A surface plasmon enhanced FLIM-FRET imaging approach based on Au nanoparticles

In this report we have demonstrated a fluorescence resonant energy transfer (FRET)-fluorescence lifetime imaging microscopy (FLIM) combined approach to study the intracellular pathway of gold nanoparticles. The detected energy transfer between gold nanorods (GNRs) and green fluorescence protein (GFP) labeled Hela cell earlyendosomes and the in-depth lifetime distribution analysis on the transfer process suggest an endocytotic uptake process of GNRs. Furthermore, the FRET-FLIM method profits from a surface plasmon enhanced energy transfer mechanism when taking into consideration of GNRs and two photon excitation, and is effective in biological imaging, sensing, and even in single molecular tracing in both in vivo and in vitro studies

On Resource-bounded versions of the van Lambalgen theorem

The van Lambalgen theorem is a surprising result in algorithmic information theory concerning the symmetry of relative randomness. It establishes that for any pair of infinite sequences $A$ and $B$, $B$ is Martin-L\"of random and $A$ is Martin-L\"of random relative to $B$ if and only if the interleaved sequence $A \uplus B$ is Martin-L\"of random. This implies that $A$ is relative random to $B$ if and only if $B$ is random relative to $A$ \cite{vanLambalgen}, \cite{Nies09}, \cite{HirschfeldtBook}. This paper studies the validity of this phenomenon for different notions of time-bounded relative randomness. We prove the classical van Lambalgen theorem using martingales and Kolmogorov compressibility. We establish the failure of relative randomness in these settings, for both time-bounded martingales and time-bounded Kolmogorov complexity. We adapt our classical proofs when applicable to the time-bounded setting, and construct counterexamples when they fail. The mode of failure of the theorem may depend on the notion of time-bounded randomness

Metalloporphyrin-incorporated diphosphine ligands for metal ion-binding

Poster: no. P48Diphosphine ligands have been widely used in organometallic chemistry and catalysis.1 By incorporation of functional units such as metallomacrocycles, the resulting functionalized diphosphines could exhibit unusual properties or binding behavior. In this study, we prepared several examples of ruthenium porphyrin phosphine complexes [RuII(Por)(dppm)2] (1; Por = TTP, 4-MeO-TPP, F20-TPP; dppm = bis(diphenylphosphino)methane) by a similar method to that previously reported for their congeners.2 Reaction of complexes 1 with a number of metal …published_or_final_versio

Elucidation of the RamA Regulon in Klebsiella pneumoniae Reveals a Role in LPS Regulation

Klebsiella pneumoniae is a significant human pathogen, in part due to high rates of multidrug resistance. RamA is an intrinsic regulator in K. pneumoniae established to be important for the bacterial response to antimicrobial challenge; however, little is known about its possible wider regulatory role in this organism during infection. In this work, we demonstrate that RamA is a global transcriptional regulator that significantly perturbs the transcriptional landscape of K. pneumoniae, resulting in altered microbe-drug or microbe-host response. This is largely due to the direct regulation of 68 genes associated with a myriad of cellular functions. Importantly, RamA directly binds and activates the lpxC, lpxL-2 and lpxO genes associated with lipid A biosynthesis, thus resulting in modifications within the lipid A moiety of the lipopolysaccharide. RamA-mediated alterations decrease susceptibility to colistin E, polymyxin B and human cationic antimicrobial peptide LL-37. Increased RamA levels reduce K. pneumoniae adhesion and uptake into macrophages, which is supported by in vivo infection studies, that demonstrate increased systemic dissemination of ramA overexpressing K. pneumoniae. These data establish that RamA-mediated regulation directly perturbs microbial surface properties, including lipid A biosynthesis, which facilitate evasion from the innate host response. This highlights RamA as a global regulator that confers pathoadaptive phenotypes with implications for our understanding of the pathogenesis of Enterobacter, Salmonella and Citrobacter spp. that express orthologous RamA proteins

Multifactorial Remodeling of the Cancer Immunopeptidome by IFNγ.

UNLABELLED: IFNγ alters the immunopeptidome presented on HLA class I (HLA-I), and its activity on cancer cells is known to be important for effective immunotherapy responses. We performed proteomic analyses of untreated and IFNγ-treated colorectal cancer patient-derived organoids and combined this with transcriptomic and HLA-I immunopeptidomics data to dissect mechanisms that lead to remodeling of the immunopeptidome through IFNγ. IFNγ-induced changes in the abundance of source proteins, switching from the constitutive to the immunoproteasome, and differential upregulation of different HLA alleles explained some, but not all, observed peptide abundance changes. By selecting for peptides which increased or decreased the most in abundance, but originated from proteins with limited abundance changes, we discovered that the amino acid composition of presented peptides also influences whether a peptide is upregulated or downregulated on HLA-I through IFNγ. The presence of proline within the peptide core was most strongly associated with peptide downregulation. This was validated in an independent dataset. Proline substitution in relevant core positions did not influence the predicted HLA-I binding affinity or stability, indicating that proline effects on peptide processing may be most relevant. Understanding the multiple factors that influence the abundance of peptides presented on HLA-I in the absence or presence of IFNγ is important to identify the best targets for antigen-specific cancer immunotherapies such as vaccines or T-cell receptor engineered therapeutics. SIGNIFICANCE: IFNγ remodels the HLA-I-presented immunopeptidome. We showed that peptide-specific factors influence whether a peptide is upregulated or downregulated and identified a preferential loss or downregulation of those with proline near the peptide center. This will help selecting immunotherapy target antigens which are consistently presented by cancer cells

PI3K/mTORC2 regulates TGF-β/Activin signalling by modulating Smad2/3 activity via linker phosphorylation

Crosstalk between the phosphatidylinositol 3-kinase (PI3K) and the transforming growth factor-β signalling pathways play an important role in regulating many cellular functions. However, the molecular mechanisms underpinning this crosstalk remain unclear. Here, we report that PI3K signalling antagonizes the Activin-induced definitive endoderm (DE) differentiation of human embryonic stem cells by attenuating the duration of Smad2/3 activation via the mechanistic target of rapamycin complex 2 (mTORC2). Activation of mTORC2 regulates the phosphorylation of the Smad2/3-T220/T179 linker residue independent of Akt, CDK and Erk activity. This phosphorylation primes receptor-activated Smad2/3 for recruitment of the E3 ubiquitin ligase Nedd4L, which in turn leads to their degradation. Inhibition of PI3K/mTORC2 reduces this phosphorylation and increases the duration of Smad2/3 activity, promoting a more robust mesendoderm and endoderm differentiation. These findings present a new and direct crosstalk mechanism between these two pathways in which mTORC2 functions as a novel and critical mediator

Antioxidant, antibacterial and α-glucosidase inhibitory activities of different extracts of Cortex Moutan

Different extracts of Cortex Moutan (CM) were investigated for their antioxidant, antibacterial and α- glucosidase inhibitory activities. The content of paeonol was quantified by high performance liquid chromatography (HPLC). The results show that the yield of acetone extract (57.14%) was significantly higher than those of other solvents. The ethyl-acetate extract exhibited maximum paeonol concentration (60.69 μg/ml), good antibacterial activities (MIC = 100 μg/ml) against Escherichia coli and possessed significant α-glucosidase inhibitory activity. In addition, among all of the extracts, ethylacetate extract demonstrated a high total phenolic value of 127.12 ± 1.42 mg GAE/g, high DPPH radical scavenging activity with an IC50 of 19.88 ± 0.26 μg/ml, and significant reducing power, suggesting that CM is a potential source of natural antioxidants.Key words: Cortex Moutan, antioxidant, 11-diphenyl-2-picrylhydrazyl hydrate (DPPH), reducing power, antibacterial, α-glucosidas

Microscopic coexistence of superconductivity and antiferromagnetism in underdoped Ba(Fe1-xRux)2As2

We use $^{75}$As nuclear magnetic resonance (NMR) to investigate the local electronic properties of Ba(Fe$_{1-x}$Ru$_{x}$)$_2$As$_2$ ($x =$ 0.23). We find two phase transitions, to antiferromagnetism at $T_N \approx$ 60 K and to superconductivity at $T_C \approx$ 15 K. Below $T_N$, our data show that the system is fully magnetic, with a commensurate antiferromagnetic structure and a moment of 0.4 $\mu_B$/Fe. The spin-lattice relaxation rate $1/^{75}T_1$ is large in the magnetic state, indicating a high density of itinerant electrons induced by Ru doping. On cooling below $T_C$, $1/^{75}T_1$ on the magnetic sites falls sharply, providing unambiguous evidence for the microscopic coexistence of antiferromagnetism and superconductivity.Comment: Accepted in Phys. Rev. Let

Ultra-strong Adhesion of Graphene Membranes

As mechanical structures enter the nanoscale regime, the influence of van der Waals forces increases. Graphene is attractive for nanomechanical systems because its Young's modulus and strength are both intrinsically high, but the mechanical behavior of graphene is also strongly influenced by the van der Waals force. For example, this force clamps graphene samples to substrates, and also holds together the individual graphene sheets in multilayer samples. Here we use a pressurized blister test to directly measure the adhesion energy of graphene sheets with a silicon oxide substrate. We find an adhesion energy of 0.45 \pm 0.02 J/m2 for monolayer graphene and 0.31 \pm 0.03 J/m2 for samples containing 2-5 graphene sheets. These values are larger than the adhesion energies measured in typical micromechanical structures and are comparable to solid/liquid adhesion energies. We attribute this to the extreme flexibility of graphene, which allows it to conform to the topography of even the smoothest substrates, thus making its interaction with the substrate more liquid-like than solid-like.Comment: to appear in Nature Nanotechnolog

Cyclin B1-Cdk1 facilitates MAD1 release from the nuclear pore to ensure a robust spindle checkpoint.

How the cell rapidly and completely reorganizes its architecture when it divides is a problem that has fascinated researchers for almost 150 yr. We now know that the core regulatory machinery is highly conserved in eukaryotes, but how these multiple protein kinases, protein phosphatases, and ubiquitin ligases are coordinated in space and time to remodel the cell in a matter of minutes remains a major question. Cyclin B1-Cdk is the primary kinase that drives mitotic remodeling; here we show that it is targeted to the nuclear pore complex (NPC) by binding an acidic face of the kinetochore checkpoint protein, MAD1, where it coordinates NPC disassembly with kinetochore assembly. Localized cyclin B1-Cdk1 is needed for the proper release of MAD1 from the embrace of TPR at the nuclear pore so that it can be recruited to kinetochores before nuclear envelope breakdown to maintain genomic stability