Acetohydrazone: A Transient Directing Group for Arylation of Unactivated C(sp³)–H Bonds

A straightforward and efficient method has been developed for the synthesis of 2-benzylbenzaldehyde derivatives from 2-methylbenzaldehyde and iodobenzene via a C­(sp³)–H activation process. In the course of the activation reaction, acetohydrazone is formed between 2-benzylbenzaldehyde and acetohydrazine as a transient directing group. As a new kind of transient directing group, acetohydrazone exhibits a remarkable directing effect to give corresponding products in good to excellent yields

Ultrasensitive Detection of Transcription Factors Using Transcription-Mediated Isothermally Exponential Amplification-Induced Chemiluminescence

Transcription factors (TFs) are important cellular components that modulate gene expression, and the malregulation of transcription will lead to a variety of diseases such as cancer and developmental syndromes. However, the conventional methods for transcription factor assay are generally cumbersome and costly with low sensitivity. Here, we develop a label-free strategy for ultrasensitive detection of transcription factors using a cascade signal amplification of RNA transcription, dual isothermally exponential amplification reaction (EXPAR), and G-quadruplex DNAzyme-driven chemiluminescence. Briefly, the specific binding of TF with the detecting probe prevents the cleavage of the detecting probe by exonuclease and subsequently facilitates the conversion of TF signal to abundant RNA triggers in the presence of T7 RNA polymerase. The obtained RNA triggers can initiate the strand displacement amplification to yield abundant DNAzymes and DNA triggers, and the released DNA triggers can further initiate the next rounds of EXPAR reaction. The synergistic operation of dual EXPAR reaction can produce large amounts of DNAzymes, which subsequently catalyze the oxidation of luminol by H₂O₂ to yield an enhanced chemiluminescence signal with the assistance of cofactor hemin. Conversely, in the absence of target TF, the naked detecting probes will be completely digested by exonucleases, leading to neither the transcription-mediated EXPAR nor the DNAzyme-driven chemiluminescence signal. This method has a low detection limit of as low as 6.03 × 10^–15 M and a broad dynamic range from 10 fM to 1 nM and can even measure the NF-κB p50 of crude cell nuclear extracts. Moreover, this method can be used to measure a variety of DNA-binding proteins by simply substituting the target-specific binding sequence in the detecting probes

Application of Nano Fe^III–Tannic Acid Complexes in Modifying Aqueous Acrylic Latex for Controlled-Release Coated Urea

Acrylic latexes are valuable waterborne materials used in controlled-release fertilizers. Controlled-release urea coated with these latexes releases a large amount of nutrients, making it difficult to meet the requirement of plants. Herein, Fe^III–tannic acid (TA) complexes were blended with acrylic latex and subsequently reassembled on a surface of polyacrylate particles. These complexes remarkably retarded the release of urea (the preliminary solubility was decreased from 22.3 to 0.8%) via decreasing the coating tackiness (<i>T</i>_g was increased from 4.17 to 6.42 °C), increasing the coating strength (tensile stress was improved from 3.88 to 4.45 MPa), and promoting the formation of denser structures (surface tension was decreased from 37.37 to 35.94 mN/m). Overall, our findings showed that a simple blending of Fe^III–TA complexes with acrylic latex produces excellent coatings that delay the release of urea, which demonstrates great potential for use in controlled-release fertilizers coated with waterborne polymers

Base-Excision-Repair-Induced Construction of a Single Quantum-Dot-Based Sensor for Sensitive Detection of DNA Glycosylase Activity

DNA glycosylase is an initiating enzyme of cellular base excision repair pathway which is responsible for the repair of various DNA lesions and the maintenance of genomic stability, and the dysregulation of DNA glycosylase activity is associated with a variety of human pathology. Accurate detection of DNA glycosylase activity is critical to both clinical diagnosis and therapeutics, but conventional methods for the DNA glycosylase assay are usually time-consuming with poor sensitivity. Here, we demonstrate the base-excision-repair-induced construction of a single quantum dot (QD)-based sensor for highly sensitive measurement of DNA glycosylase activity. We use human 8-oxoguanine-DNA glycosylase 1 (hOGG1), which is responsible for specifically repairing the damaged 8-hydroxyguanine (8-oxoG, one of the most abundant and widely studied DNA damage products), as a model DNA glycosylase. In the presence of biotin-labeled DNA substrate, the hOGG1 may catalyze the removal of 8-oxo G from 8-oxoG·C base pairs to generate an apurinic/apyrimidinic (AP) site. With the assistance of apurinic/apyrimidinic endonuclease (APE1), the cleavage of the AP site results in the generation of a single-nucleotide gap. Subsequently, DNA polymerase β incorporates a Cy5-labeled dGTP into the DNA substrate to fill the gap. With the addition of streptavidin-coated QDs, a QD-DNA-Cy5 nanostructure is formed via specific biotin–streptavidin binding, inducing the occurrence of fluorescence resonance energy transfer (FRET) from the QD to Cy5. The resulting Cy5 signal can be simply monitored by total internal reflection fluorescence (TIRF) imaging. The proposed method enables highly sensitive measurement of hOGG1 activity with a detection limit of 1.8 × 10^–6 U/μL. Moreover, it can be used to measure the enzyme kinetic parameters and detect the hOGG1 activity in crude cell extracts, offering a powerful tool for biomedical research and clinical diagnosis

Supplementary Material from Photoprotection through ultrafast charge recombination in photochemical reaction centres under oxidizing conditions

Materials and Methods; Figures S1-S10

Intensity-Modulated Scanning Kelvin Probe Microscopy for Probing Recombination in Organic Photovoltaics

We study surface photovoltage decays on sub-millisecond time scales in organic solar cells using intensity-modulated scanning Kelvin probe microscopy (SKPM). Using polymer/fullerene (poly[<i>N</i>-9″-heptadecanyl-2,7-carbazole-<i>alt</i>-5,5-(4′,7′-di-2-thienyl-2′,1′,3′-benzothiadiazole)]/[6,6]-phenyl C₇₁-butyric acid methyl ester, PCDTBT/PC₇₁BM) bulk heterojunction devices as a test case, we show that the decay lifetimes measured by SKPM depend on the intensity of the background illumination. We propose that this intensity dependence is related to the well-known carrier-density-dependent recombination kinetics in organic bulk heterojunction materials. We perform transient photovoltage (TPV) and charge extraction (CE) measurements on the PCDTBT/PC₇₁BM blends to extract the carrier-density dependence of the recombination lifetime in our samples, and we find that the device TPV and CE data are in good agreement with the intensity and frequency dependence observed <i>via</i> SKPM. Finally, we demonstrate the capability of intensity-modulated SKPM to probe local recombination rates due to buried interfaces in organic photovoltaics (OPVs). We measure the differences in photovoltage decay lifetimes over regions of an OPV cell fabricated on an indium tin oxide electrode patterned with two different phosphonic acid monolayers known to affect carrier lifetime

Direct Observation of Energy Detrapping in LH1-RC Complex by Two-Dimensional Electronic Spectroscopy

The purple bacterial core light harvesting antenna-reaction center (LH1-RC) complex is the simplest system able to achieve the entire primary function of photosynthesis. During the past decade, a variety of photosynthetic proteins were studied by a powerful technique, two-dimensional electronic spectroscopy (2DES). However, little attention has been paid to LH1-RC, although its reversible uphill energy transfer, trapping, and backward detrapping processes, represent a crucial step in the early photosynthetic reaction dynamics. Thus, in this work, we employed 2DES to study two LH1-RC complexes of Thermochromatium (Tch.) tepidum. By direct observation of detrapping, the complex reversible process was clearly identified and an overall scheme of the excitation evolution in LH1-RC was obtained

Controllable Mismatched Ligation for Bioluminescence Screening of Known and Unknown Mutations

Single-nucleotide polymorphisms (SNPs) are closely related to human diseases and individual drug responses, and the accurate detection of SNPs is crucial to both clinical diagnosis and development of personalized medicine. Among various SNPs detection methods, ligase detection reaction (LDR) has shown great potential due to its low detection limit and excellent specificity. However, frequent involvement of expensive labels increases the experimental cost and compromises the assay efficiency, and the requirement of careful predesigned probes limits it to only known SNPs assays. In this research, we develop a controllable mismatched ligation for bioluminescence screening of both known and unknown mutations. Especially, the ligation specificity of <i>E. coli</i> ligase is tunable under different experimental conditions. The mismatches locating on the 3′-side of the nick cannot be ligated efficiently by <i>E. coli</i> ligase, whereas all mismatches locating on the 5′-side of the nick can be ligated efficiently by <i>E. coli</i> ligase. We design a 3′-discriminating probe (3′-probe) for the discrimination of known mutation and introduce a T7 Endo I for the detection of unknown mutation. With the integration of bioluminescence monitoring of ligation byproduct adenosine 5'-monophosphate (AMP), both known and unknown SNPs can be easily detected without the involvement of any expensive labels and labor-intensive separation. This method is simple, homogeneous, label-free, and cost-effective and may provide a valuable complement to current sequencing technologies for disease diagnostics, personalized medicine, and biomedical research

Vibronic Coherence in the Charge Separation Process of the <i>Rhodobacter sphaeroides</i> Reaction Center

Two-dimensional electronic spectroscopy was applied to a variant of the reaction center (RC) of purple bacterium <i>Rhodobacter sphaeroides</i> lacking the primary acceptor ubiquinone in order to understand the ultrafast separation and transfer of charge between the bacteriochlorin cofactors. For the first time, characteristic 2D spectra were obtained for the participating excited and charge-transfer states, and the electron-transfer cascade (including two different channels, the P* and B* channels) was fully mapped. By analyzing quantum beats using 2D frequency maps, excited-state vibrational modes at 153 and 33 cm^–1 were identified. We speculate that these modes couple to the charge separation (CS) process and collectively optimize the CS and are responsible for the superhigh efficiency

Excitonic and Vibrational Coherence in the Excitation Relaxation Process of Two LH1 Complexes as Revealed by Two-Dimensional Electronic Spectroscopy

Ultrafast excitation relaxation within a manifold exciton state and long-lived vibrational coherence are two universal characteristics of photosynthetic antenna complexes. In this work, we studied the two-dimensional electronic spectra of two core light-harvesting (LH1) complexes of <i>Thermochromatium</i> (<i>Tch.</i>) <i>tepidum</i>, native Ca²⁺-LH1 and modified Ba²⁺-LH1. The role of the vibrational coherence in the exciton relaxation was revealed by comparing the two LH1 with similar structures but different electronic properties and by the evolution of the exciton and vibrational coherence as a function of temperature