Increase of Calcium Levels at Interphase in NIH 3T3 Cells

The fluorescent calcium ion indicator dye Fluo-3 and DNA-binding dye Hoechst 33342 were employed to determine, in a quantitative microspectrofluorometric study, the intracellular calcium ion concentration ([Ca~(2+)]_i) and the DNA content of individual living NIH3T3 cells. The well-separated excitation and emission properties of these dyes allowed us to establish for each cell both the phase of the cell cycle using DNA content and [Ca~(2+)]_i. We found that the transition from G_1, through S, to the G_2 phase is accompanied by a two-fold increase in [Ca~(2+)]_i. The [Ca~(2+)]_i was inhomologous in each phase of the interphase (G_1, S and G_2) although [Ca~(2+)]_i in the S and G_2 phases was never lower than certain threshold values in the G_1 and S phases respectively. [Ca~(2+)]_i in G_0 cells was lower than that in G_2 cells. These changes in [Ca~(2+)]_i suggest that [Ca~(2+)]_i may be an import regulator of cell cycle progression.     

Nanoreactors for Chemical Synthesis and Biomedical Applications

With the rapid development of nanoscience and nanotechnology, various types of functional nanoreactors have been designed for diverse applications. Here, the recent evolution of the rational design of nanoreactors for chemical synthesis and biomedical applications are briefly summarized and discussed. The presence of nanoreactors provides constrained space isolated from the surrounding environment. Scientists are committed to studying changes in chemical reactions when the reaction system is confined to the nanosized space. Nanoreactors accelerate the reaction rate and even change mechanism of some chemical reactions. Cells and organelles as natural nanoreactors are also discussed. The development of intracellular synthesis makes it possible to realize various applications in biomedicine. The challenges on the rational design of nanoreactors and perspectives are also discussed.     

Lactose as a “Trojan Horse” for Quantum Dot Cell Transport

A series of glycan‐coated quantum dots were prepared to probe the effect of glycan presentation in intracellular localization in HeLa and SV40 epithelial cells. We show that glycan density mostly impacts on cell toxicity, whereas glycan type affects the cell uptake and intracellular localization. Moreover, we show that lactose can act as a “Trojan horse” on bi‐functionalized QDs to help intracellular delivery of other non‐internalizable glycan moieties and largely avoid the endosomal/lysosomal degradative pathway.     

A Bioorthogonal Small‐Molecule‐Switch System for Controlling Protein Function in Live Cells

Chemically induced dimerization (CID) has proven to be a powerful tool for modulating protein interactions. However, the traditional dimerizer rapamycin has limitations in certain in vivo applications because of its slow reversibility and its affinity for endogenous proteins. Described herein is a bioorthogonal system for rapidly reversible CID. A novel dimerizer with synthetic ligand of FKBP′ (SLF′) linked to trimethoprim (TMP). The SLF′ moiety binds to the F36V mutant of FK506‐binding protein (FKBP) and the TMP moiety binds to E. coli dihydrofolate reductase (eDHFR). SLF′‐TMP‐induced heterodimerization of FKBP(F36V) and eDHFR with a dissociation constant of 0.12 μM . Addition of TMP alone was sufficient to rapidly disrupt this heterodimerization. Two examples are presented to demonstrate that this system is an invaluable tool, which can be widely used to rapidly and reversibly control protein function in vivo.     

Visualization and In Situ Ablation of Intracellular Bacterial Pathogens through Metabolic Labeling

Protected by the host cells, the hidden intracellular bacteria are typically difficult to kill by common antibiotics and cannot be visualized without complex cellular pretreatments. Herein, we successfully developed a bacteria‐metabolizable dual‐functional probe TPEPy‐d ‐Ala, which is based on d ‐alanine and a photosensitizer with aggregation‐induced emission for fluorescence turn‐on imaging of intracellular bacteria in living host cells and photodynamic ablation in situ. Once metabolically incorporated into bacterial peptidoglycan, the intramolecular motions of TPEPy‐d ‐Ala are inhibited, leading to an enhanced fluorescent signal, which allows the clear visualization of the intracellular bacteria. Moreover, TPEPy‐d ‐Ala can effectively ablate the labeled intracellular bacteria in situ owing to covalent ligation to peptidoglycan, yielding a low intracellular minimum inhibitory concentration (MIC) of 20±0.5 μg mL^?1, much more efficient than that of a commonly used antibiotic, vancomycin.     

Polymer Nanocontainers for Intracellular Delivery

Carriers for intracellular delivery are required to overcome limitations of therapeutic agents such as low specificity, systemic toxicity, high clearance rate, and low therapeutic index. Nanocontainers comprised of an aqueous core and a polymer shell have received increasing attention because they readily combine stimuli response to improve intracellular payload release and surface modification to enhance selectivity towards the desired region of action. This Minireview summarizes the design and properties of polymer nanocontainers for intracellular delivery, classified according to the polymer architecture.     

Intracellular Ruthenium‐Promoted (2+2+2) Cycloadditions

Metal‐mediated intracellular reactions are becoming invaluable tools in chemical and cell biology, and hold promise for strongly impacting the field of biomedicine. Most of the reactions reported so far involve either uncaging or redox processes. Demonstrated here for the first time is the viability of performing multicomponent alkyne cycloaromatizations inside live mammalian cells using ruthenium catalysts. Both fully intramolecular and intermolecular cycloadditions of diynes with alkynes are feasible, the latter providing an intracellular synthesis of appealing anthraquinones. The power of the approach is further demonstrated by generating anthraquinone AIEgens (AIE=aggregation induced emission) that otherwise do not go inside cells, and by modifying the intracellular distribution of the products by simply varying the type of ruthenium complex.     

Theil Entropy as a Non-Lineal Analysis for Spectral Inequality of Physiological Oscillations

Theil entropy is a statistical measure used in economics to quantify income inequalities. However, it can be applied to any data distribution including biological signals. In this work, we applied different spectral methods on heart rate variability signals and cellular calcium oscillations previously to Theil entropy analysis. The behavior of Theil entropy and its decomposable property was investigated using exponents in the range of [−1, 2], on the spectrum of synthetic and physiological signals. Our results suggest that the best spectral decomposition method to analyze the spectral inequality of physiological oscillations is the Lomb–Scargle method, followed by Theil entropy analysis. Moreover, our results showed that the exponents that provide more information to describe the spectral inequality in the tested signals were zero, one, and two. It was also observed that the intra-band component is the one that contributes the most to total inequality for the studied oscillations. More in detail, we found that in the state of mental stress, the inequality determined by the Theil entropy analysis of heart rate increases with respect to the resting state. Likewise, the same analytical approach shows that cellular calcium oscillations present on developing interneurons display greater inequality distribution when inhibition of a neurotransmitter system is in place. In conclusion, we propose that Theil entropy is useful for analyzing spectral inequality and to explore its origin in physiological signals.