Conductance Evolution of Photoisomeric Single-Molecule Junctions under Ultraviolet Irradiation and Mechanical Stretching.

A comprehensive understanding of carrier transport in photoisomeric molecular junctions is crucial for the rational design and delicate fabrication of single-molecule functional devices. It has been widely recognized that the conductance of azobenzene (a class of photoisomeric molecules) based molecular junctions is mainly determined by photoinduced conformational changes. In this study, it is demonstrated that the most probable conductance of amine-anchored azobenzene-based molecular junctions increases continuously upon UV irradiation. In contrast, the conductance of pyridyl-anchored molecular junctions with an identical azobenzene core exhibits a contrasting trend, highlighting the pivotal role that anchoring groups play, potentially overriding (even reversing) the effects of photoinduced conformational changes. It is further demonstrated that the molecule with cis-conformation cannot be fully mechanically stretched into the trans-conformation, clarifying that it is a great challenge to realize a reversible molecular switch by purely mechanical operation. Additionally, it is revealed that the coupling strength of pyridyl-anchored molecules is dramatically weakened when the UV irradiation time is prolonged, whereas it is not observed for amine-anchored molecules. The mechanisms for these observations are elucidated with the assistance of density functional theory calculations and UV-Vis spectra combined with flicker noise measurements which confirm the photoinduced conformational changes, providing insight into understanding the charge transport in photoisomeric molecular junctions and offering a routine for logical designing synchro opto-mechanical molecular switches.

Identification of seven-gene marker to predict the survival of patients with lung adenocarcinoma using integrated multi-omics data analysis.

BACKGROUND: The mechanism of cancer occurrence and development could be understood with multi-omics data analysis. Discovering genetic markers is highly necessary for predicting clinical outcome of lung adenocarcinoma (LUAD). METHODS: Clinical follow-up information, copy number variation (CNV) data, single nucleotide polymorphism (SNP), and RNA-Seq were acquired from The Cancer Genome Atlas (TCGA). To obtain robust biomarkers, prognostic-related genes, genes with SNP variation, and copy number differential genes in the training set were selected and further subjected to feature selection using random forests. Finally, a gene-based prediction model for LUAD was validated in validation datasets. RESULTS: The study filtered 2071 prognostic-related genes and 230 genomic variants, 1878 copy deletions, and 438 significant mutations. 218 candidate genes were screened through integrating genomic variation genes and prognosis-related genes. 7 characteristic genes (RHOV, CSMD3, FBN2, MAGEL2, SMIM4, BCKDHB, and GANC) were identified by random forest feature selection, and many genes were found to be tumor progression-related. A 7-gene signature constructed by Cox regression analysis was an independent prognostic factor for LUAD patients, and at the same time a risk factor in the test set, external validation set, and training set. Noticeably, the 5-year AUC of survival in the validation set and training set was all ˃ 0.67. Similar results were obtained from multi-omics validation datasets. CONCLUSIONS: The study builds a novel 7-gene signature as a prognostic marker for the survival prediction of patients with LUAD. The current findings provided a set of new prognostic and diagnostic biomarkers and therapeutic targets.

Imaging NF-κB activity in a murine model of early stage diabetes.

Early pro-inflammatory signaling in the endocrine pancreas involves activation of NF-κB, which is believed to be important for determining the ultimate fate of ß-cells and hence progression of type 1 diabetes (T1D). Thus, early non-invasive detection of NF-κB in pancreatic islets may serve as a potential strategy for monitoring early changes in pancreatic endocrine cells eventually leading to T1D. We investigated the feasibility of optical imaging of NF-κB transcription factor activation induced by low-dose streptozocin (LD-STZ) treatment in the immunocompetent SKH1 mouse model of early stage diabetes. In this model, we showed that the levels of NF-κB may be visualized and measured by fluorescence intensity of specific near-infrared (NIR) fluorophore-labeled oligodeoxyribonucleotide duplex (ODND) probes. In addition, NF-κB activation following LD-STZ treatment was validated using immunofluorescence and transgenic animals expressing NF-κB inducible imaging reporter. We showed that LD-STZ-treated SKH1 mice had significantly higher (2-3 times, P < .01) specific NIR FI in the nuclei and cytoplasm of islets cells than in non-treated control mice and this finding was corroborated by immunoblotting and electrophoretic mobility shift assays. Finally, using semi-quantitative confocal analysis of non-fixed pancreatic islet microscopy we demonstrated that ODND probes may be used to distinguish between the islets with high levels of NF-κB transcription factor and control islet cells.

What county-level factors influence COVID-19 incidence in the United States? Findings from the first wave of the pandemic.

Effective control of the COVID-19 pandemic via appropriate management of the built environment is an urgent issue. This study develops a research framework to explore the relationship between COVID-19 incidence and influential factors related to protection of vulnerable populations, intervention in transmission pathways, and provision of healthcare resources. Relevant data for regression analysis and structural equation modeling is collected during the first wave of the pandemic in the United States, from counties with over 100 confirmed cases. In addition to confirming certain factors found in the existing literature, we uncover six new factors significantly associated with COVID-19 incidence. Furthermore, incidence during the lockdown is found to significantly affect incidence after the reopening, highlighting that timely quarantining and treating of patients is essential to avoid the snowballing transmission over time. These findings suggest ways to mitigate the negative effects of subsequent waves of the pandemic, such as special attention of infection prevention in neighborhoods with unsanitary and overcrowded housing, minimization of social activities organized by neighborhood associations, and contactless home delivery service of healthy food. Also worth noting is the need to provide support to people less capable of complying with the stay-at-home order because of their occupations or socio-economic disadvantage.

Synthesis and Testing of Modular Dual-Modality Nanoparticles for Magnetic Resonance and Multispectral Photoacoustic Imaging.

Magnetic resonance (MR) and photoacoustic (PA) imaging are currently being investigated as complementing strategies for applications requiring sensitive detection of cells in vivo. While combined MR/PAI detection of cells requires biocompatible cell labeling probes, water-based synthesis of dual-modality MR/PAI probes presents significant technical challenges. Here we describe facile synthesis and characterization of hybrid modular dextran-stabilized gold/iron oxide (Au-IO) multimetallic nanoparticles (NP) enabling multimodal imaging of cells. The stable association between the IO and gold NP was achieved by priming the surface of dextran-coated IO with silver NP resulting from silver(I) reduction by aldehyde groups, which are naturally present within the dextran coating of IO at the level of 19-23 groups/particle. The Au-IO NP formed in the presence of silver-primed Au-IO were stabilized by using partially thiolated MPEG5-gPLL graft copolymer carrying residual amino groups. This stabilizer served as a carrier of near-infrared fluorophores (e.g., IRDye 800RS) for multispectral PA imaging. Dual modality imaging experiments performed in capillary phantoms of purified Au-IO-800RS NPs showed that these NPs were detectible using 3T MRI at a concentration of 25 µM iron. PA imaging achieved approximately 2.5-times higher detection sensitivity due to strong PA signal emissions at 530 and 770 nm, corresponding to gold plasmons and IRDye integrated into the coating of the hybrid NPs, respectively, with no "bleaching" of PA signal. MDA-MB-231 cells prelabeled with Au-IO-800RS retained plasma membrane integrity and were detectable by using both MR and dual-wavelength PA at 49 ± 3 cells/imaging voxel. We believe that modular assembly of multimetallic NPs shows promise for imaging analysis of engineered cells and tissues with high resolution and sensitivity.

Gold nanoparticles stabilized with MPEG-grafted poly(l-lysine): in vitro and in vivo evaluation of a potential theranostic agent.

As the number of diagnostic and therapeutic applications utilizing gold nanoparticles (AuNPs) increases, so does the need for AuNPs that are stable in vivo, biocompatible, and suitable for bioconjugation. We investigated a strategy for AuNP stabilization that uses methoxypolyethylene glycol-graft-poly(l-lysine) copolymer (MPEG-gPLL) bearing free amino groups as a stabilizing molecule. MPEG-gPLL injected into water solutions of HAuCl4 with or without trisodium citrate resulted in spherical (Zav = 36 nm), monodisperse (PDI = 0.27), weakly positively charged nanoparticles (AuNP3) with electron-dense cores (diameter: 10.4 ± 2.5 nm) and surface amino groups that were amenable to covalent modification. The AuNP3 were stable against aggregation in the presence of phosphate and serum proteins and remained dispersed after their uptake into endosomes. MPEG-gPLL-stabilized AuNP3 exhibited high uptake and very low toxicity in human endothelial cells, but showed a high dose-dependent toxicity in epithelioid cancer cells. Highly stable radioactive labeling of AuNP3 with (99m)Tc allowed imaging of AuNP3 biodistribution and revealed dose-dependent long circulation in the blood. The minor fraction of AuGNP3 was found in major organs and at sites of experimentally induced inflammation. Gold analysis showed evidence of a partial degradation of the MPEG-gPLL layer in AuNP3 particles accumulated in major organs. Radiofrequency-mediated heating of AuNP3 solutions showed that AuNP3 exhibited heating behavior consistent with 10 nm core nanoparticles. We conclude that PEG-pPLL coating of AuNPs confers "stealth" properties that enable these particles to exist in vivo in a nonaggregating, biocompatible state making them suitable for potential use in biomedical applications such as noninvasive radiofrequency cancer therapy.

Release of PAHs from sediments to seawater under wave: Indoor microcosms and level IV fugacity models.

Estuaries and coasts are located at the land-sea interface, where sediment liquefaction due to strong wave action results in significant material exchange at the sediment-seawater system. Polycyclic aromatic hydrocarbons (PAHs), as organic pollutants, are distributed across various media. Herein, the impact of wave was studied on the release of PAHs through indoor microcosmic experiments combined with a level IV fugacity model. Comparison revealed that the release amount and rate of PAHs during static consolidation stage were minimal, whereas wave action substantially enhanced the release. Particularly the sediments in a liquefied state, the PAHs release in Stage III was 1.55-1.86 times that in Stage II, reaching 84.73 µg/L. The loss of soil strength and strong hydrodynamic effects resulted in a substantial release of PAHs into seawater along with suspended solids. Due to volatility of 2-ring PAHs and difficult desorption of 6-ring PAHs, 3-5-ring PAHs are the main contributors to releases into seawater. The model results also indicated that the three PAHs had different fates in the sediment-seawater system, with sediment serving as an important "reservoir" for benzo[a]pyrene entering seawater, while functioning as both a "sink" and a "source" for pyrene.

The effect of bioturbation on the release behavior of polycyclic aromatic hydrocarbons from sediments: A sediment-seawater microcosm experiment combined with a fugacity model.

The effects of two benthonic species, Perinereis aibuhitensis and Matuta planipes Fabricius, on the release of polycyclic aromatic hydrocarbons (PAHs) from sediments were investigated using a sediment-seawater microcosm. A Level IV fugacity model was used to simulate the behavior and fate of PAHs in the environment. This study revealed that both benthos significantly influenced the release of PAHs, and Matuta planipes Fabricius had a stronger disturbance effect than another. The final concentrations of Matuta planipes Fabricius group, Perinereis aibuhitensis group and the control group in the seawater phase reached 10.8, 9.94 and 7.90 µg/L, respectively. There were certain differences in the behaviour of the two benthonic species. Matuta planipes Fabricius caused more sediment resuspension, while Perinereis aibuhitensis increased the total organic carbon (TOC) content in the environment. The vertical concentration distribution of sediment indicated that vertical mixing was slightly stronger in the Matuta planipes Fabricius group than that in the Perinereis aibuhitensis group. The fugacity model effectively simulated the release behavior of PAHs, providing insight into PAH transport and distribution. The results demonstrated that bioturbation could promote the release of PAHs from seawater. The amount of PAHs released was significantly correlated with the biological habits of the benthos.

Governing public health emergencies during the coronavirus disease outbreak: Lessons from four Chinese cities in the first wave.

The ongoing coronavirus disease (COVID-19) pandemic has had a far-reaching impact on urban living, prompting emergency preparedness and response from public health governance at multiple levels. The Chinese government has adopted a series of policy measures to control infectious disease, for which cities are the key spatial units. This research traces and reports analyses of those policy measures and their evolution in four Chinese cities: Zhengzhou, Hangzhou, Shanghai and Chengdu. The theoretical framework stems from conceptualisations of urban governance and its role in public health emergencies, wherein crisis management and emergency response are highlighted. In all four cities, the trend curves of cumulative diagnosed cases, critical policies launched in key time nodes and local governance approaches in the first wave were identified and compared. The findings suggest that capable local leadership is indispensable for controlling the coronavirus epidemic, yet local governments' approaches are varied, contributing to dissimilar local epidemic control policy pathways and positive outcomes in the fight against COVID-19. The effectiveness of disease control is determined by how local governments' measures have adapted to geospatial and socioeconomic heterogeneity. The coordinated actions from central to local governments also reveal an efficient, top-down command transmission and execution system for coping with the pandemic. This article argues that effective control of pandemics requires both a holistic package of governance strategies and locally adaptive governance measures/processes, and concludes with proposals for both a more effective response at the local level and identification of barriers to achieving these responses within diverse subnational institutional contexts.

A study combining a sediment-seawater microcosm with multimedia fugacity model to evaluate the effect of tidal cycles on polycyclic aromatic hydrocarbon release from sediments.

Estuarine sediments are key storage sites for persistent organic pollutants (POPs), and estuaries are strongly influenced by tides throughout the year. Although much work has been done concerning on POPs release, related questions on tidal action have not been considered in the release process. Herein, polycyclic aromatic hydrocarbons (PAHs) release from sediment to seawater was investigated under tidal action by combining a tidal microcosm with level IV fugacity model. The results showed that PAHs release with tidal action was 2.0-3.5 times the accumulation of that without tidal action. Tidal action was confirmed to influence strongly PAHs release from sediment to seawater. We also quantified suspended substance (SS) in the overlying water, and an obvious positive correlation between the PAHs concentration and SS content was found. In addition, an increase in seawater depth enhanced the intensity of tidal action, and more PAHs were released, especially dissolved PAHs. Moreover, the fugacity model results showed a good fit with the experimental results. The simulated values demonstrated that the PAHs release was accomplished via two processes, "rapid release" and "slow release". And the sediment played a key role in the fate of PAHs and was a major sink in the sediment-seawater system.

Examining the association between the built environment and active travel using GPS data: A study of a large residential area (Daju) in Shanghai.

Automobile dependence and physical inactivity have become common health challenges for residents in large suburban residential areas. Limited literature has examined the associations between the built environment and active travel in such residential areas and the differences in these associations among residents from different neighborhoods. To avoid inaccurate results potentially derived from residence-based measures, we adopt a mobility-based approach for environmental exposure assessment. Using GPS data from 530 trips made by 98 participants in a large residential area in Shanghai, we investigate the relationships between neighborhood types, pollution perceptions, built environment features and active travel. The results indicate that residents in affordable and relocation housing make fewer active trips than those in market-rate housing, while the built environment seems to mitigate this difference. Sports facilities promote active travel while commercial facilities and road intersections discourage it. We identify significant interactions between the percentage of green space and neighborhood type, as well as floor area ratio and air pollution perception. Interventions promoting active travel include active-travel-friendly design for commercial facilities and road intersections, the provision of more sports facilities, a careful increase in floor area ratio, and the provision of more green space that is attractive to residents from different neighborhoods.

Spatiotemporal variability of PAHs and their derivatives in sediments of the Laizhou Bay in the eastern China: Occurrence, source, and ecological risk assessment.

To understand the pollution status and risk levels in the Laizhou Bay, the spatiotemporal distribution, source, and ecological risk of 16 polycyclic aromatic hydrocarbons (PAHs) and 20 substituted PAHs (SPAHs) were studied in surface sediments in 2022. The findings indicated significant seasonal differences in the concentrations of PAHs and SPAHs under the influences of precipitation, temperature, light, and human activities, with higher storage levels in summer than in spring, and there was also a spatial distribution trend of estuary > coast > offshore. 2-Nitrofluorene (2-NF) and 2-methylnaphthalene (2-MN) were the most abundant components of SPAHs in both spring and summer, with levels of 21.44 ng/g and 17.89 ng/g in spring, 43.22 ng/g and 25.51 ng/g in summer, respectively. The results of the diagnostic ratio and principal component analysis - multiple linear regression identified sources of PAHs and SPAHs as combustion sources, including petroleum, coal, and biomass. The risk level of PAHs was low-to-moderate according to the toxicity equivalent quotient (TEQ) and risk quotient. A novel calculation method based on TEQ was proposed to assess the ecological risk of SPAHs, and the results indicated that the risk level of SPAHs was moderate-to-high.

Lamina Variation and Its Relationship with Sedimentary Facies in Alkaline Lacustrine, Permian Fengcheng Formation, Junggar Basin, Northwest China.

The Permian Fengcheng Formation in the Mahu Sag was deposited in a volcanic-alkaline lacustrine evaporative environment and contains a unique variety of fine-grained sediments. This study examines, at a millimeter-scale, the influence of sedimentary microfacies on variability of lamina quality in fine-grained sediments in the second member of the Fengcheng Formation (P1f2). The methods used include thin-section identification, X-ray diffraction (XRD), X-ray fluorescence (XRF), scanning electron microscopy (SEM), nitrogen adsorption, and nuclear magnetic resonance (NMR). Six types of lamina were identified in two different lithofacies: fan-delta front facies (FDFF) and semideep/deep lacustrine facies (SDDLF). The laminae in FDFF are predominantly feldspar-quartz laminae (FQL), reedmergnerite laminae (RL), shortite laminae (SL), alkaline mineral laminae (AML), and chert laminae (CL). The laminae in SDDLF are predominantly FQL, RL, SL, CL, and dolomite laminae (DOL). Variations in reservoir quality, oil-bearing properties, and the fracability of laminae in different sedimentary facies are determined by the combined effects of lamina density, mineral composition, rock structure, organic matter abundance, and microfractures. Analysis of these factors indicates superior reservoir qualities in FDFF. In SDDLF, the pore structure is limited by high lamina density, chert content, and fine grain size with the NMR porosities of FQL, RL, SL, and CL being 1.32, 0.18, 0.84, and 0.39%, respectively. However, in FDFF, the combination of high organic matter content, feldspar, pyrite, and clay minerals has a superior effect on the organic matter and minerals deposited resulting in better pore structure and more storage space for shale oil. The NMR porosities of FQL, RL, SL, and CL are 2.81, 2.53, 1.80, and 1.12%, respectively. Overall, analysis of lamina variations and their relationships with sedimentary facies indicates that the reservoir in FDFF may offer more favorable targets for "sweet spot" evaluation.

Plasmon-Assisted Trapping of Single Molecules in Nanogap.

The manipulation of single molecules has attracted extensive attention because of their promising applications in chemical, biological, medical, and materials sciences. Optical trapping of single molecules at room temperature, a critical approach to manipulating the single molecule, still faces great challenges due to the Brownian motions of molecules, weak optical gradient forces of laser, and limited characterization approaches. Here, we put forward localized surface plasmon (LSP)-assisted trapping of single molecules by utilizing scanning tunneling microscope break junction (STM-BJ) techniques, which could provide adjustable plasmonic nanogap and characterize the formation of molecular junction due to plasmonic trapping. We find that the plasmon-assisted trapping of single molecules in the nanogap, revealed by the conductance measurement, strongly depends on the molecular length and the experimental environments, i.e., plasmon could obviously promote the trapping of longer alkane-based molecules but is almost incapable of acting on shorter molecules in solutions. In contrast, the plasmon-assisted trapping of molecules can be ignored when the molecules are self-assembled (SAM) on a substrate independent of the molecular length.

In Situ Adjustable Nanogaps and In-Plane Break Junctions.

The ability to precisely regulate the size of a nanogap is essential for establishing high-yield molecular junctions, and it is crucial for the control of optical signals in extreme optics. Although remarkable strategies for the fabrication of nanogaps are proposed, wafer-compatible nanogaps with freely adjustable gap sizes are not yet available. Herein, two approaches for constructing in situ adjustable metal gaps are proposed which allow Ångstrom modulation resolution by employing either a lateral expandable piezoelectric sheet or a stretchable membrane. These in situ adjustable nanogaps are further developed into in-plane molecular break junctions, in which the gaps can be repeatedly closed and opened thousands of times with self-assembled molecules. The conductance of the single 1,4-benzenediamine (BDA) and the BDA molecular dimer is successfully determined using the proposed strategy. The measured conductance agreeing well with the data by employing another well-established scanning tunneling microscopy break junction technique provides insight into the formation of molecule dimer via hydrogen bond at single molecule level. The wafer-compatible nanogaps and in-plane dynamical break-junctions provide a potential approach to fabricate highly compacted devices using a single molecule as a building block and supply a promising in-plane technique to address the dynamical properties of single molecules.

Deciphering the impact of urban built environment density on respiratory health using a quasi-cohort analysis of 5495 non-smoking lung cancer cases.

INTRODUCTION: Lung cancer is a major health concern and is influenced by air pollution, which can be affected by the density of urban built environment. The spatiotemporal impact of urban density on lung cancer incidence remains unclear, especially at the sub-city level. We aimed to determine cumulative effect of community-level density attributes of the built environment on lung cancer incidence in high-density urban areas. METHODS: We selected 78 communities in the central city of Shanghai, China as the study site; communities included in the analysis had an averaged population density of 313 residents per hectare. Using data from the city cancer surveillance system, an age-period-cohort analysis of lung cancer incidence was performed over a five-year period (2009-2013), with a total of 5495 non-smoking/non-secondhand smoking exposure lung cancer cases. Community-level density measures included the density of road network, facilities, buildings, green spaces, and land use mixture. RESULTS: In multivariate models, built environment density and the exposure time duration had an interactive effect on lung cancer incidence. Lung cancer incidence of birth cohorts was associated with road density and building coverage across communities, with a relative risk of 1·142 (95 % CI: 1·056-1·234, P = 0·001) and 1·090 (95 % CI: 1·053-1·128, P < 0·001) at the baseline year (2009), respectively. The relative risk increased exponentially with the exposure time duration. As for the change in lung cancer incidence over the five-year period, lung cancer incidence of birth cohorts tended to increase faster in communities with a higher road density and building coverage. CONCLUSION: Urban planning policies that improve road network design and building layout could be important strategies to reduce lung cancer incidence in high-density urban areas.

Hairpin-like fluorescent probe for imaging of NF-κB transcription factor activity.

Three oligodeoxyribonucleotides (ODN) covalently labeled with near-infrared (NIR) fluorochromes were synthesized and characterized with a goal of comparing in vitro a hairpin-based and a duplex-based FRET probe designed for the detection of human recombinant NF-κB p50/p65 heterodimer binding to DNA. Using deoxyguanosine phosphoramidite with a phosphorus-linked aminoethylene (diethylene glycol) hydrophilic linker, we synthesized ODNs with internucleoside reactive sites. The hairpin loop amino linker was modified with IRDye 800CW (FRET acceptor), and the 3'-end was modified with Cy5.5 (FRET donor) using a dithio-linker. To obtain a duplex probe, we conjugated Cy5.5 and 800CW to complementary strands at the distance of ten base pairs in the resultant duplex. No quenching of dyes was observed in either probe. The FRET efficiency was higher in the duplex (71%) than in the hairpin (56%) due to a more favorable distance between the donor and the acceptor. However, the hairpin design allowed more precise ratiometric measurement of fluorescence intensity changes as a result of NF-κB p50/p65 binding to the probe. We determined that as a result of binding there was a statistically significant increase of fluorescence intensity of Cy5.5 (donor) due to a decrease of FRET if normalized by 800CW intensity measured independently of FRET. We conclude that the hairpin based probe design allows for the synthesis of a dual fluorescence imaging probe that renders signal changes that are simple to interpret and stoichiometrically correct for detecting transcription factor-DNA interactions.

Fluorescence resonance energy transfer in near-infrared fluorescent oligonucleotide probes for detecting protein-DNA interactions.

Optical imaging in the near-infrared (NIR) range enables detecting ligand-receptor interactions and enzymatic activity in vivo due to lower scattering and absorption of NIR photons in the tissue. We designed and tested prototype NIR fluorescent oligodeoxyribonucleotide (ODN) reporters that can sense transcription factor NF-kappaB p50 protein binding. The reporter duplexes included donor NIR Cy5.5 indodicarbocyanine fluorochrome linked to the 3' end of the first ODN and NIR acceptor fluorochromes (indodicarbocyanine Cy7 or, alternatively, a heptamethine cyanine IRDye 800CW) that were linked at the positions +8 and +12 to the complementary ODN that encoded p50 binding sites. Both Cy7 and 800CW fluorochromes were linked by using hydrophilic internucleoside phosphate linkers that enable interaction between the donor and the acceptor with no base-pairing interference. We observed efficient fluorescence resonance energy transfer (FRET) both in the case of Cy5.5-Cy7 and Cy5.5-800CW pairs of fluorochromes, which was sensitive to the relative position of the dyes. Higher FRET efficiency observed in the case of Cy5.5-Cy7 pair was due to a larger overlap between the ODN-linked Cy5.5 emission and Cy7 excitation spectra. Fluorescent mobility shift assay showed that the addition of human recombinant p50 to ODN duplexes resulted in p50 binding and measurable increase of Cy5.5 emission. In addition, p50 binding provided a concomitant protection of FRET effect from exonuclease-mediated hydrolysis. We conclude that NIR FRET effect can be potentially used for detecting protein-DNA interactions and that the feasibility of detection depends on FRET efficacy and relative fluorochrome positions within ODN binding sites.

In situ photoconductivity measurements of imidazole in optical fiber break-junctions.

We developed a method based on the mechanically controllable break junction technique to investigate the electron transport properties of single molecular junctions upon fiber waveguided light. In our strategy, a metal-coated tapered optical fiber is fixed on a flexible substrate, and this tapered fiber serves as both the optical waveguide and metal electrodes after it breaks. For an imidazole bridged single-molecule junction, two probable conductance values below 1G0 are observed. The higher value shows an approximately 40% enhancement under illumination, while the lower one does not show distinguishable difference under illumination. Theoretical calculations reveal these two conductance values resulting from the imidazole monomer junction and the imidazole dimer junction linked via a hydrogen bond, respectively. In imidazole monomer junctions, the absorption of a single photon strongly shifts the transmission function resulting in optical-induced conductance enhancement. In contrast, the transmission function of imidazole dimer junctions remains at the same level in the bias window despite the light illumination. This work provides a robust experimental framework for studying the underlying mechanisms of photoconductivity in single-molecule junctions and offers tools for tuning the optoelectronic performance of single-molecule devices in situ.

Affinity enhancement pretargeting: synthesis and testing of a 99mTc-labeled bivalent MORF.

Pretargeting with bivalent effectors capable of bridging antitumor antibodies (affinity enhancement pretargeting) has been reported to provide superior results by affinity enhancement. Phosphorodiamidate morpholinos (MORFs) and other DNA analogues used for pretargeting are ideally suited as bivalent effectors since they are easily synthesized and the distance between binding regions, a determinant of binding, may be adjusted simply by lengthening the chain. We have shown by surface plasmon resonance that bivalent MORFs will provide superior affinity enhancement provided that suitable spacing exists between the binding regions. The goals of this study were to synthesize a bivalent MORF with a MAG(3) group attached for technetium-99m ((99m)Tc) radiolabeling, investigate whether the bivalent MORF showed improved cell accumulation in culture compared to its corresponding monovalent MORF and compare biodistributions in normal mice and in pretargeted tumored mice. An excess of an amine derivatized 18 mer MORF with 6 nonbinding bases for spacing was reacted with Fmoc-l-beta-homoglutamic acid to form duplexes via their carboxylate groups and, after deprotection, conjugated with NHS-MAG(3) to attach the chelator. The anti-CEA antibody MN14 was conjugated with a 12 mer complementary MORF (i.e., cMORF). The binding behavior between radiolabeled monovalent and bivalent MORFs was compared in LS174T tumor cells at 4 degrees C pretargeted with MN14-cMORF. Biodistributions of radiolabeled monovalent and bivalent MORFs at 3 h postadministration were measured in normal mice and in tumor mice pretargeted with MN14-cMORF. In the pretargeted cells in culture, the accumulation of the bivalent MORF was significantly higher than the monovalent MORF (p = 0.002), thus providing strong evidence for affinity enhancement. In normal mice, whole body clearance of the bivalent and monovalent MORFs was equally rapid. In tumored mice, tumor accumulation of the radiolabeled bivalent MORF was significantly higher than that of the monovalent MORF. In conclusion, a bivalent MAG(3)-MORF was successfully synthesized and radiolabeled with (99m)Tc. While a pharmacokinetic effect for the higher tumor accumulations in pretargeted mice of the radiolabeled bivalent MORF cannot be excluded, the results may be best explained by affinity enhancement. Thus two monovalent MORFs were covalently conjugated into a bivalent MORF effector to improve tumor targeting by both pharmacokinetics and affinity enhancement influences.