Radioactive Smart Probe for Potential Corrected Matrix Metalloproteinase Imaging

Although various activatable optical probes have been developed to visualize metalloproteinase (MMP) activities in vivo, precise quantification of the enzyme activity is limited due to the inherent scattering and attenuation (limited depth penetration) properties of optical imaging. In this investigation, a novel activatable peptide probe ⁶⁴Cu-BBQ650-PLGVR-K­(Cy5.5)-E-K­(DOTA)-OH was constructed to detect tumor MMP activity in vivo. This agent is optically quenched in its native form, but releases strong fluorescence upon cleavage by selected enzymes. MMP specificity was confirmed both in vitro and in vivo by fluorescent imaging studies. The use of a single modality to image biomarkers/processes may lead to erroneous interpretation of imaging data. The introduction of a quantitative imaging modality, such as PET, would make it feasible to correct the enzyme activity determined from optical imaging. In this proof of principle report, we demonstrated the feasibility of correcting the activatable optical imaging data through the PET signal. This approach provides an attractive new strategy for accurate imaging of MMP activity, which may also be applied for other protease imaging

Three Stages of Dynamic Assembly Process of Dipeptide-Based Supramolecular Gel Revealed by In Situ Infrared Spectroscopy

The exploration of short peptide-based assembly is vital for understanding protein-misfolding-associated diseases and seeking strategies to attenuate aggregate formation. While, the molecular mechanism of their structural evolution remains poorly studied in view of the dynamic and unpredictable assembly process. Herein, infrared (IR) spectroscopy, which serves as an in situ and real-time analytical technique, was intelligently employed to investigate the mechanism of phase transition and aggregate formation during the dynamic assembly process of diphenylalanine. Combined with other spectroscopy and electron microscopy technologies, three stages of gel formation and the main driving forces in different stages were revealed. A variety of stoichiometric methods such as continuous wavelet transform, principal component analysis, and two-dimensional correlation spectroscopy techniques were conducted to analyze the original time-dependent IR spectra to obtain detailed information on the changes in the amide bands and hydration layer. The microenvironment of hydrogen bonding among amide bands was significantly changed with the addition of pyridine derivatives, resulting in great differences in the properties of co-assembled gels. This work not only provides a universal analytical way to reveal the dynamic assembly process of dipeptide-based supramolecular gel but also expands their applications in supramolecular regulation and high-throughput screens in situ

Molecular Imaging of P‑glycoprotein in Chemoresistant Tumors Using a Dual-Modality PET/Fluorescence Probe

Overexpression of P-glycoprotein (Pgp) has been considered a primary cause for multidrug resistance in a variety of cancers for three decades. However, clinical translation of Pgp targeted therapeutics has been hindered by lack of patient preselection based on the Pgp presence in tumors. We aim to develop a molecularly targeted probe for imaging tumoral Pgp <i>in vivo</i> with positron emission tomography (PET) and fluorescence, and to provide a tool for preselecting the patients with tumoral Pgp expression. Thus, a Pgp monoclonal antibody 15D3 was chemically modified with IRDye800 (IR800) and DOTA chelator. The specificity of the antibody conjugates DOTA-Pab-IR800 was verified in Pgp-expressing 3T3-MDR1 and control 3T3 cells. After radiolabeling with ⁶⁴Cu, the probe was applied in small animal PET imaging of Pgp in a mouse xenograft model of NCI/ADR-Res cells, which are chemoresistant through overexpression of Pgp. Quantification analysis of the PET images demonstrated that the tumor uptake of the radioactive probe was 9.9 ± 1.4, 12.1 ± 1.2, and 10.5 ± 1.0%ID/g at 4, 24, and 48 h post injection. The tumor-to-muscle ratio was 20.9 at 48 h post injection based on biodistribution studies. Fluorescence imaging was performed following PET experiments, and it demonstrated excellent tumor accumulation of this dual-modality probe in the NCI/ADR-Res tumors. Further, an image-guided surgery was successfully performed using the fluorescence modality of the probe, demonstrating potential utility of this probe in image-guided surgical removal of Pgp-positive drug resistant tumors in the patients. In conclusion, this study clearly demonstrated that the Pgp-targeted antibody probe, ⁶⁴Cu-DOTA-Pab-IR800, could provide a promising diagnosis tool for detection of Pgp-expressing tumors <i>in vivo</i>

Data_Sheet_1_Activation of pheromone-sensitive olfactory neurons by plant volatiles in the moth Agrotis ipsilon does not occur at the level of the pheromone receptor protein.docx

In moths, mate finding relies on female-emitted sex pheromones that the males have to decipher within a complex environmental odorant background. Previous studies have shown that interactions of both sex pheromones and plant volatiles can occur in the peripheral olfactory system, and that some plant volatiles can activate the pheromone-specific detection pathway. In the noctuid moth Agrotis ipsilon, plant volatiles such as heptanal activate the receptor neurons tuned to the pheromone component (Z)7-12:OAc. However, the underlying mechanisms remain totally unknown. Following the general rule that states that one olfactory receptor neuron usually expresses only one type of receptor protein, a logic explanation would be that the receptor protein expressed in (Z)7-12:OAc-sensitive neurons recognizes both pheromone and plant volatiles. To test this hypothesis, we first annotated odorant receptor genes in the genome of A. ipsilon and we identified a candidate receptor putatively tuned to (Z)7-12:OAc, named AipsOR3. Then, we expressed it in Drosophila olfactory neurons and determined its response spectrum to a large panel of pheromone compounds and plant volatiles. Unexpectedly, the receptor protein AipsOR3 appeared to be very specific to (Z)7-12:OAc and was not activated by any of the plant volatiles tested, including heptanal. We also found that (Z)7-12:OAc responses of Drosophila neurons expressing AipsOR3 were not affected by a background of heptanal. As the Drosophila olfactory sensilla that house neurons in which AipsOR3 was expressed contain other olfactory proteins – such as odorant-binding proteins – that may influence its selectivity, we also expressed AipsOR3 in Xenopus oocytes and confirmed its specificity and the lack of activation by plant volatiles. Altogether, our results suggest that a still unknown second odorant receptor protein tuned to heptanal and other plant volatiles is expressed in the (Z)7-12:OAc-sensitive neurons of A. ipsilon.</p

Table_1_Activation of pheromone-sensitive olfactory neurons by plant volatiles in the moth Agrotis ipsilon does not occur at the level of the pheromone receptor protein.docx

In moths, mate finding relies on female-emitted sex pheromones that the males have to decipher within a complex environmental odorant background. Previous studies have shown that interactions of both sex pheromones and plant volatiles can occur in the peripheral olfactory system, and that some plant volatiles can activate the pheromone-specific detection pathway. In the noctuid moth Agrotis ipsilon, plant volatiles such as heptanal activate the receptor neurons tuned to the pheromone component (Z)7-12:OAc. However, the underlying mechanisms remain totally unknown. Following the general rule that states that one olfactory receptor neuron usually expresses only one type of receptor protein, a logic explanation would be that the receptor protein expressed in (Z)7-12:OAc-sensitive neurons recognizes both pheromone and plant volatiles. To test this hypothesis, we first annotated odorant receptor genes in the genome of A. ipsilon and we identified a candidate receptor putatively tuned to (Z)7-12:OAc, named AipsOR3. Then, we expressed it in Drosophila olfactory neurons and determined its response spectrum to a large panel of pheromone compounds and plant volatiles. Unexpectedly, the receptor protein AipsOR3 appeared to be very specific to (Z)7-12:OAc and was not activated by any of the plant volatiles tested, including heptanal. We also found that (Z)7-12:OAc responses of Drosophila neurons expressing AipsOR3 were not affected by a background of heptanal. As the Drosophila olfactory sensilla that house neurons in which AipsOR3 was expressed contain other olfactory proteins – such as odorant-binding proteins – that may influence its selectivity, we also expressed AipsOR3 in Xenopus oocytes and confirmed its specificity and the lack of activation by plant volatiles. Altogether, our results suggest that a still unknown second odorant receptor protein tuned to heptanal and other plant volatiles is expressed in the (Z)7-12:OAc-sensitive neurons of A. ipsilon.</p

Development of [¹⁸F]AlF-NOTA-NT as PET Agents of Neurotensin Receptor‑1 Positive Pancreatic Cancer

Several studies have suggested that neurotensin receptors (NTRs) and neurotensin (NT) greatly affect the growth and survival of pancreatic ductal adenocarcinoma (PDAC). Developing NTR-targeted PET probes could therefore be important for the management of a pancreatic cancer patient by providing key information on the NTR expression profile noninvasively. Despite the initial success on the synthesis of ¹⁸F-labeled NT PET probes, the labeling procedure generally requires lengthy steps including azeotropic drying of ¹⁸F. Using a straightforward chelation method, here we report the simple preparation of aluminum-¹⁸F-NOTA-NT starting from aqueous ¹⁸F. The cell binding test demonstrated that [¹⁹F]­AlF-NOTA-NT maintained high receptor-binding affinity to NTR1. This probe was then further evaluated in NTR1 positive pancreatic tumor models (AsPC-1 and PANC-1). After the administration of [¹⁸F]­AlF-NOTA-NT, small animal PET studies showed a high contrast between tumor and background in both models at 1 and 4 h time points. A blocking experiment was performed to demonstrate the receptor specificity: the tumor uptake in AsPC1 without and with blocking agent was 1.0 ± 0.2 and 0.1 ± 0.0%ID/g, respectively, at 4 h post injection. In summary, a NTR specific PET agent, [¹⁸F]­AlF-NOTA-NT, was prepared through the simple chelation method. This NTR-targeted PET probe may not only be used to detect NTR1 positive pancreatic tumors (diagnosis), but also it may be fully integrated to NTR target therapy leading to personalized medicine (theranostic)

Tunable Polymer Brush/Au NPs Hybrid Plasmonic Arrays Based on Host–guest Interaction

The fabrication of versatile gold nanoparticle (Au NP) arrays with tunable optical properties by a novel host–guest interaction are presented. The gold nanoparticles were incorporated into polymer brushes by host–guest interaction between β-cyclodextrin (β-CD) ligand of gold nanoparticles and dimethylamino group of poly­(2-(dimethylamino)­ethyl methacrylate) (PDMAEMA). The gold nanoparticle arrays were prepared through the template of PDMAEMA brush patterns which were fabricated combining colloidal lithography and surface-initiated atom-transfer radical polymerization (SI-ATRP). The structure parameters of gold nanoparticle patterns mediated by polymer brushes such as height, diameters, periods and distances, could be easily tuned by tailoring the etching time or size of colloidal spheres in the process of colloidal lithography. The change of optical properties induced by different gold nanoparticle structures was demonstrated. The direct utilization of PDMAEMA brushes as guest avoids a series of complicated modification process and the PDMAEMA brushes can be grafted on various substrates, which broaden its applications. The prepared gold naoparticle arrays are promising in applications of nanosensors, memory storage and surface enhanced spectroscopy

Scatter plot of the relationship between the PCC-right medial superior frontal gyrus functional connectivity and hemoglobin levels in the patients.

<p>Scatter plot of the relationship between the PCC-right medial superior frontal gyrus functional connectivity and hemoglobin levels in the patients.</p

Controlling Flow Behavior of Water in Microfluidics with a Chemically Patterned Anisotropic Wetting Surface

We report the flow behavior of water in microfluidic systems based on a chemically patterned anisotropic wetting surface. When water flows inside a microchannel on top of a micropatterned surface with alternating hydrophilic/hydrophobic stripes, it exhibits an anisotropic flowing characteristic owing to the anisotropic wettability; thus, the patterned surface acts as a microvalve for the microfluidic system. The anisotropic flow of water is influenced by the microscale features of the patterns and the dimensions of the microchannels. Furthermore, by reasonably combining the patterned surface and microchannel together, we realize the transportation of water in a microchannel along a “virtual” wall, which is the boundary of the hydrophilic and hydrophobic area. We believe that the chemically patterned surfaces could be an alternative strategy to control the flow behavior of water in microfluidic channels

Radiolabeling Diaminosarcophagine with Cyclotron-Produced Cobalt-55 and [⁵⁵Co]Co-NT-Sarcage as a Proof of Concept in a Murine Xenograft Model

Cobalt–sarcophagine complexes exhibit high kinetic inertness under various stringent conditions, but there is limited literature on radiolabeling and in vivo positron emission tomography (PET) imaging using no carrier added 55Co. To fill this gap, this study first investigates the radiolabeling of DiAmSar (DSar) with 55Co, followed by stability evaluation in human serum and EDTA, pharmacokinetics in mice, and a direct comparison with [55Co]CoCl2 to assess differences in pharmacokinetics. Furthermore, the radiolabeling process was successfully used to generate the NTSR1-targeted PET agent [55Co]Co-NT-Sarcage (a DSar-functionalized SR142948 derivative) and administered to HT29 tumor xenografted mice. The [55Co]Co-DSar complex can be formed at 37 °C with purity and stability suitable for preclinical in vivo radiopharmaceutical applications, and [55Co]Co-NT-Sarcage demonstrated prominent tumor uptake with a low background signal. In a direct comparison with [64Cu]Cu-NT-Sarcage, [55Co]Co-NT-Sarcage achieved a higher tumor-to-liver ratio but with overall similar biodistribution profile. These results demonstrate that Sar would be a promising chelator for constructing Co-based radiopharmaceuticals including 55Co for PET and 58mCo for therapeutic applications