A novel imaging tool for hepatic portal system using phase contrast technique with hydrogen peroxide‐generated O2 gas

The objective of this study was to investigate the potential of hydrogen peroxide‐generated oxygen gas‐based phase contrast imaging (PCI) for visualizing mouse hepatic portal veins. The O₂ gas was made from the reaction between H₂O₂ and catalase. The gas production was imaged by PCI in real time. The H₂O₂ was injected into the enteric cavity of the lower sigmoid colon to produce O₂ in the submucosal venous plexus. The generated O₂ gas could be finally drained into hepatic portal veins. Absorption contrast imaging (ACI) and PCI of O₂‐filled portal veins were performed and compared. PCI offers high resolution and real‐time visualization of the O₂ gas production. Compared with O₂‐based ACI, O₂‐based PCI significantly enhanced the revealing of the portal vein in vivo. It is concluded that O₂‐based PCI is a novel and promising imaging modality for future studies of portal venous disorders in mice models.     

Fast Photochemical Oxidation of Proteins (FPOP) Maps the Epitope of EGFR Binding to Adnectin

Epitope mapping is an important tool for the development of monoclonal antibodies, mAbs, as therapeutic drugs. Recently, a class of therapeutic mAb alternatives, adnectins, has been developed as targeted biologics. They are derived from the 10th type III domain of human fibronectin (¹⁰Fn3). A common approach to map the epitope binding of these therapeutic proteins to their binding partners is X-ray crystallography. Although the crystal structure is known for Adnectin 1 binding to human epidermal growth factor receptor (EGFR), we seek to determine complementary binding in solution and to test the efficacy of footprinting for this purpose. As a relatively new tool in structural biology and complementary to X-ray crystallography, protein footprinting coupled with mass spectrometry is promising for protein–protein interaction studies. We report here the use of fast photochemical oxidation of proteins (FPOP) coupled with MS to map the epitope of EGFR-Adnectin 1 at both the peptide and amino-acid residue levels. The data correlate well with the previously determined epitopes from the crystal structure and are consistent with HDX MS data, which are presented in an accompanying paper. The FPOP-determined binding interface involves various amino-acid and peptide regions near the N terminus of EGFR. The outcome adds credibility to oxidative labeling by FPOP for epitope mapping and motivates more applications in the therapeutic protein area as a stand-alone method or in conjunction with X-ray crystallography, NMR, site-directed mutagenesis, and other orthogonal methods. Figure

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Two mononuclear molybdenum(VI) oxo complexes with tridentate hydrazone ligands: Synthesis and crystal structures

Reaction of [MoO₂(Acac)₂] (Acac = acetylacetonate) with two similar hydrazone ligands in methanol yielded two mononuclear molybdenum(VI) oxocomplexes with general formula [MoO₂(L)(CH₃OH)], where L = L¹ = (4-nitrophenoxy)acetic acid [1-(3-ethoxy-2-hydroxyphenyl)methylidene]hydrazide (H₂L¹) and L = L² = (4-nitrophenoxy)acetic acid [1-(5-bromo-2-hydroxyphenyl)methylidene]hydrazide (H₂L²). Crystal and molecular structures of the complexes were determined by single crystal X-ray diffraction method. All investigated compounds were further characterized by elemental analysis and FT-IR spectra. Single crystal X-ray structural studies indicate that the hydrazone ligands coordinate to the MoO₂ cores through enolate oxygen, phenolate oxygen, and azomethine nitrogen. The Mo atoms in both complexes are in octahedral coordination.     

A hydrogen peroxide biosensor with high stability based on gelatin-multiwalled carbon nanotubes modified glassy carbon electrode

A biosensor with high stability was prepared to determine hydrogen peroxide (H₂O₂). This hydrogen peroxide biosensor was obtained by modifying glassy carbon electrode (GCE) with a composite film composed of gelatin-multiwalled carbon nanotubes. Catalase (Cat) was covalently immobilized into gelatin-multiwalled carbon nanotubes modified GCE through the well-known glutaraldehyde (GAD) chemistry in order to enhance the stability of electrodes. The enzyme sensor can achieve direct electrochemical response of hydrogen peroxide. The cyclic voltammograms at different scan rates, electrochemical impedance spectroscopy (EIS), and scanning electron microscope (SEM) tests indicate that the enzyme sensor performs positively on increasing permeability, reducing the electron transfer resistance, and improving the electrode performance. The linear response of standard curve for H₂O₂ is in the range of 0.2 to 5.0 mM with a correlation coefficient of 0.9972, and the detection limit of 0.001 mM. A high operational and storage stability is demonstrated for the biosensor. The peak potential at room temperature in two consecutive weeks stays almost consistent, and the enzyme activity is kept stable even after 30 days in further study.     

A Supercharged Fluorescent Protein as a Versatile Probe for Homogeneous DNA Detection and Methylation Analysis

Supercharged proteins are a new class of functional proteins with exceptional stability and potent ability to deliver bio‐macromolecules into cells. As a proof‐of‐principle, a novel application of supercharged proteins as a versatile biosensing platform for nucleic acid detection and epigenetics analysis is presented. Taking supercharged green fluorescent protein (ScGFP) as the signal reporter, a simple turn‐on homogenous method for DNA detection has been developed based on the polyionic nanoscale complex of ScGFP/DNA and toehold strand displacement. This assay shows high sensitivity and potent ability to detect single‐base mismatch. Furthermore, combined with bisulfite conversion, this ScGFP‐based assay was further applied to analyze site‐specific DNA methylation status of genomic DNA extracted from real human colon carcinoma tissue sample with ultrahigh sensitivity (4 amol methylated DNA).     

Pretreatment Methods for Bioethanol Production

Lignocellulosic biomass, such as wood, grass, agricultural, and forest residues, are potential resources for the production of bioethanol. The current biochemical process of converting biomass to bioethanol typically consists of three main steps: pretreatment, enzymatic hydrolysis, and fermentation. For this process, pretreatment is probably the most crucial step since it has a large impact on the efficiency of the overall bioconversion. The aim of pretreatment is to disrupt recalcitrant structures of cellulosic biomass to make cellulose more accessible to the enzymes that convert carbohydrate polymers into fermentable sugars. This paper reviews several leading acidic, neutral, and alkaline pretreatments technologies. Different pretreatment methods, including dilute acid pretreatment (DAP), steam explosion pretreatment (SEP), organosolv, liquid hot water (LHW), ammonia fiber expansion (AFEX), soaking in aqueous ammonia (SAA), sodium hydroxide/lime pretreatments, and ozonolysis are intensively introduced and discussed. In this minireview, the key points are focused on the structural changes primarily in cellulose, hemicellulose, and lignin during the above leading pretreatment technologies.