytic performance than parent Hβ zeolite due to its larger surface area and smaller amount of weak acid sites, confirmed by the Brunauer–Emmett–Teller (BET) method, NH 3 temperature-programmed desorption (NH 3 -TPD) and Fourier Transform Infrared Spectrometer after adsorption of pyridine (Py-IR). Furthermore, it is proposed that citric acid treatment is appropriate for dealumination of Hβ zeolite, which results in the CA-Hβ zeolite having a larger number of active centers for the desired transformation. Significantly, CA-Hβ has shown good stability during a 100 h test.

Our goal was to gain a better understanding of the contribution of hydrophobic interactions to protein stability. We measured the change in conformational stability, Δ(ΔG), for hydrophobic mutants of four proteins: villin headpiece subdomain (VHP) with 36 residues, a surface protein from Borrelia burgdorferi (VlsE) with 341 residues, and two proteins previously studied in our laboratory, ribonucleases Sa and T1. We compared our results with those of previous studies and reached the following conclusions: (1) Hydrophobic interactions contribute less to the stability of a small protein, VHP (0.6±0.3 kcal/mol per -CH(2)- group), than to the stability of a large protein, VlsE (1.6±0.3 kcal/mol per -CH(2)- group). (2) Hydrophobic interactions make the major contribution to the stability of VHP (40 kcal/mol) and the major contributors are (in kilocalories per mole) Phe18 (3.9), Met13 (3.1), Phe7 (2.9), Phe11 (2.7), and Leu21 (2.7). (3) Based on the Δ(ΔG) values for 148 hydrophobic mutants in 13 proteins, burying a -CH(2)- group on folding contributes, on average, 1.1±0.5 kcal/mol to protein stability. (4) The experimental Δ(ΔG) values for aliphatic side chains (Ala, Val, Ile, and Leu) are in good agreement with their ΔG(tr) values from water to cyclohexane. (5) For 22 proteins with 36 to 534 residues, hydrophobic interactions contribute 60±4% and hydrogen bonds contribute 40±4% to protein stability. (6) Conformational entropy contributes about 2.4 kcal/mol per residue to protein instability. The globular conformation of proteins is stabilized predominantly by hydrophobic interactions.

Increasing the conformational stability of proteins is an important goal for both basic research and industrial applications. In vitro selection has been used successfully to increase protein stability, but more often site-directed mutagenesis is used to optimize the various forces that contribute to protein stability. In previous studies, we showed that improving electrostatic interactions on the protein surface and improving the beta-turn sequences were good general strategies for increasing protein stability, and used them to increase the stability of RNase Sa. By incorporating seven of these mutations in RNase Sa, we increased the stability by 5.3 kcal/mol. Adding one more mutation, D79F, gave a total increase in stability of 7.7 kcal/mol, and a melting temperature 28 degrees C higher than the wild-type enzyme. Surprisingly, the D79F mutation lowers the change in heat capacity for folding, DeltaC(p), by 0.6 kcal/mol/K. This suggests that this mutation stabilizes structure in the denatured state ensemble. We made other mutants that give some insight into the structure present in the denatured state. Finally, the thermodynamics of folding of these stabilized variants of RNase Sa are compared with those observed for proteins from thermophiles.

The goal of this article is to gain a better understanding of the denatured state ensemble (DSE) of proteins through an experimental and computational study of their denaturation by urea. Proteins unfold to different extents in urea and the most hydrophobic proteins have the most compact DSE and contain almost as much secondary structure as folded proteins. Proteins that unfold to the greatest extent near pH 7 still contain substantial amounts of secondary structure. At low pH, the DSE expands due to charge-charge interactions and when the net charge per residue is high, most of the secondary structure is disrupted. The proteins in the DSE appear to contain substantial amounts of polyproline II conformation at high urea concentrations. In all cases considered, including staph nuclease, the extent of unfolding by urea can be accounted for using the data and approach developed in the laboratory of Wayne Bolen (Auton et al., Proc Natl Acad Sci 2007; 104:15317-15323).

Our goal was to gain a better understanding of how protein stability can be increased by improving β-turns. We studied 22 β-turns in nine proteins with 66 to 370 residues by replacing other residues with proline and glycine and measuring the stability. These two residues are statistically preferred in some β-turn positions. We studied: Cold shock protein B (CspB), Histidine-containing phosphocarrier protein (HPr), Ubiquitin, Ribonucleases Sa2, Sa3, T1, and HI, Tryptophan synthetase α-subunit (TSα), and Maltose binding protein (MBP). Of the fifteen single proline mutations, 11increased stability (Average = 0.8 ± 0.3; Range = 0.3 – 1.5 kcal/mol), and the stabilizing effect of double proline mutants was additive. Based on this and our previous work, we conclude that proteins can generally be stabilized by replacing non-proline residues with proline residues at the i + 1 position of Type I and II β-turns and at the i position in Type II β-turns. Other turn positions can sometimes be used if the φ angle is near −60° for the residue replaced. It is important that the side chain of the residue replaced is less than 50% buried. Identical substitutions in β-turns in related proteins give similar results. Proline substitutions increase stability mainly by decreasing the entropy of the denatured state. In contrast, the large, diverse group of proteins considered here had almost no residues in β-turns that could be replaced by Gly to increase protein stability. Improving β-turns by substituting Pro residues is a generally useful way of increasing protein stability.

The behavior of phase coherence is studied in two-dimensional hole gas through the integer quantum Hall plateau-to-plateau transition. From the plateau transition as a function of temperature, scaling properties of multiple transitions are analyzed. Our results are in good agreement with the assumption of the zero-point fluctuations of the coherent holes, and support the intrinsic saturation of the coherence time at low temperature limit. The critical exponent $p$ can also be determined under the scheme of the zero-point fluctuations. The similarity and difference in experimental observations between quantum Griffiths singularity and plateau transition is discussed. The spin-orbit coupling effect's influence on the plateau transition is explored by comparing the results from different transitions.

FGF10 is a member of fibroblast growth factors (FGFs). We previously showed that FGF10 protects neuron against oxygen-glucose deprivation injury in vitro; however, the effect of FGF10 in ischemic stroke in vivo is unknown. In the present study, we showed that FGF10 was mainly expressed in neurons but not astrocytes and detected FGF10 in mouse cerebrospinal fluid. The FGF10 levels in neurons culture medium and cell lysate were much higher than those in astrocytes. FGF10 expression in brain tissue and FGF10 level in CSF were increased in mouse middle cerebral artery occlusion (MCAO) model. Administration of FGF10 into lateral cerebroventricle not only decreased MCAO-induced brain infarct volume and neurological deficit, but also reduced the number of TUNEL-positive cells and activities of Caspases. Moreover, FGF10 treatment depressed the triggered inflammatory factors (TNF-α and IL-6) and NF-κB signaling pathway and increased phosphorylation of PI3K/Akt signaling pathway. Blockade of PI3K/Akt signaling pathway by wortmannin and Akt1/2-kinase inhibitor, partly compromised the neuroprotection of FGF10. However, blockade of PI3K/Akt signaling pathway did not impair the anti-inflammation action of FGF10. Collectively, our results demonstrate that neuron-derived FGF10 ameliorates cerebral ischemia injury via inhibiting NF-κB-dependent neuroinflammation and activating PI3K/Akt survival signaling pathway in mice.

The tumor–stroma interaction is critical for the development and progression of cancer. Cancer-associated fibroblasts (CAFs), one of the major components of the tumor stroma, can promote tumor growth and metastasis. Exosomes are secreted microvesicles that mediate cell-to-cell communication. Exosomal contents, including proteins, nucleic acids, and lipids, can be shuttled from donor cells to target cells. Recent studies suggest that exosomes play important roles in the tumor–stroma interaction. Herein, we review the multifaceted roles of exosomes in the tumor–stroma interaction and the underlying molecular mechanisms. Literature search for all relevant publications was performed on PubMed databases. The keywords of exosomes, tumor, stroma, CAFs, mesenchymal stem cells (MSCs) and other closely related terms were used for searching. Tumor cell-derived exosomes induce the differentiation of fibroblasts and MSCs into CAFs. In turn, exosomes secreted by CAFs promote tumor growth, metastasis, and drug resistance through distinct mechanisms. Moreover, exosomes from stromal cells can be used as therapeutic vehicles for the delivery of anticancer drugs. Tumor cells communicate with CAFs through exosomes, which establishes a bidirectional cross talk to promote tumor growth, metastasis, and drug resistance. Targeting exosomes in tumor–stroma interaction may have important implications for anticancer therapy.

The selective detection of ultra trace amounts of aflatoxin B1 (AF1) is extremely important for food safety, since it is the most toxic mycotoxin class that is allowed to be present in edible food and agricultural products with strictly Maximum Residue Limit (MRL). Sensitive detection of AFB1 residues requires time-consuming techniques and expensive instruments. An aptasensor for AFB1 detection, using unmodified gold nanoparticles (AuNPs) indicator, was developed in the present study, based on the salt-induced AuNPs aggregation phenomenon. Its linear dynamic range and detection sensitivity were found to be 0.025 ng/mL-100 ng/mL and 0.025 ng/mL of AFB1 respectively, which were lower than the maximum residue limit (MRL) in edible food, as set by China and the European Commission. Our study provides a simple, fast, and visible method for AFB1 analysis, with high sensitivity, which can be applied in future on-site detection for food and agricultural products.

We report on the observation of a helical Luttinger-liquid in the edge of InAs/GaSb quantum spin Hall insulator, which shows characteristic suppression of conductance at low temperature and low bias voltage. Moreover, the conductance shows power-law behavior as a function of temperature and bias voltage. The results underscore the strong electron-electron interaction effect in transport of InAs/GaSb edge states. Because of the fact that the Fermi velocity of the edge modes is controlled by gates, the Luttinger parameter can be fine tuned. Realization of a tunable Luttinger-liquid offers a one-dimensional model system for future studies of predicted correlation effects., Comment: 23 pages, 9 figures

Background: Metabolic syndrome (MS) increases a risk of developing cardiovascular disease (CVD) and type 2 diabetes mellitus (T2DM). The Vitamin D Receptor gene (VDR) may be important for developing MS. The aim of this study is to investigate the correlation between the VDR gene polymorphisms and MS in North China. Methods: A case–control study included 391 participants with MS according to the MS diagnostic criteria of International Diabetes Federation 2005 (IDF2005) and 400 controls was conducted on the basis of a cross sectional study which was performed from 2008 to 2012 in Ningxia Hui Autonomous Region, China. Anthropometric data, blood pressure and blood samples were collected in the field investigation. Blood biochemistry analyses were carried out in the laboratory. Two single-nucleotide polymorphisms (SNPs) in the VDR gene, BsmI (rs1544410 A>G) and FokI (rs 2228570 C > T), were genotyped. Results: The difference in the occurrence of genotypes in BsmI between individuals with MS and the control group was significant. Compared with genotype Bb/bb and allele b, genotype BB and allele B showed higher frequencies in MS cases than controls, which suggested they were risk factors. In addition, the genotype BB carriers with MS presented a higher waist circumference, while genotype FF for the FokI polymorphism was correlated with lower BMI in subjects with MS. Conclusion: Our study suggests that the VDR gene polymorphisms appear to be associated with MS in the Northern Chinese population. Allele B and BB genotype for BsmI are risk factors for MS. The BsmI polymorphism seems to influence waist circumference, while the FokI polymorphism influence BMI in subjects with MS.

Our goal was to gain a better understanding of the contribution of the burial of polar groups and their hydrogen bonds to the conformational stability of proteins. We measured the change in stability, Δ(ΔG), for a series of hydrogen bonding mutants in four proteins: villin headpiece subdomain (VHP) containing 36 residues, a surface protein from Borrelia burgdorferi (VlsE) containing 341 residues, and two proteins previously studied in our laboratory, ribonucleases Sa (RNase Sa) and T1 (RNase T1). Crystal structures were determined for three of the hydrogen bonding mutants of RNase Sa: S24A, Y51F, and T95A. The structures are very similar to wild type RNase Sa and the hydrogen bonding partners form intermolecular hydrogen bonds to water in all three mutants. We compare our results with previous studies of similar mutants in other proteins and reach the following conclusions. (1) Hydrogen bonds contribute favorably to protein stability. (2) The contribution of hydrogen bonds to protein stability is strongly context dependent. (3) Hydrogen bonds by side chains and peptide groups make similar contributions to protein stability. (4) Polar group burial can make a favorable contribution to protein stability even if the polar groups are not hydrogen bonded. (5) The contribution of hydrogen bonds to protein stability is similar for VHP, a small protein, and VlsE, a large protein.

While in the lowest Landau level the electron-electron interaction leads to the formation of the Wigner crystal, in higher Landau levels a solid phase with multiple electrons in a lattice site of crystal was predicted, which was called the bubble phase. Reentrant integer quantum Hall states are believed to be the insulating bubble phase pinned by disorder. We carry out nonlinear transport measurements on the reentrant states and study the de-pinning of the bubble phase, which is complementary to previous microwave measurements and provides unique information. In this study, conductivity is directly measured with Corbino geometry. Based on the threshold electric field of de-pinning, a phase diagram of the reentrant state is mapped. We discuss an interaction-driven topological phase transition between the integer quantum Hall state and the reentrant integer quantum Hall state., Comment: 11 pages, 3 figures

Hydrogen sulfide (H2S) is the third most common endogenously produced gaseous signaling molecule, but its impact on hepatic ischemia/reperfusion (I/R) injury, especially on mitochondrial function, remains unclear. In this study, rats were randomized into Sham, I/R, ischemia preconditioning (IPC) or sodium hydrosulfide (NaHS, an H2S donor) preconditioning groups. To establish a model of segmental (70%) warm hepatic ischemia, the hepatic artery, left portal vein and median liver lobes were occluded for 60 min and then unclamped to allow reperfusion. Preconditioning with 12.5, 25 or 50 μmol/kg NaHS prior to the I/R insult significantly increased serum H2S levels, and, similar to IPC, NaHS preconditioning decreased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the plasma and prevented hepatocytes from undergoing I/R-induced necrosis. Moreover, a sub-toxic dose of NaHS (25 μmol/kg) did not disrupt the systemic hemodynamics but dramatically inhibited mitochondrial permeability transition pore (MPTP) opening and thus prevented mitochondrial-related cell death and apoptosis. Mechanistic studies revealed that NaHS preconditioning markedly increased the expression of phosphorylated protein kinase B (p-Akt), phosphorylated glycogen synthase kinase-3 beta (p-GSK-3β) and B-cell lymphoma-2 (Bcl-2) and decreased the release of mitochondrial cytochrome c and cleaved caspase-3/9 levels. Therefore, NaHS administration prior to hepatic I/R ameliorates mitochondrial and hepatocellular damage through the inhibition of MPTP opening and the activation of Akt-GSK-3β signaling. Furthermore, this study provides experimental evidence for the clinical use of H2S to reduce liver damage after perioperative I/R injury.

Flurbiprofen acts as a nonselective inhibitor for cyclooxygenases (COX-1 and COX-2), but its impact on hepatic ischemia/reperfusion (I/R) injury remains unclear. Mice were randomized into sham, I/R and flurbiprofen (Flurb) groups. The hepatic artery and portal vein to the left and median liver lobes were occluded for 90 min and unclamped for reperfusion to establish a model of segmental (70%) warm hepatic ischemia. Pretreatment of animals with flurbiprofen prior to I/R insult significantly decreased serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH), and prevented hepatocytes from I/R-induced apoptosis/necrosis. Moreover, flurbiprofen dramatically inhibited mitochondrial permeability transition (MPT) pore opening, and thus prevented mitochondrial-related cell death and apoptosis. Mechanistic studies revealed that flurbiprofen markedly inhibited glycogen synthase kinase (GSK)-3β activity and increased phosphorylation of GSK-3β at Ser9, which, consequently, could modulate the adenine nucleotide translocase (ANT)-cyclophilin D (CyP-D) complex and the susceptibility to MPT induction. Therefore, administration of flurbiprofen prior to hepatic I/R ameliorates mitochondrial and hepatocellular damage through inhibition of MPT and inactivation of GSK-3β, and provides experimental evidence for clinical use of flurbiprofen to protect liver function in surgical settings in addition to its conventional use for pain relief.

Interleukin-6 (IL-6) has emerged as an important cytokine involved in the regulation of metabolism. However, the role of IL-6 in the etiology of obesity and insulin resistance is not fully understood. Mitochondria are key organelles of energy metabolism, and there is growing evidence that mitochondrial dysfunction plays a crucial role in the pathogenesis of obesity-associated insulin resistance. In this study, we determined the direct effect of IL-6 on lipolysis in adipocytes, and the effects of IL-6 on mitochondrial function were investigated. We found that cells treated with IL-6 displayed fewer lipids and an elevated glycerol release rate. Further, IL-6 treatment led to decreased mitochondrial membrane potential, decreased cellular ATP production, and increased intracellular ROS levels. The mitochondria in IL-6-treated cells became swollen and hollow with reduced or missing cristae. However, insulin-stimulated glucose transport was unaltered. PGC-1α, NRF1, and mtTFA mRNA levels were markedly increased, and the mitochondrial contents were also increased. Our results demonstrate that IL-6 can exert a direct lipolytic effect and induce mitochondrial dysfunction. However, IL-6 did not affect insulin sensitivity in adipocytes in vitro. We deduce that in these cells, enhanced mitochondrial biogenesis might play a compensatory role in glucose transport.