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Cell membrane (CM) is a phospholipid bilayer that maintains integrity of a whole cell and relates to many physiological and pathological processes. Developing CM imaging tools is a feasible method for visualizing membrane-related events. In recent decades, small-molecular fluorescent probes in the near-infrared (NIR) region have been pursued extensively for CM staining to investigate its functions and related events. In this review, we summarize development of such probes from the aspect of design principles, CM-targeting mechanisms and biological applications. Moreover, at the end of this review, the challenges and future research directions in designing NIR CM-targeting probes are discussed. This review indicates that more efforts are required to design activatable NIR CM-targeting probes, easily prepared and biocompatible probes with long retention time regarding CM, super-resolution imaging probes for monitoring CM nanoscale organization and multifunctional probes with imaging and phototherapy effects.

Lanthanide doping is an effective strategy for modulating the emission of emitters. Herein, by changing the cluster composition to control the energy transfer pathway, the application potential of high-nuclearity lanthanide cluster (HLC) as white-light emitter has been confirmed for the first time. Specifically, by precisely controlling the proportion of Gd III , Tb III , and Eu III ions in reactants, we obtained a spherical heterotrimetallic nanocluster Gd 10 Tb 12 Eu 10 , a white-light emitter with quantum yield (QY) of 12.58% and lifetime of 327.14 µs. High-resolution electrospray ionization mass spectrometry (HRESI-MS) demonstrates that homometallic nanoclusters Ln 32 (Ln = Gd, Tb, and Eu) are tetracationic clusters and are highly stable in solution. The peripheral dense organic ligands provide a protective layer for the cluster core, which improves the stability of Ln 32 in aqueous solution, avoids the contact between metal centers and bioactive molecules, and greatly reduces the biological toxicity. In cell imaging experiments, cationic clusters Ln 32 are mainly localized on the cell membrane with negative charge distribution. As far as we know, this is the first time that spherical lanthanide nanoclusters have been used for membrane imaging of living cell, opening the door for the application of HLCs in biological imaging.

Plant metabolism is altered in response to various environmental changes. In vegetable crops such as tomato (Solanum lycopersicum), the metabolic composition of fruits varies depending on the variety or cultivar as well as the cultivation method used. Few studies have examined the metabolic fluctuations in fruits under stress conditions, such as drought. We previously examined the metabolomes of mature green tomato fruits, which undergo drastic changes in chemical composition during ripening, and mature red fruits in response to drought stress. We detected or predicted fluctuations in the levels of fatty acids and phospholipid constituents, such as inositol and ethanolamine. In this study, we determined the localizations of these metabolites in fruits using mass spectrometry imaging. The accumulation patterns of stearic acid and palmitic acid were similar, but unlike these fatty acids, oleic acid accumulated to high levels in the placenta. Inositol is involved in various physiological processes; under drought conditions, this metabolite is synthesized by a different pathway compared to under normal conditions. The biosynthesis of pectin, a component of the gel surrounding the seeds, was suppressed under drought stress but increased in seeds. We propose that under drought conditions, a shift to phospholipid biosynthesis occurs that protects seeds from dehydration.

Dopamine released by amphetamine decreases the in vivo binding of PET radioligands to the dopamine D2 receptor. Although concentrations of extracellular dopamine largely return to baseline within 1 to 2 h after amphetamine treatment, radioligand binding remains decreased for several hours. The purpose of this study was to determine whether the prolonged decrease of radioligand binding after amphetamine administration is caused by receptor internalization. To distinguish dopamine displacement from receptor internalization, we used wild-type and arrestin3 (arr3) knockout mice, which are incapable of internalizing D2 receptors. In addition, we used both the D2 selective agonist [11C]MNPA (which is thought to bind to the high affinity state of the receptor) and the D2 selective antagonist [18F]fallypride (which does not differentiate between high and low affinity state). After an initial baseline scan, animals were divided in three groups for a second scan: either 30 min or 4 h after amphetamine administration (3 mg/kg, i.p.) or as retest. At 30 min, [11C]MNPA showed greater displacement than [18F]fallypride, but each radioligand gave similar displacement in knockout and wild-type mice. At 4 h, the binding of both radioligands returned to baseline in arr3 knockout mice, but remained decreased in wild-type mice. Radioligand binding was unaltered on retest scanning. Our results suggest that the prolonged decrease of radioligand binding after amphetamine is mainly due to internalization of the D2 receptor rather than dopamine displacement. In addition, this study demonstrates the utility of small animal PET to study receptor trafficking in vivo in genetically modified mice.

Demobesin 1 is a potent new GRP-R-selective bombesin (BN) analogue containing an open chain tetraamine chelator for stable technetium-99m binding. Following a convenient labelling protocol, the radiopeptide, [(99m)Tc]Demobesin 1, formed in nearly quantitative yields and with high specific activities. Both unlabelled and labelled peptide demonstrated high-affinity binding in membrane preparations of the human androgen-independent prostate adenocarcinoma PC-3 cell line. The IC(50) values determined for Demobesin 1 and [Tyr(4)]BN were 0.70+/-0.08 n M and 1.5+/-0.20 n M, respectively, while the K(d) defined for [(99m)Tc/(99g)Tc]Demobesin 1 was 0.67+/-0.10 n M. [(99m)Tc]Demobesin 1 was rather stable in murine plasma, whereas it degraded rapidly in kidney and liver homogenates. After injection in healthy Swiss albino mice, [(99m)Tc]Demobesin 1 accumulated very efficiently in the target organs (pancreas, intestinal tract) via a GRP-R-mediated process, as shown by in vivo receptor blocking experiments. An equally high and GRP-R-mediated uptake was exhibited by [(99m)Tc]Demobesin 1 after injection in PC-3 tumour-bearing athymic mice. The initial high radioligand uptake of 16.2+/-3.1%ID/g in the PC-3 xenografts at 1 h p.i. remained at a similar level (15.61+/-1.19%ID/g) at 4 h p.i. Even after 24 h p.i., when the radioactivity had cleared from all other tissues, a value of 5.24+/-0.67%ID/g was still observed in the tumour. The high and prolonged localization of [(99m)Tc]Demobesin 1 at the tumour site and its rapid background clearance are very promising qualities for GRP-R-targeted tumour imaging in man.

Growth of alkaliphilic Bacillus halodurans C-125 both on agar plates and in liquid culture was inhibited by methyl-β-cyclodextrin (CD). Furthermore, resting cells of the strain were lysed by contact with methyl-β-CD higher than 10 mM. α-CD also showed lysis activity against Bacillus and related strains. The activity was not observed with Gram-negative and Gram-positive bacteria except for Bacillus strains. Fluorescence staining and scanning electron microscopy of cells revealed that methyl-β-CD disrupted cell membranes, and consequently, the cells were lysed. This is a novel physiological property of CDs.

Images recorded in a transmission electron microscope are \"projection\" images, meaning that they contain information from all regions of the specimen through which the beam was transmitted. [...]each image contains information from all heights of the specimen collapsed into a single plane. By recording a series of images in which the orientation of the specimen is varied relative to the incident beam, it is possible to obtain a series of projection views of the object (Figure 1), which can be converted into a 3-D volume using methods that generally rely on the use of weighted back-projection algorithms. Since the first use of these methods nearly four decades ago [10], they have been applied to a variety of biological objects to describe 3-D structures at varying resolutions.

Purpose: The biologic effects and the underlying mechanisms of Ehrlich ascitic tumor (EAT) cells induced by ultrasound were investigated in this study. Methods: Cells were subjected to ultrasonic irradiation with a frequency of 2.17 MHz and an intensity of 3 W/cm2 for variable periods of time. Trypan blue exclusion was used to detect the integrity of cellular membrane; the membrane permeability was investigated by the incorporation of fluorescein isothiocyanate dextran during ultrasound exposure; and the cell membrane ultrastructure changes were observed under a scanning electron microscope. The potential mechanism was estimated from the generation of hydroxyl radicals, the lipid peroxidation levels, and intracellular reactive oxygen radicals production. Results: The cell membrane damage effects induced by ultrasound increased with a prolonged exposure time; the fluorescent rates of the cells irradiated with ultrasound for 30 and 60 seconds were 11.46% and 18.50%, respectively; the amount of hydroxyl radicals in 30 (26.10 U/mL) and 60 seconds (28.47 U/mL) were significantly enhanced, compared with the control group (24.44 U/mL); then, the level of lipid peroxidation was also changed from 0.27 to 0.54 (30 seconds) and 1.21 nmol/mL (60 seconds). Conclusions: Shear forces and free radicals produced by acoustic cavitation may play important roles in these actions.

The peritoneal contact surface area (PCSA), which represents the area parameter in the mass transfer area coefficient (MTAC), is a crucial marker in the evaluation of peritoneal dialysis effectiveness. However, the capacity to recruit a larger PCSA has only been rarely demonstrated in vivo and, in most cases, changes in MTAC are interpreted as permeability changes and not as surface area variations. Here, we report the use of micro-computerized tomography (μCT) for the measurement of PCSA changes to various fill volumes. Using this three-dimensional imaging method, PCSA was measured in vivo in 26 healthy Wistar rats receiving intraperitoneally increasing fill volumes of peritoneal dialysis solutions: 5 mL (group 1, n  = 8), 10 mL (group 2, n  = 8) and 15 mL (group 3, n  = 10) per 100 g of body weight. A non-ionic iodinated contrast agent was added to the dialysis solution in order to distinguish the intraperitoneal dialysis solutions from soft tissues. The normalized PCSA/weight ratio (cm 2 /g) increased with fill volume: 1.12 ± 0.10 cm 2 /g (range 0.98–1.25) in group 1; 1.74 ± 0.08 cm 2 /g (range 1.64–1.87) in group 2; 2.13 ± 0.09 cm 2 /g(range 1.90–2.30) in group 3. With this μCT method, PCSA recruited in vivo with a 10 mL/100 g fill volume was in the range 94–107%) of ex vivo total peritoneal surface area (evPSA), as calculated with the Kuzlan’s formula. With a 15 mL/100 g fill volume, the in vivo-measured PCSA, the exchange surface area, surpassed the evPSA (range 113–139%).

Mitochondrial dysfunction has been attributed a critical role in the etiology and pathogenesis of numerous diseases, and is manifested by alterations of the organelle's membrane potential (Deltapsi(m)). This suggests that Deltapsi(m) measurement can be highly useful for diagnostic purposes. In the current study, we characterized the capability of the novel PET agent (18)F-fluorobenzyl triphenylphosphonium ((18)F-FBnTP) to assess Deltapsi(m), compared with the well-established voltage sensor (3)H-tetraphenylphosphonium ((3)H-TPP). (18)F-FBnTP and (3)H-TPP uptake under conditions known to alter Deltapsi(m) and plasma membrane potential (Deltapsi(p)) was assayed in the H345 lung carcinoma cell line. (18)F-FBnTP biodistribution was assessed in CD1 mice using dynamic PET and ex vivo gamma well counting. (18)F-FBnTP and (3)H-TPP demonstrated similar uptake kinetics and plateau concentrations in H345 cells. Stepwise membrane depolarization resulted in a linear decrease in (18)F-FBnTP cellular uptake, with a slope (-0.58+/-0.06) and correlation coefficient (0.94+/-0.07) similar (p>0.17) to those measured for (3)H-TPP (-0.63+/-0.06 and 0.96+/-0.05, respectively). Selective collapse of Deltapsi(m) caused a substantial decrease in cellular uptake for (18)F-FBnTP (81.6+/-8.1%) and (3)H-TPP (85.4+/-6.7%), compared with control. Exposure to the proapoptotic staurosporine, known to collapse Deltapsi(m), resulted in a decrease of 68.7+/-10.1% and 71.5+/-8.4% in (18)F-FBnTP and (3)H-TPP cellular uptake, respectively. (18)F-FBnTP accumulated mainly in kidney, heart and liver. (18)F-FBnTP is a mitochondria-targeting PET radiopharmaceutical responsive to alterations in membrane potential with voltage-dependent performance similar to that of (3)H-TPP. (18)F-FBnTP is a promising new voltage sensor for detection of physiological and pathological processes associated with mitochondrial dysfunction, such as apoptosis, using PET.