A novel nanoparticle fluorescent probe based on a water-soluble conjugated polymer for real-time monitoring of ATP fluctuation and configuration of the Golgi apparatus during the inhibition of glycolysis.

BACKGROUND: Adenosine 5'-triphosphate (ATP) plays an important role in cell metabolism and has been regarded as an indicator of cell survival and damage. Golgi apparatus participates in the signal transduction processes of substance transport, ion homeostasis and stress when extracellular substances enter cells. Till now, there is no fluorescent probe for monitoring Golgi ATP level fluctuation and visualizing the configuration change of the Golgi apparatus during the inhibition of glycolysis. RESULTS: Herein, we report the synthesis of a novel water-soluble cationic polythiophene derivative (PEMTEA) that can be employed as a fluorescent sensor for measuring ATP in the Golgi apparatus. PEMTEA self-assembles into PT-NP nanoparticles in aqueous solution with a diameter of approximately 2 nm. PT-NP displays high sensitivity and superb selectivity towards ATP with a detection limit of 90 nM and a linear detection range from 0 to 3.0 µM. The nanoparticles show low toxicity to HepG2 cells and good photostability in the Golgi apparatus. With the stimulation of Ca2+, PT-NP was practically applied to real-time monitor of endogenous ATP levels in the Golgi apparatus through fluorescence microscopy. Finally, we studied the relationship between the concentration of ATP and configuration of the Golgi apparatus during the inhibition of glycolysis using PT-NP. SIGNIFICANCE: We have demonstrated that PT-NP can not only indicate the fluctuation and distribution of ATP in the Golgi apparatus, but also give the information of the configuration change of the Golgi apparatus at the single-cell level during the inhibition of glycolysis.

ALT-803 in the treatment of non-muscle-invasive bladder cancer: Preclinical and clinical evidence and translational potential.

Bladder cancer (BCa) is one of the most common malignant tumors that cause death. Approximately 75%-85% of BCa develop into non-muscle-invasive bladder cancer (NMIBC). Bacillus Calmette-Guérin (BCG) is the gold standard for avoiding cystectomy in the treatment of NMIBC. Unfortunately, up to 30% of patients do not respond to BCG treatment, and up to 70% of BCG responders relapse. The United States Food and Drug Administration (FDA) approved valrubicin (1998) and pembrolizumab (2020) for the treatment of BCG-unresponsive (BCGu) NMBIC. However, the complete remission rate for valrubicin and pembrolizumab was only 16% and 40.6%, respectively. ALT-803 (N-803) is an IL-15 superagonist and reduces tumor burden by promoting the proliferation and activation of NK cells and CD8+ T cells. The FDA received (23 May 2022) and accepted to review (28 July 2022) the marketing submission of ALT-803 plus BCG for the treatment of BCGu NMIBC. However, the FDA previously rejected the application for oportuzumab monatox (OM) due to a lack of data comparing it with pembrolizumab on August 20, 2021. Interestingly, the clinical efficacy and safety of ALT-803 were higher than that of pembrolizumab and OM, suggesting that ALT-803 may be approved by FDA. This review aims to further knowledge of the preclinical and clinical evidence of ALT-803 in the treatment of NMIBC and discuss its translational potential.

Advances in Trop2-targeted therapy: Novel agents and opportunities beyond breast cancer.

Trop2 is a transmembrane glycoprotein and calcium signal transducer with limited expression in normal human tissues. It is consistently overexpressed in a variety of malignant tumors and participates in several oncogenic signaling pathways that lead to tumor development, invasion, and metastasis. As a result, Trop2 has become an attractive therapeutic target in cancer treatment. The anti-Trop2 antibody-drug conjugate (Trodelvy™, sacituzumab govitecan) has been approved to treat metastatic triple-negative breast cancer. However, it is still unclear whether the success observed in Trop2-positive breast cancer could be replicated in other tumor types, owing to the differences in the expression levels and functions of Trop2 across cancer types. In this review, we summarize the recent progress on the structures and functions of Trop2 and highlight the potential diagnostic and therapeutic value of Trop2 beyond breast cancer. In addition, the promising novel Trop2-targeted agents in the clinic were discussed, which will likely alter the therapeutic landscape of Trop2-positive tumors in the future.

Targeting NAD+: is it a common strategy to delay heart aging?

Heart aging is the main susceptible factor to coronary heart disease and significantly increases the risk of heart failure, especially when the aging heart is suffering from ischemia-reperfusion injury. Numerous studies with NAD+ supplementations have suggested its use in anti-aging treatment. However, systematic reviews regarding the overall role of NAD+ in cardiac aging are scarce. The relationship between NAD+ signaling and heart aging has yet to be clarified. This review comprehensively summarizes the current studies on the role of NAD+ signaling in delaying heart aging from the following aspects: the influence of NAD+ supplementations on the aging heart; the relationship and cross-talks between NAD+ signaling and other cardiac aging-related signaling pathways; Importantly, the therapeutic potential of targeting NAD+ in delaying heart aging will be discussed. In brief, NAD+ plays a vital role in delaying heart aging. However, the abnormalities such as altered glucose and lipid metabolism, oxidative stress, and calcium overload could also interfere with NAD+ function in the heart. Therefore, the specific physiopathology of the aging heart should be considered before applying NAD+ supplementations. We believe that this article will help augment our understanding of heart aging mechanisms. In the meantime, it provides invaluable insights into possible therapeutic strategies for preventing age-related heart diseases in clinical settings.

Numerical Investigation on the Optimum Thermal Design of the Shape and Geometric Parameters of Microchannel Heat Exchangers with Cavities.

Due to the large surface-area-to-volume ratio, microchannel heat exchangers have a higher heat transfer rate compared with traditional scale heat exchangers. In this study, the optimum microchannel cavity with high heat transfer and low flow resistance is designed to further improve microchannel exchangers' thermal performance. A three-dimensional laminar flow model, consisting of Navier-Stokes equations and an energy conservation equation is solved and the conjugate heat transfer between the silicon basement and deionized water is taken into consideration. The impact of the shape, aspect ratio, size and spacing of the cavity on the thermal performance of microchannel exchangers are numerically investigated, respectively. The results indicated that the cavity on the sidewall can enhance heat transfer and reduce flow resistance simultaneously, and cavities with a relatively small expansion angle and streamlined edge could enhance thermal performance the most. Based on the conclusions, a new cavity shape is proposed, and the simulation results verify its excellent thermal performance as expected. Furthermore, investigation is performed to figure out the optimum design of the new cavity and the optimal geometric parameters of the cavity under different flow conditions have been obtained in principle for microchannel exchangers' design.

Experimental Investigation of the Flow and Heat Transfer Characteristics in Microchannel Heat Exchangers with Reentrant Cavities.

The application of microchannel heat exchangers is of great significance in industrial fields due to their advantages of miniaturized scale, large surface-area-to-volume ratio, and high heat transfer rate. In this study, microchannel heat exchangers with and without fan-shaped reentrant cavities were designed and manufactured, and experiments were conducted to investigate the flow and heat-transfer characteristics. The impact rising from the radius of reentrant cavities, as well as the Reynolds number on the heat transfer and the pressure drop, is also analyzed. The results indicate that, compared with straight microchannels, microchannels with reentrant cavities could enhance the heat transfer and, more importantly, reduce the pressure drop at the same time. For the ranges of parameters studied, increasing the radius of reentrant cavities could augment the effect of pressure-drop reduction, while the corresponding variation of heat transfer is complicated. It is considered that adding reentrant cavities in microchannel heat exchangers is an ideal approach to improve performance.

Flow Characteristics of the Entrance Region with Roughness Effect within Rectangular Microchannels.

We conducted systematic numerical investigations of the flow characteristics within the entrance region of rectangular microchannels. The effects of the geometrical aspect ratio and roughness on entrance lengths were analyzed. The incompressible laminar Navier-Stokes equations were solved using finite volume method (FVM). In the simulation, hydraulic diameters (Dh) ranging from 50 to 200 µm were studied, and aspect ratios of 1, 1.25, 1.5, 1.75, and 2 were considered as well. The working fluid was set as water, and the Reynolds number ranged from 0.5 to 100. The results showed a good agreement with the conducted experiment. Correlations are proposed to predict the entrance lengths of microchannels with respect to different aspect ratios. Compared with other correlations, these new correlations are more reliable because a more practical inlet condition was considered in our investigations. Instead of considering the influence of the width and height of the microchannels, in our investigation we proved that the critical role is played by the aspect ratio, representing the combination of the aforementioned parameters. Furthermore, the existence of rough elements obviously shortens the entrance region, and this effect became more pronounced with increasing relative roughness and Reynolds number. A similar effect could be seen by shortening the roughness spacing. An asymmetric distribution of rough elements decreased the entrance length compared with a symmetric distribution, which can be extrapolated to other irregularly distributed forms.

Experimental and Numerical Investigations on the Flow Characteristics within Hydrodynamic Entrance Regions in Microchannels.

Flow characteristics within entrance regions in microchannels are important due to their effect on heat and mass transfer. However, relevant research is limited and some conclusions are controversial. In order to reveal flow characteristics within entrance regions and to provide empiric correlation estimating hydrodynamic entrance length, experimental and numerical investigations were conducted in microchannels with square cross-sections. The inlet configuration was elaborately designed in a more common pattern for microdevices to diminish errors caused by separation flow near the inlet and fabrication faults so that conclusions which were more applicable to microchannels could be drawn. Three different microchannels with hydraulic diameters of 100 µm, 150 µm, and 200 µm were investigated with Reynolds (Re) number ranging from 0.5 to 50. For the experiment, deionized water was chosen as the working fluid and microscopic particle image velocimetry (micro-PIV) was adopted to record and analyze velocity profiles. For numerical simulation, the test-sections were modeled and incompressible laminar Navier-Stokes equations were solved with commercial software. Strong agreement was achieved between the experimental data and the simulated data. According to the results of both the experiments and the simulations, new correlations were proposed to estimate entrance length. Re numbers ranging from 12.5 to 15 was considered as the transition region where the relationship between entrance length and Re number converted. For the microchannels and the Reynolds number range investigated compared with correlations for conventional channels, noticeable deviation was observed for lower Re numbers (Re < 12.5) and strong agreement was found for higher Re numbers (Re > 15).

Dual-target cancer theranostic for glutathione S-transferase and hypoxia-inducible factor-1α inhibition.

We developed a dual-target theranostic F671, which could exhibit synergetic anticancer effects for inhibiting the activities of glutathione S-transferase and the accumulation of hypoxia inducible factor-1α. F671 undergoes self-immolative cleavage when exposed to GSTP1-1 in live cancer cells, facilitating the visualization of molecule release and distribution, as well as confirming the autophagy-induced apoptosis.

Water-Soluble Conjugated Polymer as a Fluorescent Probe for Monitoring Adenosine Triphosphate Level Fluctuation in Cell Membranes during Cell Apoptosis and in Vivo.

Adenosine triphosphate (ATP) is used as the energy source in cells and plays crucial roles in various cellular events. The cellular membrane is the protective barrier for the cytoplasm of living cells and involved in many essential biological processes. Many fluorescent probes for ATP have been successfully developed, but few of these probes were appropriate for visualizing ATP level fluctuation in cell membranes during the apoptotic cell death process. Herein, we report the synthesis of a new water-soluble cationic polythiophene derivative that can be utilized as a fluorescent sensor for detecting ATP in cell membranes. Poly((3-((4-methylthiophen-3-yl)oxy)propyl)triphenylphosphonium chloride) (PMTPP) exhibits high sensitivity and good selectivity to ATP, and the detection limit is 27 nM. The polymer shows low toxicity to live cells and excellent photostability in cell membranes. PMTPP was practically utilized for real-time monitoring of ATP levels in the cell membrane through fluorescence microscopy. We have demonstrated that the ATP levels in cell membranes increased during the apoptotic cell death process. The probe was also capable of imaging ATP levels in living mice.

Molecular visualizing and quantifying immune-associated peroxynitrite fluxes in phagocytes and mouse inflammation model.

Reactions of peroxynitrite (ONOO-) with biomolecules can lead to cytotoxic and cytoprotective events. Due to the difficulty of directly and unambiguously measuring its levels, most of the beneficial effects associated with ONOO- in vivo remain controversial or poorly characterized. Recently, optical imaging has served as a powerful noninvasive approach to studying ONOO- in living systems. However, ratiometric probes for ONOO- are currently lacking. Herein, we report the design, synthesis, and biological evaluation of F482, a novel fluorescence indicator that relies on ONOO--induced diene oxidation. The remarkable sensitivity, selectivity, and photostability of F482 enabled us to visualize basal ONOO- in immune-stimulated phagocyte cells and quantify its generation in phagosomes by high-throughput flow cytometry analysis. With the aid of in vivo ONOO- imaging in a mouse inflammation model assisted by F482, we envision that F482 will find widespread applications in the study of the ONOO- biology associated with physiological and pathological processes in vitro and in vivo.

Lysosomal ATP imaging in living cells by a water-soluble cationic polythiophene derivative.

Lysosomes in astrocytes and microglia can release ATP as the signaling molecule for the cells through ca(2+)-dependent exocytosis in response to various stimuli. At present, fluorescent probes that can detect ATP in lysosomes have not been reported. In this work, we have developed a new water-soluble cationic polythiophene derivative that can be specifically localized in lysosomes and can be utilized as a fluorescent probe to sense ATP in cells. PEMTEI exhibits high selectivity and sensitivity to ATP at physiological pH values and the detection limit of ATP is as low as 10(-11)M. The probe has low cytotoxicity, good permeability and high photostability in living cells and has been applied successfully to real-time monitoring of the change in concentrations of ATP in lysosomes though fluorescence microscopy. We also demonstrated that lysosomes in Hela cells can release ATP through Ca(2+)-dependent exocytosis in response to drug stimuli.

Halochromism of a polythiophene derivative induced by conformational changes and its sensing application of carbon dioxide.

We report the design and synthesis of a halochromic polythiophene derivative, whose conformation can be alternated between random coil and rodlike phase by adjusting the pH of the solution. Distinct solution color changes associated with the pH-induced conformational transitions can be used to construct a colorimetric probe for sensing carbon dioxide. This probe can be recovered by bubbling nitrogen gas into carbon dioxide-treated solutions for over 20 cycles.

Colorimetric detection of copper ions based on a supramolecular complex of water-soluble polythiophene and ATP.

A colorimetric probe for the detection of copper(II) ions in aqueous media by the naked eye has been developed based on a supramolecular complex comprised of a cationic polythiophene derivative and ATP with a detection limit as low as 0.05 mM.