The discovery and expansion of Hippo signaling pathway in Drosophila model.

The discovery of Hippo signaling pathway is another breakthrough of fly genetics. Similar to the other signaling pathways, Hippo pathway also functions crucially in tremendous physiological and pathological conditions, like organ size control and cancer. There are three main stages of Hippo pathway study: Firstly, identifications of core components by fly genetic screens; secondly, regulations by versatile upstream cues, like cytoskeleton, mechanical tension, and nutrition; thirdly, functions in different biological processes, like cell proliferation regulation, stem cell biology, and immunology. In this review, we summarize the current understanding of Hippo pathway and highlight its regulations and transcriptional complex assembly. We also discuss the potential future directions in Drosophila model system.

[Investigation of interaction between riboflavin and riboflavin binding protein by fluorescence spectroscopy].

The interaction between riboflavin and egg white riboflavin binding protein (RBP) was investigated using fluorescence spectroscopy. The binding mode, binding constants, thermodynamic parameters between riboflavin and RBP and energy transfer were studied. The experimental results showed that riboflavin has the ability to quench the intrinsic fluorescence of RBP because of a complex formed, and the quenching mechanism is static quenching. The binding constants were 5.35 x 10(8), 1.54 x 10(8), 0.56 x 10(8) L x mol(-1) at 298, 308 and 318 K, respectively. The thermodynamic parameters were calculated, which suggested hydrogen bonds and Van der Waals played a major role in the interaction. The distance and efficiency of energy transfer between riboflavin and RBP were 0.70 nm and 0.39, respectively, based on the theory of Forster nonradiative energy transfer. Furthermore, the synchronous fluorescence spectroscopy was utilized to investigate the conformational transformation.

[Effect of S-configuration transformation on the microstructure of ovalbumin].

The effect of S-configuration transformation on the microstructure of ovalbumin was studied by CD spectra, XRD spectra, ANS fluorescence probe emission spectra and UV absorption spectra. CD spectra was used to examine the changes in the secondary structure of the ovalbumin during S-ovalbumin information process. When the induction time was prolonged, the mutual transformation between alpha-helix, beta-sheet, beta-turn and the random coil was observed, and the orderliness of the secondary structure was increased with alpha-helix decreasing slightly and beta-sheet increasing correspondingly. XRD spectra analysis showed that the crystal structure content of the ovalbumin increased with prolonging the induction time and the largest data was observed at 72 h, indicating that the orderliness of the secondary structure was increased. The results were similar to CD spectra analysis. The ANS fluorescence probe emission spectra analysis demonstrated that S-configuration transformation induced an increase in surface hydrophobicity with prolonging the induction time, and the largest data was also observed at 72 h. In addition, UV absorption spectra analysis indicated that S-configuration transformation resulted in a decrease in the UV-absorption maximum value with prolonging the induction time, indicating that the aromatic amino acid was buried in the molecular interior. The results indicated that the changes in the microstructure of ovalbumin were relevant to S-configuration transformation.

An efficient TP-FRET-based lysosome-targetable fluorescent probe for imaging peroxynitrite with two well-resolved emission channels in living cells, tissues and zebrafish.

Peroxynitrite (ONOO-), an important role of reactive oxygen species (ROS) in vivo, and studies showed abnormal of ROS can induce lysosomal membrane permeabilization (LMP) and lead to the death of cells. Thus, it is of great significance for designing an effective method for investigating relationship between physiology and pathology between ONOO- and lysosome. Herein, for the first time, we adopted a Förster resonance energy transfer (FRET) strategy to construct a lysosome-targetable small molecular ratiometric two-photon (TP) fluorescent probe (NpRh-ONOO) for detecting ONOO- in living cells, tissues and zebrafish. Specifically, a TP fluorophore and a rhodamine B fluorophore are directly connected by a flexible piperidine linker to form the TP-FRET-scaffold, a hydrazide as ONOO- reactive set, and a dimethylamino as lysosome targeting-group, which shows a target-modulated ratiometric TP fluorescence response, two well-resolved emission peaks separated by 73 nm, rapid response (<10 s), high selectivity and sensitivity with the detection limit is as low as 3.3 nM for ONOO-. These prominent features of probe were then applied for ratiometric bioimaging both exogenous and endogenous peroxynitrite in living cells, tissues and zebrafish, demonstrating it can be used as a powerful tool for biological research of lysosomal peroxynitrite in biological systems.